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Sample records for goat preantral follicles

  1. Assessment of DNA damage in goat preantral follicles after vitrification of the ovarian cortex.

    PubMed

    Faustino, Luciana R; Carvalho, Adeline A; Silva, Cleidson M G; Rossetto, Rafael; Lopes, Cláudio A P; van Tilburg, Maurício F; Carneiro, Pedro B M; Báo, Sônia N; Moura, Arlindo A A; Bordignon, Vilceu; Figueiredo, José R; Rodrigues, Ana Paula R

    2015-03-01

    Effective methods for gamete preservation should have low impact on DNA integrity. The present study investigated the effects of vitrification of goat ovarian tissues on the occurrence of DNA fragmentation and DNA double-stand breaks using the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) assay and detection of phosphorylated histone H2AX (γH2AX), respectively. Goat ovaries were collected at a local abattoir and 12 tissue fragments were prepared from each ovarian pair. Tissue fragments were used as fresh control samples or were cultured in vitro, vitrified or vitrified and cultured. Vitrification was performed using the Ovarian Tissue Cryosystem. Fragments from all groups (control and treatments) were processed for histology, transmission electron microscopy, TUNEL assay and immunofluorescence. Compared with fresh control samples, a lower percentage of morphologically normal follicles was detected in the vitrification followed by culture treatment group (P<0.05). Normal follicular ultrastructure was observed in all groups. Immunofluorescence revealed the presence of γH2AX foci in few oocytes and ovarian stromal cells. TUNEL-positive follicles were found in samples without significant differences among groups (P>0.05). In conclusion, the vitrification protocol used in the present study did not increase DNA damage in preantral follicles enclosed in goat ovarian tissues. PMID:25481978

  2. Effect of sequential medium with fibroblast growth factor-10 and follicle stimulating hormone on in vitro development of goat preantral follicles.

    PubMed

    Almeida, A P; Magalhães-Padilha, D M; Araújo, V R; Costa, S L; Chaves, R N; Lopes, C A P; Donato, M A M; Peixoto, C A; Campello, C C; Junior, J Buratini; Figueiredo, J R

    2015-01-01

    A sequential medium with fibroblast growth factor-10 (FGF-10) and follicle stimulating hormone (FSH) was evaluated on the survival, ultrastructure, activation and growth rate of caprine preantral follicles submitted to long-term culture, aiming to establish an ideal in vitro culture system. Ovarian fragments were cultured for 16 days in α-MEM(+) alone or supplemented with FGF-10 and/or FSH added sequentially on different days of culture. Ovarian fragments were cultured during the first (days 0-8) and second (days 8-16) halves of the culture period, generating 10 treatments: α-MEM(+)/α-MEM(+) (cultured control), FSH/FSH, FSH/FGF-10, FSH/FSH+FGF-10, FGF-10/FGF-10, FGF-10/FSH, FGF-10/FSH+FGF-10, FSH+FGF-10/FSH+FGF-10, FSH+FGF-10/FSH and FSH+FGF-10/FGF-10. Follicle morphology, viability and ultrastructure were analyzed. The FSH/FGF-10 treatment showed a higher (P<0.05) percentage of normal follicles compared to all other treatments. In addition, follicles from the FSH/FGF-10 treatment maintained ultrastructural integrity after the culture period. After 16 days of culture, the FSH/FGF-10 and FSH/FSH treatments showed a higher percentage of activation compared to the cultured control (α-MEM(+)/α-MEM(+)). Moreover, the FSH/FGF-10 treatment promoted greater follicular and oocyte diameters compared to the fresh control. In conclusion, this study showed that a sequential medium with FSH followed by FGF-10 (FSH/FGF-10 and FSH/FSH) maintains follicular viability and ultrastructure and promotes transition from the primordial to primary stage (activation) and growth in goat preantral follicles cultured in vitro.

  3. Supplemented base medium containing Amburana cearensis associated with FSH improves in vitro development of isolated goat preantral follicles.

    PubMed

    Gouveia, B B; Macedo, T J S; Santos, J M S; Barberino, R S; Menezes, V G; Müller, M C; Almeida, J R G S; Figueiredo, J R; Matos, M H T

    2016-09-15

    The effects of Amburana cearensis ethanolic extract, with or without addition of a mix of supplements associated or not with FSH, on in vitro morphology and development of caprine secondary follicles were evaluated. In experiment 1, isolated follicles (250 μm in diameter) were cultured for 12 days in alpha-modified minimal essential medium (α-MEM) alone (control) or in medium composed of different concentrations of A. cearensis extract (Amb 0.1; 0.2, or 0.4 mg/mL). In experiment 2, culture media were α-MEM or Amb 0.2 mg/mL (both without supplements), or these same media supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred as α-MEM(+) and Amb 0.2(+), respectively), or these last groups also supplemented with sequential FSH (100 ng/mL from Day 0 to Day 6; 500 ng/mL from Day 6 to Day 12), constituting groups α-MEM(+) + FSH and Amb 0.2(+) + FSH. At the end of culture in experiment 1, control medium (α-MEM) and Amb 0.2 mg/mL had higher percentages (P < 0.05) of morphologically normal follicles and percentage of fully grown oocytes, i.e., oocyte greater than 110 μm, compared to the other A. cearensis extract concentrations. In experiment 2, all supplemented media had higher percentages (P < 0.05) of normal follicles and antrum formation than nonsupplemented media. In addition, follicles cultured in Amb 0.2(+) + FSH showed an average increase in diameter higher (P < 0.05) than the other treatments. Oocytes cultured in both treatments supplemented with FSH showed greater glutathione and active mitochondria levels than nonsupplemented media but similar to the other treatments. In conclusion, A. cearensis extract (0.2 mg/mL) added by supplements and FSH improves follicular growth. Therefore, it can be an alternative culture medium for goat preantral follicle development. PMID:27287468

  4. Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

    PubMed Central

    Cunha, E.V.; Costa, J.J.N.; Rossi, R.O.D.S.; Silva, A.W.B.; Passos, J.R.S.; Portela, A.M.L.R.; Pereira, D.C.S.T.; Donato, M.A.M.; Campello, C.C.; Saraiva, M.V.A.; Peixoto, C.A.; Silva, J.R.V.; Santos, R.P.

    2013-01-01

    The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1). In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro. PMID:23558855

  5. Catalase addition to vitrification solutions maintains goat ovarian preantral follicles stability.

    PubMed

    Carvalho, A A; Faustino, L R; Silva, C M G; Castro, S V; Lobo, C H; Santos, F W; Santos, R R; Campello, C C; Bordignon, V; Figueiredo, J R; Rodrigues, A P R

    2014-08-01

    The aim of this study was to verify whether the addition of catalase (20 IU/mL) at different steps of goat ovarian tissue vitrification affects ROS levels, follicular morphology and viability, stromal cell density, apoptosis and the expression of proteins related to DNA-damage signaling (γH2AX) and repair (53BP1). Goat ovarian tissues were analyzed fresh (control) or after vitrification: without catalase (VS-/WS-), with catalase in vitrification solutions (VS+/WS-), with catalase in washing solutions (VS-/WS+) or with catalase in both solutions (VS+/WS+). The vitrification without catalase had higher ROS levels than the control. The catalase, regardless the step of addition, maintained ROS levels similar to the control. There were no difference between treatments regarding follicular viability, stromal cell density and detection of γH2AX and 53BP1. There was no difference in follicular morphology and DNA fragmentation between groups vitrified. In conclusion, catalase addition to vitrification solutions prevents ROS formation in cryopreserved goat ovarian tissues.

  6. Catalase addition to vitrification solutions maintains goat ovarian preantral follicles stability.

    PubMed

    Carvalho, A A; Faustino, L R; Silva, C M G; Castro, S V; Lobo, C H; Santos, F W; Santos, R R; Campello, C C; Bordignon, V; Figueiredo, J R; Rodrigues, A P R

    2014-08-01

    The aim of this study was to verify whether the addition of catalase (20 IU/mL) at different steps of goat ovarian tissue vitrification affects ROS levels, follicular morphology and viability, stromal cell density, apoptosis and the expression of proteins related to DNA-damage signaling (γH2AX) and repair (53BP1). Goat ovarian tissues were analyzed fresh (control) or after vitrification: without catalase (VS-/WS-), with catalase in vitrification solutions (VS+/WS-), with catalase in washing solutions (VS-/WS+) or with catalase in both solutions (VS+/WS+). The vitrification without catalase had higher ROS levels than the control. The catalase, regardless the step of addition, maintained ROS levels similar to the control. There were no difference between treatments regarding follicular viability, stromal cell density and detection of γH2AX and 53BP1. There was no difference in follicular morphology and DNA fragmentation between groups vitrified. In conclusion, catalase addition to vitrification solutions prevents ROS formation in cryopreserved goat ovarian tissues. PMID:24972862

  7. Xenobiotic Effects on Ovarian Preantral Follicles1

    PubMed Central

    Mark-Kappeler, Connie J.; Hoyer, Patricia B.; Devine, Patrick J.

    2011-01-01

    Women are born with a finite population of ovarian follicles, which are slowly depleted during their reproductive years until reproductive failure (menopause) occurs. The rate of loss of primordial follicles is determined by genetic and environmental influences, but certain toxic exposures can accelerate this process. Ionizing radiation reduces preantral follicle numbers in rodents and humans in a dose-dependent manner. Cigarette smoking is linked to menopause occurring 1–4 yr earlier than with nonsmokers, and components of smoke, polycyclic aromatic hydrocarbons, can cause follicle depletion in rodents or in ovaries in vitro. Chemotherapeutic agents, such as alkylating drugs and cisplatin, also cause loss of preantral ovarian follicles. Effects depend on dose, type, and reactivity of the drug, and the age of the individual. Evidence suggests DNA damage may underlie follicle loss induced by one common alkylating drug, cyclophosphamide. Occupational exposures have also been linked to ovarian damage. In an industrial setting, 2-bromopropane caused infertility in men and women, and it can induce ovarian follicle depletion in rats. Solvents, such as butadiene, 4-vinylcyclohexene, and their diepoxides, can also cause specific preantral follicle depletion. The mechanism(s) underlying effects of the latter compound may involve alterations in apoptosis, survival factors such as KIT/Kit Ligand, and/or the cellular signaling that maintains primordial follicle dormancy. Estrogenic endocrine disruptors may alter follicle formation/development and impair fertility or normal development of offspring. Thus, specific exposures are known or suspected of detrimentally impacting preantral ovarian follicles, leading to early ovarian failure. PMID:21697514

  8. In vitro culture methods of preantral follicles.

    PubMed

    Green, Lisa J; Shikanov, Ariella

    2016-07-01

    Folliculogenesis is a highly regulated, dynamic process requiring various autocrine, paracrine, and endocrine factors that need to be reproduced in vitro. The mammalian ovary consists of a finite number of follicles at birth, with the majority in a quiescent state, awaiting recruitment. In vitro culture of preantral follicles presents a unique challenge of mimicking the dynamics of the ovarian environment, a methodical sequence of cell communication, and interaction with secretory, hormonal, and growth factors. In larger mammalian species, maintenance of follicular morphology is imperative to maintain communication between the cumulus-oocyte complexes. Three-dimensional culture systems help to maintain this architecture and have been used for the successful culture of follicles in various animal models, including humans and nonhuman primates, and have resulted in live birth in rodent models. Clinically, in vitro follicular development could revolutionize assisted reproductive technology by providing a means of using the more numerous immature ovarian follicles in patient populations that are unable to undergo ovarian stimulation. PMID:27173961

  9. The Mare Model to Study the Effects of Ovarian Dynamics on Preantral Follicle Features.

    PubMed

    Alves, Kele A; Alves, Benner G; Gastal, Gustavo D A; de Tarso, Saulo G S; Gastal, Melba O; Figueiredo, José R; Gambarini, Maria L; Gastal, Eduardo L

    2016-01-01

    Ovarian tissue collected by biopsy procedures allows the performance of many studies with clinical applications in the field of female fertility preservation. The aim of the present study was to investigate the influence of reproductive phase (anestrous vs. diestrous) and ovarian structures (antral follicles and corpus luteum) on the quality, class distribution, number, and density of preantral follicles, and stromal cell density. Ovarian fragments were harvested by biopsy pick-up procedures from mares and submitted to histological analysis. The mean preantral follicle and ovarian stromal cell densities were greater in the diestrous phase and a positive correlation of stromal cell density with the number and density of preantral follicles was observed. The mean area (mm2) of ovarian structures increased in the diestrous phase and had positive correlations with number of preantral follicles, follicle density, and stromal cell density. Biopsy fragments collected from ovaries containing an active corpus luteum had a higher follicle density, stromal cell density, and proportion of normal preantral follicles. In conclusion, our results showed: (1) the diestrous phase influenced positively the preantral follicle quality, class distribution, and follicle and stromal cell densities; (2) the area of ovarian structures was positively correlated with the follicle and stromal cell densities; and (3) the presence of an active corpus luteum had a positive effect on the quality of preantral follicles, and follicle and stromal densities. Therefore, herein we demonstrate that the presence of key ovarian structures favors the harvest of ovarian fragments containing an appropriate number of healthy preantral follicles. PMID:26900687

  10. The Mare Model to Study the Effects of Ovarian Dynamics on Preantral Follicle Features

    PubMed Central

    Alves, Kele A.; Alves, Benner G.; Gastal, Gustavo D. A.; de Tarso, Saulo G. S.; Gastal, Melba O.; Figueiredo, José R.; Gambarini, Maria L.; Gastal, Eduardo L.

    2016-01-01

    Ovarian tissue collected by biopsy procedures allows the performance of many studies with clinical applications in the field of female fertility preservation. The aim of the present study was to investigate the influence of reproductive phase (anestrous vs. diestrous) and ovarian structures (antral follicles and corpus luteum) on the quality, class distribution, number, and density of preantral follicles, and stromal cell density. Ovarian fragments were harvested by biopsy pick-up procedures from mares and submitted to histological analysis. The mean preantral follicle and ovarian stromal cell densities were greater in the diestrous phase and a positive correlation of stromal cell density with the number and density of preantral follicles was observed. The mean area (mm2) of ovarian structures increased in the diestrous phase and had positive correlations with number of preantral follicles, follicle density, and stromal cell density. Biopsy fragments collected from ovaries containing an active corpus luteum had a higher follicle density, stromal cell density, and proportion of normal preantral follicles. In conclusion, our results showed: (1) the diestrous phase influenced positively the preantral follicle quality, class distribution, and follicle and stromal cell densities; (2) the area of ovarian structures was positively correlated with the follicle and stromal cell densities; and (3) the presence of an active corpus luteum had a positive effect on the quality of preantral follicles, and follicle and stromal densities. Therefore, herein we demonstrate that the presence of key ovarian structures favors the harvest of ovarian fragments containing an appropriate number of healthy preantral follicles. PMID:26900687

  11. Population estimate of the preantral follicles and frequency of multioocyte follicles in prepubertal and adult bitches.

    PubMed

    Lunardon, N T; Silva-Santos, K C; Justino, R C; Dessunti, G T; Seneda, M M; Martins, M I M

    2015-04-01

    Oocytes from preantral follicles could be an alternative for in vitro maturation because most follicles are at the preantral stage. There are few studies that have sought to estimate the number of preantral follicles in bitches. Therefore, the aims of this study were to estimate the population of preantral follicles in the ovaries of small- and medium-sized prepubertal and adult bitches and compare the population of preantral follicles between the right and left ovaries and evaluate the frequency of multioocyte follicles (MOF). Eighty ovaries were collected by elective ovariohysterectomy from 40 healthy bitches. The bitches were divided into four groups: small-size prepubertal bitches (<10 kg, n = 20), medium-size prepubertal bitches (10-20 kg, n = 20), small-size adult bitches (<10 kg, n = 20), and medium-size adult bitches (10-20 kg, n = 20). Immediately after surgery, the ovaries were fixed in Bouin's solution and processed for histology. For each specimen, 70 histologic sections were cut and mounted on slides; then, the number of preantral follicles was estimated using a correction factor. The preantral follicles were classified according to the developmental stage. The data were analyzed using the Kruskal-Wallis test followed by Dunn's test for comparison between groups, and Fisher's exact test was used to evaluate the frequency of MOF (P ≤ 0.05). Considering the population of preantral follicles from the pair of ovaries, medium-size prepubertal bitches had the highest (P < 0.05) population of preantral follicles compared with the small and medium-size adult groups. There was a large variation in the numbers of preantral follicles among individuals of the same weight and within each group. There were differences between medium-size prepubertal and adult bitches regarding the population of preantral follicles in the right ovaries (145,482 ± 110,712 vs. 49,500 ± 44,821; P = 0.02); however, no differences were observed between the

  12. Production of buffalo embryos using oocytes from in vitro grown preantral follicles.

    PubMed

    Gupta, P S P; Ramesh, H S; Manjunatha, B M; Nandi, S; Ravindra, J P

    2008-02-01

    The present study examines the use of buffalo preantral follicles as a source of oocytes for in vitro embryo production. Preantral follicles were isolated from abattoir-derived buffalo ovaries and were grown for 100 days in five different culture systems: (1) minimum essential medium (MEM); (2) coconut water; (3) MEM + ovarian mesenchymal cell (OMC) co-culture; (4) MEM + granulosa cell (GC) co-culture; or (5) MEM + cumulus cell (CC) co-culture. Low growth rates for the preantral follicles were observed when follicles were cultured in MEM or coconut water medium. Moderate growth rates were seen for OMC and GC co-cultures, and high rates of growth were observed when follicles were grown in CC co-culture. The survival of preantral follicles was low in the MEM culture (<25%), but was over 75% in the other culture systems. Oocytes were not recovered from the MEM group, while an oocyte recovery rate of 80-100% was observed when the follicles were cultured with coconut water/somatic cells. Transferable embryos could be produced only with the oocytes obtained from preantral follicles grown in the OMC and CC co-culture systems. This study demonstrates, for the first time, that it is possible to produce buffalo embryos by in vitro fertilization of oocytes derived from in vitro grown preantral follicles.

  13. Oxygen consumption by bovine granulosa cells with prediction of oxygen transport in preantral follicles.

    PubMed

    Li, Dongxing; Redding, Gabe P; Bronlund, John E

    2013-01-01

    The rate of oxygen consumption by granulosa cells is a key parameter in mathematical models that describe oxygen transport across ovarian follicles. This work measured the oxygen consumption rate of bovine granulosa cells in vitro to be in the range 2.1-3.3×10⁻¹⁶ mol cell⁻¹ s⁻¹ (0.16-0.25 mol m⁻³ s⁻¹). The implications of the rates for oxygen transport in large bovine preantral follicles were examined using a mathematical model. The results indicate that oocyte oxygenation becomes increasingly constrained as preantral follicles grow, reaching hypoxic levels near the point of antrum formation. Beyond a preantral follicle radius of 134 µm, oxygen cannot reach the oocyte surface at typical values of model parameters. Since reported sizes of large bovine preantral follicles range from 58 to 145 µm in radius, this suggests that oocyte oxygenation is possible in all but the largest preantral follicles, which are on the verge of antrum formation. In preantral bovine follicles, the oxygen consumption rate of granulosa cells and fluid voidage will be the key determinants of oxygen levels across the follicle.

  14. Effect of different culture systems and 3, 5, 3'-triiodothyronine/follicle-stimulating hormone on preantral follicle development in mice.

    PubMed

    Zhang, Cheng; Wang, Xiaoxia; Wang, Zhengpin; Niu, Wanbao; Zhu, Baochang; Xia, Guoliang

    2013-01-01

    The mechanical method to isolate preantral follicle has been reported for many years. However, the culture systems in vitro are still unstable. The aim of this study was to analyze the effect of the culture system of mice preantral follicles on the follicular development in vitro. The results showed that the 96-well plate system was the most effective method for mice follicle development in vitro (volume change: 51.71%; survival rate: 89%, at day 4). Follicle-stimulating hormone (FSH) and Thyroid hormone (TH) are important for normal follicular development and dysregulation of hormones are related with impaired follicular development. To determine the effect of hormone on preantral follicular development, we cultured follicle with hormones in the 96-well plate culture system and found that FSH significantly increased preantral follicular growth on day 4. The FSH-induced growth action was markedly enhanced by T₃ although T₃ was ineffective alone. We also demonstrated by QRT-PCR that T₃ significantly enhanced FSH-induced up-regulation of Xiap mRNA level. Meanwhile, Bad, cell death inducer, was markedly down-regulated by the combination of hormones. Moreover, QRT-PCR results were also consistent with protein regulation which detected by Western Blotting analysis. Taken together, the findings of the present study demonstrate that 96-well plate system is an effective method for preantral follicle development in vitro. Moreover, these results provide insights on the role of thyroid hormone in increasing FSH-induced preantral follicular development, which mediated by up-regulating Xiap and down-regulating Bad.

  15. Simulated Microgravity Using a Rotary Culture System Compromises the In Vitro Development of Mouse Preantral Follicles

    PubMed Central

    Wu, Yonggen; Lin, Wei; Chen, Zaichong; Meng, Luhe; Liu, Jun; Zhou, Ying

    2016-01-01

    Background Growing cells in simulated weightlessness condition might be a highly promising new technique to maintain or generate tissue constructs in a scaffold-free manner. There is limited evidence that microgravity condition may affect development of ovarian follicles. The objective of the present study was to investigate the effects of simulated microgravity on the in vitro development of mouse preantral follicles. Methods and Results Ovarian tissue from 14-day-old mice, or preantral follicles mechanically isolated from 14-day-old mouse ovaries were cultured at a simulated microgravity condition generated using a rotating wall vessel apparatus. Follicle survival was assessed quantitatively using H&E staining. Follicle diameter and oocyte diameter were measured under an inverted microscope. Ultrastructure of oocytes was evaluated using transmission electron microscopy. We observed that simulated microgravity compromised follicle survival in vitro, downregulated PCNA and GDF-9 expressions, and caused ultrastructural abnormalities in oocytes. Conclusion This study showed for the first time that three-dimensional culture condition generated by simulated microgravity is detrimental to the initial stage development of mouse preantral follicles in vitro. The experimental setup provides a model to further investigate the mechanisms involved in the in vitro developmental processes of oocytes/granulosa cells under the microgravity condition. PMID:26963099

  16. Maternal-Effect Gene Expression in Cultured Preantral Follicles Derived from Vitrified-Warmed Mouse Ovary

    PubMed Central

    Fatehi, Roya; Ebrahimi, Bita

    2015-01-01

    Objective This study was conducted to assess survival of follicles, their oocyte maturation and fertilization potential as well as expression of early embryo developmental genes in in vitro cultured pre-antral follicles derived from vitrified-warmed mouse ovary. Materials and Methods In this experimental study, ovaries of 12-day old Naval Medical Research Institute (NMRI) female mice were placed into non-vitrified and vitrifiedwarmed groups. Isolated preantral follicles from experimental groups were cultured in vitro for 12 days. On the 12th day of culture, oocyte maturation was induced and then matured oocytes were in vitro fertilized. The rates of oocyte maturation and two-cell stage embryo formation were assessed. Relative expression of Mater and Zar1 was evaluated on days 1, 6, 10 and 12 of culture. Data analysis was performed by t test and two-way ANOVA (P<0.05). Results Our data showed no significant difference between the control and vitrification groups in the rate of follicular survival, oocyte maturation and two-cell stage embryo formation. The level of gene expression was higher on the 6thand 10thdays of culture for Mater and Zar1 in vitrified-warmed group compared with non-vitrified group, however, there was no significant difference between the two groups. Conclusion It seems that the applied vitrification method did not reveal any negative effect on maturation and developmental competence of oocytes surrounded in preantral follicles and therefore could preserve follicular reserves efficiently. PMID:26199912

  17. Oxygen consumption rate of early pre-antral follicles from vitrified human ovarian cortical tissue

    PubMed Central

    ISHIKAWA, Takayuki; KYOYA, Toshihiko; NAKAMURA, Yusuke; SATO, Eimei; TOMIYAMA, Tatsuhiro; KYONO, Koichi

    2014-01-01

    The study of human ovarian tissue transplantation and cryopreservation has advanced significantly. Autotransplantation of human pre-antral follicles isolated from cryopreserved cortical tissue is a promising option for the preservation of fertility in young cancer patients. The purpose of the present study was to reveal the effect of vitrification after low-temperature transportation of human pre-antral follicles by using the oxygen consumption rate (OCR). Cortical tissues from 9 ovaries of female-to-male transsexuals were vitrified after transportation (6 or 18 h). The follicles were enzymatically isolated from nonvitrified tissue (group I, 18 h of transportation), vitrified-warmed tissue (group II, 6 and 18 h of transportation) and vitrified-warmed tissue that had been incubated for 24 h (group III, 6 and 18 h of transportation). OCR measurement and the LIVE/DEAD viability assay were performed. Despite the ischemic condition, the isolated pre-antral follicles in group I consumed oxygen, and the mean OCRs increased with developmental stage. Neither the transportation time nor patient age seemed to affect the OCR in this group. Meanwhile, the mean OCR was significantly lower (P < 0.05) in group II but was comparable to that of group I after 24 h of incubation. The integrity of vitrified-warmed primordial and primary follicles was clearly corroborated by the LIVE/DEAD viability assay. These results demonstrate that the OCR can be used to directly estimate the effect of vitrification on the viability of primordial and primary follicles and to select the viable primordial and primary follicles from vitrified-warmed follicles. PMID:25262776

  18. Mesenchymal Stem Cells Facilitate In Vitro Development of Human Preantral Follicle.

    PubMed

    Xia, Xi; Wang, Tianren; Yin, Tailang; Yan, Liying; Yan, Jie; Lu, Cuilin; Zhao, Liang; Li, Min; Zhang, Yan; Jin, Hongyan; Zhu, Xiaohui; Liu, Ping; Li, Rong; Qiao, Jie

    2015-11-01

    Biological folliculogenesis is a lengthy and complicated process, and follicle growth microenvironment is poorly understood. Mesenchymal stem cells (MSCs) have been shown to establish a supportive microenvironment for wound repair, autoimmune diseases amelioration, and tumor development. Therefore, this study is aimed to investigate whether MSCs could help to reconstruct a microenvironment to facilitate the in vitro follicle development. Here we show human MSCs significantly promote the survival rates, increase the growth velocity, and improve the viability of preantral follicles in a dose-dependent manner. Further analyses reveal that growth differentiation factor 9 and bone morphogenetic protein 15 in oocytes and inhibin βA and transforming growth factor β1 in granulose cells within the follicles cocultured with MSCs express notably higher than those in the follicles cultured without MSCs. In summary, our findings demonstrate a previously unrecognized function of MSCs in promoting preantral follicle development and provide a useful strategy to optimize fertility preservation and restoration by facilitating in vitro follicle growth.

  19. Gonadotropin ratio affects the in vitro growth of rhesus ovarian preantral follicles.

    PubMed

    Kim, Yoon Young; Yun, Jun-Won; Kim, Jong Min; Park, Chung Gyu; Rosenwaks, Zev; Liu, Hung Ching; Kang, Byeong-Cheol; Ku, Seung-Yup

    2016-04-01

    In vitro follicle growth (IVFG) strategy is critical in the fertility preservation of cancer survivors; however, its optimal protocol needs to be developed using primate models since the availability of human samples is limited. Only a few previous studies have reported the successful IVFG of rhesus monkey ovaries using low-dose follicle-stimulating hormone (FSH) (0.3 or 3 ng/mL) and long-term culture (up to 5 weeks) and it is still uncertain in regard to the optimal culture duration and effective dose of treated gonadotropins applicable to the IVFG of rhesus preantral follicles. Recently, we have reported that the FSH to luteinizing hormone (LH) ratio affects the in vitro growth of murine ovarian follicles. We aimed to investigate whether gonadotropin ratios affect the efficiency of rhesus follicular growth in vitro Ovaries were collected from six necropsied rhesus macaques (4-9 years) and preantral follicles were retrieved and cultured for 14 days using 200 mIU/mL FSH. The characteristics of follicular growth were compared between the FSH:LH=1:1 (n=24) and FSH:LH=2:1 (n=24) groups. High concentration gonadotropin treatment shortened the duration required for in vitro maturation of rhesus preantral follicles. The FSH:LH=2:1 group showed a faster follicular growth and enabled the acquisition of mature oocytes, although the expression of growth differentiation factor (GDF)-9 and anti-Müllerian hormone (AMH) did not differ significantly between the two groups. Taken together, high dose gonadotropin treatment can shorten the duration of IVFG and the gonadotropin ratio is important in the IVFG of rhesus monkey ovaries.

  20. Gonadotropin ratio affects the in vitro growth of rhesus ovarian preantral follicles

    PubMed Central

    Kim, Yoon Young; Yun, Jun-Won; Kim, Jong Min; Park, Chung Gyu; Rosenwaks, Zev; Liu, Hung Ching; Kang, Byeong-Cheol; Ku, Seung-Yup

    2016-01-01

    In vitro follicle growth (IVFG) strategy is critical in the fertility preservation of cancer survivors; however, its optimal protocol needs to be developed using primate models since the availability of human samples is limited. Only a few previous studies have reported the successful IVFG of rhesus monkey ovaries using low-dose follicle-stimulating hormone (FSH) (0.3 or 3 ng/mL) and long-term culture (up to 5 weeks) and it is still uncertain in regard to the optimal culture duration and effective dose of treated gonadotropins applicable to the IVFG of rhesus preantral follicles. Recently, we have reported that the FSH to luteinizing hormone (LH) ratio affects the in vitro growth of murine ovarian follicles. We aimed to investigate whether gonadotropin ratios affect the efficiency of rhesus follicular growth in vitro. Ovaries were collected from six necropsied rhesus macaques (4–9 years) and preantral follicles were retrieved and cultured for 14 days using 200 mIU/mL FSH. The characteristics of follicular growth were compared between the FSH:LH=1:1 (n=24) and FSH:LH=2:1 (n=24) groups. High concentration gonadotropin treatment shortened the duration required for in vitro maturation of rhesus preantral follicles. The FSH:LH=2:1 group showed a faster follicular growth and enabled the acquisition of mature oocytes, although the expression of growth differentiation factor (GDF)-9 and anti-Müllerian hormone (AMH) did not differ significantly between the two groups. Taken together, high dose gonadotropin treatment can shorten the duration of IVFG and the gonadotropin ratio is important in the IVFG of rhesus monkey ovaries. PMID:26980777

  1. Alginate hydrogel matrix stiffness influences the in vitro development of caprine preantral follicles.

    PubMed

    Brito, I R; Silva, C M G; Duarte, A B G; Lima, I M T; Rodrigues, G Q; Rossetto, R; Sales, A D; Lobo, C H; Bernuci, M P; Rosa-E-Silva, A C J S; Campello, C C; Xu, M; Figueiredo, J R

    2014-07-01

    This study examined caprine follicular development in different concentrations of alginate matrix to determine the optimal conditions for culture. Caprine preantral follicles were cultured in a two-dimensional system (control) or a three-dimensional encapsulated system in 0.25%, 0.5%, or 1% alginate (ALG 0.25, ALG 0.5, and ALG 1, respectively). A higher percentage of morphologically normal follicles developed in ALG 0.5 and ALG 1 than in ALG 0.25 or the control (P < 0.05). The rate of antrum formation, however, was higher in ALG 0.25 than in ALG 0.5 and ALG 1 conditions (P < 0.05), but similar to the control. Follicles cultured in ALG 0.25 had higher growth rates and meiotic resumption than those cultured in ALG 0.5, ALG 1, or the control (P < 0.05). Moreover, follicles cultured in ALG 0.25 had higher levels of estradiol and progesterone than those cultured in ALG 0.5, ALG 1, or the control, as well as higher levels of CYP19A1 and HSD3B mRNA. In conclusion, a three-dimensional system that uses ALG 0.25 fosters the in vitro development of caprine preantral follicles and increases the rate of meiotic resumption.

  2. Vitrification and Subsequent In Vitro Maturation of Mouse Preantral Follicles in Presence of Growth Factors

    PubMed Central

    Oryan Abkenar, Zahra; Ganji, Roya; Eghbal Khajehrahimi, Amir; Bahadori, Mohammad Hadi

    2014-01-01

    Objective Cryopreservation of ovarian tissue or follicles has been proposed as an alternative method for fertility preservation. Although successful vitrification of follicles has been reported in several mammalian species, the survival rate is generally low. The aim of this study was to investigate the effects of fibroblast growth factor (FGF) and epidermal growth factor (EGF) on in vitro preantral follicle development after vitrification. Materials and Methods In this experimental study, preantral follicles with diameter of 150-180 µm were mechanically isolated from ovaries of 18-21 days old NMRI mice. Follicles were vitrified and warmed, then cultured in a-minimal essential medium (α-MEM) without growth factor supplementation as control group (group I), while supplemented with 20 ng/ml FGF (group II), 20 ng/ml EGF (group III), and 20 ng/ml FGF +20 ng/ml EGF (group IV). After 12 days, human chorionic gonadotrophin (hCG)/EGF was added to culture medium, and after 18-20 hours, the presence of cumulus oocyte complexes (COCs) and oocyte maturation were assessed. The chi-square (Χ2) test was used to analyze survival and ovulation rates of the follicles. Results Our results showed that the rate of metaphase II (MII) oocytes in FGF group increased in comparison with control and other treatment groups (p<0.027), but there was no difference between control with EGF and EGF+FGF groups in oocyte maturation rate (p>0.05). There was a significant decrease in survival rate of follicles in EGF+FGE group in comparison with other groups (p<0.008). After in vitro ovulation induction, the follicles in EGF group showed a higher ovulation rate (p<0.008) than those cultured in other groups. Conclusion FGF has beneficial effect on oocyte maturation, and EGF increases COCs number in vitro. Combination of EGF and FGE decreases the number of survived follicles. PMID:24611145

  3. The Invasion and Reproductive Toxicity of QDs-Transferrin Bioconjugates on Preantral Follicle in vitro

    PubMed Central

    Xu, Gaixia; Lin, Suxia; Law, Wing-Cheung; Roy, Indrajit; Lin, Xiaotan; Mei, Shujiang; Ma, Hanwu; Chen, Siping; Niu, Hanben; Wang, Xiaomei

    2012-01-01

    The toxicity of QD has been extensively studied over the past decade. However, the potential toxicity of QDs impedes its use for clinical research. In this work, we established a preantral follicle in vitro culture system to investigate the effects of QD-Transferrin (QDs-Tf) bioconjugates on follicle development and oocyte maturation. The preantral follicles were cultured and exposed to CdTe/ZnTe QDs-Tf bioconjugates with various concentrations and the reproductive toxicity was assessed at different time points post-treatment. The invasion of QDs-Tf for oocytes was verified by laser scanning confocal microscope. Steroid production was evaluated by immunoassay. C-band Giemsa staining was performed to observe the chromosome abnormality of oocytes. The results showed that the QDs-Tf bioconjugates could permeate into granulosa cells and theca cells, but not into oocyte. There are no obvious changes of oocyte diameter, the mucification of cumulus-oocyte-complexes and the occurrence of aneulpoidy as compared with the control group. However, delay in the antrum formation and decrease in the ratio of oocytes with first polar body were observed in QDs-Tf-treated groups. The matured oocytes with first polar body decreased significantly by ~16% (from 79.6±10 % to 63±2.9 %) when the concentration of QDs-Tf bioconjugates exceeded 2.89 nmol·L-1 (P < 0.05). Our results implied that the CdTe/ZnTe QDs-Tf bioconjugates were reproductive toxic for follicle development, and thus also revealed that this in vitro culture system of preantral follicle is a highly sensitive tool for study on the reproductive toxicity of nanoparticles. PMID:22916073

  4. Characterization of freshly retrieved preantral follicles using a low-invasive, mechanical isolation method extended to different ruminant species.

    PubMed

    Langbeen, A; Jorssen, E P A; Fransen, E; Rodriguez, A P A; García, M Chong; Leroy, J L M R; Bols, P E J

    2015-10-01

    Due to the increased interest in preantral follicular physiology, non-invasive retrieval and morphological classification are crucial. Therefore, this study aimed: (1) to standardize a minimally invasive isolation protocol, applicable to three ruminant species; (2) to morphologically classify preantral follicles upon retrieval; and (3) to describe morphological features of freshly retrieved follicles compared with follicle characteristics using invasive methods. Bovine, caprine and ovine ovarian cortex strips were retrieved from slaughterhouse ovaries and dispersed. This suspension was filtered, centrifuged, re-suspended and transferred to a Petri dish, to which 0.025 mg/ml neutral red (NR) was added to assess the viability of the isolated follicles. Between 59 and 191 follicles per follicle class and per species were collected and classified by light microscopy, based on follicular cell morphology. Subsequently, follicle diameters were measured. The proposed isolation protocol was applicable to all three species and showed a significant, expected increase in diameter with developmental stage. With an average diameter of 37 ± 5 μm for primordial follicles, 47 ± 6.3 μm for primary follicles and 67.1 ± 13.1 μm for secondary follicles, no significant difference in diameter among the three species was observed. Bovine, caprine and ovine follicles (63, 59 and 50% respectively) were graded as viable upon retrieval. Using the same morphological characteristics as determined by invasive techniques [e.g. haematoxylin-eosin (HE) sections], cumulus cell morphology and follicle diameter could be used routinely to classify freshly retrieved follicles. Finally, we applied a mechanical, minimally invasive, follicle isolation protocol and extended it to three ruminant species, yielding viable preantral follicles without compromising further in vitro processing and allowing routine follicle characterization upon retrieval.

  5. Survival and growth of human preantral follicles after cryopreservation of ovarian tissue, follicle isolation and short-term xenografting.

    PubMed

    Paulini, Fernanda; Vilela, Janice M V; Chiti, Maria Costanza; Donnez, Jacques; Jadoul, Pascale; Dolmans, Marie-Madeleine; Amorim, Christiani A

    2016-09-01

    In women, chemotherapy and radiotherapy can be harmful to the ovaries, causing loss of endocrine and reproductive functions. When gonadotoxic treatment cannot be delayed, ovarian tissue cryobanking is the only way of preserving fertility. This technique, however, is not advisable for patients with certain types of cancer, because of the risk of reintroducing malignant cells present in the cryopreserved tissue. Our objective is therefore to develop a transplantable artificial ovary. To this end, cryopreserved human preantral follicles were isolated and embedded in fibrin formulations prepared with 50 mg/ml fibrinogen and 10 IU/ml thrombin supplemented or not with 3% hyaluronic acid, and respectively xenografted to specially created right and left peritoneal pockets in eight nude mice. On days 0 and 7, the animals were killed and the matrices retrieved. On day 7, no difference was observed in the recovery rate of follicles embedded in fibrin alone (23.4%) or fibrin-hyaluronic acid (20.5%). Ki67 staining confirmed growth of the grafted follicles and terminal deoxynucleotidyl transferase)-mediated dUDP nick-end labelling assay revealed 100% of the follicles to be viable in both groups on day 7. In conclusion, fibrin seems to be a promising material for creation of an artificial ovary, supporting follicle survival and development. PMID:27210771

  6. Short-term storage of canine preantral ovarian follicles using a powdered coconut water (ACP)-based medium.

    PubMed

    Lima, G L; Costa, L L M; Cavalcanti, D M L P; Rodrigues, C M F; Freire, F A M; Fontenele-Neto, J D; Silva, A R

    2010-07-01

    The objective was to investigate the use of powdered coconut water (ACP)-based medium for short-term preservation of canine preantral follicles. Pairs of ovaries from mongrel bitches (n=9) were divided into fragments. One ovarian fragment, treated as a fresh control, was immediately fixed for histological analysis, whereas the other six ovarian fragments were stored either in phosphate-buffered saline (PBS; control group) or ACP medium in isothermal Styrofoam boxes containing biological ice packs. The boxes were sealed and opened only after 12, 24, or 36h. After opening each box, the ovarian fragments were submitted to histological analysis. In total, 12,302 preantral follicles were evaluated, with 64.5% primordial, 33.3% primary, and 2.3% secondary follicles. There were multiple oocytes in 1.3% of the follicles analyzed. At 24h, ACP was more efficient in preserving follicular morphology than PBS (P<0.05). Compared with the fresh control group, a significant reduction in the percentage of morphologically normal ovarian follicles was observed for PBS, starting at 24h; however, the decline started only at 36h for the ACP medium. During the experiment, the temperature inside the isothermal boxes increased from 3 to 9 degrees C (P<0.05), despite a constant room temperature. In conclusion, powdered coconut water (ACP) was an appropriate medium for short-term storage of canine preantral ovarian follicles.

  7. Short-term storage of canine preantral ovarian follicles using a powdered coconut water (ACP)-based medium.

    PubMed

    Lima, G L; Costa, L L M; Cavalcanti, D M L P; Rodrigues, C M F; Freire, F A M; Fontenele-Neto, J D; Silva, A R

    2010-07-01

    The objective was to investigate the use of powdered coconut water (ACP)-based medium for short-term preservation of canine preantral follicles. Pairs of ovaries from mongrel bitches (n=9) were divided into fragments. One ovarian fragment, treated as a fresh control, was immediately fixed for histological analysis, whereas the other six ovarian fragments were stored either in phosphate-buffered saline (PBS; control group) or ACP medium in isothermal Styrofoam boxes containing biological ice packs. The boxes were sealed and opened only after 12, 24, or 36h. After opening each box, the ovarian fragments were submitted to histological analysis. In total, 12,302 preantral follicles were evaluated, with 64.5% primordial, 33.3% primary, and 2.3% secondary follicles. There were multiple oocytes in 1.3% of the follicles analyzed. At 24h, ACP was more efficient in preserving follicular morphology than PBS (P<0.05). Compared with the fresh control group, a significant reduction in the percentage of morphologically normal ovarian follicles was observed for PBS, starting at 24h; however, the decline started only at 36h for the ACP medium. During the experiment, the temperature inside the isothermal boxes increased from 3 to 9 degrees C (P<0.05), despite a constant room temperature. In conclusion, powdered coconut water (ACP) was an appropriate medium for short-term storage of canine preantral ovarian follicles. PMID:20207405

  8. Investigations of TGF-β signaling in preantral follicles of female mice reveal differential roles for bone morphogenetic protein 15.

    PubMed

    Fenwick, Mark A; Mora, Jocelyn M; Mansour, Yosef T; Baithun, Christina; Franks, Stephen; Hardy, Kate

    2013-09-01

    Bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) are 2 closely related TGF-β ligands implicated as key regulators of follicle development and fertility. Animals harboring mutations of these factors often exhibit a blockage in follicle development beyond the primary stage and therefore little is known about the role of these ligands during subsequent (preantral) stages. Preantral follicles isolated from immature mice were cultured with combinations of BMP15, GDF9, and activin receptor-like kinase (ALK) inhibitors. Individually, GDF9 and BMP15 promoted follicle growth during the first 24 hours, whereas BMP15 subsequently (48-72 h) caused follicle shrinkage and atresia with increased granulosa cell apoptosis. Inhibition of ALK6 prevented the BMP15-induced reduction in follicle size and under basal conditions promoted a rapid increase in granulosa cell proliferation, suggesting BMP15 signals through ALK6, which in turn acts to restrain follicle growth. In the presence of GDF9, BMP15 no longer promoted atresia and in fact follicle growth was increased significantly more than with either ligand alone. This cooperative effect was accompanied by differential expression of Id1-3, Smad6-7, and Has2 and was blocked by the same ALK5 inhibitor used to block GDF9 signaling. Immunostaining for SMAD2/3 and SMAD1/5/8, representing the 2 main branches of TGF-β signaling, supported the fact that both canonical pathways have the potential to be active in growing follicles, whereas primordial follicles only express SMAD2/3. Overall results highlight differential effects of the 2 main TGF-β signaling pathways during preantral follicle growth.

  9. In vitro development of secondary follicles from pre-pubertal and adult goats cultured in two-dimensional or three-dimensional systems.

    PubMed

    Silva, G M; Rossetto, R; Chaves, R N; Duarte, A B G; Araújo, V R; Feltrin, C; Bernuci, M P; Anselmo-Franci, J A; Xu, M; Woodruff, T K; Campello, C C; Figueiredo, J R

    2015-08-01

    The aim of this study was to evaluate the influence of two-dimensional (2D) and three-dimensional (3D) alginate culture systems on in vitro development of pre-antral caprine follicles. In addition, the influence of the reproductive age of the ovary donor on the in vitro culture success was investigated. Pre-antral follicles from pre-pubertal or adult goats were isolated and cultured directly on a plastic surface (2D) or encapsulated in an alginate-based matrix (3D). After 18 days, the oocytes underwent in vitro maturation (IVM) and in vitro fertilization (IVF) to produce embryos. The 3D system showed higher rates of follicle survival, lower rates of oocyte extrusion, and a greater number of recovered oocytes for IVM and IVF (P < 0.05). Only pre-antral follicles from adult animals produced MII oocytes and embryos. The estradiol concentrations increased from day 2 to day 12 of culture in all groups tested (P < 0.05). Conversely, progesterone concentrations were lower in 3D-cultured follicles than in 2D-cultured follicles, with differences on days 2 and 6 of culture (P < 0.05). We provide compelling evidence that a 2D or 3D alginate in vitro culture system offers a promising approach to achieving full in vitro development of caprine pre-antral follicles to produce mature oocytes that are capable of fertilization and viable embryos.

  10. FSH supplementation to culture medium is beneficial for activation and survival of preantral follicles enclosed in equine ovarian tissue.

    PubMed

    Aguiar, F L N; Lunardi, F O; Lima, L F; Rocha, R M P; Bruno, J B; Magalhães-Padilha, D M; Cibin, F W S; Nunes-Pinheiro, D C S; Gastal, M O; Rodrigues, A P R; Apgar, G A; Gastal, E L; Figueiredo, J R

    2016-04-01

    This study investigated the effect of adding different concentrations of bovine recombinant follicle-stimulating hormone on the IVC of equine preantral follicles enclosed in ovarian tissue fragments. Randomized ovarian fragments were fixed immediately (fresh noncultured control) or cultured for 1 or 7 days in α-MEM(+) supplemented with 0, 10, 50, and 100 ng/mL FSH and subsequently analyzed by classical histology. Culture media collected on Day 1 or Day 7 and were analyzed for steroids (estradiol and progesterone) and reactive oxygen species (ROS). After Day 1 and Day 7 of culture, 50-ng/mL FSH treatment had a greater (P < 0.05) percentage of morphologically normal follicles when compared to the other groups, except the 10-ng/mL FSH treatment at Day 1 of culture. The percentage of developing follicles (transition, primary, and secondary), and follicular and oocyte diameters were higher (P < 0.05) in the 50-ng/mL FSH treatment compared to the other groups after Day 7 of culture. Furthermore, estradiol secretion and ROS production were maintained (P > 0.05) throughout the culture in the 50-ng/mL FSH treatment. In conclusion, the addition of 50 ng/mL of FSH promoted activation of primordial follicles to developing follicles, improved survival of preantral follicles, and maintained estradiol and ROS production of equine ovarian tissue after 7 days of culture.

  11. Lower apoptosis rate in ovine preantral follicles from ovaries stored in supplemented preservation media.

    PubMed

    Gonçalves, R J S; Cavalcante, A Y P; Gouveia, B B; Lins, T L B; Barberino, R S; Menezes, V G; Barros, V R P; Macedo, T J S; Figueiredo, J R; Matos, M H T

    2015-12-01

    The aim of this study was to investigate the effect of ovarian tissue transportation conditions (medium and period of time) on the morphology, apoptosis and development of ovine preantral follicles cultured in vitro. Each ovarian pair was cut into nine slices, with one fragment being fixed immediately (fresh control). The remaining fragments were placed individually in cryotubes containing conservation medium (minimal essential medium (MEM) without supplementation or MEM+ - with supplementation) and stored at 35ºC for 6 or 12 h without (non-cultured) or with subsequent culture for 5 days. Then, the fragments were processed for histological and terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labelling (TUNEL) examination. Preservation of ovarian slices in MEM or MEM+ (non-cultured) resulted in similar percentages of normal follicles when compared with the fresh control. Nevertheless, compared with the fresh control, a decrease in the percentage of normal follicles was observed in tissues cultured for 5 days. Only for tissues preserved in supplemented medium (MEM+) for 6 h, the percentage of TUNEL positive cells was similar between non-cultured tissues and tissues cultured for 5 days. Follicular activation and growth (follicular and oocyte diameter) were higher in cultured tissues than in fresh control or non-cultured tissues, except those from fragments preserved for 6 h in MEM and then cultured for 5 days in which no growth was observed. In conclusion, ovine ovarian tissue was successfully preserved in supplemented medium (MEM+) at a temperature close to physiological values (35°C) for up to 6 h without affecting apoptosis in the ovarian follicles and their ability to develop in vitro.

  12. Growth differentiation factor-9 mediates follicle-stimulating hormone-thyroid hormone interaction in the regulation of rat preantral follicular development.

    PubMed

    Kobayashi, Noriko; Orisaka, Makoto; Cao, Mingju; Kotsuji, Fumikazu; Leader, Arthur; Sakuragi, Noriaki; Tsang, Benjamin K

    2009-12-01

    FSH regulates follicular growth in a stage-development fashion. Although preantral follicle stage is gonadotropin responsive, FSH is not required for preantral follicular growth. With the antrum, the follicles continue growing under the influence of FSH and become gonadotropin dependent. Although thyroid hormone is important for normal female reproductive function, its role and interaction with FSH in the regulation of preantral ovarian follicular growth is yet to be defined. In the present study, we have examined the action and interaction of FSH and T(3) in the regulation of the growth of preantral follicles, especially in their transition from preantral to early antral stage, using an established follicle culture system and evaluated the involvement of growth differentiation factor-9 (GDF-9) in this process in vitro. We have demonstrated that although T(3) alone had no effect on follicular development, it markedly enhanced FSH-induced preantral follicular growth. Although FSH alone significantly down-regulated FSH receptor (FSHR) mRNA abundance in the preantral follicles and T(3) alone was ineffective, expression of the message was significantly increased in the presence of both hormones. In addition, intra-oocyte injection of GDF-9 antisense oligonucleotides (GDF-9 morpholino) induced follicular cell apoptosis and suppressed follicular growth induced by FSH and T(3). These responses were attenuated by exogenous GDF-9. Our findings support the concept that thyroid hormone regulates ovarian follicular development through its direct action on the ovary and that promotes FSH-induced preantral follicular growth through up-regulation of FSHR, a mechanism dependent on the expression and action of oocyte-derived GDF-9.

  13. Effect of leptin on in vitro development of ovine preantral ovarian follicles.

    PubMed

    Kamalamma, P; Kona, S S R; Praveen Chakravarthi, V; Siva Kumar, A V N; Punyakumari, B; Rao, V H

    2016-01-15

    The influence of human or ovine leptin on in vitro culture of preantral follicles (PFs) isolated from sheep ovaries was investigated. Among the 12 different concentrations (0-1000 ng/mL) of human leptin tested, proportion of PFs exhibiting growth, mean increase in diameter, antrum formation, and maturation of the oocytes to MII stage were the best in 10 ng/mL. Culture of sheep ovarian PFs in TCM 199 supplemented with 10 ng/mL of human or ovine leptin FSH (2.5 μg/mL), thyroxine (1 μg/mL), insulinlike growth factor I (10 ng/mL), and GH (1 mIU/mL) resulted in significantly (P ≤ 0.05) greater average increase in diameter (11 and 9 vs. 6 μm), better proportions of PFs exhibiting growth (66% and 58% vs. 48%), antrum formation (51% and 51% vs. 34%), and maturation of oocytes to MII stage (24% and 22% vs. 7%) than the control medium. It is concluded that (1) the optimum dose of leptin for the growth of sheep PFs in vitro was 10 ng/mL, (2) human or ovine leptin supported similar development in vitro of PFs in sheep, (3) inclusion of leptin along with FSH, thyroxine, insulinlike growth factor I, and GH resulted in only a marginal further improvements in in vitro development of sheep PFs'.

  14. In Vitro Culture of Early Secondary Preantral Follicles in Hanging Drop of Ovarian Cell-Conditioned Medium to Obtain MII Oocytes from Outbred Deer Mice

    PubMed Central

    Choi, Jung Kyu; Agarwal, Pranay

    2013-01-01

    The ovarian follicle (each contains a single oocyte) is the fundamental functional tissue unit of mammalian ovaries. In humans, it has been long held true that females are born with a maximum number of follicles (or oocytes) that are not only nonrenewable, but also undergoing degeneration with time with a sharply decreased oocyte quality after the age of ∼35. Therefore, it is of importance to isolate and bank ovarian follicles for in vitro culture to obtain fertilizable oocytes later, to preserve the fertility of professional women who may want to delay childbearing, young and unmarried women who may lose gonadal function because of exposure to environmental/occupational hazards or aggressive medical treatments, such as radiation and chemotherapy, and even endangered species and breeds. Although they contributed significantly to the understanding of follicle science and biology, most studies reported to date on this topic were done using the man-made, unnatural inbred animal species. It was found in this study that the conventional two-dimensional microliter drop and three-dimensional hanging drop (HD) methods, reported to be effective for in vitro culture of preantral follicles from inbred mice, are not directly transferrable to outbred deer mice. Therefore, a modified HD method was developed in this study to achieve a much higher (>5 times compared to the best conventional methods) percentage of developing early secondary preantral follicles from the outbred mice to the antral stage, for which, the use of an ovarian cell-conditioned medium and multiple follicles per HD were identified to be crucial. It was further found that the method for in vitro maturation of oocytes in antral follicles obtained by in vitro culture of preantral follicles could be very different from that for oocytes in antral follicles obtained by hormone stimulation in vivo. Therefore, this study should provide important guidance for establishing effective protocols of in vitro follicle

  15. In vitro culture of early secondary preantral follicles in hanging drop of ovarian cell-conditioned medium to obtain MII oocytes from outbred deer mice.

    PubMed

    Choi, Jung Kyu; Agarwal, Pranay; He, Xiaoming

    2013-12-01

    The ovarian follicle (each contains a single oocyte) is the fundamental functional tissue unit of mammalian ovaries. In humans, it has been long held true that females are born with a maximum number of follicles (or oocytes) that are not only nonrenewable, but also undergoing degeneration with time with a sharply decreased oocyte quality after the age of ∼35. Therefore, it is of importance to isolate and bank ovarian follicles for in vitro culture to obtain fertilizable oocytes later, to preserve the fertility of professional women who may want to delay childbearing, young and unmarried women who may lose gonadal function because of exposure to environmental/occupational hazards or aggressive medical treatments, such as radiation and chemotherapy, and even endangered species and breeds. Although they contributed significantly to the understanding of follicle science and biology, most studies reported to date on this topic were done using the man-made, unnatural inbred animal species. It was found in this study that the conventional two-dimensional microliter drop and three-dimensional hanging drop (HD) methods, reported to be effective for in vitro culture of preantral follicles from inbred mice, are not directly transferrable to outbred deer mice. Therefore, a modified HD method was developed in this study to achieve a much higher (>5 times compared to the best conventional methods) percentage of developing early secondary preantral follicles from the outbred mice to the antral stage, for which, the use of an ovarian cell-conditioned medium and multiple follicles per HD were identified to be crucial. It was further found that the method for in vitro maturation of oocytes in antral follicles obtained by in vitro culture of preantral follicles could be very different from that for oocytes in antral follicles obtained by hormone stimulation in vivo. Therefore, this study should provide important guidance for establishing effective protocols of in vitro follicle

  16. Insulin improves in vitro survival of equine preantral follicles enclosed in ovarian tissue and reduces reactive oxygen species production after culture.

    PubMed

    Aguiar, F L N; Lunardi, F O; Lima, L F; Rocha, R M P; Bruno, J B; Magalhães-Padilha, D M; Cibin, F W S; Rodrigues, A P R; Gastal, M O; Gastal, E L; Figueiredo, J R

    2016-04-01

    This study investigated the effect of insulin concentration on the in vitro culture of equine preantral follicles enclosed in ovarian tissue. Ovarian tissue samples were immediately fixed (noncultured control) or cultured for 1 or 7 days in α-MEM(+) supplemented with 0 ng/mL, 10 ng/mL, or 10 μg/mL insulin. Ovarian tissues were processed and analyzed by classical histology. Culture medium samples were collected after 1 and 7 days of culture for steroid and reactive oxygen species (ROS) analyses. The percentage of morphologically normal follicles was greater (P < 0.001) in insulin-treated groups after 1 day of culture; likewise, more (P < 0.02) normal follicles were observed after 7 days of culture in medium supplemented with 10-ng/mL insulin. Furthermore, an increase (P < 0.01) in developing (transition, primary, and secondary) follicles between Days 1 and 7 of culture was observed only with the 10-ng/mL insulin treatment. ROS production after 1 or 7 days of culture was lower (P < 0.0001) in medium with 10-ng/mL insulin than the other treatments. Ovarian tissues containing preantral follicles were able to produce estradiol and progesterone after 1 and 7 days of culture; however, treatments did not differ in steroid production. In conclusion, the use of a physiological concentration (10 ng/mL) of insulin rather than the previously reported concentration (10 μg/mL) for in vitro culture of equine preantral follicles improved follicular survival and growth and lowered oxidative stress. Results from this study shed light on new perspectives for producing an appropriate medium to improve equine preantral follicle in vitro survival and growth.

  17. Improvement of development of equine preantral follicles after 6 days of in vitro culture with ascorbic acid supplementation.

    PubMed

    Gomes, R G; Lisboa, L A; Silva, C B; Max, M C; Marino, P C; Oliveira, R L; González, S M; Barreiros, T R R; Marinho, L S R; Seneda, M M

    2015-09-15

    The aim of this study was to evaluate the effects of different concentrations of ascorbic acid (25, 50, and 100 μg/mL) in supplemented minimum essential medium (MEM+) on the development of equine preantral follicles that were cultured in vitro for 2 or 6 days. The contralateral ovaries (n = 5) from five mares in seasonal anestrus were collected from a local abattoir. Nine ovarian tissue fragments of approximately 5 × 5 × 1 mm were obtained from each animal. One fragment was immediately fixed and subjected to histologic analysis (control group; Day 0), and the other eight were placed in PBS supplemented with penicillin (200 IU/mL) and streptomycin (200 mg/mL) at 4 °C for 1 hour (during transport to the laboratory). The fragments were cultured in situ for 2 days (D2) or 6 days (D6) in MEM+ or MEM+ plus ascorbic acid at three different concentrations, establishing the following nine groups: control; MEM+ (D2); MEM+ (D6); MEM+ 25 μg/mL of ascorbic acid (D2); MEM+ 25 μg/mL of ascorbic acid (D6); MEM+ 50 μg/mL of ascorbic acid (D2); MEM+ 50 μg/mL of ascorbic acid (D6); MEM+ 100 μg/mL of ascorbic acid (D2); and MEM+ 100 μg/mL of ascorbic acid (D6). The preantral follicles were classified according to their stage (primordial, primary, secondary, or antral) and their morphology (normal or abnormal). Slides (n = 951) including 4450 histologic sections were evaluated. Follicles were observed in only 4.85% (216 of 4450) of the histologic sections. Of the 407 follicles evaluated, 120 were in the primordial stage and 287 were in different developmental stages; additionally, 43.5% were morphologically normal. After 6 days of culture, the groups cultured with 50 and 100 μg/mL of ascorbic acid differed in terms of follicular development compared with the other groups. On the basis of occurrence of follicular development and the presence of viable follicles, it can be concluded that a positive effect of culture for 6 days in MEM+ supplemented with 50 and 100 μg/mL of

  18. Long-term in vitro culture of bovine preantral follicles: Effect of base medium and medium replacement methods.

    PubMed

    Araújo, V R; Gastal, M O; Wischral, A; Figueiredo, J R; Gastal, E L

    2015-10-01

    Two culture media and replacement methods were compared during long-term in vitro culture of secondary follicles of cattle using α-MEM(+) or TCM-199(+) as base media. The medium replacement methods were: Conventional - removal and subsequent addition of the same amount (60μl) in a 100μl aliquot (MEM-C and TCM-C), and Small Supplementation - addition of 5μl of fresh medium to an initial small aliquot (50μl), resulting in a final volume of 125μl on the last day of culture (MEM-S and TCM-S). A total of 207 secondary follicles were cultured individually for 32 days at 38.5°C in 5% CO2 and medium replacement was performed every other day. The MEM-S treatment resulted in a larger (P<0.01) follicular diameter, greater (P<0.02) growth rate, greater (P<0.02) antrum formation, as well as greater (P<0.0001) estradiol concentrations when compared with the MEM-C treatment. The medium change methods did not affect (P>0.05) the follicular and estradiol end points for TCM-199(+). The expression of the FSHR gene was greater (P<0.03) with the TCM-C than TCM-S treatment, while the relative amounts of mRNA for IGF1 was greater (P<0.02) with MEM-S than TCM-S treatments and for VEGF was greater (P<0.02) with MEM-C than TCM-C treatment. In conclusion, the type of base medium and the effect of periodic addition of medium differentially affected follicle development, estradiol production, and gene expression. Furthermore, α-MEM(+) can be used to replace TCM-199(+) for culture of preantral follicles of cattle if progressive addition of medium is used for medium change.

  19. Monitoring preantral follicle survival and growth in bovine ovarian biopsies by repeated use of neutral red and cultured in vitro under low and high oxygen tension.

    PubMed

    Jorssen, Ellen P A; Langbeen, An; Fransen, Erik; Martinez, Emilia L; Leroy, Jo L M R; Bols, Peter E J

    2014-08-01

    The development and optimization of preantral follicle culture methods are crucial in fertility preservation strategies. As preantral follicle dynamics are usually assessed by various invasive techniques, the need for alternative noninvasive evaluation tools exists. Recently, neutral red (NR) was put forward to visualize preantral follicles in situ within ovarian cortical fragments. However, intense light exposure of NR-stained tissues can lead to cell death because of increased reactive oxygen species production, which is also associated with elevated oxygen tension. Therefore, we hypothesize that after repeated NR staining, follicle viability and dynamics can be altered by changes in oxygen tension. In the present study, we aim (1) to determine whether NR can be used to repeatedly assess follicular growth, activation, and viability and (2) to assess the effect of a low (5% O2) or high (20% O2) oxygen tension on the viability, growth, and stage transition of preantral follicles cultured in vitro by means of repeated NR staining. Cortical slices (n = 132; six replicates) from bovine ovaries were incubated for 3 hours at 37 °C in a Leibovitz medium with 50 μg/mL NR. NR-stained follicles were evaluated in situ for follicle diameter and morphology. Next, cortical fragments were individually cultured in McCoy's 5A medium for 6 days at 37 °C, 5% CO2, and 5% or 20% O2. On Days 4 and 6, the fragments were restained by adding NR to the McCoy's medium and follicles were reassessed. In both low and high oxygen tension treatment groups, approximately 70% of the initial follicles survived a 6-day in vitro culture, but no significant difference in follicle survival on Day 4 or 6 could be observed compared with Day 0 (P > 0.05). A significant decrease in the number of primordial and increase in primary and secondary follicles was observed within 4 days of culture (P < 0.001). In addition, a significant increase of the mean follicle diameter in NR-stained follicles was

  20. Paracrine Regulation of Steroidogenesis in Theca Cells by Granulosa Cells Derived from Mouse Preantral Follicles

    PubMed Central

    Liu, Xiaoqiang; Qiao, Pengyun; Jiang, Aifang; Jiang, Junyi; Han, Haiyan; Wang, Li; Ren, Chune

    2015-01-01

    Interaction partners of follicular cells play a significant role in steroidogenesis, follicular formation, and development. Androgen secreted by theca cells (TCs) can initiate follicle development and ovulation and provide precursor materials for estrogen synthesis. Therefore, studies on ovarian microenvironment will not only lead to better understanding of the steroidogenesis but also have clinical significance for ovarian endocrine abnormalities such as hyperandrogenism in polycystic ovary syndrome (PCOS). This study applied the Transwell coculture model to investigate if the interaction between granulosa and theca cells may affect androgen production in theca cells. Concentrations of testosterone and androstenedione in the spent medium were measured by radioimmunoassay and enzyme linked immunosorbent assay, respectively. The results show that the coculture with granulosa cells (GCs) increases steroidogenesis in TCs. In addition, testosterone and androstenedione productions in response to LH stimulation were also increased in the coculture model. Significantly increased mRNA expressions of steroidogenic enzymes (Star, Cyp11a1, Cyp17a1, and Hsd3b2) were observed in the cocultured TCs. Thus, GCs were capable of promoting steroidogenesis and LH responsiveness in TCs. This study provided a basis for further exploration of ovarian endocrine mechanism and pathologies. PMID:26357661

  1. Prepubertal goat oocytes from large follicles result in similar blastocyst production and embryo ploidy than those from adult goats.

    PubMed

    Romaguera, R; Moll, X; Morató, R; Roura, M; Palomo, M J; Catalá, M G; Jiménez-Macedo, A R; Hammami, S; Izquierdo, D; Mogas, T; Paramio, M T

    2011-07-01

    Developmental competence of oocytes from prepubertal females is lower than those from adult females. Oocyte development competence is positively related to follicular diameter. Most of the follicles of prepubertal goat ovaries are smaller than 3 mm. The aim of this study was to compare oocytes of two follicle sizes (< 3 mm and ≥ 3 mm) from prepubertal goats with oocytes from adult goats in relation to their in vitro production and quality of blastocysts. Oocytes from prepubertal goats were obtained from slaughterhouse ovaries and selected according to the follicle diameter whereas oocytes from adult goats were recovered in vivo by LOPU technique without prior selection of follicle size. COCs were IVM for 27 h, IVF at the conventional conditions with fresh semen and presumptive zygotes were cultured in SOF medium for 8 days. Blastocysts obtained were vitrified and after warming their blastocoele re-expansion and the ploidy by FISH technique were assessed. We found significant differences between blastocysts yield of oocytes recovered from follicles smaller than 3 mm of prepubertal goats compared to those from adult goats (5.45% vs 20. 83%, respectively) however, these differences disappear if oocytes were recovered form large follicles (18.07%). A total of 28 blastocysts were analysed and 96.43% showed mixoploidy. Age did not affect the number of embryos with abnormal ploidy or blastocyst re-expansion after warming. Furthermore, the percentage of diploid blastomeres per embryo was similar in the 3 groups studied, adult, prepubertal from follicles ≥ 3 mm and < 3 mm (68.6%, 80.8% and 73.6%, respectively). In conclusion, IVP of blastocysts coming from follicles larger than 3 mm of goats 45 days old were not different to the blastocysts produced from adult goats, both in terms of quantity and quality.

  2. Hair follicle transcriptome profiles during the transition from anagen to catagen in Cashmere goat (Capra hircus).

    PubMed

    Fan, Y X; Wu, R B; Qiao, X; Zhang, Y J; Wang, R J; Su, R; Wu, J H; Dong, Y; Li, J Q

    2015-12-22

    Previous molecular genetic studies of the goat hair life cycle have focused primarily on a limited number of genes and proteins. To identify additional genes that may play important roles in hair follicle cycle regulation, Illumina sequencing technology was used to catalog differential gene expression profiles in the hair growth cycle (anagen to catagen) of goat, comparing the primary hair follicle with the secondary hair follicle. There were 13,769 and 12,240 unigenes assembled from the reads obtained from primary hair follicle and secondary hair follicle, respectively. Genes encoding keratin proteins and keratin-associated proteins were the most highly expressed. A total of 5899 genes were differentially expressed in anagen vs catagen primary hair follicles, with 532 genes up-regulated and 5367 genes down-regulated. A total of 5208 genes were differentially expressed in anagen vs catagen secondary hair follicle, including 545 genes that were up-regulated and 4663 genes that were down-regulated. Numerous hair growth genes are expressed in the goat hair follicle, of which 73 genes showed co-up-regulation in both hair follicles during the anagen stage. Many of these up-regulated genes, such as STC2, VEGFR, and ROR2, are known to be transfactors in the process of cell differentiation and in the cell cycle. The differential gene expression profiles between primary hair follicles and secondary hair follicles obtained provide a foundation for future studies examining the network of gene expression controlling hair growth cycle in Cashmere goat.

  3. Effect of heat stress on the survival and development of in vitro cultured bovine preantral follicles and on in vitro maturation of cumulus-oocyte complex.

    PubMed

    Paes, V M; Vieira, L A; Correia, H H V; Sa, N A R; Moura, A A A; Sales, A D; Rodrigues, A P R; Magalhães-Padilha, D M; Santos, F W; Apgar, G A; Campello, C C; Camargo, L S A; Figueiredo, J R

    2016-09-01

    The deleterious effect of heat stress (HS) on competence of oocytes from antral follicles is well recognized, but there is a lack of data regarding its impact on the viability and growth of preantral follicles. In this study, we used in vitro preantral follicle cultures to investigate the effects of HS on the following parameters: survival and development of primordial follicles after in vitro culture of ovarian fragments (experiment I); growth and antrum formation of isolated advanced secondary follicles (experiment II); and maturation rates after in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) from antral follicles (>2-6 mm) grown in vivo (experiment III). Furthermore, the following end points were evaluated in all experiments: follicle/oocyte survival, reactive oxygen species (ROS), estradiol (E2) and progesterone (P4) production, as well as mRNA expression for select genes related to stress (HSP70) and apoptosis (MCL1 and BAX). In all experiments, HS consisted of exposing the structures (ovarian fragments, isolated preantral follicles and COCs) to 41 °C for 12 hours and then to 38.5 °C until the end of the culture (7 days for experiments I and II and 24 hours for experiment III). The temperature for the control group was held at 38.5 °C for the entire culture period. Heat stress increased (P < 0.05) the percentage of developing follicles (intermediate, primary, and secondary follicles) at 12 hours and increased levels of ROS at all evaluated time points (12, 24 hours, and D7), when compared to the control (experiment I). Heat stress did not affect (P > 0.05) any identified end points when preantral follicles were cultured in their isolated form (experiment II). However, in experiment III, HS decreased (P < 0.05) both the rates of metaphase II after 24 hours and E2 production at 12 hours of IVM. Moreover, HS increased (P < 0.0001) levels of P4 after IVM and ROS production at every evaluated time point, compared with the

  4. Catalase prevents lipid peroxidation and enhances survival of caprine preantral follicles cryopreserved in a 1,2-propanediol-freezing medium.

    PubMed

    Luz, Hiedely K M; Santos, Regiane R; Wanderley, Livia S; Faustino, Luciana R; Silva, Cleidson M G; Carvalho, Adeline A; Campello, Claudio C; Santos, Francielli W; Figueiredo, José R; Rodrigues, Ana Paula R

    2012-08-01

    The objectives of this study were to determine: 1) the optimal concentration (1.0 or 1.5 M) and duration of exposure (5, 10, or 20 min) of ovarian tissue to 1,2-propanediol (PROH) on morphology and viability of caprine preantral follicles; and 2) the effect of supplementing cryopreservation medium supplementation with Trolox(®) (0.1, 0.5, or 1.0 mM) or catalase (5, 10, or 20 IU/mL) on follicular morphology, viability, and lipid peroxidation. Cryopreservation decreased (p<0.05) percentages of normal follicles relative to the control (84%). Although supplementation of the cryopreservation medium (1.0 M PROH) with catalase (10 or 20 IU/mL) or Trolox(®) (0.1 mM) resulted in follicular morphology and viability similar to that in the controls (P>0.05), lipid peroxidation was reduced only when 20 IU/mL catalase was added to the cryopreservation medium.

  5. Fiber growth initiation in hair follicles of goats treated with melatonin.

    PubMed

    Nixon, A J; Choy, V J; Parry, A L; Pearson, A J

    1993-09-15

    The sequence of structural changes in goat hair follicles was investigated using melatonin implants to advance and synchronize spring hair growth. Ten pasture fed cashmere wethers each received a controlled release formulation of 70 mg of melatonin on September 1 1989, and showed plasma melatonin elevated above physiological levels over 14 days post-treatment (914 +/- 154 pg/ml [mean +/- SEM] on day 14). In ten untreated animals, daytime plasma melatonin was 19.9 +/- 4.7 pg/ml. Histological examination of skin biopsies taken over the 14 days from the start of the experiment showed that primary hair follicles of goats with manipulated hormone levels had initiated fiber growth (entered proanagen), whereas primary follicles of untreated goats largely remained in the quiescent phase (telogen). A standardized terminology was used to describe the sequence of events during induced proanagen. Structural reorganization of follicles began in treated animals between days 6 and 12 post-treatment, and emergent fibers grew by day 24. Advancement of spring fiber growth was associated with a suppression of the normal rise in plasma prolactin concentration. Prolactin levels in untreated goats increased from 7.4 +/- 1.8 ng/ml on day 1 to 12.8 +/- 1.6 ng/ml on day 14, but declined in treated goats from 6.3 +/- 2.3 ng/ml to 2.2 +/- 0.8 ng/ml over the same period. PMID:8376951

  6. Expression of TGF-beta superfamily growth factors, their receptors, the associated SMADs and antagonists in five isolated size-matched populations of pre-antral follicles from normal human ovaries.

    PubMed

    Kristensen, Stine Gry; Andersen, Kasper; Clement, Christian Alexandro; Franks, Stephen; Hardy, Kate; Andersen, Claus Yding

    2014-04-01

    In mammals, members of the transforming growth factor-beta (TGF-β) superfamily are known to have key roles in the regulation of follicular growth and development. The aim of the study was to evaluate the expression of TGF-β superfamily growth factors, their receptors, downstream SMAD signalling molecules and TGF-β/bone morphogenetic protein (BMP) antagonists during early human folliculogenesis. Human pre-antral follicles were enzymatically isolated from surplus ovarian tissue obtained from women having ovarian cortical tissue frozen for fertility preservation. A total of 348 human pre-antral follicles, ranging from 40 to 200 µm in diameter, were isolated from ovarian tissue obtained from 15 women, aged 24-34 years. Isolated pre-antral follicles were grouped according to diameter in five size-matched populations spanning the primordial, primary and secondary stage follicles and analysed by whole-genome microarray analysis. Selected proteins/genes were analysed by immunocytochemistry and quantitative RT-PCR. TGF-β superfamily genes with overall highest mRNA expressions levels included growth differentiation factors 9 (GDF9), BMP15, BMP6, BMP-receptor-2 (BMPR2), anti-Müllerian hormone receptor 2 (AMHR2), TGFβR3, inhibin-α (INHA) and intracellular SMAD3 and SMAD4. Moreover, genes which were differentially expressed from the primordial to the late secondary stage follicles included GDF9, BMP15, AMH, INHBB, TGFβR3, SMAD4 and antagonists Follistatin (FST) and GREM1. Collectively, these data indicate that the active TGF-β superfamily pathways in early human folliculogenesis consist of primarily GDF9 combined with possible synergistic effects of BMP15 through the BMPR2 and intracellular activation of SMAD3 and SMAD4, and that AMH and INHBB are engaged in intrafollicular events from the onset of follicular growth. Moreover, the presence of multiple TGF-β/BMP antagonists imply that certain growth factors are subjected to local regulation on different levels that

  7. Effects of jacalin and follicle-stimulating hormone on in vitro goat primordial follicle activation, survival and gene expression.

    PubMed

    Ribeiro, Regislane P; Portela, Antonia M L R; Silva, Anderson W B; Costa, José J N; Passos, José R S; Cunha, Ellen V; Souza, Glaucinete B; Saraiva, Márcia V A; Donato, Mariana A M; Peixoto, Christina A; van den Hurk, Robert; Silva, José R V

    2015-08-01

    This study aims to investigate the effects of jacalin and follicle-stimulating hormone (FSH) on activation and survival of goat primordial follicles, as well as on gene expression in cultured ovarian tissue. Ovarian fragments were cultured for 6 days in minimum essential medium (MEM) supplemented with jacalin (10, 25, 50 or 100 μg/ml - Experiment 1) or in MEM supplemented with jacalin (50 μg/ml), FSH (50 ng/ml) or both (Experiment 2). Non-cultured and cultured tissues were processed for histological and ultrastructural analysis. Cultured tissues from Experiment 2 were also stored to evaluate the expression of BMP-15, KL (Kit ligand), c-kit, GDF-9 and proliferating cell nuclear antigen (PCNA) by real-time polymerase chain reaction (PCR). The results of Experiment 1 showed that, compared with tissue that was cultured in control medium, the presence of 50 μg/ml of jacalin increased both the percentages of developing follicles and viability. In Experiment 2, after 6 days, higher percentages of normal follicles were observed in tissue cultured in presence of FSH, jacalin or both, but no synergistic interaction between FSH and jacalin was observed. These substances had no significant effect on the levels of mRNA for BMP-15 and KL, but FSH increased significantly the levels of mRNA for PCNA and c-kit. On the other hand, jacalin reduced the levels of mRNA for GDF-9. In conclusion, jacalin and FSH are able to improve primordial follicle activation and survival after 6 days of culture. Furthermore, presence of FSH increases the expression of mRNA for PCNA and c-kit, but jacalin resulted in lower GDF-9 mRNA expression. PMID:24869637

  8. Oocytes from small and large follicles exhibit equal development competence following goat cloning despite their differences in meiotic and cytoplasmic maturation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Somatic cell nuclear transfer (SCNT) in animals has been around for nearly 20 years and has been successfully used for cloning of various livestock species. In this study, goat oocytes were collected from large follicles (>3mm) and small follicles (<3mm) to compare the success rate when used in goat...

  9. Transcriptome Sequencing Reveals Differences between Primary and Secondary Hair Follicle-derived Dermal Papilla Cells of the Cashmere Goat (Capra hircus)

    PubMed Central

    Yuan, Jianlong; Guo, Xudong; Liu, Dongjun

    2013-01-01

    The dermal papilla is thought to establish the character and control the size of hair follicles. Inner Mongolia Cashmere goats (Capra hircus) have a double coat comprising the primary and secondary hair follicles, which have dramatically different sizes and textures. The Cashmere goat is rapidly becoming a potent model for hair follicle morphogenesis research. In this study, we established two dermal papilla cell lines during the anagen phase of the hair growth cycle from the primary and secondary hair follicles and clarified the similarities and differences in their morphology and growth characteristics. High-throughput transcriptome sequencing was used to identify gene expression differences between the two dermal papilla cell lines. Many of the differentially expressed genes are involved in vascularization, ECM-receptor interaction and Wnt/β-catenin/Lef1 signaling pathways, which intimately associated with hair follicle morphogenesis. These findings provide valuable information for research on postnatal morphogenesis of hair follicles. PMID:24069460

  10. Insulin-like 3-induced rat preantral follicular growth is mediated by growth differentiation factor 9.

    PubMed

    Xue, Kai; Kim, Ji Young; Liu, Jia-yin; Tsang, Benjamin K

    2014-01-01

    The communication of somatic cells and oocytes by intrafollicular paracrine factors is essential for follicular growth in the ovary. Insulin-like 3 (INSL3) is a theca cell-secreted paracrine factor. Androgens and growth differentiation factor 9 (GDF9), an oocyte-derived growth factor, are essential for follicular development. Using a rat preantral follicle culture model, we examined in the present study the influence of INSL3 on preantral follicular growth and the molecular mechanisms involved. We have observed that the receptor for INSL3, relaxin/insulin-like family peptide receptor 2 (RXFP2), was exclusively expressed in oocytes. Recombinant INSL3 stimulated Gdf9 expression, preantral follicular growth, and testosterone synthesis in vitro. Inhibition of the cAMP/protein kinase A signaling pathway (with cAMP antagonist, 8-bromoadenosine 3',5'-cyclic monophosphorothioate, Rp-isomer) attenuated INSL3-induced Gdf9 expression and preantral follicular growth. Moreover, knocking down Gdf9 expression (with small interfering RNA) or inhibiting GDF9 signaling (with SB431542, an activin receptor-like kinase receptor 5 inhibitor, or specific inhibitor of mothers against decapentaplegic homolog 3) or androgen action (with flutamide, an androgen receptor antagonist) suppressed INSL3-induced preantral follicular growth. In addition, LH and DHT regulated the expression of Insl3 mRNA in preantral follicles. These observations suggest that INSL3 is a key theca cell-derived growth factor for preantral follicle and that its action is mediated by GDF9.

  11. Effect of nutrition on plasma progesterone levels, metabolic parameters and small follicles development in unstimulated goats reared under constant photoperiod regimen.

    PubMed

    Rondina, D; Freitas, V J F; Spinaci, M; Galeati, G

    2005-12-01

    Sixteen local adult goats were submitted for 9 weeks to 2.09 (high group) and 0.54 (low group) x dietary maintenance respectively. During the experimental period, goats were weighed, oestrus was detected and plasma insulin, urea, non-esterified fatty acids and progesterone concentrations were assessed. At the end of the experiment, ovarian small follicles population was studied by histological analysis. Final weight loss in low group was 18.37 +/- 2.02%, whereas weight gain of high group was 13.84 +/- 2.70%. Insulin and urea were lower in low group, while non-esterified fatty acids were significantly higher. A lower number of fasted goats was in oestrus or ovulated and an extended length of oestrus (p < 0.05) and a higher frequency of short or long cycles (p < 0.05) were also observed. Fed animals showed heavier ovaries (p < 0.01) and a lower number of primordial follicles (p < 0.05). In restricted goats a significant qualitative alteration of follicle classes involved in the initiation process of primordial pool was found. In this phase, granulosa thickness and oocyte size were the most affected (p < 0.01). However in small follicles beyond the primary stage no differences were found between the groups in either number or qualitative characteristics (p > 0.05). Collectively, these results indicate that opposite dietary intakes for a medium period induce a composite reproductive response in goats and can regulate the early onset of follicle growth. PMID:16324082

  12. Oestrous behaviour and development of preovulatory follicles in goats induced to ovulate using the male effect with and without progesterone priming.

    PubMed

    Gonzalez-Bulnes, A; Carrizosa, J A; Urrutia, B; Lopez-Sebastian, A

    2006-01-01

    The objective of the current study was to examine follicular dynamics in goats induced to ovulate by using the male effect with or without progesterone pretreatment. Ten Murcian-Granadina goats were treated with a single dose of 25 mg of progesterone in olive oil on the day of male effect induction (Day 0), whereas ten does remained untreated. Changes in the ovarian follicular population and onset of oestrous behaviour were checked daily from Day 0 to the day on which ovulation was detected by ultrasonography. All of the treated goats and only 20% of control goats (P < 0.001) showed oestrous behaviour between 72 and 96 h after the introduction of bucks. Differences in follicle growth were found between the groups. The control goats exhibiting oestrous behaviour during the first 4 days of ovarian screening and all of the goats treated with progesterone had their preovulatory follicles in growing phase. However, the largest follicles in control does not showing oestrus in these first 4 days were in regressing phase, being necessary for the emergence of new preovulatory follicles from a new follicular wave between Days 6 and 7, for exhibition of oestrous behaviour and subsequent ovulation. In conclusion, the appearance of short luteal phases and delayed ovulation in goats treated with the male effect during seasonal anoestrous depends on the stage of follicle development at the time of buck introduction. A single injection of progesterone stimulates the emergence of a new follicular wave, able to grow and give adequate ovulation after induction of the male effect. PMID:17032582

  13. Stable transfection and identification of a hair follicle-specific expression vector of IGFBP-5 in goat fetal fibroblasts.

    PubMed

    Wang, X J; Su, H M; Liang, Y; Wang, Y F; Guo, X D; Wang, Z G; Liu, D J

    2014-03-17

    The insulin-like growth factor-binding protein-5 (IGFBP-5) is one of the 6 members of the IGFBP family and is involved in the regulation of cell growth, apoptosis, and other IGF-stimulated signaling pathways. To determine the significance of IGFBP-5 in the Inner Mongolia Cashmere goat (Capra hircus), a hair follicle-specific expression vector of IGFBP-5, pCDsRed2-K-IGFBP5 (6.7 kb), was constructed by cloning IGFBP-5 downstream of the keratin-association protein (KAP)6-1 promoter and inserting this fragment into pCDsRed2, which contains a red fluorescent protein (DsRed) expression unit. Inner Mongolia Cashmere goat fetal fibroblast (GFb) cells were transfected with the expression vector by using Lipofectamine(TM) 2000. Cell clones that stably expressed red fluorescence were obtained after selection with Geneticin (G418). The transgene in the cell clones was examined by polymerase chain reaction to verify that exogenous DNA (pKAP6-1 and IGFBP-5) had integrated stably into GFb cells. These data suggest that this method can be used for the construction of a hair follicle-specific expression vector for functional genetic analyses and for obtaining stable transfection donor cells for nuclear transfer.

  14. Comparative Transcriptome Analysis of Fetal Skin Reveals Key Genes Related to Hair Follicle Morphogenesis in Cashmere Goats

    PubMed Central

    Yan, Hailong; Zeng, Jie; Ma, Sen; Niu, Yiyuan; Zhou, Guangxian; Jiang, Yu; Chen, Yulin

    2016-01-01

    Cashmere goat skin contains two types of hair follicles (HF): primary hair follicles (PHF) and secondary hair follicles (SHF). Although multiple genetic determinants associated with HF formation have been identified, the molecules that determine the independent morphogenesis of HF in cashmere goats remain elusive. The growth and development of SHF directly influence the quantity and quality of cashmere production. Here, we report the transcriptome profiling analysis of nine skin samples from cashmere goats using 60- and 120-day-old embryos (E60 and E120, respectively), as well as newborns (NB), through RNA-sequencing (RNA-seq). HF morphological changes indicated that PHF were initiated at E60, with maturation from E120, while differentiation of SHF was identified at E120 until formation of cashmere occurred after birth (NB). The RNA-sequencing analysis generated over 20.6 million clean reads from each mRNA library. The number of differentially expressed genes (DEGs) in E60 vs. E120, E120 vs. NB, and E60 vs. NB were 1,024, 0 and 1,801, respectively, indicating that no significant differences were found at transcriptomic levels between E120 and NB. Key genes including B4GALT4, TNC, a-integrin, and FGFR1, were up-regulated and expressed in HF initiation from E60 to E120, while regulatory genes such as GPRC5D, PAD3, HOXC13, PRR9, VSIG8, LRRC15, LHX2, MSX-2, and FOXN1 were up-regulated and expressed in HF keratinisation and hair shaft differentiation from E120 and NB to E60. Several genes belonging to the KRT and KRTAP gene families were detected throughout the three HF developmental stages. The transcriptional trajectory analyses of all DEGs indicated that immune privilege, glycosaminoglycan biosynthesis, extracellular matrix receptor interaction, and growth factor receptors all played dominant roles in the epithelial-mesenchymal interface and HF formation. We found that the Wnt, transforming growth factor-beta/bone morphogenetic protein, and Notch family members

  15. Comparative Transcriptome Analysis of Fetal Skin Reveals Key Genes Related to Hair Follicle Morphogenesis in Cashmere Goats.

    PubMed

    Gao, Ye; Wang, Xiaolong; Yan, Hailong; Zeng, Jie; Ma, Sen; Niu, Yiyuan; Zhou, Guangxian; Jiang, Yu; Chen, Yulin

    2016-01-01

    Cashmere goat skin contains two types of hair follicles (HF): primary hair follicles (PHF) and secondary hair follicles (SHF). Although multiple genetic determinants associated with HF formation have been identified, the molecules that determine the independent morphogenesis of HF in cashmere goats remain elusive. The growth and development of SHF directly influence the quantity and quality of cashmere production. Here, we report the transcriptome profiling analysis of nine skin samples from cashmere goats using 60- and 120-day-old embryos (E60 and E120, respectively), as well as newborns (NB), through RNA-sequencing (RNA-seq). HF morphological changes indicated that PHF were initiated at E60, with maturation from E120, while differentiation of SHF was identified at E120 until formation of cashmere occurred after birth (NB). The RNA-sequencing analysis generated over 20.6 million clean reads from each mRNA library. The number of differentially expressed genes (DEGs) in E60 vs. E120, E120 vs. NB, and E60 vs. NB were 1,024, 0 and 1,801, respectively, indicating that no significant differences were found at transcriptomic levels between E120 and NB. Key genes including B4GALT4, TNC, a-integrin, and FGFR1, were up-regulated and expressed in HF initiation from E60 to E120, while regulatory genes such as GPRC5D, PAD3, HOXC13, PRR9, VSIG8, LRRC15, LHX2, MSX-2, and FOXN1 were up-regulated and expressed in HF keratinisation and hair shaft differentiation from E120 and NB to E60. Several genes belonging to the KRT and KRTAP gene families were detected throughout the three HF developmental stages. The transcriptional trajectory analyses of all DEGs indicated that immune privilege, glycosaminoglycan biosynthesis, extracellular matrix receptor interaction, and growth factor receptors all played dominant roles in the epithelial-mesenchymal interface and HF formation. We found that the Wnt, transforming growth factor-beta/bone morphogenetic protein, and Notch family members

  16. Discovery of cashmere goat (Capra hircus) microRNAs in skin and hair follicles by Solexa sequencing

    PubMed Central

    2013-01-01

    Background MicroRNAs (miRNAs) are a large family of endogenous, non-coding RNAs, about 22 nucleotides long, which regulate gene expression through sequence-specific base pairing with target mRNAs. Extensive studies have shown that miRNA expression in the skin changes remarkably during distinct stages of the hair cycle in humans, mice, goats and sheep. Results In this study, the skin tissues were harvested from the three stages of hair follicle cycling (anagen, catagen and telogen) in a fibre-producing goat breed. In total, 63,109,004 raw reads were obtained by Solexa sequencing and 61,125,752 clean reads remained for the small RNA digitalisation analysis. This resulted in the identification of 399 conserved miRNAs; among these, 326 miRNAs were expressed in all three follicular cycling stages, whereas 3, 12 and 11 miRNAs were specifically expressed in anagen, catagen, and telogen, respectively. We also identified 172 potential novel miRNAs by Mireap, 36 miRNAs were expressed in all three cycling stages, whereas 23, 29 and 44 miRNAs were specifically expressed in anagen, catagen, and telogen, respectively. The expression level of five arbitrarily selected miRNAs was analyzed by quantitative PCR, and the results indicated that the expression patterns were consistent with the Solexa sequencing results. Gene Ontology and KEGG pathway analyses indicated that five major biological pathways (Metabolic pathways, Pathways in cancer, MAPK signalling pathway, Endocytosis and Focal adhesion) accounted for 23.08% of target genes among 278 biological functions, indicating that these pathways are likely to play significant roles during hair cycling. Conclusions During all hair cycle stages of cashmere goats, a large number of conserved and novel miRNAs were identified through a high-throughput sequencing approach. This study enriches the Capra hircus miRNA databases and provides a comprehensive miRNA transcriptome profile in the skin of goats during the hair follicle cycle. PMID

  17. Melatonin regulating the expression of miRNAs involved in hair follicle cycle of cashmere goats skin.

    PubMed

    Fu, Shaoyin; Zhao, Hongli; Zheng, Zhuqing; Li, Jinquan; Zhang, Wenguang

    2014-12-01

    Melatonin and microRNAs (miRNAs) play important roles in regulating hair follicle development. However, the effect of melatonin on the expression pattern of miRNAs in skin and follicle of cashmere goats remain largely undefined. To explore the mechanism of melatonin affecting cashmere growth mediated by miRNAs, the effect of melatonin implants administered in Nei Mongol cashmere goats was assessed. In the experiment, five yearling does were implanted with melatonin, with the remaining other five females as control group. The expression of six candidate miRNAs was quantified by reverse transcription-real time polymerase chain reaction (RT-qPCR). The results indicated that melatonin significantly altered the expression pattern of miRNAs. Except for let-7a, the expression levels of miR-203, miR-205, miR-96, miR-183 and miR-199a occur three transitions during a cashmere cycle; melatonin changed the co-expression pattern of miRNAs. The correlation coefficient between miRNAs is 0.87-0.99 in control group(P<0.01). Compared with the control group, the correlation coefficient between some miRNAs (let-7a and miR-96, miR-199a, miR-205;miR-203 and miR-96, miR-199a; miR-96 and miR-183; miR-183 and miR-199a) was significantly weakened in melatonin group. Melatonin might induce second growth of cashmere mediated by down-regulating the expression level of some miRNAs in June in melatonin implanted group. PMID:25487268

  18. Fibroblast growth factor 5-short (FGF5s) inhibits the activity of FGF5 in primary and secondary hair follicle dermal papilla cells of cashmere goats.

    PubMed

    He, Xiaolin; Chao, Yuan; Zhou, Guangxian; Chen, Yulin

    2016-01-10

    To determine the relationship between fibroblast growth factor 5 (FGF5) and FGF5-short (FGF5s) in dermal papilla cells of cashmere goat primary and secondary hair follicles. We isolated dermal papilla cells from primary hair follicle (PHF) and secondary hair follicle (SHF) of cashmere goat, and found that the FGF5 receptor, fibroblast growth factor receptor 1 (FGFR1), was expressed in these two types of dermal papilla cells. Moreover, adenovirus-mediated overexpression of FGF5 could upregulate the mRNA expression of insulin-like growth factor-1 (IGF-1), versican and noggin that were important for follicle growth maintenance, whereas downregulate the expression of anagen chalone bone morphogenetic protein 4 (BMP4) in dermal papilla cells. However, these alterations were partly reversed by FGF5s overexpression. In conclusion, our results demonstrated that FGF5s acted as an inhibitor of FGF5 in the regulation of anagen-catagen transition of cashmere goat dermal papilla cells.

  19. Molecular characterization, expression and methylation status analysis of BMP4 gene in skin tissue of Liaoning cashmere goat during hair follicle cycle.

    PubMed

    Bai, Wen L; Dang, Yun L; Wang, Jiao J; Yin, Rong H; Wang, Ze Y; Zhu, Yu B; Cong, Yu Y; Xue, Hui L; Deng, Liang; Guo, Dan; Wang, Shi Q; Yang, Shu H

    2016-08-01

    Bone morphogenetic protein 4 (BMP4) is a member of the bone morphogenetic protein family (BMPs). It is involved in the development and cycle of hair follicle, as well as, is thought to be a potential candidate gene for cashmere traits in goats. In the present study, we isolated and characterized a full-length open reading frame (ORF) of BMP4 cDNA from the skin tissue of Liaoning cashmere goat, and investigated the transcriptional pattern and methylation status of BMP4 gene in skin tissue of this breed during different stages of hair follicle cycle. The sequence analysis indicated that the isolated cDNA was 1264-bp in length containing a complete ORF of 1230-bp. It encoded a precursor peptide of 409 amino acids with a signal peptide of 19 amino acids. The structural analysis indicated that goat BMP4 contains typical TGF-β propeptide and TGF-β domains. In skin tissue, BMP4 is generally transcribed in an ascendant pattern from anagen to telogen. The methylation level of 5' flanking regulatory region of BMP4 gene might be involved in its mRNA expression in skin tissue: a higher BMP4 methylation level in skin coincides with a lower expression of BMP4 mRNA. These results from the present work provided a foundation for further insight into the functional and regulatory characteristics of BMP4 in the development and cycle of hair follicle in Liaoning Cashmere goat. PMID:27406581

  20. Molecular characterization, expression and methylation status analysis of BMP4 gene in skin tissue of Liaoning cashmere goat during hair follicle cycle.

    PubMed

    Bai, Wen L; Dang, Yun L; Wang, Jiao J; Yin, Rong H; Wang, Ze Y; Zhu, Yu B; Cong, Yu Y; Xue, Hui L; Deng, Liang; Guo, Dan; Wang, Shi Q; Yang, Shu H

    2016-08-01

    Bone morphogenetic protein 4 (BMP4) is a member of the bone morphogenetic protein family (BMPs). It is involved in the development and cycle of hair follicle, as well as, is thought to be a potential candidate gene for cashmere traits in goats. In the present study, we isolated and characterized a full-length open reading frame (ORF) of BMP4 cDNA from the skin tissue of Liaoning cashmere goat, and investigated the transcriptional pattern and methylation status of BMP4 gene in skin tissue of this breed during different stages of hair follicle cycle. The sequence analysis indicated that the isolated cDNA was 1264-bp in length containing a complete ORF of 1230-bp. It encoded a precursor peptide of 409 amino acids with a signal peptide of 19 amino acids. The structural analysis indicated that goat BMP4 contains typical TGF-β propeptide and TGF-β domains. In skin tissue, BMP4 is generally transcribed in an ascendant pattern from anagen to telogen. The methylation level of 5' flanking regulatory region of BMP4 gene might be involved in its mRNA expression in skin tissue: a higher BMP4 methylation level in skin coincides with a lower expression of BMP4 mRNA. These results from the present work provided a foundation for further insight into the functional and regulatory characteristics of BMP4 in the development and cycle of hair follicle in Liaoning Cashmere goat.

  1. Disruption of FGF5 in Cashmere Goats Using CRISPR/Cas9 Results in More Secondary Hair Follicles and Longer Fibers

    PubMed Central

    Zhu, Haijing; Niu, Yiyuan; Ma, Baohua; Yu, Honghao; Lei, Anmin; Yan, Hailong; Shen, Qiaoyan; Shi, Lei; Zhao, Xiaoe; Hua, Jinlian; Huang, Xingxu; Qu, Lei; Chen, Yulin

    2016-01-01

    Precision genetic engineering accelerates the genetic improvement of livestock for agriculture and biomedicine. We have recently reported our success in producing gene-modified goats using the CRISPR/Cas9 system through microinjection of Cas9 mRNA and sgRNAs targeting the MSTN and FGF5 genes in goat embryos. By investigating the influence of gene modification on the phenotypes of Cas9-mediated goats, we herein demonstrate that the utility of this approach involving the disruption of FGF5 results in increased number of second hair follicles and enhanced fiber length in Cas9-mediated goats, suggesting more cashmere will be produced. The effects of genome modifications were characterized using H&E and immunohistochemistry staining, quantitative PCR, and western blotting techniques. These results indicated that the gene modifications induced by the disruption of FGF5 had occurred at the morphological and genetic levels. We further show that the knockout alleles were likely capable of germline transmission, which is essential for goat population expansion. These results provide sufficient evidences of the merit of using the CRISPR/Cas9 approach for the generation of gene-modified goats displaying the corresponding mutant phenotypes. PMID:27755602

  2. Comparative Transcriptome Analysis Reveals that a Ubiquitin-Mediated Proteolysis Pathway Is Important for Primary and Secondary Hair Follicle Development in Cashmere Goats

    PubMed Central

    Zheng, Zhu-qing; Fu, Shao-yin; Tarekegn, Getinet Mekuriaw; Bai, Xue; Bai, Yong-sheng; Li, Heng; Zhang, Wen-guang

    2016-01-01

    Background The fleece of cashmere goats contains two distinct populations of fibers, a short and fine non-medullated insulating cashmere fiber and a long and coarse medullated guard hair. The former is produced by secondary follicles (SFs) and the later by primary follicles (PFs). Evidence suggests that the induction of PFs and SFs may require different signaling pathways. The regulation of BMP2/4 signaling by noggin and Edar signaling via Downless genes are essential for the induction of SFs and PFs, respectively. However, these differently expressed genes of the signaling pathway cannot directly distinguish between the PFs and SFs. Results In this study, we selected RNA samples from 11 PFs and 7 SFs that included 145,525 exons. The pathway analysis of 4512 differentially expressed exons revealed that the most statistically significant metabolic pathway was related to the ubiquitin–mediated proteolysis pathway (UMPP) (P<3.32x 10−7). In addition, the 51 exons of the UMPP that were differentially expressed between the different types of hair follicle (HFs) were compared by cluster analysis. This resulted in the PFs and SFs being divided into two classes. The expression level of two selected exons was analyzed by qRT-PCR, and the results indicated that the expression patterns were consistent with the deep sequencing results obtained by RNA-Seq. Conclusions Based on the comparative transcriptome analysis of 18 HFs from cashmere goats, a large number of differentially expressed exons were identified using a high-throughput sequencing approach. This study suggests that UMPP activation is a prominent signaling pathway for distinguishing the PFs and SFs of cashmere goats. It is also a meaningful contribution to the theoretical basis of the biological study of the HFs of cashmere goats and other mammals. PMID:27695037

  3. Relative abundance of tissue mRNA and association of the single nucleotide polymorphism of the goat NGF gene with prolificacy.

    PubMed

    Naicy, Thomas; Venkatachalapathy, R Thirupathy; Aravindakshan, T V; Raghavan, K C; Mini, M; Shyama, K

    2016-10-01

    Nerve Growth Factor (NGF) promotes the development of pre-antral ovarian follicles through ovarian innervations and regulation of ovarian response to gonadotropins. The present study was conducted to study the tissue gene expression profile, to characterize the genetic variants, find associations of the NGF gene with prolificacy in the prolific Malabari and less prolific Attappady Black goats because NGF has an important role in reproduction by augmenting ovarian folliculogenesis. Relative abundance of NGF mRNA was greatest in reproductive tissues signifying its role in reproduction. The PCR-SSCP analysis of a 251bp fragment of Exon 3 of the NGF gene from the 277 goats revealed four diplotypes (EE, EF, FF and EG) with respective frequencies of 0.76, 0.22, 0.01 and 0.01. Sequencing of the representative samples revealed one synonymous and one novel non synonymous mutations (g.705G>A and g.715C>T). Statistical analysis indicated that the SNP g.705G>A was associated with litter size in Attappady Black goats (P<0.05) and a PCR-RFLP was designed using the restriction enzyme, BpiI, for rapid screening of the SNP. The results of the present study suggest that the NGF gene is a primary candidate gene affecting prolificacy in goats and may be used for Marker Assisted Selection (MAS) in goats, especially in lowly prolific Attappady Black goats. PMID:27576174

  4. The localisation and characterisation of insulin-like growth factor-I receptors and the investigation of melatonin receptors on the hair follicles of seasonal and non-seasonal fibre-producing goats.

    PubMed

    Dicks, P; Morgan, C J; Morgan, P J; Kelly, D; Williams, L M

    1996-10-01

    To define the hormonal influences that are directly involved in the hair follicle cycles of animals with differing patterns of fibre growth and moulting, we have investigated the possible presence of IGF-I and melatonin receptors on the dermis and hair follicles of cashmere and Angora goats, sampled in February. March and June, using quantitative in vitro autoradiography. The presence of IGF-I receptors in the dermis of both breeds of goat was determined using cryostat sections incubated with 50 pM 125I-labelled IGF-I in the presence or absence of 50 nM IGF-I. Sections of the growing tip of deep antlers concerning the cartilaginous zone, a tissue known to contain high concentrations of specific IGF-I receptors, were used as a positive control. As the production of antler velvet uniquely involves the generation of hair follicles de novo, the presence of IGF-I receptors in the velvet-producing region was also investigated. In both breeds of goat, specific 125I-IGF-I binding was localised over the inner and outer root sheath, the matrix, the germinal matrix, the dermal papilla and the sebaceous glands and satisfied the basic kinetic criteria considered to be representative of a specific IGF-I receptor. Analysis of saturation isotherms using a one-site binding model revealed dissociation constants (Kd) in the range 0.1-0.9 nM and theoretical maximal numbers of binding sites (Bmax) between 21.4 and 45.6 fmol/mg tissue. Kd and Bmax values derived from cashmere and Angora goats sampled at different times of the year did not differ significantly between breeds or sampling times. Specific 125I-IGF-I binding was also localised to the developing follicles on the deer antler dermis. The presence of melatonin receptors within the goat dermis was also investigated. Sections were incubated with 100 pM 2-[125I]iodomelatonin with or without 0.1 microM melatonin, along with sections of sheep pars tuberalis which are known to contain high levels of high-affinity melatonin receptors

  5. Ultrastructure of the basal lamina of bovine ovarian follicles and its relationship to the membrana granulosa.

    PubMed

    Irving-Rodgers, H F; Rodgers, R J

    2000-03-01

    Different morphological phenotypes of follicular basal lamina and of membrana granulosa have been observed. Ten preantral follicles (< 0. 1 mm), and 17 healthy and six atretic antral follicles (0.5-12 mm in diameter) were processed for light and electron microscopy to investigate the relationship the between follicular basal lamina and membrana granulosa. Within each antral follicle, the shape of the basal cells of the membrana granulosa was uniform, and either rounded or columnar. There were equal proportions of follicles follicles had only rounded basal cells. Conventional basal laminae of a single layer adjacent to the basal granulosa cells were observed in healthy follicles at the preantral and antral stages. However, at the preantral stage, the conventional types of basal lamina were enlarged or even partially laminated. A second type of basal lamina, described as 'loopy', occurred in about half the preantral follicles and in half the antral follicles follicles. 'Loopy' basal laminae were composed of basal laminae aligning the basal surface of basal granulosa cells, but with additional layers or loops often branching from the innermost layer. Each loop was usually < 1 microm long and had vesicles (20-30 nm) attached to the inner aspect. Basal cellular processes were also common, and vesicles could be seen budding off from these processes. In antral follicles, conventional basal laminae occurred in follicles with rounded basal granulosa cells. Other follicles with columnar cells, and atretic follicles, had the 'loopy' basal lamina phenotype. Thus, follicles have different basal laminae that relate to the morphology of the membrana granulosa. PMID:10864785

  6. Morphological classification of bovine ovarian follicles.

    PubMed

    Rodgers, R J; Irving-Rodgers, H F

    2010-02-01

    Follicle classification is an important aid to the understanding of follicular development and atresia. Some bovine primordial follicles have the classical primordial shape, but ellipsoidal shaped follicles with some cuboidal granulosa cells at the poles are far more common. Preantral follicles have one of two basal lamina phenotypes, either a single aligned layer or one with additional layers. In antral follicles <5 mm diameter, half of the healthy follicles have columnar shaped basal granulosa cells and additional layers of basal lamina, which appear as loops in cross section ('loopy'). The remainder have aligned single-layered follicular basal laminas with rounded basal cells, and contain better quality oocytes than the loopy/columnar follicles. In sizes >5 mm, only aligned/rounded phenotypes are present. Dominant and subordinate follicles can be identified by ultrasound and/or histological examination of pairs of ovaries. Atretic follicles <5 mm are either basal atretic or antral atretic, named on the basis of the location in the membrana granulosa where cells die first. Basal atretic follicles have considerable biological differences to antral atretic follicles. In follicles >5 mm, only antral atresia is observed. The concentrations of follicular fluid steroid hormones can be used to classify atresia and distinguish some of the different types of atresia; however, this method is unlikely to identify follicles early in atresia, and hence misclassify them as healthy. Other biochemical and histological methods can be used, but since cell death is a part of normal homoeostatis, deciding when a follicle has entered atresia remains somewhat subjective. PMID:19786400

  7. Biology and Biotechnology of Follicle Development

    PubMed Central

    Palma, Gustavo Adolfo; Argañaraz, Martin Eduardo; Barrera, Antonio Daniel; Rodler, Daniela; Mutto, Adrian Ángel; Sinowatz, Fred

    2012-01-01

    Growth and development of ovarian follicles require a series of coordinated events that induce morphological and functional changes within the follicle, leading to cell differentiation and oocyte development. The preantral early antral follicle transition is the stage of follicular development during which gonadotropin dependence is obtained and the progression into growing or atresia of the follicle is made. Follicular growth during this period is tightly regulated by oocyte-granulosatheca cell interactions. A cluster of early expressed genes is required for normal folliculogenesis. Granulosa cell factors stimulate the recruitment of theca cells from cortical stromal cells. Thecal factors promote granulosa cell proliferation and suppress granulosa cell apoptosis. Cell-cell and cell-extracellular matrix interactions influence the production of growth factors in the different follicular compartments (oocyte, granulosa, and theca cells). Several autocrine and paracrine factors are involved in follicular growth and differentiation; their activity is present even at the time of ovulation, decreasing the gap junction communication, and stimulating the theca cell proliferation. In addition, the identification of the factors that promote follicular growth from the preantral stage to the small antral stage may provide important information for the identification for assisted reproduction techniques. PMID:22666170

  8. Evidence for ovarian granulosa stem cells: telomerase activity and localization of the telomerase ribonucleic acid component in bovine ovarian follicles.

    PubMed

    Lavranos, T C; Mathis, J M; Latham, S E; Kalionis, B; Shay, J W; Rodgers, R J

    1999-08-01

    We have previously postulated that granulosa cells of developing follicles arise from a population of stem cells. Stem cells and cancer cells can divide indefinitely partly because they express telomerase. Telomerase is a ribonucleoprotein enzyme that repairs the ends of telomeres that otherwise shorten progressively upon each successive cell division. In this study we carried out cell cycle analyses and examined telomerase expression to examine our hypothesis. Preantral (60-100 microm) and small (1 mm) follicles, as well as granulosa cells from medium-sized (3 mm) and large (6-8 mm) follicles, were isolated. Cell cycle analyses and expression of Ki-67, a cell cycle-related protein, were undertaken on follicles of each size (n = 3) by flow cytometry; 12% to 16% of granulosa cells in all follicles were in the S phase, and less than 2% were in the G(2)/M phase. Telomerase activity (n = 3) was highest in the small preantral follicles, declining at the 1-mm stage and even further at the 3-mm stage. In situ hybridization histochemistry was carried out on bovine ovaries, and telomerase RNA was detected in the granulosa cells of growing follicles but not primordial follicles. Two major patterns of staining were observed in the membrana granulosa of antral follicles: staining in the middle and antral layers, and staining in the middle and basal layers. No staining was detected in oocytes. Our results strongly support our hypothesis that granulosa cells arise from a population of stem cells. PMID:10411512

  9. Effect of growth differentiation factor-9 (GDF-9) on the progression of buffalo follicles in vitrified-warmed ovarian tissues.

    PubMed

    Abdel-Ghani, M A; El-Sherry, T M; Abdelhafeez, H H

    2016-10-01

    To improve the reproductive performance of water buffalo to level can satisfy our needs, the mechanisms controlling ovarian follicular growth and development should be thoroughly investigated. Therefore, in this study, the expressions of growth differentiation factor-9 (GDF-9) in buffalo ovaries were examined by immunohistochemistry, and the effects of GDF-9 treatment on follicle progression were investigated using a buffalo ovary organ culture system. Frozen-thawed buffalo ovarian follicles within slices of ovarian cortical tissue were cultured for 14 days in the presence or absence of GDF-9. After culture, ovarian slices were fixed, sectioned and stained. The follicles were morphologically analysed and counted. Expression pattern of GDF-9 was detected in oocytes from primordial follicles onwards, besides, also presented in granulosa cells. Moreover, GDF-9 was detected in mural granulosa cells and theca cells of pre-antral follicles. In antral follicles, cumulus cells and theca cells displayed positive expression of GDF-9. In corpora lutea, GDF-9 was expressed in both granulosa and theca lutein cells. After in vitro culture, there was no difference in the number of primordial follicles between cultured plus GDF-9 and cultured control that indicated the GDF-9 treatment has no effect on the primordial to primary follicle transition. GDF-9 treatment caused a significant decrease in the number of primary and secondary follicles compared with controls accompanied with a significant increase in pre-antral and antral follicles. These results suggest that a larger number of primary and secondary follicles were stimulated to progress to later developmental stages when treated with GDF-9. Vitrification/warming of buffalo ovarian tissue had a little remarkable effect, in contrast to culturing for 14 days, on the expression of GDF-9. In conclusion, treatment with GDF-9 was found to promote progression of primary follicle that could provide an alternative approach to

  10. Effect of growth differentiation factor-9 (GDF-9) on the progression of buffalo follicles in vitrified-warmed ovarian tissues.

    PubMed

    Abdel-Ghani, M A; El-Sherry, T M; Abdelhafeez, H H

    2016-10-01

    To improve the reproductive performance of water buffalo to level can satisfy our needs, the mechanisms controlling ovarian follicular growth and development should be thoroughly investigated. Therefore, in this study, the expressions of growth differentiation factor-9 (GDF-9) in buffalo ovaries were examined by immunohistochemistry, and the effects of GDF-9 treatment on follicle progression were investigated using a buffalo ovary organ culture system. Frozen-thawed buffalo ovarian follicles within slices of ovarian cortical tissue were cultured for 14 days in the presence or absence of GDF-9. After culture, ovarian slices were fixed, sectioned and stained. The follicles were morphologically analysed and counted. Expression pattern of GDF-9 was detected in oocytes from primordial follicles onwards, besides, also presented in granulosa cells. Moreover, GDF-9 was detected in mural granulosa cells and theca cells of pre-antral follicles. In antral follicles, cumulus cells and theca cells displayed positive expression of GDF-9. In corpora lutea, GDF-9 was expressed in both granulosa and theca lutein cells. After in vitro culture, there was no difference in the number of primordial follicles between cultured plus GDF-9 and cultured control that indicated the GDF-9 treatment has no effect on the primordial to primary follicle transition. GDF-9 treatment caused a significant decrease in the number of primary and secondary follicles compared with controls accompanied with a significant increase in pre-antral and antral follicles. These results suggest that a larger number of primary and secondary follicles were stimulated to progress to later developmental stages when treated with GDF-9. Vitrification/warming of buffalo ovarian tissue had a little remarkable effect, in contrast to culturing for 14 days, on the expression of GDF-9. In conclusion, treatment with GDF-9 was found to promote progression of primary follicle that could provide an alternative approach to

  11. The domestic dog and cat as models for understanding the regulation of ovarian follicle development in vitro.

    PubMed

    Songsasen, N; Comizzoli, P; Nagashima, J; Fujihara, M; Wildt, D E

    2012-12-01

    The culture of ovarian follicles is an important tool for understanding the mechanisms controlling follicle development and differentiation of the oocyte. The benefit of recovering meiotically and developmentally competent oocytes from early stage follicles (primordial, primary, pre-antral and early antral) also would be significant, ranging from rescue of genomes from endangered species to preserving fertility in women facing cancer treatments. This research field is at an early stage of scientific discovery. To-date, live offspring from cultured primordial follicles that produced fertilizable oocytes has occurred only in the mouse. Progress in other more complex species has been limited because larger animals have longer durations of natural folliculogenesis, thereby requiring more culture time to generate fully grown follicles and oocytes. We believe the dog and cat are excellent models for understanding more about folliculogenesis in vitro. This review highlights what is known about this topic for these two species as well as future priorities. We have discovered that it is more challenging to maintain viability of primordial follicles within ovarian tissues in vitro in the dog than the cat. Nonetheless, it is possible to grow both isolated cat and dog pre-antral follicles in culture. Although the follicles of both species have the capacity to increase in size and produce steroids, only cat oocytes appear morphologically normal. The reason for this striking difference between these two species is an area of high research priority. While much more fundamental data are required, we envision advanced technology that will allow harvesting oocytes from the vast, unused follicle stores sequestered within carnivore ovaries. These gametes have utility for reproducing genetically valuable dogs and cats that are 'companions' or biomedical models for investigating human disorders as well as for salvaging the genomes of rare canid and felid species that die before

  12. Kit ligand promotes the transition from primordial to primary follicles after in vitro culture of ovine ovarian tissue.

    PubMed

    Cavalcante, A Y P; Gouveia, B B; Barberino, R S; Lins, T L B G; Santos, L P; Gonçalves, R J S; Celestino, J J H; Matos, M H T

    2016-08-01

    This study evaluated the effects of kit ligand (KL) on the morphology and development of ovine preantral follicles (fresh control) and after 7 days of in vitro culture in α-Minimal Essential Medium (α-MEM; control medium) or the presence of KL (1, 10, 50, 100 or 200 ng/ml). There was an increase in the percentage of primary follicles at the concentration of 100 ng/ml KL, compared with the fresh control, control medium (α-MEM) and the other KL concentrations. Follicle diameter was significantly higher than the control medium only at concentrations of 50 and 100 ng/ml KL. In conclusion, 100 ng/ml KL promoted the transition from primordial to primary follicles (follicular activation) after in vitro culture of ovine ovarian tissue.

  13. Lipopolysaccharide reduces the primordial follicle pool in the bovine ovarian cortex ex vivo and in the murine ovary in vivo.

    PubMed

    Bromfield, John J; Sheldon, I Martin

    2013-04-01

    Infections of the uterus or mammary gland with Gram-negative bacteria cause infertility in cattle, not only during disease but also for some time afterward. Even though these infections are in organs distant from the ovary, metritis and mastitis perturb antral follicle development and function in vivo. Although granulosa cells from antral follicles express toll-like receptor 4 (TLR4), and detect and mount an inflammatory response to lipopolysaccharide (LPS) from Gram-negative bacteria, it is not known whether LPS impacts preantral follicle development. The present study tested the hypothesis that LPS perturbs the development of primordial ovarian follicles. Exposure of bovine ovarian cortex ex vivo to LPS reduced the primordial follicle pool associated with increased primordial follicle activation. Ovarian cortex culture supernatants accumulated the inflammatory mediators IL-1beta, IL-6, and IL-8 in an LPS concentration-dependent manner. In addition, LPS exposure modulated key intracellular regulators of follicle activation with loss of the primordial follicle PTEN and cytoplasmic translocation of FOXO3. Acute exposure of mice in vivo to LPS also reduced the primordial follicle pool associated with increased follicle atresia. The increased follicle atresia was TLR4-dependent as Tlr4-deficient mice were insensitive to LPS-mediated follicle atresia. However, LPS did not affect the diameter of individually cultured bovine secondary follicles or their enclosed oocytes. In conclusion, LPS reduced the primordial ovarian follicle pool in the bovine ovarian cortex ex vivo and in the murine ovary in vivo. These observations provide an insight into how bacterial infections distant from the ovary have long term effects on fertility.

  14. Promotion of ovarian follicle growth following mTOR activation: synergistic effects of AKT stimulators.

    PubMed

    Cheng, Yuan; Kim, Jaehong; Li, Xiao Xiao; Hsueh, Aaron J

    2015-01-01

    Mammalian target of rapamycin (mTOR) is a serine/threonine kinase and mTOR signaling is important in regulating cell growth and proliferation. Recent studies using oocyte- and granulosa cell-specific deletion of mTOR inhibitor genes TSC1 or TSC2 demonstrated the important role of mTOR signaling in the promotion of ovarian follicle development. We now report that treatment of ovaries from juvenile mice with an mTOR activator MHY1485 stimulated mTOR, S6K1 and rpS6 phosphorylation. Culturing ovaries for 4 days with MHY1485 increased ovarian explant weights and follicle development. In vivo studies further demonstrated that pre-incubation of these ovaries with MHY1485 for 2 days, followed by allo-grafting into kidney capsules of adult ovariectomized hosts for 5 days, led to marked increases in graft weights and promotion of follicle development. Mature oocytes derived from MHY1485-activated ovarian grafts could be successfully fertilized, leading the delivery of healthy pups. We further treated ovaries with the mTOR activator together with AKT activators (PTEN inhibitor and phosphoinositol-3-kinase stimulator) before grafting and found additive enhancement of follicle growth. Our studies demonstrate the ability of an mTOR activator in promoting follicle growth, leading to a potential strategy to stimulate preantral follicle growth in infertile patients.

  15. SEM cryofracture study of ovarian follicles of immature rats.

    PubMed

    Apkarian, R; Curtis, J C

    1981-01-01

    Follicles were examined with the scanning electron microscope (SEM) following preparation of cryofractured specimens of rat ovaries. After perfusion fixation with 2.5% glutaraldehyde buffered at pH 7.4 with 0.1M cacodylate, ovaries were trimmed of fat and decapsulated, fixed for an additional 12 hours, washed in distilled H2O for 20 hours and postfixed in 1% OsO4 in distilled H2O for 30 minutes. Following a brief wash, ovaries were dehydrated in a linear gradient of dist. H2O/ethanol, frozen in Freon 22, cryofractured under liquid nitrogen, brought to 1 degree C in ethanol, and critical point dried from CO2. Follicles cleaved through their oocytes were examined with SEM to determine the ultrastructural characteristics of developing and atretic follicles revealed by cryofracture. Cytoplasmic arrays of parallel sheets, nucleoli and microvilli were prominent structures of cryofractured oocytes. Numerous cytoplasmic projections of granulosa cells and oocyte microvilli penetrated the zonae pellucida of preantral follicles. The numbers of oocyte microvilli were greatly diminished in large antral follicles. Other structures made prominent by cryofracture included cell nuclei, a microvillous border at the granulosa-theca boundary, and numerous holes in the cytoplasm of thecal cells which correspond in size and distribution to the liquid droplets in these cells. The soluble proteins and glycoproteins of the follicular fluid were visualized as a homogeneous granular precipitate. The disappearance of oocyte microvilli and granulosa cell projections from the zonae pellucida and the blebbing of granulosa cells at the surface of the antral cavity appeared characteristic of an early stage of follicular atresia. Increased numbers of holes in thecal cells, oocyte fragmentation, and extensive dissociation and fragmentation of granulosa cells were typical of more advanced stages of atresia. PMID:7347419

  16. Dynamics of extracellular matrix in ovarian follicles and corpora lutea of mice.

    PubMed

    Irving-Rodgers, Helen F; Hummitzsch, Katja; Murdiyarso, Lydia S; Bonner, Wendy M; Sado, Yoshikazu; Ninomiya, Yoshifumi; Couchman, John R; Sorokin, Lydia M; Rodgers, Raymond J

    2010-03-01

    Despite the mouse being an important laboratory species, little is known about changes in its extracellular matrix (ECM) during follicle and corpora lutea formation and regression. Follicle development was induced in mice (29 days of age/experimental day 0) by injections of pregnant mare's serum gonadotrophin on days 0 and 1 and ovulation was induced by injection of human chorionic gonadotrophin on day 2. Ovaries were collected for immunohistochemistry (n=10 per group) on days 0, 2 and 5. Another group was mated and ovaries were examined on day 11 (n=7). Collagen type IV alpha1 and alpha2, laminin alpha1, beta1 and gamma1 chains, nidogens 1 and 2 and perlecan were present in the follicular basal lamina of all developmental stages. Collagen type XVIII was only found in basal lamina of primordial, primary and some preantral follicles, whereas laminin alpha2 was only detected in some preantral and antral follicles. The focimatrix, a specialised matrix of the membrana granulosa, contained collagen type IV alpha1 and alpha2, laminin alpha1, beta1 and gamma1 chains, nidogens 1 and 2, perlecan and collagen type XVIII. In the corpora lutea, staining was restricted to capillary sub-endothelial basal laminas containing collagen type IV alpha1 and alpha2, laminin alpha1, beta1 and gamma1 chains, nidogens 1 and 2, perlecan and collagen type XVIII. Laminins alpha4 and alpha5 were not immunolocalised to any structure in the mouse ovary. The ECM composition of the mouse ovary has similarities to, but also major differences from, other species with respect to nidogens 1 and 2 and perlecan. PMID:20033213

  17. Oocytes lacking O-glycans alter follicle development and increase fertility by increasing follicle FSH sensitivity, decreasing apoptosis, and modifying GDF9:BMP15 expression.

    PubMed

    Grasa, Patricia; Ploutarchou, Panayiota; Williams, Suzannah A

    2015-02-01

    The number of eggs ovulated varies within and between species and is influenced by many variables. However, the regulatory mechanisms remain poorly understood. We previously demonstrated a key role for the oocyte because mice generating oocytes deficient in core 1-derived O-glycans ovulate ∼40-50% more eggs than Controls. Here we analyze the basis of this phenotype using Mutant [core 1 β1,3-galactosyltransferase 1 (C1galt1)(FF):zona pellucida glycoprotein 3 Cre (ZP3Cre)] and Control (C1galt1(FF)) female mice. In culture, Mutant follicles exhibited delayed antrum formation [indicative of follicle stimulant hormone (FSH) dependence] and increased sensitivity to FSH. Although the Mutant estrous cycle was extended, comprehensive endocrine changes were not observed; rather FSH, LH, inhibin B, and anti-Mullerian hormone were temporally altered, revealing estrous cycle stage-specific modifications to the hypothalamic-pituitary-gonadal axis. At proestrus, when FSH levels were decreased in Mutants, ovaries contained more, smaller, preantral follicles. Mutant follicles exhibited reduced levels of apoptosis, and both B-cell lymphoma 2 (Bcl-2) and BCL-2-associated X protein (Bax) were altered compared with Controls. Mutant ovaries also had an increase in the expression ratio of growth differentiation factor 9 (GDF9):bone morphogenetic protein 15 (BMP15) at diestrus. On the basis of these data, we propose that modified oocyte glycoproteins alter GDF9:BMP15 expression modifying follicle development resulting in the generation of more follicles. Thus, the oocyte is a key regulator of follicle development and has a crucial role in determining ovulation rate.

  18. Modulation of ovarian follicle maturation and effects on apoptotic cell death in Holtzman rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in utero and lactationally.

    PubMed

    Heimler, I; Trewin, A L; Chaffin, C L; Rawlins, R G; Hutz, R J

    1998-01-01

    Recent reports have described the reproduction-modulating and endocrine-disrupting effects following exposure to toxic substances such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Herein, we set out (1) to determine whether TCDD exposure exerts detrimental effects on follicle maturation in the Holtzman rat ovary and (2) to determine whether the effects of TCDD are mediated in part via apoptotic cell death. In certain species, dioxin exposure is correlated with reduced fecundity, reduced ovulatory rate, an increased incidence of endometriosis, and various reproductive cancers. Although some of the effects of TCDD are mediated via the hypothalamic-pituitary axis, direct effects on the ovary have also been observed. In the present study, an oral dose of 1 microgram TCDD/kg maternal body weight was administered on Day 15 of gestation. Female pups were sacrificed on Postnatal Day 21/22, and the ovaries were excised, fixed for histologic analysis, and analyzed in a double-blind paradigm. The analysis included a count and measurement and classification of preantral and antral follicles throughout the entire ovary. The contralateral ovary from each animal was analyzed for DNA fragmentation indicative of apoptotic cell death. The results indicate that TCDD treatment significantly reduced the number of antral follicles in the size classes 50,000 to 74,999 microns2 and > 100,000 microns2. We also observed a reduction in the number of preantral follicles less than 50,000 microns2. No difference was observed in the degree of apoptotic cell death in antral (50,000 to > 100,000 microns2) and preantral follicles (50,000 microns2 to > 75,000 microns2) between TCDD-treated and control-treated tissues. These data support the hypothesis that TCDD results in a diminution in the number of antral and preantral follicles of certain size classes in animals exposed during critical periods of development, but that apoptosis does not appear to be the underlying mechanism in these particular

  19. Quantitative expression patterns of GDF9 and BMP15 genes in sheep ovarian follicles grown in vivo or cultured in vitro.

    PubMed

    Kona, S S R; Praveen Chakravarthi, V; Siva Kumar, A V N; Srividya, D; Padmaja, K; Rao, V H

    2016-01-15

    Quantitative patterns of expression of the growth differentiation factor 9 (GDF9) and bone morphogenic protein 15 (BMP15) genes in different development stages of in vivo and in vitro grown ovarian follicles in sheep were studied for the first time. Both GDF9 and BMP15 were expressed in the cumulus cells and oocytes at all the development stages of in vivo and in vitro grown ovarian follicles. Growth differentiation factor 9 and bone morphogenic protein 15 exhibited stage-specific undulations in the expression in the cumulus cells and oocytes isolated from in vivo grown ovarian follicles. These undulations could be related to discrete development events during the ovarian follicle development. The expression of GDF9 and BMP15 was highest (3.38 ± 0.02 and 2.69 ± 0.06, respectively; P ≤ 0.05) in the primordial follicles compared with preantral, early antral, antral, and large antral stages. Similarly, GDF9 and BMP15 expression in the cumulus cells (0 ± 0.16 and 0 ± 0.07) and oocytes (1.47 ± 0.07 and 1.32 ± 0.03) was lowest (P ≤ 0.05) in the in vivo grown antral follicles. In the cultured follicles, the stage-specific undulations observed in the expression of GDF9 and BMP15 in the in vivo grown follicles were either different or abolished. For example, in the oocytes from in vitro grown follicles, the expression of BMP15 did not change as the development progressed all the way from preantral to large antral follicle stage although in the oocytes from in vivo grown follicles BMP15 expression exhibited stage-specific variations. It is concluded that GDF9 and BMP15 follow a stage-specific pattern of expression during the in vivo development of ovarian follicles in sheep, and in vitro culture altered the stage-specific changes in the expression of these two genes.

  20. The role of androgens in follicle maturation and ovulation induction: friend or foe of infertility treatment?

    PubMed Central

    2011-01-01

    Background Effects of androgens on follicle maturation have been controversial for some time. Here, we review the potential of their applications in improving human ovulation induction, based on human and animal data, reported in the literature. Methods We reviewed the published literature for the years 2005-2011, using relevant key words, in PubMed, Medline and Cochrane reviews, and then performed secondary reviews of referenced articles, which previously had not been known or preceded the searched time period. A total of 217 publications were reviewed. Results Contrary to widely held opinion, recent data, mostly developed in the mouse, convincingly demonstrate essential contribution of androgens to normal follicle maturation and, therefore, female fertility. Androgens appear most engaged at preantral and antral stages, primarily affect granulosa cells, and exert effects via androgen receptors (AR) through transcriptional regulation but also in non-genomic ways, with ligand-activated AR modulating follicle stimulating hormone (FSH) activity in granulosa cells. While some androgens, like testosterone (T) and dehydroepiandrosterone (DHEA), appear effective in improving functional ovarian reserve (FOR) in women with diminished ovarian reserve (DOR), others may even exert opposite effects. Such differences in androgens may, at least partially, reflect different levels of agonism to AR. Discussion Selective androgens appear capable of improving early stages of folliculogenesis. They, therefore, may represent forerunners of a completely new class of ovulation-inducing medications, which, in contrast to gonadotropins, affect follicle maturation at much earlier stages. PMID:21849061

  1. Antiequine chorionic gonadotropin (eCG) antibodies generated in goats treated with eCG for the induction of ovulation modulate the luteinizing hormone and follicle-stimulating hormone bioactivities of eCG differently.

    PubMed

    Hervé, Virginie; Roy, François; Bertin, Jean; Guillou, Florian; Maurel, Marie-Christine

    2004-01-01

    In dairy goats, treatments associating a progestogen and the equine chorionic gonadotropin (eCG) are the easiest way to induce and synchronize estrus and ovulation and to permit artificial insemination (AI) and/or out of season breeding. From the first treatment, the injection of eCG induces, in some females, the production of anti-eCG antibodies (Abs) that will interfere with the effectiveness of subsequent treatments. These anti-eCG Abs delay the preovulatory LH surge and the ovulation time, leading to poor fertility of the treated females. In this study, by in vitro bioassays, we show that anti-eCG Abs can positively or negatively modulate the LH and/or FSH bioactivities of eCG. Moreover, the modulation level of eCG bioactivity does not depend on the anti-eCG Ab affinity for eCG, as shown by surface plasmon resonance technology. The specificity of anti-eCG Abs tested by competitive ELISA highlighted the importance of a glycan environment in the recognition mechanism, especially the sialic acids specific to eCG. The different effects of anti-eCG Abs on eCG bioactivities could be explained by two hypotheses. First, steric hindrance preventing the interaction of eCG with its receptors would explain the inhibitory effect of some anti-eCG Abs; second, a conformational change in eCG by anti-eCG Abs could induce inhibition or potentiation of eCG bioactivities. It is significant that these modulations of eCG bioactivities by anti-eCG Abs impact mainly on the FSH bioactivity of eCG, which is essential for ovarian stimulation and subsequent fertility after treatment and AI, and to a lesser extent on LH bioactivity.

  2. Knockout of RNA Binding Protein MSI2 Impairs Follicle Development in the Mouse Ovary: Characterization of MSI1 and MSI2 during Folliculogenesis

    PubMed Central

    Sutherland, Jessie M.; Sobinoff, Alexander P.; Gunter, Kara M.; Fraser, Barbara A.; Pye, Victoria; Bernstein, Ilana R.; Boon, Evan; Siddall, Nicole A.; De Andres, Luisa I.; Hime, Gary R.; Holt, Janet E.; Graf, Thomas; McLaughlin, Eileen A.

    2015-01-01

    Characterizing the mechanisms underlying follicle development in the ovary is crucial to understanding female fertility and is an area of increasing research interest. The RNA binding protein Musashi is essential for post-transcriptional regulation of oocyte maturation in Xenopus and is expressed during ovarian development in Drosophila. In mammals Musashi is important for spermatogenesis and male fertility, but its role in the ovary has yet to be characterized. In this study we determined the expression of mammalian Musashi proteins MSI1 and MSI2 during mouse folliculogenesis, and through the use of a MSI2-specific knockout mouse model we identified that MSI2 is essential for normal follicle development. Time-course characterization of MSI1 and MSI2 revealed distinct differences in steady-state mRNA levels and protein expression/localization at important developmental time-points during folliculogenesis. Using a gene-trap mouse model that inactivates Msi2, we observed a significant decrease in ovarian mass, and change in follicle-stage composition due to developmental blocking of antral stage follicles and pre-antral follicle loss through atresia. We also confirmed that hormonally stimulated Msi2-deficient mice produce significantly fewer MII oocytes (60.9% less than controls, p < 0.05). Furthermore, the majority of these oocytes are of poor viability (62.2% non-viable/apoptotic, p < 0.05), which causes a reduction in female fertility evidenced by decreased litter size in Msi2-deficient animals (33.1% reduction to controls, p < 0.05). Our findings indicate that MSI1 and MSI2 display distinct expression profiles during mammalian folliculogenesis and that MSI2 is required for pre-antral follicle development. PMID:26131972

  3. Combined effect of follicle-follicle interactions and declining follicle-stimulating hormone on murine follicle health in vitro.

    PubMed

    Baker, S J; Srsen, V; Lapping, R; Spears, N

    2001-10-01

    Follicle selection occurs throughout an adult female's reproductive life, with selected, dominant follicle(s) developing to the preovulatory stage whereas the remaining, subordinate follicles within the growing cohort instead undergo atresia and die. To date, most research into follicle dominance has concentrated on its endocrine regulation, although it seems likely that intraovarian mechanisms are also involved in its regulation. We demonstrate here that the response of singly cultured murine follicles to declining concentrations of FSH depends on their developmental stage, with follicles at an earlier stage of development being much more susceptible than mature follicles to a lowering of FSH levels. We then extrapolate this information to follicle cocultures, in which a large dominant follicle was grown with a small subordinate follicle in a manner that maintained a dominant/subordinate relationship, with follicle health assessed by a terminal transferase-mediated 2'-deoxyuracil 5'-triphosphate nick end-labeled reaction on whole-follicle mounts. Our investigations show a combined negative effect of coculture and FSH withdrawal on small subordinate follicles, such that subordinate follicles cocultured with dominant follicles and subjected to a lowering of FSH levels during the culture period exhibit a greatly increased incidence of apoptosis in the granulosa cells (750% increase) compared with that exhibited by the dominant follicles (97% increase). We suggest that a similar interaction between endocrine and intraovarian factors regulates follicular dominance in vivo, such that dominant follicles, in addition to bringing about a fall in FSH levels via the hypothalamic-pituitary axis, exert local, direct effects on subordinate follicles, with both of these influences combining to induce atresia in subordinate follicles.

  4. Tamoxifen prevents apoptosis and follicle loss from cyclophosphamide in cultured rat ovaries.

    PubMed

    Piasecka-Srader, Joanna; Blanco, Fernando F; Delman, Devora H; Dixon, Dan A; Geiser, James L; Ciereszko, Renata E; Petroff, Brian K

    2015-05-01

    Recent studies documented that the selective estrogen receptor modulator tamoxifen prevents follicle loss and promotes fertility following in vivo exposure of rodents to irradiation or ovotoxic cancer drugs, cyclophosphamide and doxorubicin. In an effort to characterize the ovarian-sparing mechanisms of tamoxifen in preantral follicle classes, cultured neonatal rat ovaries (Day 4, Sprague Dawley) were treated for 1-7 days with active metabolites of cyclophosphamide (i.e., 4-hydroxycyclophosphamide; CTX) (0, 1, and 10 μM) and tamoxifen (i.e., 4-hydroxytamoxifen; TAM) (0 and 10 μM) in vitro, and both apoptosis and follicle numbers were measured. CTX caused marked follicular apoptosis and follicular loss. TAM treatment decreased follicular loss and apoptosis from CTX in vitro. TAM alone had no effect on these parameters. IGF-1 and IGF-1 receptor were assessed in ovarian tissue showing no impact of TAM or CTX on these endpoints. Targeted mRNA analysis during follicular rescue by TAM revealed decreased expression of multiple genes related to inflammation, including mediators of lipoxygenase and prostaglandin production and signaling (Alox5, Pla2g1b, Ptgfr), cytokine binding (Il1r1, Il2rg ), apoptosis (Tnfrsf1a), second messenger signaling (Mapk1, Mapk14, Plcg1), as well as tissue remodeling and vasodilation (Bdkrb2, Klk15). The results suggest that TAM protects the ovary from CTX-mediated toxicity through direct ovarian actions that oppose follicular loss. PMID:25833159

  5. The Protective Effects of Exogenous Melatonin on Nicotine-induced Changes in Mouse Ovarian Follicles

    PubMed Central

    Mohammadghasemi, Fahimeh; Jahromi, Sina Khajeh; Hajizadeh, Hadi; Homafar, Mohammad Amin; Saadat, Nazanin

    2012-01-01

    Background Nicotine exposure causes impaired fertility and ovarian dysfunction. The aim of this study was to investigate the possible protective role of melatonin, which is known as an antioxidant agent on altered ovarian functions upon nicotine exposure. Methods A total of 32 female adult NMRI mice were divided randomly into four groups (n = 8). The control group received vehicle, while group 2 received nicotine (40 µg/kg) for 15 days and group 3 melatonin (10 mg/kg) for 5 days. Group 4 received both nicotine (40 µg/kg) and melatonin (10 mg/kg) for the same periods. All animals were treated intraperitoneally. After autopsy on the 16th day, histopathological and morphometrical examinations were performed and serum estradiol concentrations were measured. The data were analyzed using ANOVA and Tukey post hoc test. A value of p < 0.05 was considered significant. Results Nicotine significantly reduced the number of pre-antral and antral follicles, as well as estradiol concentration compared to the control group (p < 0.05). However, the decrease in the number of primordial follicles was not significant in the nicotine treated group. A significant increase in the atretic follicles were observed in group 2 compared to the control group (p < 0.05). Moreover, melatonin caused a marked normalization in the number of ovarian follicles and estradiol levels in group 4 compared to group 2. Conclusion The results from this study suggest that melatonin may have a protective effect against nicotine-induced ovarian changes on the number of different stages of follicle growth. PMID:23926539

  6. Amburana cearensis leaf extract maintains survival and promotes in vitro development of ovine secondary follicles.

    PubMed

    Barberino, R S; Barros, V R P; Menezes, V G; Santos, L P; Araújo, V R; Queiroz, M A A; Almeida, J R G S; Palheta, R C; Matos, M H T

    2016-04-01

    The antioxidant properties of Amburana cearensis extract may be a useful substitute for standard cell culture medium. Thus, the aim of this study was to evaluate the effect of this extract, with or without supplementation, on in vitro survival and development of sheep isolated secondary follicles. After collection of the ovaries, secondary follicles were isolated and cultured for 18 days in α-MEM+ supplemented with bovine serum albumin, insulin, transferrin, selenium, glutamine, hypoxanthine and ascorbic acid (control medium) or into medium composed of different concentrations of A. cearensis extract without supplements (Amb 0.1; 0.2 or 0.4 mg/ml) or A. cearensis extract supplemented with the same substances described above for α-MEM+ supplementation. The A. cearensis supplemented medium was named Amb 0.1+; 0.2+ or 0.4+ mg/ml. There were more morphologically normal follicles in Amb 0.1 or Amb 0.4 mg/ml than in the control medium (α-MEM+) after 18 days of culture. Moreover, the percentage of antrum formation was significantly higher in Amb 0.1 or Amb 0.2 mg/ml than in α-MEM+ and Amb 0.1+ mg/ml, and similar to the other treatments. All A. cearensis extract media induced a progressive and significant increase in follicular diameter throughout the culture period. In conclusion, this study showed that 0.1 mg/ml of this extract, without supplementation, maintains follicular survival and promotes the development of ovine isolated secondary follicles in vitro. This extract can be an alternative culture medium for preantral follicle development.

  7. Horny Goat Weed

    MedlinePlus

    Horny goat weed is an herb. The leaves are used to make medicine. As many as 15 horny goat weed species are known as “yin yang huo” ... Herbs and supplements that might lower blood pressureHorny goat weed might lower blood pressure. Taking it along ...

  8. Bortezomib Prevents Acute Doxorubicin Ovarian Insult and Follicle Demise, Improving the Fertility Window and Pup Birth Weight in Mice

    PubMed Central

    Roti Roti, Elon C.; Ringelstetter, Ashley K.; Kropp, Jenna; Abbott, David H.; Salih, Sana M.

    2014-01-01

    Increasing numbers of female patients survive cancer, but succumb to primary ovarian insufficiency after chemotherapy. We tested the hypothesis that Bortezomib (Bort) protects ovaries from doxorubicin (DXR) chemotherapy by treating female mice with Bort 1 hour prior to DXR. By preventing DXR accumulation in the ovary, Bort attenuated DXR-induced DNA damage in all ovarian cell types, subsequent γH2AFX phosphorylation, and resulting apoptosis in preantral follicles. Bort pretreatment extended the number of litters per mouse, improved litter size and increased pup weight following DXR treatment, thus increasing the duration of post-chemotherapy fertility and improving pup health. As a promising prophylactic ovoprotective agent, Bort does not interfere with cancer treatment, and is currently used as a chemotherapy adjuvant. Bort-based chemoprotection may preserve ovarian function in a non-invasive manner that avoids surgical ovarian preservation, thus diminishing the health complications of premature menopause following cancer treatment. PMID:25251158

  9. Expression and regulation of SNAP-25 and synaptotagmin VII in developing mouse ovarian follicles via the FSH receptor.

    PubMed

    Choi, Sung Sik; Jung, Joo Young; Lee, Dong Ho; Kang, Ji Yoon; Lee, Sang Ho

    2013-02-01

    Soluble-NSF attachment protein receptor (SNARE) proteins play a role in vesicle fusion, exocytosis, and intracellular trafficking in neuronal cells as well as in fertilization and embryogenesis. We investigated the expression patterns of two SNARE proteins, SNAP-25 and synaptotagmin VII (SytVII), and their regulation by pregnant mare serum gonadotropin (PMSG) during mouse ovarian follicular development. Ovaries were obtained at 0, 12, 24, 36, and 48 h post-PMSG injection of immature mice. SNAP-25 and SytVII mRNA expression levels increased gradually in a time-dependant manner. However, protein levels revealed different patterns of expression, suggesting different translational regulation following PMSG stimulation. SNAP-25 and SytVII expression was closely associated with thickening of the granulosa cell (GC) layer and follicle morphological changes from a flattened to a cuboidal shape. To explore follicle stimulating hormone receptor (FSHR)-mediated regulation of their expression, GCs from preantral follicles were cultured to examine the effects of FSHR siRNA knockdown. FSHR siRNA abolished upregulation of the SNAREs in both PMSG and FSH-stimulated GCs. This abolished gene expression was rescued by adding dibutyryl cyclic AMP to the cultures. These results suggest that SNAP-25 and SytVII expression is regulated via the FSHR-cAMP pathway during follicular development.

  10. Effects of ionizing radiation and pretreatment with (D-Leu6,des-Gly10) luteinizing hormone-releasing hormone ethylamide on developing rat ovarian follicles

    SciTech Connect

    Jarrell, J.; YoungLai, E.V.; McMahon, A.; Barr, R.; O'Connell, G.; Belbeck, L.

    1987-10-01

    To assess the effects of a gonadotropin-releasing hormone agonist, (D-Leu6,des-Gly10) luteinizing hormone-releasing hormone ethylamide, in ameliorating the damage caused by ionizing radiation, gonadotropin-releasing hormone agonist was administered to rats from day 22 to 37 of age in doses of 0.1, 0.4, and 1.0 microgram/day or vehicle and the rats were sacrificed on day 44 of age. There were no effects on estradiol, progesterone, luteinizing, or follicle-stimulating hormone, nor an effect on ovarian follicle numbers or development. In separate experiments, rats treated with gonadotropin-releasing hormone agonist in doses of 0.04, 0.1, 0.4, or 1.0 microgram/day were either irradiated or sham irradiated on day 30 and all groups sacrificed on day 44 of age. Irradiation produced a reduction in ovarian weight and an increase in ovarian follicular atresia. Pretreatment with the agonist prevented the reduction in ovarian weight and numbers of primordial and preantral follicles but not healthy or atretic antral follicles. Such putative radioprotection should be tested on actual reproductive performance.

  11. [Variation of plasma INH B, ACT A and FSH concentrations during an estrus cycle in Dazu black goat and Sannen dairy goat].

    PubMed

    Zhang, Chao; Luo, Yan-Mei; Zhang, Jia-Hua; He, Jing-Jing; Jin, Lu; Zhao, Zhong-Quan

    2011-06-01

    To study the relationship between the concentrations of INH B (Inhibin B), ACT A (Activin A), and FSH (Follicle stimulating hormone) in blood plasma and fecundity, Dazu black goat with high productivity and Sannen dairy goat with low productivity were used as experiment objects in this research. The concentrations of INH B, ACT A, and FSH in blood plasma were measured by enzyme-linked immunosorbent assay (ELISA) in order to study the secretion rule of INH B, ACT A, and FSH during an estrus cycle of two goat breeds. The results indicated that the secretion of FSH showed a positive correlation with ACT A and a negative correlation with INH B. The mean concentration of FSH in Dazu black goat was higher than that in Sanen dairy goat during a estrous cycle. However, during the time from obviously estrus to ovulation, the mean concentration of FSH in Dazu black goat was significantly higher than that in Sannen dairy goat (0.01goats. The main effect of ACT A may be extention of follicular stage. Inhibin B indirectly influences ovulation by regulation of FSH level.

  12. [Hair follicle regeneration].

    PubMed

    Itami, Satoshi

    2008-05-01

    Hair growth cycle is coordinated with complex processes that are dependent on the interactions of follicular stem cells and dermal papilla cells (DPCs). For the past 10 years, the developmental mechanism of hair follicles has been extensively studied, and spatial and temporal expressions of many molecules are required for the hair morphogenesis. These molecules are also required for hair cycle progression. Androgen receptor, which is a ligand dependent transcription factor, plays an important role in human hair cycle. Frontal scalp DPCs from androgenetic alopecia (AGA) are the target cells of androgen action. Minoxidil and Finasteride were recently introduced for the treatment of AGA, and cell therapy using DPCs is a next strategy for the innovative treatment. PMID:18464507

  13. Extracellular matrix in ovarian follicles.

    PubMed

    Rodgers, R J; Irving-Rodgers, H F; van Wezel, I L

    2000-05-25

    A lot is known about the control of the development of ovarian follicles by growth factors and hormones, but less is known about the roles of extracellular matrix in the control of follicular growth and development. In this review we focus on the specialized extracellular matrix of the basal laminas that are present in ovarian follicles. These include the follicular basal lamina itself, the Call-Exner bodies of the membrana granulosa, the subendothelial and arteriole smooth muscle basal laminas in the theca, and the basal lamina-like material of the thecal matrix. We discuss the evidence that during follicle development the follicular basal lamina changes in composition, that many of its components are produced by the granulosa cells, and that the follicular basal laminas of different follicles have different ultrastructural appearances, linked to the shape of the aligning granulosa cells. All these studies suggest that the follicular basal lamina is extremely dynamic during follicular development. PMID:10963877

  14. [Cashmere goat bacterial artificial chromosome recombination and cell transfection system].

    PubMed

    Huang, Tian; Cao, Zhongyang; Yang, Yaohui; Cao, Gengsheng

    2016-03-01

    The Cashmere goat is mainly used to produce cashmere, which is very popular for its delicate fiber, luscious softness and natural excellent warm property. Keratin associated protein (KAP) and bone morphogenetic protein (BMP) of the Cashmere goat play an important role in the proliferation and development of cashmere fiber follicle cells. Bacterial artificial chromosome containing kap6.3, kap8.1 and bmp4 genes were used to increase the production and quality of Cashmere. First, we constructed bacterial artificial chromosomes by homology recombination. Then Tol2 transposon was inserted into bacterial artificial chromosomes that were then transfected into Cashmere goat fibroblasts by Amaxa Nucleofector technology according to the manufacture's instructions. We successfully constructed the BAC-Tol2 vectors containing target genes. Each vector contained egfp report gene with UBC promoter, Neomycin resistant gene for cell screening and two loxp elements for resistance removing after transfected into cells. The bacterial artificial chromosome-Tol2 vectors showed a high efficiency of transfection that can reach 1% to 6% with a highest efficiency of 10%. We also obtained Cashmere goat fibroblasts integrated exogenous genes (kap6.3, kap8.1 and bmp4) preparing for the clone of Cashmere goat in the future. Our research demonstrates that the insertion of Tol2 transposons into bacterial artificial chromosomes improves the transfection efficiency and accuracy of bacterial artificial chromosome error-free recombination.

  15. [Cashmere goat bacterial artificial chromosome recombination and cell transfection system].

    PubMed

    Huang, Tian; Cao, Zhongyang; Yang, Yaohui; Cao, Gengsheng

    2016-03-01

    The Cashmere goat is mainly used to produce cashmere, which is very popular for its delicate fiber, luscious softness and natural excellent warm property. Keratin associated protein (KAP) and bone morphogenetic protein (BMP) of the Cashmere goat play an important role in the proliferation and development of cashmere fiber follicle cells. Bacterial artificial chromosome containing kap6.3, kap8.1 and bmp4 genes were used to increase the production and quality of Cashmere. First, we constructed bacterial artificial chromosomes by homology recombination. Then Tol2 transposon was inserted into bacterial artificial chromosomes that were then transfected into Cashmere goat fibroblasts by Amaxa Nucleofector technology according to the manufacture's instructions. We successfully constructed the BAC-Tol2 vectors containing target genes. Each vector contained egfp report gene with UBC promoter, Neomycin resistant gene for cell screening and two loxp elements for resistance removing after transfected into cells. The bacterial artificial chromosome-Tol2 vectors showed a high efficiency of transfection that can reach 1% to 6% with a highest efficiency of 10%. We also obtained Cashmere goat fibroblasts integrated exogenous genes (kap6.3, kap8.1 and bmp4) preparing for the clone of Cashmere goat in the future. Our research demonstrates that the insertion of Tol2 transposons into bacterial artificial chromosomes improves the transfection efficiency and accuracy of bacterial artificial chromosome error-free recombination. PMID:27349114

  16. Exposure of female macaques to Western-style diet with or without chronic T in vivo alters secondary follicle function during encapsulated 3-dimensional culture.

    PubMed

    Xu, Jing; McGee, Whitney K; Bishop, Cecily V; Park, Byung S; Cameron, Judy L; Zelinski, Mary B; Stouffer, Richard L

    2015-03-01

    Increased adiposity and hyperandrogenemia alter reproductive parameters in both animal models and women, but their effects on preantral follicles in the ovary remain unknown. We recently reported that Western-style diet (WSD) consumption over 1 year, with or without chronic exposure to elevated circulating T, increased the body fat percentage, elicited insulin resistance, suppressed estradiol and progesterone production, as well as altered the numbers, size, and dynamics of antral follicles in the ovary during the menstrual cycle in female macaques. Therefore, experiments were designed to compare the WSD and WSD+T effects to age-matched controls on the survival, growth, and function of isolated secondary follicles during 5 weeks of encapsulated 3-dimensional culture. Follicle survival significantly declined in the WSD and WSD+T groups compared with the control (CTRL) group. Although media progesterone levels were comparable among groups, androstenedione and estradiol levels were markedly reduced in the WSD and WSD+T groups compared with the CTRL group at week 5. Anti-Müllerian hormone levels peaked at week 3 and were lower in the WSD+T group compared with the WSD or CTRL group. Vascular endothelial growth factor levels also decreased at week 5 in the WSD+T group compared with the WSD or CTRL group. After human chorionic gonadotropin exposure, only antral follicles developed from the CTRL group yielded metaphase II oocytes. Thus, WSD with or without T exposure affects the cohort of secondary follicles in vivo, suppressing their subsequent survival, production of steroid hormones and local factors, as well as oocyte maturation in vitro.

  17. Exposure of Female Macaques to Western-Style Diet With or Without Chronic T In Vivo Alters Secondary Follicle Function During Encapsulated 3-Dimensional Culture

    PubMed Central

    McGee, Whitney K.; Bishop, Cecily V.; Park, Byung S.; Cameron, Judy L.; Zelinski, Mary B.; Stouffer, Richard L.

    2015-01-01

    Increased adiposity and hyperandrogenemia alter reproductive parameters in both animal models and women, but their effects on preantral follicles in the ovary remain unknown. We recently reported that Western-style diet (WSD) consumption over 1 year, with or without chronic exposure to elevated circulating T, increased the body fat percentage, elicited insulin resistance, suppressed estradiol and progesterone production, as well as altered the numbers, size, and dynamics of antral follicles in the ovary during the menstrual cycle in female macaques. Therefore, experiments were designed to compare the WSD and WSD+T effects to age-matched controls on the survival, growth, and function of isolated secondary follicles during 5 weeks of encapsulated 3-dimensional culture. Follicle survival significantly declined in the WSD and WSD+T groups compared with the control (CTRL) group. Although media progesterone levels were comparable among groups, androstenedione and estradiol levels were markedly reduced in the WSD and WSD+T groups compared with the CTRL group at week 5. Anti-Müllerian hormone levels peaked at week 3 and were lower in the WSD+T group compared with the WSD or CTRL group. Vascular endothelial growth factor levels also decreased at week 5 in the WSD+T group compared with the WSD or CTRL group. After human chorionic gonadotropin exposure, only antral follicles developed from the CTRL group yielded metaphase II oocytes. Thus, WSD with or without T exposure affects the cohort of secondary follicles in vivo, suppressing their subsequent survival, production of steroid hormones and local factors, as well as oocyte maturation in vitro. PMID:25545382

  18. Methoxychlor induces atresia of antral follicles in ERalpha-overexpressing mice.

    PubMed

    Tomic, Dragana; Frech, Maria Silvina; Babus, Janice K; Gupta, Rupesh K; Furth, Priscilla A; Koos, Robert D; Flaws, Jodi A

    2006-09-01

    Methoxychlor (MXC) is a pesticide that is known to bind to estrogen receptor alpha (ERalpha) and to induce atresia of antral ovarian follicles. Although studies have shown that MXC is toxic to the ovary, we hypothesize that perturbation to the estrogen-signaling system (i.e., increase or decrease in estrogen sensitivity) might alter ovarian responsiveness to MXC. Thus, we examined whether ERalpha overexpression alters the ability of MXC to increase follicle atresia. To do so, we employed a transgenic mouse model in which ERalpha can be inducibly overexpressed in animal tissues (ERalpha overexpressors). We dosed female controls and ERalpha overexpressors with sesame oil (vehicle control) or MXC (32 and 64 mg/kg/day) for 20 days. After dosing, the ovaries were collected for histological evaluation of follicle numbers and follicle atresia, while blood was collected for measurements of hormones. Estrous cycles were determined in all animals to ensure that all were terminated during estrus. Although there were no significant effects of MXC on the numbers of primordial, primary, and preantral follicles in both controls and ERalpha overexpressors, there was an effect on antral follicles. Specifically, our data indicate that 32 and 64 mg/kg MXC increased the percentage of atretic follicles compared to vehicle in both control and ERalpha overexpressor groups. Moreover, there was a clear trend toward greater sensitivity to 64 mg/kg MXC in ERalpha-overexpressing mice compared to control animals. Specifically, at the 64-mg/kg MXC dose, ERalpha-overexpressing mice had a significantly higher percentage of atretic follicles compared to control animals (controls = 21.5 +/- 3%, n = 5; ERalpha overexpressors = 37 +/- 23%, n = 9, p < or = 0.05 vs. controls). After 20 days of dosing, there were no differences in estradiol levels between controls and ERalpha-overexpressing mice in all treatment groups. Follicle-stimulating hormone (FSH) levels were similar in sesame oil-treated control

  19. Differential ovarian morphometry and follicular expression of BMP15, GDF9 and BMPR1B influence the prolificacy in goat.

    PubMed

    Pramod, R K; Sharma, S K; Singhi, A; Pan, S; Mitra, A

    2013-10-01

    The variation in the kidding size of Black Bengal and Sirohi breed of goats makes them an interesting genetic material to study the underlying genetic mechanism of prolificacy. Accordingly, we studied the comparative ovarian morphometry including disparity in numbers of antral follicles of different sizes between these two breeds. Further, we evaluated the differential expression of the important candidate genes (viz., BMP15, GDF9 and BMPR1B) known to influence the ovulation rates and the prolificacy. The ovaries of Black Bengal (n = 20) goat were lighter (p < 0.01) in weight and smaller (p < 0.01) in diameter than those of Sirohi (n = 19) goats but possessed more numbers (p < 0.05) of corpus luteum (CL), large and small antral follicles. Quantitative real-time PCR (RT-qPCR) analysis revealed differential expression of mRNAs encoding for the BMP15 and GDF9. Small antral follicles of Black Bengal goats expressed 2.78-fold more (p < 0.05) of BMP 15 than those of Sirohi goat. Expression of BMP15 (p < 0.01) and GDF9 (p < 0.05) mRNAs was more abundant in the small than the large antral follicles of Black Bengal goat. The more numbers of antral follicles per unit of ovarian mass and differential expression of BMP15 and GDF9 may serve as an important clue for higher prolificacy.

  20. Relation between progesterone concentrations during the early luteal phase and follicular dynamics in goats.

    PubMed

    Menchaca, Alejo; Rubianes, Edgardo

    2002-03-15

    We studied the relationship between progesterone (P4) concentrations early in the estrus cycle and follicular dynamics in dairy goats. We used seven untreated goats (control group) and six progesterone treated goats (P group) with a controlled internal drug release device from Days 0 to 5 (Day 0: day of ovulation). We performed daily ultrasonograph during the interovulatory interval to determine ovarian change and took daily blood samples to determine serum estradiol 17beta (E2) and P4 concentrations by RIA. We divided the control goats into 3- (n = 4) and 4-wave goats (n = 3), according to the number of follicular waves recorded during the ovulatory cycle. Mean progesterone concentrations between Days I and 5 were higher and mean estradiol concentrations between Days 3 and 5 were lower in 4-wave goats (P4: 3.8+/-0.2 ng/ml; E2: 1.6+/-0.2 pg/ml) than in 3-wave goats (P4: 2.0+/-0.5 ng/ml, P < 0.05; E2: 4.4+/-0.9 pg/ml, P < 0.05). Wave 2 emerged earlier in 4-wave (Day 4.2+/-0.3) than in 3-wave goats (Day 7.3+/-0.3, P < 0.05). Three out of six of the progesterone-treated goats had short cycles (mean 8.0+/-0.0 days) and ovulated from Wave 1. The other three goats had shorter cycles (mean 18.3+/-0.3 days) than the control group (20.0+/-0.2 days; P < 0.05), although they were within the normal range of control cycles (shortened cycles). In the three treated goats with shortened cycles (two with four waves, one with three waves), mean progesterone concentrations between Days I and 5 were higher (4.7+/-0.6 ng/ml) than in the 3-wave control goats. In these goats, Wave 2 emerged at Day 4.3+/-0.3, similar to the time observed in 4-wave goats but earlier (P < or = 0.05) than in 3-wave control goats. Overall results confirm a relationship between the progesterone levels and the follicular wave turnover during the early luteal phase in the goat. Higher progesterone concentrations may accelerate follicular turnover probably by an early decline of the negative feedback action of

  1. Thermal equilibrium of goats.

    PubMed

    Maia, Alex S C; Nascimento, Sheila T; Nascimento, Carolina C N; Gebremedhin, Kifle G

    2016-05-01

    The effects of air temperature and relative humidity on thermal equilibrium of goats in a tropical region was evaluated. Nine non-pregnant Anglo Nubian nanny goats were used in the study. An indirect calorimeter was designed and developed to measure oxygen consumption, carbon dioxide production, methane production and water vapour pressure of the air exhaled from goats. Physiological parameters: rectal temperature, skin temperature, hair-coat temperature, expired air temperature and respiratory rate and volume as well as environmental parameters: air temperature, relative humidity and mean radiant temperature were measured. The results show that respiratory and volume rates and latent heat loss did not change significantly for air temperature between 22 and 26°C. In this temperature range, metabolic heat was lost mainly by convection and long-wave radiation. For temperature greater than 30°C, the goats maintained thermal equilibrium mainly by evaporative heat loss. At the higher air temperature, the respiratory and ventilation rates as well as body temperatures were significantly elevated. It can be concluded that for Anglo Nubian goats, the upper limit of air temperature for comfort is around 26°C when the goats are protected from direct solar radiation.

  2. Thermal equilibrium of goats.

    PubMed

    Maia, Alex S C; Nascimento, Sheila T; Nascimento, Carolina C N; Gebremedhin, Kifle G

    2016-05-01

    The effects of air temperature and relative humidity on thermal equilibrium of goats in a tropical region was evaluated. Nine non-pregnant Anglo Nubian nanny goats were used in the study. An indirect calorimeter was designed and developed to measure oxygen consumption, carbon dioxide production, methane production and water vapour pressure of the air exhaled from goats. Physiological parameters: rectal temperature, skin temperature, hair-coat temperature, expired air temperature and respiratory rate and volume as well as environmental parameters: air temperature, relative humidity and mean radiant temperature were measured. The results show that respiratory and volume rates and latent heat loss did not change significantly for air temperature between 22 and 26°C. In this temperature range, metabolic heat was lost mainly by convection and long-wave radiation. For temperature greater than 30°C, the goats maintained thermal equilibrium mainly by evaporative heat loss. At the higher air temperature, the respiratory and ventilation rates as well as body temperatures were significantly elevated. It can be concluded that for Anglo Nubian goats, the upper limit of air temperature for comfort is around 26°C when the goats are protected from direct solar radiation. PMID:27157333

  3. Inhibition of cytochrome P-450 C17 enzyme by a GnRH agonist in ovarian follicles from gonadotropin-stimulated rats.

    PubMed

    Irusta, Griselda; Parborell, Fernanda; Tesone, Marta

    2007-05-01

    Our objective was to study the direct action of a GnRH-I agonist, leuprolide acetate (LA), on ovarian steroidogenesis in preovulatory follicles obtained from equine chorionic gonadotropin (eCG)-treated rats. Previously, we have demonstrated an inhibitory effect of LA on steroidogenesis and follicular development. In this study, we tested the hypothesis that gonadotropin-releasing hormone (GnRH) exerts its negative effect on follicular development by inhibiting thecal cytochrome P-450 C17 (P450C17) alpha-hydroxylase expression and, consequently, androgen synthesis. Studies were carried out in prepubertal female rats injected with either eCG (control) or eCG plus LA (LA) and killed at different time points. Immunohistochemical studies indicated that LA induced steroidogenic acute regulatory protein (StAR) expression mainly in theca cells of preantral and antral follicles. In addition, serum progesterone levels increased significantly (P < 0.05), whereas those of androsterone decreased (P < 0.05) after 8 h of LA treatment. This inhibition caused by LA seemed to be a consequence of the decreased expression of follicular P450C17 alpha-hydroxylase, as demonstrated by Western blot and RT-PCR techniques. In vitro studies using follicles isolated from 48-h-eCG-treated rats and cultured with LA showed a significant (P < 0.05) inhibition of FSH-induced androsterone follicular content as well as P450C17 alpha-hydroxylase protein levels, as determined by Western analysis. However, LA increased StAR protein expression in these follicles without significant changes in P450scc enzyme levels. Taking all these findings into account, we suggest that GnRH-I exerts a direct inhibitory action on gonadotropin-induced follicular development by decreasing the temporal expression of the P450C17 enzyme and, consequently, androgen production, thus reducing the supply of estrogens available to developing follicles. PMID:17468395

  4. In vitro development of bovine secondary follicles in two- and three-dimensional culture systems using vascular endothelial growth factor, insulin-like growth factor-1, and growth hormone.

    PubMed

    Araújo, V R; Gastal, M O; Wischral, A; Figueiredo, J R; Gastal, E L

    2014-12-01

    The aim of this study was to evaluate the development and estradiol production of isolated bovine secondary follicles in two-dimensional (2D, experiment 1) and three-dimensional (3D using alginate, experiment 2) long-term culture systems in the absence (control group; only α-MEM(+)) or presence of vascular endothelial growth factor (VEGF), insulin-like growth factor-1, or GH alone, or a combination of all. A total of 363 isolated secondary follicles were cultured individually for 32 days at 38.5 °C in 5% CO2 in a humidified incubator with addition of medium (5 μL) every other day. In 2D culture system, follicular growth and antrum formation rates were higher (P < 0.05) in VEGF treatment compared with the other treatments. In 3D culture system, only estradiol concentration was greater (P < 0.05) in the GH than in the control group, whereas the other end points were similar (P > 0.05). In summary, this study demonstrated that the benefits of using a certain type of medium supplement depended on the culture system (2D vs. 3D). Vascular endothelial growth factor was an effective supplement for the in vitro culture of bovine secondary follicles when the 2D culture system was used, whereas GH only affected estradiol production using the 3D culture system. This study sheds light on advancements in methodology to facilitate subsequent studies on bovine preantral follicle development.

  5. Bioengineering the Ovarian Follicle Microenvironment

    PubMed Central

    Shea, Lonnie D.; Woodruff, Teresa K.; Shikanov, Ariella

    2014-01-01

    Chemo- and radiation therapies used to treat cancer can have the unintended effect of making patients infertile. Clinically established fertility preservation methods, such as egg and embryo cryopreservation, are not applicable to all patients, which has motivated the development of strategies that involve ovarian tissue removal and cryopreservation before the first sterilizing treatment. To restore fertility at a later date, the early-stage follicles present in the tissue must be matured to produce functional oocytes, a process that is not possible using existing cell culture technologies. This review describes the application of tissue engineering principles to promote ovarian follicle maturation and produce mature oocytes through either in vitro culture or transplantation. The design principles for these engineered systems are presented, along with identification of emerging opportunities in reproductive biology. PMID:24849592

  6. Bioengineering the ovarian follicle microenvironment.

    PubMed

    Shea, Lonnie D; Woodruff, Teresa K; Shikanov, Ariella

    2014-07-11

    Chemo- and radiation therapies used to treat cancer can have the unintended effect of making patients infertile. Clinically established fertility preservation methods, such as egg and embryo cryopreservation, are not applicable to all patients, which has motivated the development of strategies that involve ovarian tissue removal and cryopreservation before the first sterilizing treatment. To restore fertility at a later date, the early-stage follicles present in the tissue must be matured to produce functional oocytes, a process that is not possible using existing cell culture technologies. This review describes the application of tissue engineering principles to promote ovarian follicle maturation and produce mature oocytes through either in vitro culture or transplantation. The design principles for these engineered systems are presented, along with identification of emerging opportunities in reproductive biology.

  7. Effect of high progesterone concentrations during the early luteal phase on the length of the ovulatory cycle of goats.

    PubMed

    Menchaca, A; Rubianes, E

    2001-10-31

    The effect of exogenous progesterone exposure early in the oestrous cycle on the duration of the interovulatory interval was studied in dairy goats. A controlled intravaginal drug release (CIDR-G) device was inserted for 5 days starting at day 0 (D0 group, n=6) or day 3 (D3 group, n=5) postovulation. A third group was composed of untreated control goats (control group, n=7). Daily transrectal ultrasound was carried out during the interovulatory interval to assess the ovarian dynamics. Oestrous behaviour was checked twice a day and serum progesterone levels were assayed in daily jugular blood samples. Treated goats showed two different responses. In three D0 goats and one D3 goat, progesterone concentrations fell immediately after CIDR withdrawal and this was followed by oestrus and ovulation between days 8 and 11 (short cycles). In the other three D0 goats and in four D3 goats the treatment significantly reduced the interovulatory interval (18.3+/-0.3 and 18.5+/-0.3 days, respectively) (shortened cycles) compared with the control group (20.0+/-0.2 days; P<0.05), but the intervals with progesterone concentrations over 1 ng/ml were not different (15.7+/-0.3, 15.8+/-0.7 and 16.0+/-0.5 days for D0, D3 and control goats, respectively). In all D0 goats with a short cycle response, the ovulatory follicle arose from the first follicular wave but in the D3 goat with a short cycle it arose from the second follicular wave. These results showed that premature progesterone exposure early in the ovulatory cycle of the goat affected its length inducing short or shortened cycles. The effect of progesterone could either affect luteotropic support of the corpus luteum (CL) and/or stimulate a premature release of the luteolysin. PMID:11600275

  8. Expression of growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), and BMP receptors in the ovaries of goats.

    PubMed

    Silva, J R V; van den Hurk, R; van Tol, H T A; Roelen, B A J; Figueiredo, J R

    2005-01-01

    The process of ovarian folliculogenesis is composed of proliferation and differentiation of the constitutive cells in developing follicles. In goats, relatively little information is available on the local factors that regulate this process. We studied the presence and distribution of growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), and BMP receptors types 2 (BMPR2), 1A (BMPR1A), and 1B (BMPR1B) in goat ovaries to find evidence for their possible roles in folliculogenesis. Ovaries of cyclic goats were collected and fixed in paraformaldehyde for immunohistochemical localization of GDF9 and BMP15 proteins or used to collect follicles and luteal tissue to study the mRNA expression of GDF9, BMP15, and BMP receptors using reverse transcriptase polymerase chain reaction (RT-PCR). GDF9 and BMP15 proteins were found in oocytes of all types of follicles and granulosa cells of primary, secondary, and antral but not primordial follicles. The mRNAs for GDF9, BMP15, BMPR2, BMPR1A, and BMPR1B were detected in primordial, primary, and secondary follicles as well as in oocyte and granulosa cells of antral follicles. Transcripts for BMPR2, BMPR1A, BMPR1B, and GDF9, and GDF9 protein were furthermore found in corpora lutea. It is concluded that the mRNAs and proteins of GDF9 and BMP15 and the mRNAs of BMP receptors are expressed in goat ovarian follicles at all stages of their development, and that they form a complex intrafollicular regulatory system during folliculogenesis. Expression of all BMP receptor mRNAs and GDF9 mRNA and protein in luteal tissue additionally points to a role of GDF9 in corpus luteum function.

  9. Expression of cumulus-oocyte complex genes and embryonic development in goats subjected to progestogen-based estrus synchronization.

    PubMed

    Thammasiri, Jiratti; Navanukraw, Chainarong; Uriyapongson, Suthipong; Khanthusaeng, Vilaivan; Kamollirt, Chuchat

    2016-07-15

    The objective was to investigate the effect of short-term (7 days) and long-term (14 days) progesterone-based estrus synchronization on number of follicles, progesterone concentrations, cumulus-oocyte complex (COC) gene expression, and embryonic development in goats. Nulliparous Thai-native goats (n = 45) were randomly assigned to one of two estrus synchronization treatments. Goats were treated with intravaginal sponges containing 60-mg medroxyprogesterone acetate (MAP; Synchrogest esponjas, Spain) during 7 or 14 days (short-term or long-term protocol, respectively). Multiple follicular development was induced by intramuscularly injections of 300-IU eCG in both groups (1 day before sponge withdrawal). An ovariectomy was performed at 24 hours after sponge removal to evaluate number of follicle and collect oocyte for IVF. Oocyte quality (healthy or nonhealthy) was determined by morphology of COCs before IVM. Recovery of COCs and total cellular RNA isolation were applied to determine apoptosis-related gene expression. After IVF, embryos were evaluated during the eight-day culture as numbers of cleaved oocyte, morula, and blastocyst embryo. Total numbers of follicles and oocytes were similar for both treatments. Plasma progesterone concentrations were not different during MAP insertion period (P > 0.05). However, goats that received the short-term protocol had a greater number of 4 to 6-mm follicle, healthy oocytes, cleaved oocytes, and morula embryos than goats that received the long-term protocol (P < 0.01). In addition, the expression of B-cell lymphoma 2 messenger RNA was greater (P < 0.05) in COCs derived from the 7 days MAP-treated when compared to the 14 days MAP-treated goats. These data highlight that the 7-day progestin-based treatment may contribute to quality of oocytes and embryonic development in goats.

  10. 7 CFR 65.150 - Goat.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 3 2014-01-01 2014-01-01 false Goat. 65.150 Section 65.150 Agriculture Regulations of..., PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.150 Goat. Goat means meat produced from goats....

  11. 7 CFR 65.150 - Goat.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Goat. 65.150 Section 65.150 Agriculture Regulations of..., PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.150 Goat. Goat means meat produced from goats....

  12. 7 CFR 65.150 - Goat.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 3 2011-01-01 2011-01-01 false Goat. 65.150 Section 65.150 Agriculture Regulations of..., PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.150 Goat. Goat means meat produced from goats....

  13. 7 CFR 65.150 - Goat.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 3 2012-01-01 2012-01-01 false Goat. 65.150 Section 65.150 Agriculture Regulations of..., PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.150 Goat. Goat means meat produced from goats....

  14. 7 CFR 65.150 - Goat.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Goat. 65.150 Section 65.150 Agriculture Regulations of..., PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.150 Goat. Goat means meat produced from goats....

  15. Brock Cole's The Goats.

    ERIC Educational Resources Information Center

    Scales, Pat

    1997-01-01

    Summarizes Brock Cole's novel for young adolescents: "The Goats." Provides discussion questions and classroom activities in language arts, drama, research; mathematics, creative writing, similes; and presents an annotated bibliography of fiction for young adolescents dealing with runaways, self-reliance, family, friendship, courage, overweight,…

  16. Immunohistochemical study of hair follicle stem cells in regenerated hair follicles induced by Wnt10b

    PubMed Central

    Zhang, Yiming; Xing, Yizhan; Guo, Haiying; Ma, Xiaogen; Li, Yuhong

    2016-01-01

    The regulation of the periodic regeneration of hair follicles is complicated. Although Wnt10b has been reported to induce hair follicle regeneration, the characteristics of induced hair follicles, especially the target cells of Wnt10b, have not yet been clearly elucidated. Thus, we systematically evaluated the expression and proliferation patterns of Wnt10b-induced hair follicles. We found that Wnt10b promoted the proliferation of hair follicle stem cells from 24 hours after AdWnt10b injection. Seventy-two hours after AdWnt10b injection, cells outside of bulge area began to proliferate. When the induced hair follicle entered full anagen, although the hair follicle stem cells were normal, canonical Wnt signaling was maintained in the hair precortex cells. Our results reveal that the target cells that overexpressed Wnt10b included hair follicle stem cells, hair precortex cells, and matrix cells. PMID:27766026

  17. Does exogenous progesterone and oestradiol treatment from the mid-luteal phase induce follicular cysts in goats?

    PubMed

    Tanaka, Tomomi; Sawai, Rie; Kumai, Ryoko; Kim, Seungjoon; Kuroiwa, Takenobu; Kamomae, Hideo

    2007-02-01

    The purpose of the present study was to investigate the effects of exogenous ovarian steroid treatment, which is known to induce follicular cyst experimentally in cows, on ovarian activity in goats. Eleven female Shiba goats with the length of the normal oestrous cycle (approximately 21 days) received subcutaneously either 1 ml of ethanol (control group, n=4) or 4 mg of progesterone and 2mg of oestradiol (treatment group, n=7) daily for 7 days beginning on day 14 of the oestrous cycle (day 0=ovulation). Ultrasonographic images of the ovary and blood samples were collected daily to monitor the ovarian activity. Ovulation was observed before 1 day after the end of treatment in the control group. In the treatment group, no detectable structures of follicles or corpus luteum (static ovarian condition) were found for 6.0+/-1.4 days (mean+/-S.D.) after the end of treatment. Then, detectable follicles appeared and ovulation was observed in all animals of the treatment group. There was no significant difference in the maximum diameter of the ovulatory follicle between the control and treatment group (4.7+/-0.4mm versus 5.1+/-0.7 mm). The large non-ovulatory follicles, which grew more than 10mm in diameter were observed after the static ovarian condition in one goat of the treatment group, whereas no turnover of the cystic follicular structures was found. The length of the inter-ovulatory intervals in the treatment group was significantly longer than that in the control group (38.4+/-7.4 days versus 20.3+/-0.5 days, P<0.05). The present results demonstrated that the exogenous treatment of progesterone and oestradiol, which was adapted from the follicular cyst model in cows, did not induce follicular cysts in goats, suggesting that there is/are different mechanism(s) mediating the occurrence of follicular cysts between cows and goats. PMID:16542801

  18. Microinjection of follicle-enclosed mouse oocytes

    PubMed Central

    Jaffe, Laurinda A.; Norris, Rachael P.; Freudzon, Marina; Ratzan, William J.; Mehlmann, Lisa M.

    2011-01-01

    Summary The mammalian oocyte develops within a complex of somatic cells known as a follicle, within which signals from the somatic cells regulate the oocyte, and signals from the oocyte regulate the somatic cells. Because isolation of the oocyte from the follicle disrupts these communication pathways, oocyte physiology is best studied within an intact follicle. Here we describe methods for quantitative microinjection of follicle-enclosed mouse oocytes, thus allowing the introduction of signaling molecules as well as optical probes into the oocyte within its physiological environment. PMID:19085139

  19. Microinjection of Follicle-Enclosed Mouse Oocytes

    NASA Astrophysics Data System (ADS)

    Jaffe, Laurinda A.; Norris, Rachael P.; Freudzon, Marina; Ratzan, William J.; Mehlmann, Lisa M.

    The mammalian oocyte develops within a complex of somatic cells known as a follicle, within which signals from the somatic cells regulate the oocyte, and signals from the oocyte regulate the somatic cells. Because isolation of the oocyte from the follicle disrupts these communication pathways, oocyte physiology is best studied within an intact follicle. Here we describe methods for quantitative microinjection of follicle-enclosed mouse oocytes, thus allowing the introduction of signaling molecules as well as optical probes into the oocyte within its physiological environment.

  20. Reflectance spectroscopy for evaluating hair follicle cycle

    NASA Astrophysics Data System (ADS)

    Liu, Caihua; Guan, Yue; Wang, Jianru; Zhu, Dan

    2014-02-01

    Hair follicle, as a mini-organ with perpetually cycling of telogen, anagen and catagen, provides a valuable experimental model for studying hair and organ regeneration. The transition of hair follicle from telogen to anagen is a significant sign for successful regeneration. So far discrimination of the hair follicle stage is mostly based on canonical histological examination and empirical speculation based on skin color. Hardly a method has been proposed to quantitatively evaluate the hair follicle stage. In this work, a commercial optical fiber spectrometer was applied to monitor diffuse reflectance of mouse skin with hair follicle cycling, and then the change of reflectance was obtained. Histological examination was used to verify the hair follicle stage. In comparison with the histological examination, the skin diffuse reflectance was relatively high for mouse with telogen hair follicles; it decreased once hair follicles transited to anagen stage; then it increased reversely at catagen stage. This study provided a new method to quantitatively evaluate the hair follicle stage, and should be valuable for the basic and therapeutic investigations on hair regeneration.

  1. Follicle Structure Influences the Availability of Oxygen to the Oocyte in Antral Follicles

    PubMed Central

    Clark, A. R.; Stokes, Y. M.

    2011-01-01

    The ability of an oocyte to successfully mature is highly dependent on intrafollicular conditions, including the size and structure of the follicle. Here we present a mathematical model of oxygen transport in the antral follicle. We relate mean oxygen concentration in follicular fluid of bovine follicles to the concentration in the immediate vicinity of the cumulus-oocyte complex (COC). The model predicts that the oxygen levels within the antral follicle are dependent on the size and structure of the follicle and that the mean level of dissolved oxygen in follicular fluid does not necessarily correspond to that reaching the COC. PMID:22162722

  2. Meat goat production and marketing.

    PubMed

    Glimp, H A

    1995-01-01

    Production opportunities, management strategies, and marketing options for meat goats in the United States are reviewed in this manuscript. The basis for any expansion must be goat production systems that are biologically and economically sustainable, meeting both producer and consumer needs. Meat goats historically have been kept for brush control. Their use to control noxious plants and in vegetation management will continue to be their primary role in the future. Meat goats are rarely the primary animal production enterprise in the United States, but they are becoming increasingly important contributors to the income of many producers. Meat goat marketing is highly unstructured in the United States, yet prices are generally higher on a per unit of weight basis than other red meat-producing species. Efforts to organize marketing have had only limited success. Over 90% of the world's goats are in developing countries. Goats are increasingly important in these countries as subsistence food producers. Production systems range from goats being a part of nomadic multispecies herds on arid desert rangelands, in agropastoral production systems, to goats being the primary animal enterprise in smallholder farming systems.

  3. Ion beam microanalysis of human hair follicles

    NASA Astrophysics Data System (ADS)

    Kertész, Zs.; Szikszai, Z.; Pelicon, P.; Simčič, J.; Telek, A.; Bíró, T.

    2007-07-01

    Hair follicle is an appendage organ of the skin which is of importance to the survival of mammals and still maintains significance for the human race - not just biologically, but also through cosmetic and commercial considerations. However data on composition of hair follicles are scarce and mostly limited to the hair shaft. In this study we provide detailed information on the elemental distribution in human hair follicles in different growth phases (anagen and catagen) using a scanning proton microprobe. The analysis of skin samples obtained from human adults undergoing plastic surgery and of organ-cultured human hair follicles may yield a new insight into the function, development and cyclic activity of the hair follicle.

  4. Reduced quality and accelerated follicle loss with female reproductive aging - does decline in theca dehydroepiandrosterone (DHEA) underlie the problem?

    PubMed Central

    2013-01-01

    Infertility, spontaneous abortion and conception of trisomic offspring increase exponentially with age in mammals but in women there is an apparent acceleration in the rate from about age 37. The problems mostly commonly occur when the ovarian pool of follicles is depleted to a critical level with age but are also found in low follicular reserve of other etiologies. Since recent clinical studies have indicated that dehydroepiandrosterone (DHEA) supplementation may reverse the problem of oocyte quality, this review of the literature was undertaken in an attempt to find an explanation of why this is effective? In affected ovaries, oxygenation of follicular fluid is low, ultrastructural disturbances especially of mitochondria, occur in granulosa cells and oocytes, and considerable disturbances of meiosis occur. There is, however, no evidence to date that primordial follicles are compromised. In females with normal fertility, pre-antral ovarian theca cells respond to stimulation by inhibin B to provide androgen-based support for the developing follicle. With depletion of follicle numbers, inhibin B is reduced with consequent reduction in theca DHEA. Theca cells are the sole ovarian site of synthesis of DHEA, which is both a precursor of androstenedione and an essential ligand for peroxisome proliferator-activated receptor alpha (PPARα), the key promoter of genes affecting fatty acid metabolism and fat transport and genes critical to mitochondrial function. As well as inducing a plethora of deleterious changes in follicular cytoplasmic structure and function, the omega 9 palmitate/oleate ratio is increased by lowered activity of PPARα. This provides conditions for increased ceramide synthesis and follicular loss through ceramide-induced apoptosis is accelerated. In humans critical theca DHEA synthesis occurs at about 70 days prior to ovulation thus effective supplementation needs to be undertaken about four months prior to intended conception; timing which is also

  5. Depression of ovarian function and plasma progesterone and estradiol-17 beta in female goats chronically infected with Trypanosoma congolense.

    PubMed

    Mutayoba, B M; Gombe, S; Waindi, E N; Kaaya, G P

    1988-04-01

    Adult normocyclic female goats experimentally infected with Trypanosoma congolense developed irregular and shorter estrous cycles before complete cessation at the fourth cycle post-infection. This was followed within a month by a decline in the mean plasma progesterone and estradiol-17 beta levels. The peak luteal progesterone as well as pre-ovulatory estradiol-17 beta level declined progressively from the second to the fourth cycle post-infection. The ovaries became atretic with reduced numbers of primordial and primary follicles. The larger follicles became atretic at the tertiary stage with subsequent lack of corpora lutea formation. The rapidity of ovarian dysfunction appeared to be related to the degree of susceptibility of the individual infected goats. PMID:3389040

  6. The Goat in the Rug.

    ERIC Educational Resources Information Center

    Blood, Charles L.; Link, Martin

    Based on the activities of the real Window Rock weaver, Glenmae, and her goat, Geraldine, this illustrated story incorporates authentic details relative to the Navajo art of rug weaving and is designed for children aged four to eight. Capitalizing on the humor inherent in Geraldine's point of view, the story centers on the goat's observation of…

  7. Cutaneous pythiosis in a goat.

    PubMed

    do Carmo, P M S; Portela, R A; Silva, T R; Oliveira-Filho, J C; Riet-Correa, F

    2015-01-01

    Pythiosis is reported for the first time in a goat. The affected goat had daily access to an aquatic environment and had developed an ulcerative lesion on the skin of the left hindlimb. Microscopically, there were dermal pyogranulomas with 'negatively stained' hyphae, which were identified immunohistochemically as Pythium insidiosum. PMID:25555632

  8. Biotechnological advances in goat reproduction.

    PubMed

    Amoah, E A; Gelaye, S

    1997-02-01

    Goat selection and reproduction have resulted from using conventional methods of natural mating and artificial insemination. Genetic improvements resulting from these are usually slow. Innovative developments in biotechnology rapidly propagate superior genes, offering hope for modeling and designing animals to fit market and environmental requirements. Use of Tris, citric acid, fructose, egg yolk, and glycerol extender has enabled goat sperm to be stored successfully for several years before being used in cervical or laparoscopic insemination. Laparoscopic recovery of goat embryos to reduce adhesions from repeat surgeries has great potential in improving embryo production for direct transfer or for future transfer after cryopreservation. Goat kids have been produced, as a result of experiments to refine techniques of in vitro maturation and fertilization of recovered oocytes, with successful culture and transfer of embryos. In vitro fertilization technology is also essential for cloning goat embryos and for gene transfer. Transgenic goats have already been produced due to new genes being expressed from biologically active molecules altering the phenotype of the transferred goat. The introduced gene is capable of transmission between generations. The goat's diversified commercial value and convenient size make it a benefactor to new technology for rapid genetic improvement as a supplement to conventional selection methods. PMID:9051483

  9. 7 CFR 65.165 - Ground goat.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Ground goat. 65.165 Section 65.165 Agriculture... OF BEEF, PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.165 Ground goat. Ground goat means...

  10. 7 CFR 65.165 - Ground goat.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Ground goat. 65.165 Section 65.165 Agriculture... OF BEEF, PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.165 Ground goat. Ground goat means...

  11. 7 CFR 65.165 - Ground goat.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 3 2012-01-01 2012-01-01 false Ground goat. 65.165 Section 65.165 Agriculture... OF BEEF, PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.165 Ground goat. Ground goat means...

  12. 7 CFR 65.165 - Ground goat.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 3 2014-01-01 2014-01-01 false Ground goat. 65.165 Section 65.165 Agriculture... OF BEEF, PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.165 Ground goat. Ground goat means...

  13. 7 CFR 65.165 - Ground goat.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 3 2011-01-01 2011-01-01 false Ground goat. 65.165 Section 65.165 Agriculture... OF BEEF, PORK, LAMB, CHICKEN, GOAT MEAT, PERISHABLE AGRICULTURAL COMMODITIES, MACADAMIA NUTS, PECANS, PEANUTS, AND GINSENG General Provisions Definitions § 65.165 Ground goat. Ground goat means...

  14. A Microarray-Based Analysis Reveals that a Short Photoperiod Promotes Hair Growth in the Arbas Cashmere Goat

    PubMed Central

    Guo, Jun; Wu, Dubala; Hao, Bayasihuliang; Li, Yurong; Zhao, Cunfa

    2016-01-01

    Many animals exhibit different behaviors in different seasons. The photoperiod can have effects on migration, breeding, fur growth, and other processes. The cyclic growth of the fur and feathers of some species of mammals and birds, respectively, is stimulated by the photoperiod as a result of hormone-dependent regulation of the nervous system. To further examine this phenomenon, we evaluated the Arbas Cashmere goat (Capra hircus), a species that is often used in this type of research. The goats were exposed to an experimentally controlled short photoperiod to study the regulation of cyclic cashmere growth. Exposure to a short photoperiod extended the anagen phase of the Cashmere goat hair follicle to increase cashmere production. Assessments of tissue sections indicated that the short photoperiod significantly induced cashmere growth. This conclusion was supported by a comparison of the differences in gene expression between the short photoperiod and natural conditions using gene chip technology. Using the gene chip data, we identified genes that showed altered expression under the short photoperiod compared to natural conditions, and these genes were found to be involved in the biological processes of hair follicle growth, structural composition of the hair follicle, and the morphogenesis of the surrounding skin appendages. Knowledge about differences in the expression of these genes as well as their functions and periodic regulation patterns increases our understanding of Cashmere goat hair follicle growth. This study also provides preliminary data that may be useful for the development of an artificial method to improve cashmere production by controlling the light cycle, which has practical significance for livestock breeding. PMID:26814503

  15. Effects of repetition, interval between treatments and season on the results from laparoscopic ovum pick-up in goats.

    PubMed

    Pierson, J; Wang, B; Neveu, N; Sneek, L; Côté, F; Karatzas, C N; Baldassarre, H

    2004-01-01

    The present study was conducted to evaluate the follicular response and oocyte yield following repeated gonadotrophin stimulation and laparoscopic aspiration in goats and to assess the effects of the time interval between procedures and season. A total of 98 adult goats were subjected to laparoscopic ovum pick-up (LOPU) five consecutive times in a transgenic production programme. Oestrus was synchronised by means of intravaginal sponges inserted for 10 days coupled with 125 microg cloprostenol 36 h before sponge removal and LOPU, and follicular development was stimulated with 80 mg follicle stimulating hormone and 300 IU equine chorionic gonadotrophin administered 36 h before LOPU. No difference was detected in the response for LOPUs 1, 2, 3 and 4. Although a small decrease in response was detected at LOPU 5 (P < 0.05), the numbers of follicles aspirated and oocytes recovered were not different from those at LOPU 1 and LOPUs 1 and 4, respectively. With respect to time interval between LOPU and season, all intervals and seasons produced acceptable responses, with no difference in follicles aspirated and oocytes recovered between intervals and seasons. These results indicate that LOPU may be repeated up to five times in goats at different intervals and in different seasons with little or no important change in overall response.

  16. Molecular cloning, expression analysis, and function of decorin in goat ovarian granulosa cells.

    PubMed

    Peng, J Y; Gao, K X; Xin, H Y; Han, P; Zhu, G Q; Cao, B Y

    2016-10-01

    Decorin (DCN), a component of the extracellular matrix (ECM), participates in ECM assembly and influences cell proliferation and apoptosis in many mammalian tissues and cells. However, expression and function of DCN in the ovary remain unclear. This study cloned the full-length cDNA of goat DCN obtained from the ovary of an adult goat. Sequence analysis revealed that the putative DCN protein shared a highly conserved amino acid sequence with known mammalian homologs. The tissue distribution of DCN mRNA expression was evaluated by real-time PCR, and the results showed that DCN was widely expressed in the tissues of adult goat. Immunohistochemistry results suggested that DCN protein existed in the granulosa cells and oocytes from all types of follicles and theca cells of antral follicles. Moreover, hCG-induced DCN mRNA expression was significantly reduced by the inhibitors of protein kinase A, PI3K, or p38 kinase (P < 0.05), which are key mediators involved in hCG-induced DCN expression. Overexpression of DCN significantly increased apoptosis and blocked cell cycle progression in cultured granulosa cells (P < 0.05). Western blot analysis also showed that overexpression of DCN upregulated the expression levels of p21 protein (P < 0.05), whereas no effects were observed on the expression of Bax and Bcl-2 and on Bcl-2/Bax ratio (P > 0.05). These findings suggested that DCN regulates the apoptosis and cell cycle of granulosa cells. PMID:27565237

  17. Proteomic Analysis of Hair Follicles

    NASA Astrophysics Data System (ADS)

    Ishioka, Noriaki; Terada, Masahiro; Yamada, Shin; Seki, Masaya; Takahashi, Rika; Majima, Hideyuki J.; Higashibata, Akira; Mukai, Chiaki

    2013-02-01

    Hair root cells actively divide in a hair follicle, and they sensitively reflect physical conditions. By analyzing the human hair, we can know stress levels on the human body and metabolic conditions caused by microgravity environment and cosmic radiation. The Japan Aerospace Exploration Agency (JAXA) has initiated a human research study to investigate the effects of long-term space flight on gene expression and mineral metabolism by analyzing hair samples of astronauts who stayed in the International Space Station (ISS) for 6 months. During long-term flights, the physiological effects on astronauts include muscle atrophy and bone calcium loss. Furthermore, radiation and psychological effects are important issue to consider. Therefore, an understanding of the effects of the space environment is important for developing countermeasures against the effects experienced by astronauts. In this experiment, we identify functionally important target proteins that integrate transcriptome, mineral metabolism and proteome profiles from human hair. To compare the protein expression data with the gene expression data from hair roots, we developed the protein processing method. We extracted the protein from five strands of hair using ISOGEN reagents. Then, these extracted proteins were analyzed by LC-MS/MS. These collected profiles will give us useful physiological information to examine the effect of space flight.

  18. Extinction of Harrington's mountain goat.

    PubMed

    Mead, J I; Martin, P S; Euler, R C; Long, A; Jull, A J; Toolin, L J; Donahue, D J; Linick, T W

    1986-02-01

    Keratinous horn sheaths of the extinct Harrington's mountain goat, Oreamnos harringtoni, were recovered at or near the surface of dry caves of the Grand Canyon, Arizona. Twenty-three separate specimens from two caves were dated nondestructively by the tandem accelerator mass spectrometer (TAMS). Both the TAMS and the conventional dates indicate that Harrington's mountain goat occupied the Grand Canyon for at least 19,000 years prior to becoming extinct by 11,160 +/- 125 radiocarbon years before present. The youngest average radiocarbon dates on Shasta ground sloths, Nothrotheriops shastensis, from the region are not significantly younger than those on extinct mountain goats. Rather than sequential extinction with Harrington's mountain goat disappearing from the Grand Canyon before the ground sloths, as one might predict in view of evidence of climatic warming at the time, the losses were concurrent. Both extinctions coincide with the regional arrival of Clovis hunters.

  19. Extinction of Harrington's mountain goat

    SciTech Connect

    Mead, J.I.; Martin, P.S.; Euler, R.C.; Long, A.; Jull, A.J.T.; Toolin, L.J.; Donahue, D.J.; Linick, T.W.

    1986-02-01

    Keratinous horn sheaths of the extinct Harrington's mountain goat, Oreamnos harringtoni, were recovered at or near the surface of dry caves of the Grand Canyon, Arizona. Twenty-three separate specimens from two caves were dated nondestructively by the tandem accelerator mass spectrometer (TAMS). Both the TAMS and the conventional dates indicate that Harrington's mountain goat occupied the Grand Canyon for at least 19,000 years prior to becoming extinct by 11,160 +/- 125 radiocarbon years before present. The youngest average radiocarbon dates on Shasta ground sloths, Nothrotheriops shastensis, from the region are not significantly younger than those on extinct mountain goats. Rather than sequential extinction with Harrington's mountain goat disappearing from the Grand Canyon before the ground sloths, as one might predict in view of evidence of climatic warming at the time, the losses were concurrent. Both extinctions coincide with the regional arrival of Clovis hunters.

  20. Extinction of Harrington's Mountain Goat

    NASA Astrophysics Data System (ADS)

    Mead, Jim I.; Martin, Paul S.; Euler, Robert C.; Long, Austin; Jull, A. J. T.; Toolin, Laurence J.; Donahue, Douglas J.; Linick, T. W.

    1986-02-01

    Keratinous horn sheaths of the extinct Harrington's mountain goat, Oreamnos harringtoni, were recovered at or near the surface of dry caves of the Grand Canyon, Arizona. Twenty-three separate specimens from two caves were dated nondestructively by the tandem accelerator mass spectrometer (TAMS). Both the TAMS and the conventional dates indicate that Harrington's mountain goat occupied the Grand Canyon for at least 19,000 years prior to becoming extinct by 11,160 ± 125 radiocarbon years before present. The youngest average radiocarbon dates on Shasta ground sloths, Nothrotheriops shastensis, from the region are not significantly younger than those on extinct mountain goats. Rather than sequential extinction with Harrington's mountain goat disappearing from the Grand Canyon before the ground sloths, as one might predict in view of evidence of climatic warming at the time, the losses were concurrent. Both extinctions coincide with the regional arrival of Clovis hunters.

  1. Ovarian dynamics, serum estradiol and progesterone concentrations during the interovulatory interval in goats.

    PubMed

    de Castro, T; Rubianes, E; Menchaca, A; Rivero, A

    1999-08-01

    Ovarian changes determined by daily transrectal ultrasonic scanning, and its correlation with serum progesterone (P4) and estradiol (E2) concentrations were studied in seven cyclic Saanen goats. Estrous cycles were synchronized with 2 injections of a PGF2 alpha analogue 9 d apart. All follicles > or = 2 mm in diameter and CL were measured each day. One goat showed a longer interestrous interval, associated with development of a cystic-luteinized structure. The mean interovulatory interval for the other 6 goats was 20.8 +/- 0.4 d. The incidence of goats with 4, 3, and 2 follicular waves was 3, 1 and 2 respectively; follicular waves emerged on Days 0.5 +/- 0.6, 7.2 +/- 0.7, 10.7 +/- 0.5 and 13.7 +/- 0.8 for Wave 1, 2, 3 and the Ovulatory wave, respectively. The largest follicle of Wave 2 was smaller (4.9 +/- 0.1 mm) than the largest follicles of Wave 3 (6.2 +/- 0.1 mm; P < or = 0.01) and of the Ovulatory wave (7.0 +/- 0.5 mm; P < or = 0.01), and tended to be smaller than the largest follicle of Wave 1 (6.3 +/- 0.6 mm; P < or = 0.09). Interval between emergence of Wave 1 and Wave 2 was longer than interval between emergence of Wave 2 and Wave 3 (7.3 +/- 0.9 d vs 4.0 +/- 0.4 d; P < or = 0.01), and between Wave 3 and the Ovulatory wave (3.8 +/- 1.1 d; P < or = 0.05). Two days before ovulation, the diameter of the ovulatory follicle was larger (P < or = 0.01) than the first subordinate follicle. Serum E2 concentrations increased from the day of ovulation (2.7 +/- 0.3 pg/mL) to Day 2 (7.6 +/- 0.9 pg/mL; P < or = 0.01), associated with the early-mid growing phase of the largest follicle of Wave 1, and then decreased to basal levels on Day 5 (P < or = 0.01) and peaked again (16.5 +/- 2.4 pg/mL) 2 d before ovulation. The CL were detected ultrasonically on Day 3 post ovulation and attained a mean maximum diameter of 13.5 +/- 0.8 mm between Days 8 and 14. The following characteristics were observed: 1) ovarian follicular development in goats is wave-like; 2) increased P4

  2. GOATS Image Projection Component

    NASA Technical Reports Server (NTRS)

    Haber, Benjamin M.; Green, Joseph J.

    2011-01-01

    When doing mission analysis and design of an imaging system in orbit around the Earth, answering the fundamental question of imaging performance requires an understanding of the image products that will be produced by the imaging system. GOATS software represents a series of MATLAB functions to provide for geometric image projections. Unique features of the software include function modularity, a standard MATLAB interface, easy-to-understand first-principles-based analysis, and the ability to perform geometric image projections of framing type imaging systems. The software modules are created for maximum analysis utility, and can all be used independently for many varied analysis tasks, or used in conjunction with other orbit analysis tools.

  3. Expression and regulative function of tissue inhibitor of metalloproteinase 3 in the goat ovary and its role in cultured granulosa cells.

    PubMed

    Peng, Jiayin; Xin, Haiyun; Han, Peng; Gao, Kexin; Gao, Teyang; Lei, Yingnan; Ji, Shengyue; An, Xiaopeng; Cao, Binyun

    2015-09-01

    Tissue inhibitor of metalloproteinase 3 (TIMP3) played a key role in female reproduction. However, its expression and function in goat are still unclear. In the present study, the full-length cDNA of goat TIMP3 was cloned from adult goat ovary; meanwhile, we demonstrated that putative TIMP3 protein shared a highly conserved amino acid sequence with known mammalian homologs. Real-time PCR results showed that TIMP3 was widely expressed in the tissues of adult goat. In the ovary, increasing expression of TIMP3 mRNA was discovered during the growth process of follicle and corpus luteum. Immunohistochemistry results suggested that TIMP3 protein existed in oocytes of all types of follicles, corpus luteum and granulosa and theca cells of primary, secondary, and antral but not primordial follicles. In vitro, human chorionic gonadotropin (hCG) stimulated the expression of TIMP3 in goat granulosa cells. hCG-induced TIMP3 mRNA expression was reduced by the inhibitors of protein kinase A, protein kinase C, MAPK kinase, or p38 kinase. Functionally, over-expression of TIMP3 significantly increased apoptosis and decreased the viability of cultured granulosa cells. Knockdown of TIMP3 could decrease hCG-induced progesterone secretion and the mRNA abundance of key steroidogenic enzymes (StAR, p450scc and HSD3B) as well as ECM proteins (DCN and FN). These findings provided evidence that the hCG induced expression of TIMP3 may play an important role in regulating goat granulosa cell survival and steroidogenesis.

  4. Pathogenesis of caprine arthritis encephalitis virus. Cellular localization of viral transcripts in tissues of infected goats.

    PubMed Central

    Zink, M. C.; Yager, J. A.; Myers, J. D.

    1990-01-01

    Pathologic specimens of 18 goats with classical lesions of caprine arthritis-encephalitis (CAE) virus infection were examined morphologically and by in situ hybridization using molecularly cloned CAEV deoxyribonucleic acid (DNA) to determine which tissues and cells of naturally infected goats supported virus replication. Large numbers of cells with viral transcripts were detected in inflamed brain, spinal cord, lung, joints, and mammary gland. These cells were morphologically compatible with macrophages. Fewer cells with viral transcripts were seen in noninflamed tissues. Viral RNA was identified in macrophagelike cells in lung, liver, spleen, and lymph nodes, in cells lining the vessels of brain and synovium, and in epithelial cells of intestinal crypts, renal tubules, and thyroid follicles. These data suggest that the cell tropism of lentiviruses may extend beyond the narrow boundaries of lymphocytes and macrophages. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 PMID:2327471

  5. Biological basis of follicle growth.

    PubMed

    Hodgen, G D

    1989-11-01

    Ovarian stimulation therapy is notoriously difficult to manage, partly because of the individual variability in response to exogenous hormones and hormone antagonists. The varied underlying pathophysiological conditions necessitating ovulation induction therapy were in great part causative of the different responses between patients. However, in-vitro fertilization therapy has provided a wider experience, particularly with ovarian stimulation in endocrinologically normal women. From these data, there is much inherent variability of response to exogenous gonadotrophins evident, even in these 'normal patients'. Furthermore, individual response types tend toward a consistent pattern from cycle to cycle, suggesting a constancy of physiological status, as opposed to a stoichastic response. In this study, we are seeking ways of reducing individual variability of responses to gonadotrophin therapy based on understanding the physiological origin. Our first objective is to identify the source of individual variability in the hypothalamic--pituitary ovarian axis. We developed a new experimental primate model, the reversible 'medical hypophysectomy', achieved by administration of a gonadotrophin-releasing hormone (GnRH) antagonist to negate endogenous gonadotrophin secretion. The second objective is to assess the validity of our model for medical hypophysectomy. Effective doses and regimens of the GnRH antagonist applications are determined. The third objective is to study the differential actions of follicle-stimulating hormone (FSH) treatment alone versus combined FSH/luteinizing hormone (LH) therapy on the ovary. In summary, it is concluded: (i) individual variation in ovarian response to gonadotrophin therapy in monkeys is, in large part, supra-ovarian in origin, although failure to respond to gonadotrophins was not negated by GnRH antagonist treatment. Furthermore, a milieu approaching 'medical hypophysectomy' using a potent GnRH antagonist deserves consideration as a

  6. Fertility and blood progesterone levels following LHRH-induced superovulation in FSH-treated anestrous goats.

    PubMed

    Akinlosotu, B A; Wilder, C D

    1993-11-01

    Twenty mature, mixed-breed, seasonally anestrous female goats were used to study the effects of luteinizing hormone releasing hormone (LHRH) on ovulation rate, fertility, and blood progesterone levels following norgestomet-induced estrus and follicle stimulating hormone (FSH) treatments. Each goat received 6 mg norgestomet by subcutaneous (sc) implant and 3 mg intramuscularly, along with an intramuscular (im) injection of 5 mg estradiol valerate. Four injections of FSH were given for 2 d in divided doses of 10, 10, 5 and 5 mg im every 12 h, starting at 24 h before implant removal. The goats were randomly assigned to 1 of 2 equal treatment groups, and were treated with 2 intravenous (iv) injections of either 0.9% saline (control) or 300 ug LHRH at 24 and 48 h after the removal of the implants. All the goats exhibited estrus within 24 or 36 h of implant withdrawal and were mated to bucks of proven fertility. At laparotomy on Day 7 or 8 after the removal of the implants, the mean number of unovulated follicles was higher (P<0.05) in Group I than in Group II. The mean number of corpora lutea (ovulation rate), the total number of embryos and the number of normal embryos recovered were higher (P<0.05) in LHRH-treated does than in the controls. Treatment with LHRH resulted in 72.14% fertility (mean number of CL = 14) as compared with the controls with 64.29% fertility (mean number of CL = 2.8). The embryos obtained from goats in Group II were of more uniform developmental age regardless of the day of embryo collection, as compared with those of the controls. Plasma progesterone levels were significantly increased on Days 4 to 6 in both treatment groups. The results of this study have demonstrated that the FSH and LHRH treatment regimen increased follicular development, ovulation rate and blood progesterone levels in norgestomet-treated anestrous goats. Moreover, LHRH treatment enhanced fertility, and improved embryo quality as indicated by the significantly higher total

  7. Ovarian response to oestrous synchronization protocol based on use of reduced doses of cloprostenol in cyclic goats.

    PubMed

    Duque-Bonisoli, C; Salvador, A; Díaz, T; Contreras-Solis, I

    2012-12-01

    The objective of this study was to assess the efficacy of two reduced doses vs a high/luteolytic dose of cloprostenol on luteolytic activity and synchronization of oestrus in cyclic goats. Experiment 1, included 24 goats randomly allocated to three groups: control group (group H) received a single high dose of cloprostenol (87.5 μg; 1.0 ml; i.m.) and M and L groups, which received half (43.75 μg; 0.5 ml) and a third (26.25 μg; 0.3 ml) of the highest dose, respectively. Experiment 2, included 24 goats randomly assigned to the same experimental groups. Each group was treated using two injections of cloprostenol administered 10 days apart to synchronize oestrus. Transrectal ultrasonographic scanning (US) was performed to detect the presence, size and development of corpora lutea and ovarian follicles. Furthermore, detection of oestrus was performed every 12 h between 24 and 72 h after the second injection of cloprostenol, and the luteolytic effect was verified by US. In Experiment 1, all goats that had corpora lutea at timing of treatment regressed their corpora lutea. In Experiment 2, the occurrence of oestrus and the interval between treatment to onset of oestrus were: 100%, 49.5 ± 3.0 h; 100%, 51.0 ± 3.0 h; and 75%, 56.0 ± 3.5 h for H, M and L groups, respectively. The development of preovulatory follicles and occurrence of subsequent corpora lutea were similar among groups. In summary, the use of 26.25 μg of cloprostenol is effective for the synchronization of oestrus in cyclic goats.

  8. GOATS - Orbitology Component

    NASA Technical Reports Server (NTRS)

    Haber, Benjamin M.; Green, Joseph J.

    2010-01-01

    The GOATS Orbitology Component software was developed to specifically address the concerns presented by orbit analysis tools that are often written as stand-alone applications. These applications do not easily interface with standard JPL first-principles analysis tools, and have a steep learning curve due to their complicated nature. This toolset is written as a series of MATLAB functions, allowing seamless integration into existing JPL optical systems engineering modeling and analysis modules. The functions are completely open, and allow for advanced users to delve into and modify the underlying physics being modeled. Additionally, this software module fills an analysis gap, allowing for quick, high-level mission analysis trades without the need for detailed and complicated orbit analysis using commercial stand-alone tools. This software consists of a series of MATLAB functions to provide for geometric orbit-related analysis. This includes propagation of orbits to varying levels of generalization. In the simplest case, geosynchronous orbits can be modeled by specifying a subset of three orbit elements. The next case is a circular orbit, which can be specified by a subset of four orbit elements. The most general case is an arbitrary elliptical orbit specified by all six orbit elements. These orbits are all solved geometrically, under the basic problem of an object in circular (or elliptical) orbit around a rotating spheroid. The orbit functions output time series ground tracks, which serve as the basis for more detailed orbit analysis. This software module also includes functions to track the positions of the Sun, Moon, and arbitrary celestial bodies specified by right ascension and declination. Also included are functions to calculate line-of-sight geometries to ground-based targets, angular rotations and decompositions, and other line-of-site calculations. The toolset allows for the rapid execution of orbit trade studies at the level of detail required for the

  9. GOAT induced ghrelin acylation regulates hedonic feeding.

    PubMed

    Davis, J F; Perello, M; Choi, D L; Magrisso, I J; Kirchner, H; Pfluger, P T; Tschoep, M; Zigman, J M; Benoit, S C

    2012-11-01

    Ghrelin is an orexigenic hormone that regulates homeostatic and reward-related feeding behavior. Recent evidence indicates that acylation of ghrelin by the gut enzyme ghrelin O-acyl transferase (GOAT) is necessary to render ghrelin maximally active within its target tissues. Here we tested the hypothesis that GOAT activity modulates food motivation and food hedonics using behavioral pharmacology and mutant mice deficient for GOAT and the ghrelin receptor (GHSR). We evaluated operant responding following pharmacological administration of acyl-ghrelin and assessed the necessity of endogenous GOAT activity for operant responding in GOAT and GHSR-null mice. Hedonic-based feeding behavior also was examined in GOAT-KO and GHSR-null mice using a "Dessert Effect" protocol in which the intake of a palatable high fat diet "dessert" was assessed in calorically-sated mice. Pharmacological administration of acyl-ghrelin augmented operant responding; notably, this effect was dependent on intact GHSR signaling. GOAT-KO mice displayed attenuated operant responding and decreased hedonic feeding relative to controls. These behavioral results correlated with decreased expression of the orexin-1 receptor in reward-related brain regions in GOAT-KO mice. In summary, the ability of ghrelin to stimulate food motivation is dependent on intact GHSR signaling and modified by endogenous GOAT activity. Furthermore, GOAT activity is required for hedonic feeding behavior, an effect potentially mediated by forebrain orexin signaling. These data highlight the significance of the GOAT-ghrelin system for the mediation of food motivation and hedonic feeding.

  10. Gene and stem cell therapy of the hair follicle.

    PubMed

    Hoffman, Robert M

    2005-01-01

    The hair follicle is a highly complex appendage of the skin containing a multiplicity of cell types. The follicle undergoes constant cycling through the life of the organism including growth and resorption with growth dependent on specific stem cells. The targeting of the follicle by genes and stem cells to change its properties, in particular, the nature of the hair shaft is discussed. Hair follicle delivery systems are described such as liposomes and viral vectors for gene therapy. The nature of the hair follicle stem cells is discussed, in particular, its pluripotency.

  11. 4-H Club Goat Guide.

    ERIC Educational Resources Information Center

    Brown, R. Kipp

    This guide provides information for 4-H Club members who have decided on a club goat project. Topics include general information in the following areas: show rules; facilities and equipment (barns/sheds, fences, feeders, water containers, and equipment); selection (structural correctness, muscle, volume and capacity, style and balance, and growth…

  12. Size-specific follicle selection improves mouse oocyte reproductive outcomes.

    PubMed

    Xiao, Shuo; Duncan, Francesca E; Bai, Lu; Nguyen, Catherine T; Shea, Lonnie D; Woodruff, Teresa K

    2015-09-01

    Encapsulated in vitro follicle growth (eIVFG) has great potential to provide an additional fertility preservation option for young women and girls with cancer or other reproductive health threatening diseases. Currently, follicles are cultured for a defined period of time and analyzed as a cohort. However, follicle growth is not synchronous, and culturing follicles for insufficient or excessive times can result in compromised gamete quality. Our objective is to determine whether the selection of follicles based on size, rather than absolute culture time, better predict follicle maturity and oocyte quality. Multilayer secondary mouse follicles were isolated and encapsulated in 0.25% alginate. Follicles were cultured individually either for defined time periods or up to specific follicle diameter ranges, at which point several reproductive endpoints were analyzed. The metaphase II (MII) percentage after oocyte maturation on day 6 was the highest (85%) when follicles were cultured for specific days. However, if follicles were cultured to a terminal diameter of 300-350 μm irrespective of absolute time in culture, 93% of the oocytes reached MII. More than 90% of MII oocytes matured from follicles with diameters of 300-350 μm showed normal spindle morphology and chromosome alignment, 85% of oocytes showed two pronuclei after IVF, 81% developed into the two-cell embryo stage and 38% developed to the blastocyst stage, all significantly higher than the percentages in the other follicle size groups. Our study demonstrates that size-specific follicle selection can be used as a non-invasive marker to identify high-quality oocytes and improve reproductive outcomes during eIVFG. PMID:26116002

  13. Expression of receptors for BMP15 is differentially regulated in dominant and subordinate follicles during follicle deviation in cattle.

    PubMed

    Gasperin, Bernardo G; Ferreira, Rogério; Rovani, Monique T; Bordignon, Vilceu; Duggavathi, Raj; Buratini, José; Oliveira, João F C; Gonçalves, Paulo B D

    2014-01-30

    Bone morphogenetic proteins are known to be involved in determining ovulation rate in mammals. The mechanisms through which these proteins determine follicle fate are incompletely understood. In the present study, we used cattle as a model to evaluate the regulation of BMP15 and GDF9 receptors in granulosa cells during dominant follicle (DF) selection. Before follicular deviation (day 2 of the follicular wave), BMPR2 mRNA abundance tended to be higher in the second largest follicles (F2; P<0.1) compared to the future dominant follicle (F1). At the expected time of follicular deviation (day 3), BMPR2 and BMPR1B mRNA levels were higher in subordinate follicles (SFs; P<0.05) compared to dominant follicles (DFs). After deviation (on day 4), BMPR1B mRNA and protein were significantly more abundant in atretic SFs (as assessed by cleaved caspase 3) than in DFs. The fact that BMPR1B is more expressed in atretic follicles was further confirmed by using intrafollicular treatment with two agents known to induce atresia, namely an estradiol receptor antagonist (fulvestrant) and FGF10. In conclusion, the fact that BMPR-1B and -2 are more expressed in the second largest follicles before and at the expected time of follicular deviation is indicative of their inhibitory role in follicle differentiation and steroidogenesis. BMPR1B also seems to have a pivotal role during follicle regression since it is upregulated in advanced atretic follicles.

  14. Extracellular matrix of the developing ovarian follicle.

    PubMed

    Irving-Rodgers, Helen F; Rodgers, Raymond J

    2006-09-01

    There are many different types of extracellular matrices in the different follicle compartments. These have different roles in follicle development and atresia, and they change in composition during these processes. This review focuses on basal lamina matrix in particular, and considers follicular fluid, the newly identified focimatrix, and thecal matrices. When follicles commence growing, the follicular basal lamina changes in its composition from containing all six alpha chains of type IV collagen to only alpha1 and alpha2. Perlecan and nidogen-1 and -2 subsequently become components of the follicular basal lamina, and there is an increase in the amount of laminin chains alpha1, beta2, and gamma1, in the bovine at least. Late in follicular development and on atresia some follicles contain laminin alpha2. On atresia the follicular basal lamina is not degraded, as occurs in ovulation, but can be breached by cells from the thecal layer when it is not aligned by granulosa cells. A novel type of basal lamina-like matrix, called focimatrix (abbreviated from focal intraepithelial matrix), develops between the cells of the membrana granulosa as aggregates of basal lamina material. It does not envelop cells and so cannot perform functions of basal lamina as currently understood. It is hypothesized that focimatrix assists or initiates depolarization of the membrana granulosa necessary for the transformation into luteal cells. The largest osmotically active molecules in follicular fluid are hyaluronan and chondroitin sulfate proteoglycans, including versican and inter-alpha trypsin inhibitor. It has been suggested that these might be responsible for the formation of follicular fluid by creating an osmotic gradient across the follicular wall. The formation, development, and then either ovulation or regression of follicles requires considerable tissue remodeling, cellular replication, and specialization. The expectation of researchers is that extracellular matrix will be

  15. Isolation and characterization of goat ovarian aromatase cDNA: assessment of the activity using an intact cell system and placental expression.

    PubMed

    Bobes, Raúl José; Miranda, Carolina; Pérez-Martinez, Mario; Luu-The, Van; Romano, Marta C

    2004-08-01

    Goat ovarian follicles produce estrone and estradiol from androgens. The synthesis of C18 estrogens from C19 androgens requires cytochrome P450 aromatase, but little information about this key enzyme is available in the goat. We report here for the first time the cDNA sequence of the goat ovarian aromatase, the activity of the enzyme in a cell system, and its expression in the term goat placenta. A cDNA library from goat ovarian poly(A)+ RNA was constructed. Human aromatase cDNA was selected as probe to screen the library; several clones were isolated, but none was complete. The longest clone was 3.1 kb long, but it lacked the sequence coding for a few amino acids in the NH(2)-terminal. To obtain the missing sequence, we performed reverse amplification of the cDNA end (RACE). Sequence analysis indicated that goat aromatase possessed a very long 3'-untranslated region ( approximately 1790 bp), and a polyadenylation signal (AATAAA) located at position 3320 downstream from the ATG start codon. The coding region of goat cDNA was inserted in an expression vector and transfected into HEK-293 cells that were cultured in presence of [14C]-androstenedione, steroids extracted and further separated by TLC. The transfected cells efficiently transformed [14C]-androstenedione into estrone. This activity was inhibited by 4-hydroxyandrostenedione. We also investigated the presence of mRNA for P450 aromatase in the goat placenta, using reverse transcription-polymerase chain reaction (RT-PCR) and primers derived from the cDNA ovarian sequence and confirmed the expression of the mRNA in term placenta.

  16. Effect of temperature variation on hormonal concentration at various gestation stages in black Bengal goat

    PubMed Central

    Kumar, Binod; Ishwar, Ajay Kumar; Choudhary, Pankaj Kumar; Akhatar, Tanveer

    2015-01-01

    Aim: The present study was conducted to evaluate the effect of risingtemperature on the metabolic as well as the reproductive performance of the black Bengal goat. Materials and Methods: A total 27 numbers of non-pregnant black Bengal goats of the same parity comprised the experimental animals. The selected goats were randomly assigned to 3 groups of 9 each, maintaining uniformity in body weight (average 14-18 kg). Goats in Group-I were kept between the temperature ranges of 35-40°C, in Group-II between 20°Cand 27°C, and Group-III were kept under loose housing system and serve as a control. Goats in all the groups were bred naturally. Blood was collected prior to feeding in the morning on the day 1 (estrus), 20, 45, 90, and 135, expected day of parturition and also 2 days after parturition from goats of all the three groups. Results: It was observed that the level of plasma estrogen decreased (p<0.05) up to day 45 of gestation, then after increased up to 135 days of gestation and was maximum on expected day of parturition which was significantly (p<0.05) higher than all the values. Plasma progesterone level increased from day 20 and was the highest on day 90 and then decreased significantly (p<0.05) on expected date of parturition. The luteinizing hormone value decreased significantly (p<0.05) on expected day of parturition and day 2 after parturition in all the groups. Follicle stimulating hormone concentration showed a significant (p<0.05) decrease from day 1 to 2 days after parturition in all the groups. The plasma triiodothyronine (T3) level did not vary between and within the treatment groups at any stage of the experiment. The plasma thyroxine (T4) level varied significantly (p<0.01) within and (p<0.05) between groups at all stages of reproduction. A significant (p<0.05) variation in plasma cortisol concentration in all the groups increased significantly until the day of parturition and dropped significantly (p<0.01) in 2 days after parturition in all the

  17. Distribution of yolk polypeptides in the follicle cells during the differentiation of the follicular epithelium in Sarcophaga bullata egg follicles.

    PubMed

    Geysen, J; Cardoen, J; De Loof, A

    1987-09-01

    In S. bullata, the ovaries contribute to the synthesis of yolk polypeptides. A specific antiserum for yolk polypeptides was used to visualize the presence of yolk polypeptides in the follicle cells during their differentiation. After vitellogenesis has started, all follicle cells contain yolk polypeptides. The squamous follicle cells covering the nurse cells and the border cells lose yolk polypeptides before mid-vitellogenesis, whereas the follicle cells over the oocyte contain yolk polypeptides until after late vitellogenesis. All follicle cells are immunonegative afterwards. In vitro translation of poly(A)+ RNA demonstrated that the presence of yolk polypeptide mRNA correlates well with follicle cell immunopositivity for yolk polypeptides. This suggests that the follicle cells synthesize the ovarian yolk polypeptides. Differences in cellular and nuclear morphology, total and poly(A)+ RNA synthesis and the rate of yolk polypeptide synthesis were shown to be correlated with the presence or absence of yolk polypeptides in the differentiating follicular epithelium. The possible relationship between these different aspects of follicle cell differentiation, follicle cell polyploidy and the extracellular current pattern around follicles are discussed.

  18. Molecular characterization of Ethiopian indigenous goat populations.

    PubMed

    Hassen, Halima; Lababidi, Samer; Rischkowsky, Barbara; Baum, Michael; Tibbo, Markos

    2012-08-01

    Six Ethiopian indigenous goat populations viz. Gumuz, Agew, Begia-Medir, Bati, Abergelle, and Central Abergelle were genotyped for 15 microsatellite markers recommended by Food and Agriculture Organization of the United Nations and International Society for Animal Genetics. A total of 158 individual goats were tested to assess genetic variations within and between the goat populations in the Amhara Region of Ethiopia. The markers revealed 100% polymorphism across six goat populations indicating the presence of genetic diversity, which is an important variable to measure genetic variability within and between populations. The mean observed and expected heterozygosity values ranged from 0.56 (Central Abergelle) to 0.68 (Bati) and 0.59 (Abergelle) to 0.69 (Agew goat), respectively. The lowest genetic distance was observed between Begia-Medir and Central Abergelle (0.039), and the largest distances between Agew and Abergelle (0.140) and Gumuz and Abergelle (0.169). Neighbor-joining and the unweighted pair group method with arithmetic mean methods with bootstrap value of 1,000 was used which grouped the six goat populations into two major groups viz. the Abergelle goat cluster as one group and the Agew, Gumuz, Bati, Begia-Medir, and Central Abergelle goats as the second group. In our study, the obtained higher total variation within the goat populations (95%) confirms a close relatedness of the studied goat ecotypes, which might have happened due to the existence of uncontrolled animal breeding strategies resulting from uncontrolled movement of animals through various market routes and agricultural extension systems. The study contributed to the genetic characterization of Ethiopian indigenous goat populations and demonstrated the usefulness of the 15 microsatellite makers for biodiversity studies in goats.

  19. Reflectance spectroscopy for noninvasive evaluation of hair follicle stage

    NASA Astrophysics Data System (ADS)

    Liu, Caihua; Guan, Yue; Wang, Jianru; Zhong, Xiewei; Liu, Xiuli; Zhu, Dan

    2015-05-01

    Hair follicle offers an excellent model for systems biology and regenerative medicine. So far, the stages of hair follicle growth have been evaluated by histological examination. In this work, a noninvasive spectroscopy was proposed by measuring the diffuse reflectance of mouse skin and analyzing the melanin value. Results show that the skin diffuse reflectance was relatively high when hair follicles were at the telogen stage and at the beginning of the anagen stage, and decreased with the progression of the anagen stage. When the hair follicle entered into the catagen stage, the diffuse reflectance gradually increased. The changes in the melanin content of skin had contrary dynamics. Substages of the hair follicle cycle could be distinguished by comparing the changes in melanin value with the histological examination. This study provided a new method for noninvasive evaluation of the hair follicle stage, and should be valuable for basic and therapeutic investigations on hair regeneration.

  20. Reflectance spectroscopy for noninvasive evaluation of hair follicle stage.

    PubMed

    Liu, Caihua; Guan, Yue; Wang, Jianru; Zhong, Xiewei; Liu, Xiuli; Zhu, Dan

    2015-05-01

    Hair follicle offers an excellent model for systems biology and regenerative medicine. So far, the stages of hair follicle growth have been evaluated by histological examination. In this work, a noninvasive spectroscopy was proposed by measuring the diffuse reflectance of mouse skin and analyzing the melanin value. Results show that the skin diffuse reflectance was relatively high when hair follicles were at the telogen stage and at the beginning of the anagen stage, and decreased with the progression of the anagen stage. When the hair follicle entered into the catagen stage, the diffuse reflectance gradually increased. The changes in the melanin content of skin had contrary dynamics. Substages of the hair follicle cycle could be distinguished by comparing the changes in melanin value with the histological examination. This study provided a new method for noninvasive evaluation of the hair follicle stage, and should be valuable for basic and therapeutic investigations on hair regeneration. PMID:25428579

  1. Inheritance of goat coat colors.

    PubMed

    Adalsteinsson, S; Sponenberg, D P; Alexieva, S; Russel, A J

    1994-01-01

    Goat color inheritance was evaluated based on color description of 218 kids and their parents (10 sires, 178 dams) from mixed crosses between several goat populations in an experiment on cashmere fiber production. Altogether 10 color patterns were observed. They were postulated to be caused by 10 alleles at the Agouti locus, with the allele for white or tan color being the top dominant allele, and the nine others codominant. The bottom recessive allele, for nonagouti color, was the 11th allele at this locus. The postulated alleles are white or tan (A(wt)), black mask (A(blm)), bezoar (A(bz)), badgerface (A(b)), grey (A(g)), lightbelly (A(lb)), swiss markings (A(sm)), lateral stripes (A(ls)), mahogany (A(mh)), red cheek (A(rc)), and nonagouti (Aa). Two types of eumelanin pigment were observed, black and light brown, the latter being dominant. Recessive brown was not observed.

  2. Ovarian follicle dynamics of female Greater Scaup during egg production

    USGS Publications Warehouse

    Gorman, K.B.; Flint, P.L.; Esler, Daniel; Williams, T.D.

    2007-01-01

    Studies of female waterfowl nutrient reserve use during egg production require a precise understanding of ovarian follicle dynamics to correctly interpret breeding status, and, therefore, derive proper inference. Concerns over numerical declines of North American scaup have increased the need to better understand the role of female condition in reproductive performance. We quantified ovarian follicle dynamics of female Greater Scaup (Aythya marila) breeding on the Yukon–Kuskokwim Delta, Alaska, using a method that accounts for within day variation in follicle size. We considered several models for describing changes in follicle growth with the best supported model estimating the duration of rapid follicle growth (RFG) to be 5.20 ± 0.52 days (±95% confidence intervals) for each developing follicle. Average diameter and dry mass of preovulatory follicles were estimated to be 9.36 mm and 0.26 g, respectively, at the onset of RFG, and these follicle characteristics were 41.47 mm and 15.57 g, respectively, at ovulation. The average diameter of postovulatory follicles immediately following ovulation was estimated to be 17.35 mm, regressing quickly over several days. In addition, we derived predictive equations using diameter and dry mass to estimate the number of days before, and after, ovulation for pre- and postovulatory follicles, as well as an equation to estimate dry mass of damaged follicles. Our results allow precise definition of RFG and nest initiation dates, clutch size, and the daily energetic and nutritional demands of egg production at the individual level. This study provides the necessary foundation for additional work on Greater Scaup reproductive energetics and physiology, and offers an approach for quantifying ovarian follicle dynamics in other species.

  3. Introduction to Hair-Follicle-Associated Pluripotent Stem Cells.

    PubMed

    Hoffman, Robert M

    2016-01-01

    Nestin-expressing stem cells of the hair follicle, discovered by our laboratory, have been shown to be able to form outer-root sheaths of the follicle as well as neurons and many other non-follicle cell types. We have termed the nestin-expressing stem cells of the hair follicle as hair-follicle-associated pluripotent (HAP) stem cells. We have shown that the HAP stem cells from the hair follicle can effect the repair of peripheral nerve and spinal cord injury. The hair follicle stem cells differentiate into neuronal and glial cells after transplantation to the injured peripheral nerve and spinal cord, and enhance injury repair and locomotor recovery. When the excised hair follicle with its nerve stump was placed in Gelfoam(®) 3D histoculture, HAP stem cells grew and extended the hair follicle nerve which consisted of βIII-tubulin-positive fibers with F-actin expression at the tip. These findings indicate that βIII-tubulin-positive fibers elongating from the whisker follicle sensory nerve stump were growing axons. The growing whisker sensory nerve was highly enriched in HAP stem cells, which appeared to play a major role in its elongation and interaction with other nerves in 3D Gelfoam(®) histoculture, including the sciatic nerve, the trigeminal nerve, and the trigeminal nerve ganglion. These results suggest that a major function of the HAP stem cells in the hair follicle is for growth of the follicle sensory nerve. Recently, we have shown that HAP stem cells can differentiate into beating cardiac muscle cells. HAP stem cells have critical advantages for regenerative medicine over embryonic stem (ES) cells and induced pluripotent stem (iPS) cells in that they are highly accessible from each patient, thereby eliminating immunological issues since they are autologous, require no genetic manipulation, are non-tumorigenic, and do not present ethical issues.

  4. Cow's milk and goat's milk.

    PubMed

    Turck, Dominique

    2013-01-01

    Cow's milk is increasingly suggested to play a role in the development of chronic degenerative, non-communicable disorders whereas goat's milk is advocated as having several health benefits. Cow's milk is a rich and cheap source of protein and calcium, and a valuable food for bone health. Despite their high content in saturated fats, consumption of full-fat dairy products does not seem to cause significant changes in cardiovascular disease risk variables. Early introduction of cow's milk is a strong negative determinant of iron status. Unmodified cow's milk does not meet nutritional requirements of infants although it is acceptable to add small volumes of cow's milk to complementary foods. Cow's milk protein allergy has a prevalence ranging from 2 to 7%, and the age of recovery is usually around 2-3 years. The evidence linking cow's milk intake to a later risk of type 1 diabetes or chronic degenerative, non-communicable disorders (obesity, metabolic syndrome, type 2 diabetes, hypertension) is not convincing. Milk probably protects against colorectal cancer, diets high in calcium are a probable cause of prostate cancer, and there is limited evidence suggesting that high consumption of milk and dairy products increases the risk for prostate cancer. There is no evidence to support the use of a cow's milk-free diet as a primary treatment for individuals with autistic spectrum disorders. Unmodified goat's milk is not suitable for infants because of the high protein and minerals content and of a low folate content. Goat's milk has no clear nutritional advantage over cow's milk and is not less allergenic. The European Food Safety Authority recently stated that proteins from goat's milk can be suitable as a protein source for infant and follow-on formula, provided the final product complies with the compositional criteria laid down in Directive 2006/141/EC.

  5. Prepubertal propagation of transgenic cloned goats by laparoscopic ovum pick-up and in vitro embryo production.

    PubMed

    Baldassarre, H; Wang, B; Pierson, J; Neveu, N; Sneek, L; Lapointe, J; Cote, F; Kafidi, N; Keefer, C L; Lazaris, A; Karatzas, C N

    2004-01-01

    The use of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production was evaluated in the early propagation of cloned goats. Ten kinder goats produced by somatic cell nuclear transfer technology were used as oocyte donors. Half of the donor animals were subjected to LOPU at 2-3 months of age (prior to induction of lactation), whereas the other five goats were subjected to LOPU at 6-7 months of age (following induction to lactation). They were stimulated with 80 mg NIH-FSH-P1 (Folltropin, Vetrepharm, Canada) together with 300 IU eCG (Novormon, Vetrepharm, Canada) administered intramuscularly 36 h prior to LOPU. The number of follicles aspirated and oocytes recovered was higher in the younger group of donors (57 +/- 7 and 41 +/- 4 vs. 28 +/- 2 and 25.8 +/- 2, p < 0.05), however, oocytes from animals in the late prepubertal age showed higher developmental capacity resulting in higher transferable embryo yield (81.4% vs. 67.8%, p < 0.01), pregnancy rate (80% vs. 40%, p < 0.05) and total kids born (27 vs. 15, p < 0.01). In conclusion, LOPU in combination with in vitro embryo production techniques is an efficient method for the early propagation of valuable goats produced by somatic cell nuclear transfer.

  6. Regulation and 3 dimensional culture of tertiary follicle growth.

    PubMed

    Cheon, Yong-Pil

    2012-09-01

    It has been revealed that multiple cohorts of tertiary follicles develop during some animal estrous cycle and the human menstrual cycle. To reach developmental competence, oocytes need the support of somatic cells. During embryogenesis, the primordial germ cells appear, travel to the gonadal rudiments, and form follicles. The female germ cells develop within the somatic cells of the ovary, granulosa cells, and theca cells. How the oocyte and follicle cells support each other has been seriously studied. The latest technologies in genes and proteins and genetic engineering have allowed us to collect a great deal of information about folliculogenesis. For example, a few web pages (http://www.ncbi.nlm.nih.gov; http://mrg.genetics.washington.edu) provide access to databases of genomes, sequences of transcriptomes, and various tools for analyzing and discovering genes important in ovarian development. Formation of the antrum (tertiary follicle) is the final phase of folliculogenesis and the transition from intraovarian to extraovian regulation. This final step coordinates with the hypothalamic-pituitary-ovarian axis. On the other hand, currently, follicle physiology is under intense investigation, as little is known about how to overcome women's ovarian problems or how to develop competent oocytes from in vitro follicle culture or transplantation. In this review, some of the known roles of hormones and some of the genes involved in tertiary follicle growth and the general characteristics of tertiary follicles are summarized. In addition, in vitro culture of tertiary follicles is also discussed as a study model and an assisted reproductive technology model.

  7. The hair follicle as a target for gene therapy.

    PubMed

    Gupta, S; Domashenko, A; Cotsarelis, G

    2001-01-01

    The hair follicle possesses progenitor cells for continued hair follicle cycling and for epidermal keratinocytes, melanocytes and Langerhans cells. These different cell types can be targeted by topical gene delivery to mouse skin. Using a combination of liposomes and DNA, we demonstrated the feasibility of targeting hair follicle cells in human scalp xenografts as well. We defined liposome composition and stage of the hair cycle as important parameters influencing transfection of human hair follicles. Transfection occurred only during anagen onset. Considerations and obstacles for using gene therapy to treat alopecias and skin disease are discussed. A theoretical framework for future gene therapy treatments for cutaneous and systemic disorders is presented.

  8. A sightability model for mountain goats

    USGS Publications Warehouse

    Rice, C.G.; Jenkins, K.J.; Chang, W.-Y.

    2009-01-01

    Unbiased estimates of mountain goat (Oreamnos americanus) populations are key to meeting diverse harvest management and conservation objectives. We developed logistic regression models of factors influencing sightability of mountain goat groups during helicopter surveys throughout the Cascades and Olympic Ranges in western Washington during summers, 20042007. We conducted 205 trials of the ability of aerial survey crews to detect groups of mountain goats whose presence was known based on simultaneous direct observation from the ground (n 84), Global Positioning System (GPS) telemetry (n 115), or both (n 6). Aerial survey crews detected 77 and 79 of all groups known to be present based on ground observers and GPS collars, respectively. The best models indicated that sightability of mountain goat groups was a function of the number of mountain goats in a group, presence of terrain obstruction, and extent of overstory vegetation. Aerial counts of mountain goats within groups did not differ greatly from known group sizes, indicating that under-counting bias within detected groups of mountain goats was small. We applied HorvitzThompson-like sightability adjustments to 1,139 groups of mountain goats observed in the Cascade and Olympic ranges, Washington, USA, from 2004 to 2007. Estimated mean sightability of individual animals was 85 but ranged 0.750.91 in areas with low and high sightability, respectively. Simulations of mountain goat surveys indicated that precision of population estimates adjusted for sightability biases increased with population size and number of replicate surveys, providing general guidance for the design of future surveys. Because survey conditions, group sizes, and habitat occupied by goats vary among surveys, we recommend using sightability correction methods to decrease bias in population estimates from aerial surveys of mountain goats.

  9. Suppression of sheep and goat lymphocyte proliferation by sheep, goat, and sheep x goat hybrid trophoblast tissue cultures.

    PubMed

    Roth, T L; White, K L; Horohov, D W

    1991-11-01

    Immunosuppressive activity of conditioned medium from cultured ovine, caprine, and hybrid trophoblast tissue was examined. Conceptuses were obtained from naturally mated donor ewes and does at d 20 of gestation and trophoblast tissue was cultured for 24 h in medium supplemented with 15% calf serum and 1% antibiotic/antimycotic. Conditioned medium was added to pokeweed mitogen-stimulated sheep and goat lymphocyte cultures. Quantification of [3H]thymidine uptake by cells was used to measure lymphocyte proliferation. Ovine, caprine, and hybrid conditioned medium effectively suppressed sheep and goat lymphocyte proliferation (P less than .01). There were no differences (P greater than .05) between the immunosuppressive activity of the three tissue types on either sheep or goat lymphocytes. For all treatment groups, sheep lymphocytes were suppressed more than goat lymphocytes (P less than .05). These results indicate that, at d 20 of gestation, sheep x goat hybrid trophoblast tissue is capable of suppressing pokeweed mitogen-stimulated lymphocyte proliferation. PMID:1752830

  10. The immature human ovary shows loss of abnormal follicles and increasing follicle developmental competence through childhood and adolescence

    PubMed Central

    Anderson, R.A.; McLaughlin, M.; Wallace, W.H.B.; Albertini, D.F.; Telfer, E.E.

    2014-01-01

    STUDY QUESTION Do the ovarian follicles of children and adolescents differ in their morphology and in vitro growth potential from those of adults? SUMMARY ANSWER Pre-pubertal ovaries contained a high proportion of morphologically abnormal non-growing follicles, and follicles showed reduced capacity for in vitro growth. WHAT IS KNOWN ALREADY The pre-pubertal ovary is known to contain follicles at the early growing stages. How this changes over childhood and through puberty is unknown, and there are no previous data on the in vitro growth potential of follicles from pre-pubertal and pubertal girls. STUDY DESIGN, SIZE, DURATION Ovarian biopsies from five pre-pubertal and seven pubertal girls and 19 adult women were analysed histologically, cultured in vitro for 6 days, with growing follicles then isolated and cultured for a further 6 days. PARTICIPANTS/MATERIALS, SETTING, METHODS Ovarian biopsies were obtained from girls undergoing ovarian tissue cryopreservation for fertility preservation, and compared with biopsies from adult women. Follicle stage and morphology were classified. After 6 days in culture, follicle growth initiation was assessed. The growth of isolated secondary follicles was assessed over a further 6 days, including analysis of oocyte growth. MAIN RESULTS AND THE ROLE OF CHANCE Pre-pubertal ovaries contained a high proportion of abnormal non-growing follicles (19.4 versus 4.85% in pubertal ovaries; 4004 follicles analysed; P = 0.02) characterized by indistinct germinal vesicle membrane and absent nucleolus. Follicles with this abnormal morphology were not seen in the adult ovary. During 6 days culture, follicle growth initiation was observed at all ages; in pre-pubertal samples there was very little development to secondary stages, while pubertal samples showed similar growth activation to that seen in adult tissue (pubertal group: P = 0.02 versus pre-pubertal, ns versus adult). Isolated secondary follicles were cultured for a further 6 days. Those

  11. Cryopreservation of the Hair Follicle Maintains Pluripotency of Nestin-Expressing Hair Follicle-Associated Pluripotent Stem Cells.

    PubMed

    Kajiura, Satoshi; Mii, Sumiyuki; Aki, Ryoichi; Hamada, Yuko; Arakawa, Nobuko; Kawahara, Katsumasa; Li, Lingna; Katsuoka, Kensei; Hoffman, Robert M; Amoh, Yasuyuki

    2015-08-01

    Hair follicles contain nestin-expressing pluripotent stem cells, the origin of which is above the bulge area, below the sebaceous gland. We have termed these cells hair follicle-associated pluripotent (HAP) stem cells. In the present study, we established efficient cryopreservation methods of the hair follicle that maintained the pluripotency of HAP stem cells. We cryopreserved the whole hair follicle from green fluorescent protein transgenic mice by slow-rate cooling in TC-Protector medium and storage in liquid nitrogen. After thawing, the upper part of the hair follicle was isolated and cultured in Dulbecco's Modified Eagle's Medium (DMEM) with fetal bovine serum (FBS). After 4 weeks of culture, cells from the upper part of the hair follicle grew out. The growing cells were transferred to DMEM/F12 without FBS. After 1 week of culture, the growing cells formed hair spheres, each containing ∼1×10(2) HAP stem cells. The hair spheres contained cells that differentiated to neurons, glial cells, and other cell types. The thawed and cultured upper part of the hair follicle produced almost as many pluripotent hair spheres as fresh follicles. The hair spheres derived from slow-cooling cryopreserved hair follicles were as pluripotent as hair spheres from fresh hair follicles. In contrast, rapid-cooling (vitrification) cryopreservation poorly preserved the pluripotency of the hair follicle stem cells. Stem cell marker genes (nestin, Sox2, and SSEA-1) were as highly expressed in slow-rate cooled cryopreserved follicles, after thawing, as in fresh follicles. However, in the vitrification cryopreserved follicles, the expression of the stem cell marker genes was greatly reduced. Direct cryopreservation of hair spheres by either the rapid-cooling, or slow-cooling method, resulted in loss of pluripotency. These results suggest that the slow-rate cooling cryopreservation of the whole hair follicle is effective to store HAP stem cells. Stored HAP stem cells would be very useful

  12. 21 CFR 522.1002 - Follicle stimulating hormone.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Follicle stimulating hormone. 522.1002 Section 522....1002 Follicle stimulating hormone. (a)(1) Specifications. Each package contains 2 vials. One vial... hormone. The other vial contains 10 milliliters of aqueous diluent. (2) Sponsor. See No. 052923 in §...

  13. 21 CFR 522.1002 - Follicle stimulating hormone.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Follicle stimulating hormone. 522.1002 Section 522....1002 Follicle stimulating hormone. (a)(1) Specifications. Each package contains 2 vials. One vial... hormone. The other vial contains 10 milliliters of aqueous diluent. (2) Sponsor. See No. 052923 in §...

  14. 21 CFR 522.1002 - Follicle stimulating hormone.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Follicle stimulating hormone. 522.1002 Section 522....1002 Follicle stimulating hormone. (a)(1) Specifications. Each package contains 2 vials. One vial... hormone. The other vial contains 10 milliliters of aqueous diluent. (2) Sponsor. See 059521 in §...

  15. 21 CFR 522.1002 - Follicle stimulating hormone.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Follicle stimulating hormone. 522.1002 Section 522....1002 Follicle stimulating hormone. (a)(1) Specifications. Each package contains 2 vials. One vial... hormone. The other vial contains 10 milliliters of aqueous diluent. (2) Sponsor. See No. 052923 in §...

  16. 21 CFR 522.1002 - Follicle stimulating hormone.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Follicle stimulating hormone. 522.1002 Section 522....1002 Follicle stimulating hormone. (a)(1) Specifications. Each package contains 2 vials. One vial... hormone. The other vial contains 10 milliliters of aqueous diluent. (2) Sponsor. See No. 052923 in §...

  17. Toxoplasma-induced abortion in dairy goats.

    PubMed

    Dubey, J P

    1981-04-01

    Fifteen pregnant goats serologically negative for Toxoplasma gondii were inoculated orally with 10, 100, or 1,000 infective oocysts of the GT-1 strain of T gondii. Four goats received 10 oocysts, 6 received 100 oocysts, and 5 received 1,000 oocysts. Six pregnant goats served as noninoculated controls. All inoculated goats became pyretic (40-41 C) and were dyspneic between 4 and 12 days after inoculation (DAI). Two goats inoculated in early pregnancy (68 and 70 days) with 100 oocysts had enlarged uteri; fetal membranes but no fetuses were found when they were killed 86 and 88 DAI. Three goats aborted or had a stillborn kid 9, 10, and 11 DAI. The remaining 10 goats delivered kids infected with T gondii; 8 goats had at least 1 stillborn kid, and 2 goats had infected but otherwise normal kids at birth. Placenta and tissues of fetuses and kids were inoculated into mice for determination of Toxoplasma infectivity. Toxoplasma gondii was isolated from the fetal placenta of 6 of 7 goats as early as 10 DAI and as late as 62 DAI. The organism was isolated from every fetus or kid that was born 18 DAI but not from fetuses or kids born 9, 10, and 11 DAI. Two kids were born 15 DAI; T gondii was isolated from 1 kid but not from the other, suggesting that T gondii invaded fetal tissues between 11 and 15 DAI. Of the 20 transplacentally infected kids, T gondii was isolated more frequently from the skeletal muscle, heart, lung, and brain than from other tissues.

  18. Effect of hormonal synchronisation and/or short-term supplementation with maize on follicular dynamics and hormone profiles in goats during the non-breeding season.

    PubMed

    Nogueira, D M; Cavalieri, J; Fitzpatrick, L A; Gummow, B; Blache, D; Parker, A J

    2016-08-01

    This study aimed to evaluate the reproductive response of anoestrous goats that were either hormonally treated and/or supplemented with maize for 9days to determine which treatment combination was the most effective in enhancing follicular development and ovulation rate, and whether these responses were associated with increases in metabolic hormones. The experiment was carried out using 28 does, using a 2×2 factorial design with seven does in each group to test the effect of synchronisation of oestrus, supplementation with maize and their interactions. Synchronisation of oestrous cycles (P<0.001) but not supplementation with maize or the interaction between the two (P>0.05) increased the number of codominant follicles, the diameter of the largest follicle on Day 9 and growth rate of follicles during the period of supplementation. Compared with non-supplemented animals, supplementation with maize increased the total number of follicles observed between Days 7 and 9 (P=0.039). In addition, nutritional supplementation with maize in combination with synchronisation of oestrus increased the ovulation rate by 43% (P=0.074). Interactions between time and supplementation with maize showed that plasma concentrations of insulin, leptin and IGF-1 were greater in does supplemented with maize compared with non-supplemented does (P<0.001). The findings show that hormonal synchronisation had the most influence on modifying follicular development and ovulation in anoestrous goats. Supplementation with maize increased the concentrations of insulin, leptin and IGF-1, which could potentially modify the sensitivity of follicles to gonadotrophins and reduce rates of atresia.

  19. The placenta shed from goats with classical scrapie is infectious to goat kids and lambs.

    PubMed

    Schneider, David A; Madsen-Bouterse, Sally A; Zhuang, Dongyue; Truscott, Thomas C; Dassanayake, Rohana P; O'Rourke, Katherine I

    2015-08-01

    The placenta of domestic sheep plays a key role in horizontal transmission of classical scrapie. Domestic goats are frequently raised with sheep and are susceptible to classical scrapie, yet potential routes of transmission from goats to sheep are not fully defined. Sparse accumulation of disease-associated prion protein in cotyledons casts doubt about the role of the goat's placenta. Thus, relevant to mixed-herd management and scrapie-eradication efforts worldwide, we determined if the goat's placenta contains prions orally infectious to goat kids and lambs. A pooled cotyledon homogenate, prepared from the shed placenta of a goat with naturally acquired classical scrapie disease, was used to orally inoculate scrapie-naïve prion genotype-matched goat kids and scrapie-susceptible lambs raised separately in a scrapie-free environment. Transmission was detected in all four goats and in two of four sheep, which importantly identifies the goat's placenta as a risk for horizontal transmission to sheep and other goats.

  20. Mesenchymal-epithelial interactions during hair follicle morphogenesis and cycling

    PubMed Central

    Sennett, Rachel; Rendl, Michael

    2012-01-01

    Embryonic hair follicle induction and formation are regulated by mesenchymal-epithelial interactions between specialized dermal cells and epidermal stem cells that switch to a hair fate. Similarly, during postnatal hair growth, communication between mesenchymal dermal papilla cells and surrounding epithelial matrix cells coordinates hair shaft production. Adult hair follicle regeneration in the hair cycle again is thought to be controlled by activating signals originating from the mesenchymal compartment and acting on hair follicle stem cells. Although many signaling pathways are implicated in hair follicle formation and growth, the precise nature, timing, and intersection of these inductive and regulatory signals remains elusive. The goal of this review is to summarize our current understanding and to discuss recent new insights into mesenchymal-epithelial interactions during hair follicle morphogenesis and cycling. PMID:22960356

  1. [Pulmonary blood vessels in goats].

    PubMed

    Roos, H; Hegner, K; Vollmerhaus, B

    1999-05-01

    The blood vessels in the lung of the goat, which until now have received little attention, are described in detail for the first time. With regard to the segments of the lung, blood vessels are bronchovascular units in the lobi craniales, lobus medius and lobus accessorius, but bronchoartery units in the lobi caudales. We investigated the types of branches of the Aa. pulmonales dextra et sinistra, the inter- and intraspecific principles of the outlet of the pulmonary veins and the importance of bronchopulmonary segmentation of the lungs. PMID:10386009

  2. [A Christmas straw goat astray].

    PubMed

    Bloch, Sune Land; Nielsen, Hans Ulrik Kjærem

    2012-12-01

    A 22-year-old otherwise healthy man presented to our clinic with suspected acute epiglottitis. The patient had a 1-week history of pain in the throat and fever for the latest 24 hours. During the physical examination, the patient mentioned that he had been eating a Christmas straw goat at a party one week previously. Direct fiberoptic laryngoscopy showed a red and swollen lingual surface of the epiglottis, but no foreign bodies were identified. After inhalation of adrenalin, a 4 cm straw became visible in the epiglottic vallecula. Mimic of acute epiglottitis from a straw in the vallecula has to our knowledge never been described in the literature.

  3. Monohaloacetic acid drinking water disinfection by-products inhibit follicle growth and steroidogenesis in mouse ovarian antral follicles in vitro.

    PubMed

    Jeong, Clara H; Gao, Liying; Dettro, Tyler; Wagner, Elizabeth D; Ricke, William A; Plewa, Michael J; Flaws, Jodi A

    2016-07-01

    Water disinfection greatly reduced the incidence of waterborne diseases, but the reaction between disinfectants and natural organic matter in water leads to the formation of drinking water disinfection by-products (DBPs). DBPs have been shown to be toxic, but their effects on the ovary are not well defined. This study tested the hypothesis that monohalogenated DBPs (chloroacetic acid, CAA; bromoacetic acid, BAA; iodoacetic acid, IAA) inhibit antral follicle growth and steroidogenesis in mouse ovarian follicles. Antral follicles were isolated and cultured with either vehicle or DBPs (0.25-1.00mM of CAA; 2-15μM of BAA or IAA) for 48 and 96h. Follicle growth was measured every 24h and the media were analyzed for estradiol levels at 96h. Exposure to DBPs significantly inhibited antral follicle growth and reduced estradiol levels compared to controls. These data demonstrate that DBP exposure caused ovarian toxicity in vitro. PMID:27151372

  4. Hydatidosis in goats in Jammu, India.

    PubMed

    Godara, R; Katoch, R; Yadav, Anish

    2014-03-01

    The prevalence of hydatidosis was carried out in slaughtered goats (n = 177) by liver and lung examination at Jammu. The prevalence rate of hydatidosis was 19.8 %. A total of 14.1 % goats had cysts in both the livers and lungs while 2.3 and 3.4 % goats had cysts in the livers or lungs, respectively. A total of 9.1 % goats had fertile cysts. The adult goats (>4 years) had a significantly higher (p = 0.01) prevalence rate as compared to the young goats (<2 years). Sex had no significant effect on the prevalence of hydatidosis in goats. The histopathological section of the affected lungs revealed a thick coat of granulation tissue causing fibrosis. The cysts caused progressive focal pressure atrophic lesions at the sites of predilection, resulting in atelectasis, desquamation of bronchial epithelium and interalveolar haemorrhages. It may be concluded that the slaughter of food animals at an early age would help to reduce the infection prevalence in dogs and subsequently in the food animals. PMID:24505182

  5. Di(2-ethylhexyl) phthalate inhibits antral follicle growth, induces atresia, and inhibits steroid hormone production in cultured mouse antral follicles

    SciTech Connect

    Hannon, Patrick R. Brannick, Katherine E. Wang, Wei Gupta, Rupesh K. Flaws, Jodi A.

    2015-04-01

    Di(2-ethylhexyl) phthalate (DEHP) is a ubiquitous environmental toxicant found in consumer products that causes ovarian toxicity. Antral follicles are the functional ovarian units and must undergo growth, survival from atresia, and proper regulation of steroidogenesis to ovulate and produce hormones. Previous studies have determined that DEHP inhibits antral follicle growth and decreases estradiol levels in vitro; however, the mechanism by which DEHP elicits these effects is unknown. The present study tested the hypothesis that DEHP directly alters regulators of the cell cycle, apoptosis, and steroidogenesis to inhibit antral follicle functionality. Antral follicles from adult CD-1 mice were cultured with vehicle control or DEHP (1–100 μg/ml) for 24–96 h to establish the temporal effects of DEHP on the follicle. Following 24–96 h of culture, antral follicles were subjected to gene expression analysis, and media were subjected to measurements of hormone levels. DEHP increased the mRNA levels of cyclin D2, cyclin dependent kinase 4, cyclin E1, cyclin A2, and cyclin B1 and decreased the levels of cyclin-dependent kinase inhibitor 1A prior to growth inhibition. Additionally, DEHP increased the mRNA levels of BCL2-associated agonist of cell death, BCL2-associated X protein, BCL2-related ovarian killer protein, B-cell leukemia/lymphoma 2, and Bcl2-like 10, leading to an increase in atresia. Further, DEHP decreased the levels of progesterone, androstenedione, and testosterone prior to the decrease in estradiol levels, with decreased mRNA levels of side-chain cleavage, 17α-hydroxylase-17,20-desmolase, 17β-hydroxysteroid dehydrogenase, and aromatase. Collectively, DEHP directly alters antral follicle functionality by inhibiting growth, inducing atresia, and inhibiting steroidogenesis. - Highlights: • DEHP inhibits antral follicle growth by dysregulating cell cycle regulators. • DEHP induces antral follicle atresia by dysregulating apoptosis regulators. • DEHP

  6. Follicle Online: an integrated database of follicle assembly, development and ovulation.

    PubMed

    Hua, Juan; Xu, Bo; Yang, Yifan; Ban, Rongjun; Iqbal, Furhan; Cooke, Howard J; Zhang, Yuanwei; Shi, Qinghua

    2015-01-01

    Folliculogenesis is an important part of ovarian function as it provides the oocytes for female reproductive life. Characterizing genes/proteins involved in folliculogenesis is fundamental for understanding the mechanisms associated with this biological function and to cure the diseases associated with folliculogenesis. A large number of genes/proteins associated with folliculogenesis have been identified from different species. However, no dedicated public resource is currently available for folliculogenesis-related genes/proteins that are validated by experiments. Here, we are reporting a database 'Follicle Online' that provides the experimentally validated gene/protein map of the folliculogenesis in a number of species. Follicle Online is a web-based database system for storing and retrieving folliculogenesis-related experimental data. It provides detailed information for 580 genes/proteins (from 23 model organisms, including Homo sapiens, Mus musculus, Rattus norvegicus, Mesocricetus auratus, Bos Taurus, Drosophila and Xenopus laevis) that have been reported to be involved in folliculogenesis, POF (premature ovarian failure) and PCOS (polycystic ovary syndrome). The literature was manually curated from more than 43,000 published articles (till 1 March 2014). The Follicle Online database is implemented in PHP + MySQL + JavaScript and this user-friendly web application provides access to the stored data. In summary, we have developed a centralized database that provides users with comprehensive information about genes/proteins involved in folliculogenesis. This database can be accessed freely and all the stored data can be viewed without any registration. Database URL: http://mcg.ustc.edu.cn/sdap1/follicle/index.php PMID:25931457

  7. Follicle Online: an integrated database of follicle assembly, development and ovulation

    PubMed Central

    Hua, Juan; Xu, Bo; Yang, Yifan; Ban, Rongjun; Iqbal, Furhan; Zhang, Yuanwei; Shi, Qinghua

    2015-01-01

    Folliculogenesis is an important part of ovarian function as it provides the oocytes for female reproductive life. Characterizing genes/proteins involved in folliculogenesis is fundamental for understanding the mechanisms associated with this biological function and to cure the diseases associated with folliculogenesis. A large number of genes/proteins associated with folliculogenesis have been identified from different species. However, no dedicated public resource is currently available for folliculogenesis-related genes/proteins that are validated by experiments. Here, we are reporting a database ‘Follicle Online’ that provides the experimentally validated gene/protein map of the folliculogenesis in a number of species. Follicle Online is a web-based database system for storing and retrieving folliculogenesis-related experimental data. It provides detailed information for 580 genes/proteins (from 23 model organisms, including Homo sapiens, Mus musculus, Rattus norvegicus, Mesocricetus auratus, Bos Taurus, Drosophila and Xenopus laevis) that have been reported to be involved in folliculogenesis, POF (premature ovarian failure) and PCOS (polycystic ovary syndrome). The literature was manually curated from more than 43 000 published articles (till 1 March 2014). The Follicle Online database is implemented in PHP + MySQL + JavaScript and this user-friendly web application provides access to the stored data. In summary, we have developed a centralized database that provides users with comprehensive information about genes/proteins involved in folliculogenesis. This database can be accessed freely and all the stored data can be viewed without any registration. Database URL: http://mcg.ustc.edu.cn/sdap1/follicle/index.php PMID:25931457

  8. Microsatellite instability in follicle centre cell lymphoma.

    PubMed

    Randerson, J; Cawkwell, L; Jack, A; Child, J A; Lewis, F; Hall, N; Johnson, P; Evans, P; Barrans, S; Morgan, G J

    1996-04-01

    Fluorescent polymerase chain reaction (PCR) was used to assay 12 microsatellite markers (APC x 2, DCC, P53 x 2, RB1, NM23, WT1, D6S260, D6S262, D6S281 and TNFa) to look for evidence of microsatellite instability in 40 cases of follicle centre cell lymphoma (FCC). Evidence of novel alleles seen in the tumour tissue but not the normal uninvolved tissue was seen in seven cases (17%). In only two of these cases (5%) was more than one locus involved but in these cases multiple affected loci were seen (4/12 and 7/12 respectively). The detection of microsatellite instability indicates a DNA repair defect such as that which would be predicted to occur in cells with mutated mismatch repair genes, a novel finding in FCC lymphoma. PMID:8611453

  9. Persistent infections after natural transmission of bovine viral diarrhoea virus from cattle to goats and among goats.

    PubMed

    Bachofen, Claudia; Vogt, Hans-Rudolf; Stalder, Hanspeter; Mathys, Tanja; Zanoni, Reto; Hilbe, Monika; Schweizer, Matthias; Peterhans, Ernst

    2013-05-15

    Bovine viral diarrhoea virus (BVDV) is an economically important pathogen of cattle worldwide. Infection of a pregnant animal may lead to persistent infection of the foetus and birth of a persistently infected (PI) calf that sheds the virus throughout its life. However, BVD viruses are not strictly species specific. BVDV has been isolated from many domesticated and wild ruminants. This is of practical importance as virus reservoirs in non-bovine hosts may hamper BVDV control in cattle. A goat given as a social companion to a BVDV PI calf gave birth to a PI goat kid. In order to test if goat to goat infections were possible, seronegative pregnant goats were exposed to the PI goat. In parallel, seronegative pregnant goats were kept together with the PI calf. Only the goat to goat transmission resulted in the birth of a next generation of BVDV PI kids whereas all goats kept together with the PI calf aborted. To our knowledge, this is the first report which shows that a PI goat cannot only transmit BVD virus to other goats but that such transmission may indeed lead to the birth of a second generation of PI goats. Genetic analyses indicated that establishment in the new host species may be associated with step-wise adaptations in the viral genome. Thus, goats have the potential to be a reservoir for BVDV. However, the PI goats showed growth retardation and anaemia and their survival under natural conditions remains questionable.

  10. Protein composition in the fluid of individual bovine follicles.

    PubMed

    Andersen, M M; Kroll, J; Byskov, A G; Faber, M

    1976-09-01

    The proteins in follicular fluid from individual and pooled bovine follicles were studied by gel chromatography and quantitative immunoelectrophoresis. The mean protein concentration was 86-4% of serum; very large proteins were present in only low concentrations. A minimum of 40 individual proteins was distinguished in follicular fluid, and 15 of these proteins were quantitated. A correlation between molecular weight and follicular fluid: serum concentration ratio was found. Fluid from individual follicles differed only in the relative concentrations of small and large proteins. An exception to this was IgG which was occasionally, but never in healthy growing follicles, present in concetrations above 150% of serum. Healthy growing, preovulatory and atretic follicles had higher, and cystic follicles mostly lower, concentrations of small proteins than serum. The concentration of alpha2-macroglobulin in healthy growing follicles never exceeded 16% of serum. The concentration of large proteins in follicular fluid increased with increasing follicle size. Attempts to detect proteins specific to follicular fluid by immunizing rabbits with pooled follicular samples and the follicular fluid proteins not bound by anti-bovine antiserum resulted in production of antibodies against fibrinogen and its split products D+E only.

  11. Studies on the pathogenesis of shimao zheng (fleece-eating) in sheep and goats.

    PubMed

    Youde, H; Huaitao, C

    2001-12-01

    Research on the disease called shimao zheng, which had resulted in widespread fleece-eating and shedding by sheep and goats in the Haizi area of Akesa County of Gansu Province of China, was carried out by both laboratory studies and pathological observations. There was a marked deficiency of sulfur in the wool and tissues of the affected animals. The pathological changes were mainly loss of fleece, exposure of the skin, keratinization of the epidermal cells, narrowing and reduced numbers of the fleece follicles, and fewer sebaceous glands and sweat glands. Histologically, there was atrophy of the striated and cardiac muscles and of the epithelium of the convoluted tubules of the kidney, together with a glomerulonephritis. The disease was considered to be a local nutritional and metabolic disorder, probably mainly caused by sulfur deficiency.

  12. Identification of goat allotypic determinants and generation of goat-mouse hybridomas secreting goat IgG2.

    PubMed

    Capparelli, R; Del Sorbo, G; Iannelli, D

    1990-01-01

    Five allotypic determinants controlled by independent genes have been identified in goat. Of these determinants, four have been detected with alloimmune antisera and one with monoclonal antibodies. The specificities A1, C1 and D1 are lipoproteins; B1 is possibly an alpha 2 macroglobulin and E1 and IgG2. The specificity B1 is not expressed until the age of 3-4 months. The gene controlling the specificity E1 is present at about the same frequency (0.38-0.41) in goat, sheep, cattle and water buffalo. Stable hybridomas secreting goat IgG2 have been obtained by the fusion of goat peripheral lymphocytes with mouse myeloma cells.

  13. Regulation and 3 dimensional culture of tertiary follicle growth.

    PubMed

    Cheon, Yong-Pil

    2012-09-01

    It has been revealed that multiple cohorts of tertiary follicles develop during some animal estrous cycle and the human menstrual cycle. To reach developmental competence, oocytes need the support of somatic cells. During embryogenesis, the primordial germ cells appear, travel to the gonadal rudiments, and form follicles. The female germ cells develop within the somatic cells of the ovary, granulosa cells, and theca cells. How the oocyte and follicle cells support each other has been seriously studied. The latest technologies in genes and proteins and genetic engineering have allowed us to collect a great deal of information about folliculogenesis. For example, a few web pages (http://www.ncbi.nlm.nih.gov; http://mrg.genetics.washington.edu) provide access to databases of genomes, sequences of transcriptomes, and various tools for analyzing and discovering genes important in ovarian development. Formation of the antrum (tertiary follicle) is the final phase of folliculogenesis and the transition from intraovarian to extraovian regulation. This final step coordinates with the hypothalamic-pituitary-ovarian axis. On the other hand, currently, follicle physiology is under intense investigation, as little is known about how to overcome women's ovarian problems or how to develop competent oocytes from in vitro follicle culture or transplantation. In this review, some of the known roles of hormones and some of the genes involved in tertiary follicle growth and the general characteristics of tertiary follicles are summarized. In addition, in vitro culture of tertiary follicles is also discussed as a study model and an assisted reproductive technology model. PMID:23106040

  14. Regulation and 3 dimensional culture of tertiary follicle growth

    PubMed Central

    2012-01-01

    It has been revealed that multiple cohorts of tertiary follicles develop during some animal estrous cycle and the human menstrual cycle. To reach developmental competence, oocytes need the support of somatic cells. During embryogenesis, the primordial germ cells appear, travel to the gonadal rudiments, and form follicles. The female germ cells develop within the somatic cells of the ovary, granulosa cells, and theca cells. How the oocyte and follicle cells support each other has been seriously studied. The latest technologies in genes and proteins and genetic engineering have allowed us to collect a great deal of information about folliculogenesis. For example, a few web pages (http://www.ncbi.nlm.nih.gov; http://mrg.genetics.washington.edu) provide access to databases of genomes, sequences of transcriptomes, and various tools for analyzing and discovering genes important in ovarian development. Formation of the antrum (tertiary follicle) is the final phase of folliculogenesis and the transition from intraovarian to extraovian regulation. This final step coordinates with the hypothalamic-pituitary-ovarian axis. On the other hand, currently, follicle physiology is under intense investigation, as little is known about how to overcome women's ovarian problems or how to develop competent oocytes from in vitro follicle culture or transplantation. In this review, some of the known roles of hormones and some of the genes involved in tertiary follicle growth and the general characteristics of tertiary follicles are summarized. In addition, in vitro culture of tertiary follicles is also discussed as a study model and an assisted reproductive technology model. PMID:23106040

  15. Paratuberculosis in sheep and goats.

    PubMed

    Windsor, P A

    2015-12-14

    Paratuberculosis is a chronic insidious, often serious, disease of the global small ruminant industries, mainly causing losses from mortalities and reduced productivity on-farm, interference in trading and, in Australia, profound socio-economic impacts that have periodically compromised harmony of rural communities. The pathogenesis, diagnosis, impacts and disease management options for ovine and caprine paratuberculosis are reviewed, comparing current controls in the extensive management system for sheep in wool flocks in Australia with the semi-intensive system of dairy flocks/herds in Greece. Improved understanding of the immune and cellular profiles of sheep with varying paratuberculosis outcomes and the recognition of the need for prolonged vaccination and biosecurity is considered of relevance to future control strategies. Paratuberculosis in goats is also of global distribution although the prevalence, economic impact and strategic control options are less well recognized, possibly due to the relatively meagre resources available for goat industry research. Although there have been some recent advances, more work is required on developing control strategies for goats, particularly in dairy situations where there is an important need for validation of improved diagnostic assays and the recognition of the potential impacts for vaccination. For all species, a research priority remains the identification of tests that can detect latent and subclinical infections to enhance removal of future sources of infectious material from flocks/herds and the food chain, plus predict the likely outcomes of animals exposed to the organism at an early age. Improving national paratuberculosis control programs should also be a priority to manage disease risk from trade. The importance of strong leadership and communication, building trust within rural communities confused by the difficulties in managing this insidious disease, reflects the importance of change management

  16. 21 CFR 862.1300 - Follicle-stimulating hormone test system.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Follicle-stimulating hormone test system. 862.1300... Systems § 862.1300 Follicle-stimulating hormone test system. (a) Identification. A follicle-stimulating hormone test system is a device intended to measure follicle-stimulating hormone (FSH) in plasma,...

  17. 21 CFR 862.1300 - Follicle-stimulating hormone test system.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Follicle-stimulating hormone test system. 862.1300... Systems § 862.1300 Follicle-stimulating hormone test system. (a) Identification. A follicle-stimulating hormone test system is a device intended to measure follicle-stimulating hormone (FSH) in plasma,...

  18. 21 CFR 862.1300 - Follicle-stimulating hormone test system.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Follicle-stimulating hormone test system. 862.1300... Systems § 862.1300 Follicle-stimulating hormone test system. (a) Identification. A follicle-stimulating hormone test system is a device intended to measure follicle-stimulating hormone (FSH) in plasma,...

  19. 21 CFR 862.1300 - Follicle-stimulating hormone test system.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Follicle-stimulating hormone test system. 862.1300... Systems § 862.1300 Follicle-stimulating hormone test system. (a) Identification. A follicle-stimulating hormone test system is a device intended to measure follicle-stimulating hormone (FSH) in plasma,...

  20. 21 CFR 862.1300 - Follicle-stimulating hormone test system.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Follicle-stimulating hormone test system. 862.1300... Systems § 862.1300 Follicle-stimulating hormone test system. (a) Identification. A follicle-stimulating hormone test system is a device intended to measure follicle-stimulating hormone (FSH) in plasma,...

  1. Extensive Hair Shaft Growth after Mouse Whisker Follicle Isolation, Cryopreservation and Transplantation in Nude Mice.

    PubMed

    Cao, Wenluo; Li, Lingna; Tran, Benjamin; Kajiura, Satoshi; Amoh, Yasuyuki; Liu, Fang; Hoffman, Robert M

    2015-01-01

    We previously demonstrated that whole hair follicles could be cryopreserved to maintain their stem-cells differentation potential. In the present study, we demonstrated that cryopreserved mouse whisker hair follicles maintain their hair growth potential. DMSO better cryopreserved mouse whisker follicles compared to glycerol. Cryopreserved hair follicles also maintained the hair follicle-associated-pluripotent (HAP) stem cells, evidenced by P75NTR expression. Subcutaneous transplantation of DMSO-cryopreserved hair follicles in nude mice resulted in extensive hair fiber growth over 8 weeks, indicating the functional recovery of hair shaft growth of cryopreserved hair follicles.

  2. Extensive Hair Shaft Growth after Mouse Whisker Follicle Isolation, Cryopreservation and Transplantation in Nude Mice.

    PubMed

    Cao, Wenluo; Li, Lingna; Tran, Benjamin; Kajiura, Satoshi; Amoh, Yasuyuki; Liu, Fang; Hoffman, Robert M

    2015-01-01

    We previously demonstrated that whole hair follicles could be cryopreserved to maintain their stem-cells differentation potential. In the present study, we demonstrated that cryopreserved mouse whisker hair follicles maintain their hair growth potential. DMSO better cryopreserved mouse whisker follicles compared to glycerol. Cryopreserved hair follicles also maintained the hair follicle-associated-pluripotent (HAP) stem cells, evidenced by P75NTR expression. Subcutaneous transplantation of DMSO-cryopreserved hair follicles in nude mice resulted in extensive hair fiber growth over 8 weeks, indicating the functional recovery of hair shaft growth of cryopreserved hair follicles. PMID:26716690

  3. Goat oocyte quality and competence to undergo IVM and embryo development after parthenogenetic activation from goats fed with different levels of cashew nut bran as source of dietary lipids.

    PubMed

    Fernandes, C C L; Feltrin, C; Martins, L T; Gaudêncio Neto, S; Aguiar, L H; Silva, A M; Oliveira, C H A; Silva, L M; Silva, C M G; Bertolini, M; Rondina, D

    2014-07-15

    Lipid-rich and energy-dense diets can have significant effects on the reproductive physiology, including the ovarian function and fertility. The aim of this study was to assess the effect of cashew nut bran supplementation as a lipid source on follicle development, plasma and intrafollicular concentrations of cholesterol, and developmental competence of in vitro-matured goat oocytes. The inclusion of cashew nut bran as 24% of the goats' diet for 28 days increased the percentage and number of degenerated oocytes compared with the control (P < 0.05), and also the plasma cholesterol levels and the proportion of grade IV oocytes compared with all other treatments (P < 0.05). Moreover, a significant reduction was observed in the proportion of viable oocytes compared with the control and in the percentage of grade II oocytes compared with all other treatments (P < 0.05). Oocyte maturation, cleavage, and blastocyst rates after parthenogenetic activation of viable oocytes were not affected by the type of diet. In conclusion, the inclusion of cashew nut bran as 24% of the diet of adult goats for 28 days changed plasma cholesterol levels and reduced the proportion of viable immature oocytes; however, the 12% and 24% diet supplementations with cashew nut bran did not interfere with competence of resulting viable oocytes to reach the metaphase II stage after IVM, and to develop after parthenogenetic activation.

  4. Goat oocyte quality and competence to undergo IVM and embryo development after parthenogenetic activation from goats fed with different levels of cashew nut bran as source of dietary lipids.

    PubMed

    Fernandes, C C L; Feltrin, C; Martins, L T; Gaudêncio Neto, S; Aguiar, L H; Silva, A M; Oliveira, C H A; Silva, L M; Silva, C M G; Bertolini, M; Rondina, D

    2014-07-15

    Lipid-rich and energy-dense diets can have significant effects on the reproductive physiology, including the ovarian function and fertility. The aim of this study was to assess the effect of cashew nut bran supplementation as a lipid source on follicle development, plasma and intrafollicular concentrations of cholesterol, and developmental competence of in vitro-matured goat oocytes. The inclusion of cashew nut bran as 24% of the goats' diet for 28 days increased the percentage and number of degenerated oocytes compared with the control (P < 0.05), and also the plasma cholesterol levels and the proportion of grade IV oocytes compared with all other treatments (P < 0.05). Moreover, a significant reduction was observed in the proportion of viable oocytes compared with the control and in the percentage of grade II oocytes compared with all other treatments (P < 0.05). Oocyte maturation, cleavage, and blastocyst rates after parthenogenetic activation of viable oocytes were not affected by the type of diet. In conclusion, the inclusion of cashew nut bran as 24% of the diet of adult goats for 28 days changed plasma cholesterol levels and reduced the proportion of viable immature oocytes; however, the 12% and 24% diet supplementations with cashew nut bran did not interfere with competence of resulting viable oocytes to reach the metaphase II stage after IVM, and to develop after parthenogenetic activation. PMID:24853280

  5. Copper toxicosis in a Boer goat.

    PubMed

    Cregar, Laura C; Wiedmeyer, Charles E; Ringen, Davin R; Evans, Tim J; Johnson, Gayle C; Kuroki, Keiichi

    2012-12-01

    A 1-year-old female Boer goat was presented with a 1-day history of pigmenturia, anorexia, and shivering. Anemia was not present initially, but progressive hemolytic anemia developed subsequently and was characterized by the finding of Heinz bodies in both intact RBCs and in ghost cells and the presence of atypical fusiform RBCs. Plasma biochemical analysis revealed increased activities of aspartate aminotransferase and gamma-glutamyltransferase, hyperbilirubinemia, and azotemia. Histopathologic examination of a liver biopsy revealed necrosis of individual hepatocytes and intracytoplasmic rhodamine-positive granules, consistent with copper. Copper concentration in ante-mortem hepatic tissue was increased, and a diagnosis of copper toxicosis was made. Despite supportive therapy, the goat continued to decline and was euthanized. Necropsy findings included hepatic necrosis and hemoglobinuric nephrosis. Freshly collected specimens of liver and kidney had markedly increased copper concentrations. The mineral composition of the water, grass hay, and goat chow was evaluated, and toxins and significant mineral imbalances were not found. The underlying cause of the hepatic accumulation and subsequent release of copper remains unclear in this goat. Recently, Boer goats have been recognized as being prone to copper toxicosis and may be more susceptible than other breeds; similar to sheep, Boer goats may experience a hemolytic crisis secondary to copper toxicosis.

  6. [The hair follicle as a target for gene therapy].

    PubMed

    Cotsarelis, G

    2002-05-01

    The hair follicle possesses progenitor cells required for continuous hair follicle cycling and for epidermal keratinocytes, melanocytes and Langerhans cells. These different cell types can be the target of topical gene delivery in the skin of the mouse. Using a combination of liposomes and DNA, we demonstrate the feasibility of targeting hair follicle cells in human scalp xenografts. We consider liposome composition and stage of the hair cycle as important parameters influencing transfection of human hair follicles. Transfection is possible only during the early anagen phase. Factors and obstacles for the use of gene therapy in treating alopecia and skin diseases are discussed. A theoretical framework for future treatment of cutaneous and systemic disorders using gene therapy is presented.

  7. Engineering the ovarian cycle using in vitro follicle culture

    PubMed Central

    Skory, Robin M.; Xu, Yuanming; Shea, Lonnie D.; Woodruff, Teresa K.

    2015-01-01

    STUDY QUESTION Can cultured follicles model the ovarian cycle, including follicular- and luteal-phase hormone synthesis patterns and ovulation? SUMMARY ANSWER Under gonadotrophin stimulation, murine follicles grown in an encapsulated three-dimensional system ovulate in vitro and murine and human follicle hormone synthesis mimics follicular and luteal phases expected in vivo. WHAT IS KNOWN ALREADY Studies of the human ovary and follicle function are limited by the availability of human tissue and lack of in vitro models. We developed an encapsulated in vitro follicle growth (eIVFG) culture system, which preserves 3D follicular structure. Thus far, the alginate system has supported the culture of follicles from mice, dog, rhesus macaque, baboon and human. These studies have shown that cultured follicles synthesize steroid hormones similar to those observed during the follicular phase in vivo. STUDY DESIGN, SIZE, DURATION Cultured murine follicles were treated with human chorionic gonadotrophin (hCG) and epidermal growth factor (EGF) and either assayed for luteinization or removed from alginate beads and assayed for ovulation. Human follicles were also cultured, treated with follicle-stimulating hormone (FSH), hCG and EGF to mimic gonadotrophin changes throughout the ovarian cycle, and culture medium was assayed for hormone production. PARTICIPANTS/MATERIALS, SETTING, METHODS Murine and human follicles were cultured in alginate hydrogel and hormone production [17β-estradiol, progesterone, inhibin A, inhibin B, activin A and anti-Müllerian hormone (AMH)] was quantified in medium by enzyme-linked immuno assay (ELISA). Human ovarian tissue was acquired from females between 6 and 34 years of age with a cancer diagnosis. These participants were undergoing ovarian tissue cryopreservation at National Physicians Cooperative sites as part of the Oncofertility Consortium. MAIN RESULTS AND THE ROLE OF CHANCE When grown in this system, 96% of mouse follicles ovulated in

  8. Valproate affects reproductive endocrine function, testis diameter and some semen variables in non-epileptic adolescent goat bucks.

    PubMed

    Krogenaes, A K; Taubøll, E; Stien, A; Oskam, I C; Lyche, J L; Dahl, E; Thomassen, R F; Sweeney, T; Ropstad, E

    2008-07-01

    Valproate (VPA) is a major antiepileptic drug with a broad spectrum of antiepileptic activity. There is, however, increasing concern about the possible effects of VPA on reproductive endocrine function. This study investigated the effects of valproate, on the endocrine and reproductive system of adolescent, non-epileptic, goat bucks. Nine goat bucks were orally treated with 62.5mg/kg valproate twice daily from 2 to 10 months of age in order to sustain therapeutic plasma concentrations of between 300 and 600 micromol/l. Seven bucks served as controls. Body weights and testicular diameters were recorded. Blood samples were collected for measurement of luteinising hormone (LH), follicle stimulating hormone (FSH) and testosterone three times weekly until sacrifice at approximately 40 weeks of age. Conventional reproductive endpoints were recorded and flow cytometric (FCM) analyses of spermatogenesis, including the sperm chromatin structure were conducted. Valproate-treated bucks had on average a higher body weight, but a lower testis diameter than controls. No significant differences were found for plasma FSH in comparison to controls. Valproate-treated bucks differed significantly from the control group by showing lower plasma concentrations of LH and testosterone and a later onset of puberty. A significantly higher proportion of sperm from valproate-treated bucks showed abnormal chromatin, demonstrating a harmful effect on DNA from valproate treatment. These results demonstrate that valproate was able to induce reproductive effects in goat bucks related to the hypothalamic-pituitary-axis, as well as to the testes.

  9. Toxicity of Gossypol from Cottonseed Cake to Sheep Ovarian Follicles

    PubMed Central

    Câmara, Antônio Carlos Lopes; Gadelha, Ivana Cristina Nunes; Borges, Pedro Augusto Cordeiro; de Paiva, Silvano Alves; Melo, Marília Martins; Soto-Blanco, Benito

    2015-01-01

    Gossypol, a polyphenol compound produced by cotton plant, has proven reproductive toxicity, but the effects of gossypol on sheep ovaries are unknown. This study was aimed to determine the in vitro and in vivo effects of gossypol on the ovarian follicles of sheep. This trial was divided into two experiments. In the first one, we used twelve non-pregnant, nulliparous, Santa Inês crossbred ewes, which were randomly distributed into two equal groups and fed diets with and without cottonseed cake. Feed was offered at 1.5% of the animal’s body weight for 63 days. The concentrations of total and free gossypol in the cottonseed cake were 3.28 mg/g and 0.11 mg/g, respectively. Throughout the trial period, no animal showed clinical signs of toxicity and no effects on body weight were observed. However, there was a significantly lower number of viable ovarian follicles (20.6%) and higher number of atretic follicles (79.4%) in the gossypol-fed sheep compared to the control (85.1 and 34.9%, respectively). These findings were observed at all stages of follicular development. In the second experiment, eight ovaries from slaughterhouse were cultured with different concentrations of gossypol acetic acid (0, 5, 10 and 20 μg/mL) for 24 hours or seven days. The in vitro action of gossypol resulted in a significant decrease in viable ovarian follicles, especially the primary and transition follicles, and a significant increase in the number of atretic follicles after 24 hours of culture. These follicles were greatly affected when cultured with gossypol for seven days. It is concluded that gossypol present in cotton seeds directly acts on ovarian follicles in sheep to increase atresia. PMID:26600470

  10. Putting the Human Hair Follicle Cycle on the Map.

    PubMed

    Panteleyev, Andrey A

    2016-01-01

    A detailed characterization of the normal (in situ) human hair follicle cycle, supplemented with expressional data on specific hair follicle markers, has been awaited by basic hair researchers and dermatologists. Combining this hair cycle guide, together with a thorough analysis of the human-on-mouse hair xenograft model, provides solid ground for examining human hair cycle biology and pathology and for hair cycle-related pharmacological testing.

  11. Multiple calcifying hyperplastic dental follicles: A case report

    PubMed Central

    Aydin, Ulkem; Baykul, Timucin; Yildirim, Benay; Yildirim, Derya; Karaduman, Ayse

    2013-01-01

    This report describes a 31-year-old female patient with six impacted teeth. The crowns of the impacted teeth were surrounded with cyst-like lesions with a mixed internal structure and well-defined cortical borders. Microscopic examination of the specimen obtained from the follicle of the left mandibular third molar tooth revealed loose to moderately dense collagenous connective tissue with abundant calcified material and sparse epithelial islands. A diagnosis of multiple calcifying hyperplastic dental follicles was made. PMID:24380071

  12. Human hair follicle: reservoir function and selective targeting.

    PubMed

    Blume-Peytavi, U; Vogt, A

    2011-10-01

    Penetration of topically applied compounds may occur via the stratum corneum, skin appendages and hair follicles. The follicular infundibulum increases the surface area, disrupts the epidermal barrier towards the lower parts of the follicle, and serves as a reservoir. Topical delivery of active compounds to specific targets within the skin, especially to distinct hair follicle compartments or cell populations, may help to treat local inflammatory reactions selectively, with reduced systemic side-effects. Various in vitro and in vivo methods exist for studying the hair follicle structure and follicular penetration pathways. These include cyanoacrylate skin surface stripping, confocal microscopy and cyanoacrylate scalp follicle biopsy. The complex anatomical structure as well as the cyclical activity of the hair follicle must be taken into consideration when designing delivery systems. In addition, delivery into and retention inside the infundibular reservoir are controlled by, for example, molecule or particle size, their polarity and the type of preparation. Preferred penetration depth and storage time must also be considered. Particles with release mechanisms should be preferred; however, the release of drugs from nanoparticles still requires further investigations. PMID:21919898

  13. Human hair follicle: reservoir function and selective targeting.

    PubMed

    Blume-Peytavi, U; Vogt, A

    2011-10-01

    Penetration of topically applied compounds may occur via the stratum corneum, skin appendages and hair follicles. The follicular infundibulum increases the surface area, disrupts the epidermal barrier towards the lower parts of the follicle, and serves as a reservoir. Topical delivery of active compounds to specific targets within the skin, especially to distinct hair follicle compartments or cell populations, may help to treat local inflammatory reactions selectively, with reduced systemic side-effects. Various in vitro and in vivo methods exist for studying the hair follicle structure and follicular penetration pathways. These include cyanoacrylate skin surface stripping, confocal microscopy and cyanoacrylate scalp follicle biopsy. The complex anatomical structure as well as the cyclical activity of the hair follicle must be taken into consideration when designing delivery systems. In addition, delivery into and retention inside the infundibular reservoir are controlled by, for example, molecule or particle size, their polarity and the type of preparation. Preferred penetration depth and storage time must also be considered. Particles with release mechanisms should be preferred; however, the release of drugs from nanoparticles still requires further investigations.

  14. Integral hair lipid in human hair follicle.

    PubMed

    Lee, Won-Soo

    2011-12-01

    Integral hair lipid (IHL) is bound to the keratinized cell surface to make an environmentally resistant lipid envelope. It is mainly positioned on the hair cuticle and inner root sheath. IHL in the hair follicle may regard as hair barrier to be similar to the epidermal lipid layer functioning as skin barrier. Major constituents of IHL are fatty acid, phytosphingosine, ceramide in decreasing order. Minor constituents of IHL are cholesterol, cholesterol sulfate and cholesterol oleate. Cuticle or cortical cell surface in hair are abundant in fatty acids unlike the keratinized area of epidermis or sebaceous gland, and about 30-40% of such fatty acids are composed of 18-methyl-eicosanoic acid which is known to be bound to proteins by ester or thioester bond. Various factors including moisture, solvent, oxidative damage during bleaching or permanent waving affect IHL. Photochemical changes also can occur in IHL as well as in hair protein and hair pigment. Lipid metabolism is thought to play an essential role in lipid envelope of hair, but also involvement in hair development and function.

  15. Effect of epidermal growth factor on follicle-stimulating hormone-induced proliferation of granulosa cells from chicken prehierarchical follicles.

    PubMed

    Lin, Jin-xing; Jia, Yu-dong; Zhang, Cai-qiao

    2011-11-01

    The development of ovarian follicular cells is controlled by multiple circulating and local hormones and factors, including follicle-stimulating hormone (FSH) and epidermal growth factor (EGF). In this study, the stage-specific effect of EGF on FSH-induced proliferation of granulosa cells was evaluated in the ovarian follicles of egg-laying chickens. Results showed that EGF and its receptor (EGFR) mRNAs displayed a high expression in granulosa cells from the prehierarchical follicles, including the large white follicle (LWF) and small yellow follicle (SYF), and thereafter the expression decreased markedly to the stage of the largest preovulatory follicle. SYF represents a turning point of EGF/EGFR mRNA expression during follicle selection. Subsequently the granulosa cells from SYF were cultured to reveal the mediation of EGF in FSH action. Cell proliferation was remarkably increased by treatment with either EGF or FSH (0.1-100 ng/ml). This result was confirmed by elevated proliferating cell nuclear antigen (PCNA) expression and decreased cell apoptosis. Furthermore, EGF-induced cell proliferation was accompanied by increased mRNA expressions of EGFR, FSH receptor, and the cell cycle-regulating genes (cyclins D1 and E1, cyclin-dependent kinases 2 and 6) as well as decreased expression of luteinizing hormone receptor mRNA. However, the EGF or FSH-elicited effect was reversed by simultaneous treatment with an EGFR inhibitor AG1478. In conclusion, EGF and EGFR expressions manifested stage-specific changes during follicular development and EGF mediated FSH-induced cell proliferation and retarded cell differentiation in the prehierarchical follicles. These expressions thus stimulated follicular growth before selection in the egg-laying chicken.

  16. Chitotriosidase activity in goat blood and colostrum.

    PubMed

    Argüello, A; Castro, N; Batista, M; Moreno-Indias, I; Morales-delaNuez, A; Sanchez-Macias, D; Quesada, E; Capote, J

    2008-05-01

    Chitotriosidase (ChT) activity has not been investigated in ruminants, and therefore, we studied this activity in blood and colostrum of 25 pregnant goats and 60 goat kids. Blood samples were taken from pregnant goats at 3, 2, and 1 d prepartum; at partum; and at 1, 2, 3, and 4 d postpartum. Colostrum samples were obtained by machine-milking at partum and 1, 2, 3, and 4 d postpartum. Goat kid blood was collected at birth and every 7 d thereafter until goats kids were 56 d old. The ChT activity ranged from 2,368 to 3,350 nmol/ mL per hour in goat blood serum, and no statistical differences were detected through time. However, activity tended to decrease from 3 d prepartum to 2 d post-partum. Colostrum ChT activity was 3,912 nmol/mL per hour and 465 nmol/mL per hour on the day of delivery and 4 d postpartum, respectively. Colostrum ChT activity was significantly higher at partum than at any other time. The ChT activity in colostrum was significantly greater at 1 d postpartum than at 2, 3, and 4 d postpartum. Chitotriosidase activity did not differ in colostrum collected on d 2, 3, and 4 postpartum. Chitotriosidase activity in goat kid blood serum ranged from 2,664 to 9,231 nmol/mL per hour at birth and 49 d of life, respectively. Chitotriosidase activity in the blood serum increased with age: at birth, activity was significantly less than at 28, 35, 42, 49, and 56 d postpartum. The maximum ChT activity in blood serum was observed at 49 d postpartum. Activity in 49-d-old kids was significantly greater than that observed in kids at 0, 7, and 14 d postpartum.

  17. Ultrasound characteristics of experimentally induced luteinized unruptured follicles (LUF) and naturally occurring hemorrhagic anovulatory follicles (HAF) in the mare.

    PubMed

    Cuervo-Arango, J; Newcombe, J R

    2012-02-01

    The development of hemorrhagic anovulatory follicles (HAF) involves luteinization and hemorrhage of the follicle. This is observed on ultrasound as an increase in the echogenicity of the granulosa layer and formation of echoic particles in the antrum. The inhibition of prostaglandin synthesis with flunixin meglumine (FM) during the periovulatory period induces ovulatory failure with development of luteinized unruptured follicles (LUF). These two types of anovulatory follicles appear to share similar ultrasound features but they have not been compared critically. The following endpoints: follicle diameter, follicular contents score, interval from hCG administration to beginning of follicular hemorrhage, interval from hemorrhage to organization of follicular contents, and cycle length were studied and compared in mares with HAF (n = 11) and LUF (n = 13). The objective of this study was to elucidate whether these two unruptured follicles have a consistent clinical pattern of development and therefore can be considered as part of the same anovulatory syndrome. None of the endpoints analyzed differed significantly between HAF and LUF. However, there was a greater individual variation in HAF as compared with LUF in regards to interval from hCG to hemorrhage, follicular diameter at the administration of hCG, and beginning of hemorrhage. In conclusion, HAF share a similar cascade of ultrasound characteristics with the experimentally induced LUF. This finding may provide new insights in elucidating the pathogenesis of HAF.

  18. 4. Light tower, interior from entrance, looking southeast Goat ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    4. Light tower, interior from entrance, looking southeast - Goat Island Light Station, Goat Island, next to entrance to Cape Porpoise Harbor, just south of Trott Island, Cape Porpoise, York County, ME

  19. 6. Boathouse, looking northwest, southwest and southeast sides Goat ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    6. Boathouse, looking northwest, southwest and southeast sides - Goat Island Light Station, Goat Island, next to entrance to Cape Porpoise Harbor, just south of Trott Island, Cape Porpoise, York County, ME

  20. Ultrasonic imaging of equine ovarian follicles and corpora lutea.

    PubMed

    Ginther, O J

    1988-08-01

    One of the most profound theriogenology applications of transrectal diagnostic ultrasonography in mares involves the imaging of ovarian follicles and corpora lutea. The resolving capabilities (frequency) and quality of the scanner directly affect the minimal size of a structure that can be imaged and the quality of the image. High-frequency scanners (5 or 7.5 MHz) of good quality can image a 2-mm follicle and the corpus luteum throughout its functional life. A low-frequency scanner (3 or 3.5 MHz) can image a 6-mm follicle and the corpus luteum for several days after ovulation. Equine follicles are excellent subjects for transrectal imaging because they are large, filled with fluid, and readily accessible. Event the small follicles (less than 10 mm) can be diagnostically important in evaluating whether ovarian infertility has occurred and whether the follicles are responding to treatment for follicular stimulation. The large, preovulatory follicles are of special interest. Averaged over a group of 79 periods, the following significant changes were found in the preovulatory follicle: increasing diameter, shape change from spherical to pear-shaped or conical, and increasing thickness of the follicular wall. No significant changes were found in the echogenicity (gray-scale value) of the wall or fluid. In retrospect, the diameter of the follicle seemed as useful for predicting impending ovulation as any of the other ultrasound criteria. The occurrence of ovulation is readily detected by the disappearance of a large follicle that was present at a recent previous examination. In addition, the ovulation site on the day of ovulation is detectable. In one study, the site was correctly identified in 24 of 24 mares. A small amount of residual follicular fluid can sometimes (7 of 10 in one study) be detected at the site of ovulation. The residual fluid usually disappears over a period of 0.5 to 20 hours. Subsequently, the developing corpus luteum may form a central nonechogenic

  1. Glycomic analyses of ovarian follicles during development and atresia

    PubMed Central

    Hatzirodos, Nicholas; Nigro, Julie; Irving-Rodgers, Helen F.; Vashi, Aditya V.; Hummitzsch, Katja; Caterson, Bruce; Sullivan, Thomas R.; Rodgers, Raymond J.

    2012-01-01

    To examine the detailed composition of glycosaminoglycans during bovine ovarian follicular development and atresia, the specialized stromal theca layers were separated from the stratified epithelial granulosa cells of healthy (n = 6) and atretic (n = 6) follicles in each of three size ranges: small (3–5 mm), medium (6-9 mm) and large (10 mm or more) (n = 29 animals). Fluorophore-assisted carbohydrate electrophoresis analyses (on a per cell basis) and immunohistochemistry (n = 14) were undertaken. We identified the major disaccharides in thecal layers and the membrana granulosa as chondroitin sulfate-derived ∆uronic acid with 4-sulfated N-acetylgalactosamine and ∆uronic acid with 6-sulfated N-acetylgalactosamine and the heparan sulfate-derived Δuronic acid with N-acetlyglucosamine, with elevated levels in the thecal layers. Increasing follicle size and atresia was associated with increased levels of some disaccharides. We concluded that versican contains 4-sulfated N-acetylgalactosamine and it is the predominant 4-sulfated N-acetylgalactosamine proteoglycan in antral follicles. At least one other non- or 6-sulfated N-acetylgalactosamine proteoglycan(s), which is not decorin or an inter-α-trypsin inhibitor family member, is present in bovine antral follicles and associated with hitherto unknown groups of cells around some larger blood vessels. These areas stained positively for chondroitin/dermatan sulfate epitopes [antibodies 7D4, 3C5, and 4C3], similar to stem cell niches observed in other tissues. The sulfation pattern of heparan sulfate glycosaminoglycans appears uniform across follicles of different sizes and in healthy and atretic follicles. The heparan sulfate products detected in the follicles are likely to be associated with perlecan, collagen XVIII or betaglycan. PMID:22057033

  2. Definition of prepartum hyperketonemia in dairy goats.

    PubMed

    Doré, V; Dubuc, J; Bélanger, A M; Buczinski, S

    2015-07-01

    A prospective cohort study was conducted on 1,081 dairy goats from 10 commercial herds in Québec (Canada) to define prepartum hyperketonemia based on optimal blood β-hydroxybutyrate acid threshold values for the early prediction of pregnancy toxemia (PT) and mortality in late-gestation dairy goats. All pregnant goats had blood sampled weekly during the last 5wk of pregnancy. The blood was analyzed directly on the farm for β-hydroxybutyrate acid quantification using a Precision Xtra meter (Abbott Diabetes Care, Saint-Laurent, QC, Canada). Body condition scores on the lumbar region and sternum were noted. Each goat was classified as being at low (n=973) or high risk (n=108) of having PT by producers based on a standardized definition. The optimal threshold for predicting a PT diagnosis or mortality for each week before kidding was determined based on the highest sum of sensitivity and specificity. The association between hyperketonemia and subsequent PT was tested using a multivariable logistic regression model considering hyperketonemia at wk 4 prepartum, litter size, and body condition score at wk 4 prepartum as covariates, and herd and parturition cohort as random effects. The association between mortality and hyperketonemia was also tested using a logistic regression model accounting for the presence or absence of treatment during the last month of pregnancy. The hyperketonemia definition based on PT varied between ≥0.4 and ≥0.9mmol/L during the last 5wk prepartum. Goats affected by hyperketonemia at wk 4 prepartum and with a large litter size (≥3 fetuses) had 2.1 and 40.5 times the odds, respectively, of subsequent PT than other goats. Hyperketonemia definitions based on mortality varied between ≥0.6 and ≥1.4mmol/L during the last 4wk prepartum, and was ≥1.7mmol/L during the first week postpartum. Goats affected by hyperketonemia and treated by producers had 3.4 and 11.8 times the odds, respectively, of subsequent mortality than did other goats

  3. 9 CFR 309.14 - Brucellosis-reactor goats.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Brucellosis-reactor goats. 309.14 Section 309.14 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... INSPECTION AND CERTIFICATION ANTE-MORTEM INSPECTION § 309.14 Brucellosis-reactor goats. Goats which...

  4. 9 CFR 309.14 - Brucellosis-reactor goats.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Brucellosis-reactor goats. 309.14 Section 309.14 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... INSPECTION AND CERTIFICATION ANTE-MORTEM INSPECTION § 309.14 Brucellosis-reactor goats. Goats which...

  5. 9 CFR 309.14 - Brucellosis-reactor goats.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 2 2011-01-01 2011-01-01 false Brucellosis-reactor goats. 309.14 Section 309.14 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... INSPECTION AND CERTIFICATION ANTE-MORTEM INSPECTION § 309.14 Brucellosis-reactor goats. Goats which...

  6. 9 CFR 309.14 - Brucellosis-reactor goats.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Brucellosis-reactor goats. 309.14 Section 309.14 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... INSPECTION AND CERTIFICATION ANTE-MORTEM INSPECTION § 309.14 Brucellosis-reactor goats. Goats which...

  7. 9 CFR 309.14 - Brucellosis-reactor goats.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Brucellosis-reactor goats. 309.14 Section 309.14 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... INSPECTION AND CERTIFICATION ANTE-MORTEM INSPECTION § 309.14 Brucellosis-reactor goats. Goats which...

  8. 21 CFR 135.115 - Goat's milk ice cream.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Goat's milk ice cream. 135.115 Section 135.115... Goat's milk ice cream. (a) Description. Goat's milk ice cream is the food prepared in the same manner... and hydrolyzed milk proteins may not be used; and paragraphs (f)(1) and (g) of § 135.110 shall...

  9. 21 CFR 135.115 - Goat's milk ice cream.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Goat's milk ice cream. 135.115 Section 135.115... Goat's milk ice cream. (a) Description. Goat's milk ice cream is the food prepared in the same manner... and hydrolyzed milk proteins may not be used; and paragraphs (f)(1) and (g) of § 135.110 shall...

  10. 21 CFR 135.115 - Goat's milk ice cream.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Goat's milk ice cream. 135.115 Section 135.115... Goat's milk ice cream. (a) Description. Goat's milk ice cream is the food prepared in the same manner... and hydrolyzed milk proteins may not be used; and paragraphs (f)(1) and (g) of § 135.110 shall...

  11. Contagious ecthyma in mountain goat of coastal British Columbia.

    PubMed

    Hebert, D M; Samuel, W M; Smith, G W

    1977-04-01

    Contagious ecthyma has been reported previously from mountain goat (Oreamnos americanus) in one restricted area of eastern British Columbia. A second focus of infection is reported for mountain goat from western British Columbia. Diagnosis was based on appearance of lesions at necropsy, histopathology and demonstration of poxvirus with the electron microscope. The epizootiology of this infection in mountain goat is discussed briefly.

  12. Traditional goat husbandry may substantially contribute to human toxoplasmosis exposure

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Raising goats in settings that are highly contaminated with oocysts of Toxoplasma gondii may contribute significantly to human exposure to this zoonotic parasite. Increasing consumption of young goats in Romania, where goats are typically reared in backyards that are also home to cats (the definitiv...

  13. 21 CFR 135.115 - Goat's milk ice cream.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Goat's milk ice cream. 135.115 Section 135.115 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD... Goat's milk ice cream. (a) Description. Goat's milk ice cream is the food prepared in the same...

  14. 21 CFR 135.115 - Goat's milk ice cream.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Goat's milk ice cream. 135.115 Section 135.115 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD... Goat's milk ice cream. (a) Description. Goat's milk ice cream is the food prepared in the same...

  15. Molecular control of ovulation and luteinization in the primate follicle.

    PubMed

    Stouffer, Richard L; Xu, Fuhua; Duffy, Diane M

    2007-01-01

    In recent years, significant progress was made, particularly through the use of the macaque monkey, in identifying three types of local factors that are induced by the midcycle LH surge and play a critical role in ovulation and/or luteinization of the primate follicle. The ovulatory gonadotropin surge increases prostaglandin (PTG, typically abbreviated PG) levels in follicles prior to rupture; although considerable attention has focused on LH stimulation of the "inducible" form of PG G/H synthase (PTGS2), other aspects of PG synthesis (notably a phospholipase A2, cPLA2, and a PGE synthase, PTGES) and metabolism (15-hydroxy PG dehydrogenase, HPGD) also appear LH-regulated and may control the timing of the PG rise in the ovulatory follicle. Local (intrafollicular) ablation and replacement of PGs suggests that PGE2 is essential for release of the oocyte; but not necessarily for follicle rupture, and not for luteinization. Novel PGE-regulated genes are being identified in macaque granulosa cells, including adipose differentiation-related protein (ADFP). Similar types of studies indicate that the rise in progesterone (P) synthesis, as well as the induction of the genomic P receptor in granulosa cells, is essential for both ovulation and luteinization of the primate follicle. Limited data suggest that P action controls cell cycle activity (via cyclin B1 and cyclin-dependent kinase inhibitor p27), cholesterol uptake and utilization (e.g., low density lipoprotein or LDL receptor), proteases and their inhibitors (matrix metalloproteinase or MMP1; tissue inhibitor of MMP or TIMP1) and cell health in the granulosa cell layer. Finally, members of two classes of angiogenic factors, originally proposed as important for embryonic and pathologic (tumorigenic) vasculogenesis, appear induced in the granulosa layer of the preovulatory follicle, i.e., vascular endothelial growth factor (VEGF) and angiopoietin (ANGPT). Local injection of antagonists to VEGF (soluble VEGF receptor) and

  16. New Jersey 4-H Goat Extravaganza: Efficiently Meeting the Educational Needs of 4-H Goat Project Members, Volunteers, and Parents

    ERIC Educational Resources Information Center

    Ripberger, Chad

    2014-01-01

    The 4-H Goat Extravaganza maximizes limited resources to help youth and adults develop knowledge and skills in goat care and management. It capitalizes on the talents and interests of volunteers to efficiently combine a goat-themed art show, team presentation contest, quiz bowl, skillathon, and adult workshop into 1 day. This article outlines the…

  17. Di(2-ethylhexyl) phthalate inhibits antral follicle growth, induces atresia, and inhibits steroid hormone production in cultured mouse antral follicles

    PubMed Central

    Hannon, Patrick R.; Brannick, Katherine E.; Wang, Wei; Gupta, Rupesh K.; Flaws, Jodi A.

    2015-01-01

    Di(2-ethylhexyl) phthalate (DEHP) is a ubiquitous environmental toxicant found in consumer products that causes ovarian toxicity. Antral follicles are the functional ovarian units and must undergo growth, survival from atresia, and proper regulation of steroidogenesis to ovulate and produce hormones. Previous studies have determined that DEHP inhibits antral follicle growth and decreases estradiol levels in vitro; however, the mechanism by which DEHP elicits these effects is unknown. The present study tested the hypothesis that DEHP directly alters regulators of the cell cycle, apoptosis, and steroidogenesis to inhibit antral follicle functionality. Antral follicles from adult CD-1 mice were cultured with vehicle control or DEHP (1-100μg/ml) for 24-96 hr to establish the temporal effects of DEHP on the follicle. Following 24-96 hr of culture, antral follicles were subjected to gene expression analysis, and media were subjected to measurements of hormone levels. DEHP increased the mRNA levels of cyclin D2, cyclin dependent kinase 4, cyclin E1, cyclin A2, and cyclin B1 and decreased the levels of cyclin-dependent kinase inhibitor 1A prior to growth inhibition. Additionally, DEHP increased the mRNA levels of BCL2-associated agonist of cell death, BCL2-associated X protein, BCL2-related ovarian killer protein, B-cell leukemia/lymphoma 2, and Bcl2-like 10, leading to an increase in atresia. Further, DEHP decreased the levels of progesterone, androstenedione, and testosterone prior to the decrease in estradiol levels, with decreased mRNA levels of side-chain cleavage, 17α-hydorxylase-17,20-desmolase, 17β-hydroxysteroid dehydrogenase, and aromatase. Collectively, DEHP directly alters antral follicle functionality by inhibiting growth, inducing atresia, and inhibiting steroidogenesis. PMID:25701202

  18. Failure of sheep-goat hybrid conceptuses to develop to term in sheep-goat chimaeras.

    PubMed

    Gustafson, R A; Anderson, G B; BonDurant, R H; Sasser, G R

    1993-09-01

    Six hybrid pregnancies were established: three in sheep-goat chimaeras, one in a sheep-(sheep-goat)hybrid chimaera and two in does. Pregnancies were monitored weekly by ultrasonography and peripheral concentrations of pregnancy specific protein B (PSPB) were measured. Placental development as detected by ultrasonography appeared to be slower in hybrid-in-goat pregnancies than in hybrid-in-chimaera pregnancies, although this difference was not reflected in PSPB concentrations. Time of fetal death could not be predicted from PSPB concentrations. Chimaeras appeared to carry hybrid pregnancies longer than ewes and does usually carry hybrid pregnancies, but none was carried to term. PMID:7506792

  19. Genetic Variation of Goat Interferon Regulatory Factor 3 Gene and Its Implication in Goat Evolution.

    PubMed

    Okpeku, Moses; Esmailizadeh, Ali; Adeola, Adeniyi C; Shu, Liping; Zhang, Yesheng; Wang, Yangzi; Sanni, Timothy M; Imumorin, Ikhide G; Peters, Sunday O; Zhang, Jiajin; Dong, Yang; Wang, Wen

    2016-01-01

    The immune systems are fundamentally vital for evolution and survival of species; as such, selection patterns in innate immune loci are of special interest in molecular evolutionary research. The interferon regulatory factor (IRF) gene family control many different aspects of the innate and adaptive immune responses in vertebrates. Among these, IRF3 is known to take active part in very many biological processes. We assembled and evaluated 1356 base pairs of the IRF3 gene coding region in domesticated goats from Africa (Nigeria, Ethiopia and South Africa) and Asia (Iran and China) and the wild goat (Capra aegagrus). Five segregating sites with θ value of 0.0009 for this gene demonstrated a low diversity across the goats' populations. Fu and Li tests were significantly positive but Tajima's D test was significantly negative, suggesting its deviation from neutrality. Neighbor joining tree of IRF3 gene in domesticated goats, wild goat and sheep showed that all domesticated goats have a closer relationship than with the wild goat and sheep. Maximum likelihood tree of the gene showed that different domesticated goats share a common ancestor and suggest single origin. Four unique haplotypes were observed across all the sequences, of which, one was particularly common to African goats (MOCH-K14-0425, Poitou and WAD). In assessing the evolution mode of the gene, we found that the codon model dN/dS ratio for all goats was greater than one. Phylogenetic Analysis by Maximum Likelihood (PAML) gave a ω0 (dN/dS) value of 0.067 with LnL value of -6900.3 for the first Model (M1) while ω2 = 1.667 in model M2 with LnL value of -6900.3 with positive selection inferred in 3 codon sites. Mechanistic empirical combination (MEC) model for evaluating adaptive selection pressure on particular codons also confirmed adaptive selection pressure in three codons (207, 358 and 408) in IRF3 gene. Positive diversifying selection inferred with recent evolutionary changes in domesticated goat IRF3

  20. Detection of Bim and Puma in mouse hair follicles using immunofluorescence and TUNEL assay double staining.

    PubMed

    Vesela, B; Matalova, E

    2015-01-01

    Apoptosis in hair follicles often is studied under pathological conditions; little is known about apoptotic mechanisms during normal hair follicle formation and maintenance. We investigated proteins of intrinsic apoptotic pathway, Bim and Puma, during hair follicle development and the first catagen stage using immunofluorescence to describe their expression patterns and to correlate them with apoptosis as determined by TUNEL assay. Both proteins were found in developing follicles. Bim and Puma overlapped apoptosis only partially during physiological apoptotic stage and they were present in non-apoptotic parts of the follicles. Our findings suggest that these primary apoptotic molecules participate in postnatal development and maintenance of hair follicles.

  1. Protocols for Cryopreservation of Intact Hair Follicle That Maintain Pluripotency of Nestin-Expressing Hair-Follicle-Associated Pluripotent (HAP) Stem Cells.

    PubMed

    Kajiura, Satoshi; Mii, Sumiyuki; Aki, Ryoichi; Hamada, Yuko; Arakawa, Nobuko; Kawahara, Katsumasa; Li, Lingna; Katsuoka, Kensei; Hoffman, Robert M; Amoh, Yasuyuki

    2016-01-01

    Hair follicles contain nestin-expressing pluripotent stem cells, the origin of which is above the bulge area, below the sebaceous gland. We have termed these cells hair-follicle-associated pluripotent (HAP) stem cells. Cryopreservation methods of the hair follicle that maintain the pluripotency of HAP stem cells are described in this chapter. Intact hair follicles from green fluorescent protein (GFP) transgenic mice were cryopreserved by slow-rate cooling in TC-Protector medium and storage in liquid nitrogen. After thawing, the upper part of the hair follicle was isolated and cultured in DMEM with fetal bovine serum (FBS). After 4 weeks culture, cells from the upper part of the hair follicles grew out. The growing cells were transferred to DMEM/F12 without FBS. After 1 week culture, the growing cells formed hair spheres, each containing approximately 1 × 10(2) HAP stem cells. The hair spheres contained cells which could differentiate to neurons, glial cells, and other cell types. The formation of hair spheres by the thawed and cultured upper part of the hair follicle produced almost as many pluripotent hair spheres as fresh follicles. The hair spheres derived from cryopreserved hair follicles were as pluripotent as hair spheres from fresh hair follicles. These results suggest that the cryopreservation of the whole hair follicle is an effective way to store HAP stem cells for personalized regenerative medicine, enabling any individual to maintain a bank of pluripotent stem cells for future clinical use. PMID:27431257

  2. Characterization of Korean native goat lactoferrin.

    PubMed

    Nam, M S; Shimazaki, K; Kumura, H; Lee, K K; Yu, D Y

    1999-06-01

    We purified lactoferrin from the colostrum of the Korean native goat (Capra hircus) by ion-exchange chromatography using CM-Toyopearl 650M followed by affinity chromatography on AF-Heparin Toyopearl 650M. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis suggested the molecular mass of Korean native goat lactoferrin is 82 kDa with an iron saturation of 30% as estimated by spectroscopic analysis. Circular dichroism analysis shows goat lactoferrin molecule contains 24.5%, alpha-helix; 36.0%, beta-structure; 13.5%, beta-turn and 26.0%, unordered structure. Heparin binding affinity is the same as that of bovine lactoferrin, but lower than that of human lactoferrin. An analysis using synthetic peptides shows that the peptide from residue 22 to 31--WQRRMRKLGA--exerts a positive heparin-binding ability.

  3. Occipitoatlantoaxial malformation in an adult goat.

    PubMed

    Seva, Juan I; Gómez, Serafin; Pallarés, Francisco J; Sánchez, Pedro; Bernabé, Antonio

    2008-09-01

    An occipitoatlantoaxial malformation was diagnosed in a 1-year-old Murciano-Granadina goat. At clinical examination, the head and cranial part of the neck were deviated to the right. Clinical signs of spinal cord or brain disease were not observed. At necropsy, morphological abnormalities were seen in the craniovertebral junction and cervical vertebrae, characterized by a firm attachment and incomplete articulation between the occipital bone and the atlas, and scoliosis in the cervical regions. The definitive diagnosis was bilateral asymmetrical occipitoatlantoaxial fusion with rotation of the atlas and atlantoaxial subluxation. To the authors' knowledge, this case report is the second occipitoatlantoaxial malformation described in a goat and the first description in an adult goat.

  4. Effect of three pFSH doses on superovulation and embryo quality in goats during two breeding seasons in north-eastern mexico.

    PubMed

    Sánchez-Dávila, F; Ledezma-Torres, R A; Padilla-Rivas, G; Del Bosque-González, A S; González Gómez, A; Bernal-Barragán, H

    2014-08-01

    The aim of this research was to evaluate the effectiveness of three pFSH doses (80 mg; 145 mg and 215 mg) on ovarian response and on quantity and quality of transferable embryos of goats during the breeding and the non-breeding seasons. Ovary structures were exposed (laparatomy under general anaesthesia) and numbers of follicles and corpora lutea were registered. Surgical embryo flushing was conducted to count and classify embryos. There were more follicles (3.4 ± 1.1) in does administered 80 mg of pFSH (p < 0.05) than in goats administered 145 mg of pFSH (2.2 ± 1.1) and 215 mg of pFSH (0.9 ± 0.6). Numbers of corpora lutea, blastocysts, and recovered and transferable embryos of goats administered 145 mg pFSH (13.4 ± 3.7, 2.42 ± 1.0, 3.4 ± 1.2 and 3.2 ± 1.1, respectively) and those of goats administered 215 mg pFSH (11.6 ± 2.6, 3.2 ± 0.9, 5.7 ± 1.5, and 5.6 ± 1.5) were greater (p < 0.05) than values obtained from goats administered 80 mg pFSH (4.0 ± 1.5, 0.5 ± 0.3, 1.0 ± 0.5, and 0.8 ± 0.5). Numbers of morula of does administered 80 and 145 mg pFSH (0.4 ± 0.4 and 0.8 ± 0.3) were lower (p < 0.05) than those obtained from animals treated with 215 mg pFSH (2.4 ± 0.9). There was no effect of season upon the analyzed variables. In conclusion, under the prevalent conditions in north-eastern Mexico, administration of 145 or 215 mg pFSH in a decreasing dose schedule over 3.5 days to bred goats provided a satisfactory superovulatory result.

  5. Induced pluripotent stem cells from goat fibroblasts.

    PubMed

    Song, Hui; Li, Hui; Huang, Mingrui; Xu, Dan; Gu, Chenghao; Wang, Ziyu; Dong, Fulu; Wang, Feng

    2013-12-01

    Embryonic stem cells (ESCs) are a powerful model for genetic engineering, studying developmental biology, and modeling disease. To date, ESCs have been established from the mouse (Evans and Kaufman, 1981, Nature 292:154-156), non-human primates (Thomson et al., , Proc Nat Acad Sci USA 92:7844-7848), humans (Thomson et al., 1998, Science 282:1145-1147), and rats (Buehr et al., , Cell 135:1287-1298); however, the derivation of ESCs from domesticated ungulates such as goats, sheep, cattle, and pigs have not been successful. Alternatively, induced pluripotent stem cells (iPSCs) can be generated by reprogramming somatic cells with several combinations of genes encoding transcription factors (OCT3/4, SOX2, KLF4, cMYC, LIN28, and NANOG). To date, iPSCs have been isolated from various species, but only limited information is available regarding goat iPSCs (Ren et al., 2011, Cell Res 21:849-853). The objectives of this study were to generate goat iPSCs from fetal goat primary ear fibroblasts using lentiviral transduction of four human transcription factors: OCT4, SOX2, KLF4, and cMYC. The goat iPSCs were successfully generated by co-culture with mitomycin C-treated mouse embryonic fibroblasts using medium supplemented with knockout serum replacement and human basic fibroblast growth factor. The goat iPSCs colonies are flat, compact, and closely resemble human iPSCs. They have a normal karyotype; stain positive for alkaline phosphatase, OCT4, and NANOG; express endogenous pluripotency genes (OCT4, SOX2, cMYC, and NANOG); and can spontaneously differentiate into three germ layers in vitro and in vivo. PMID:24123501

  6. [Occurrence of nematodiasis among sheep and goats].

    PubMed

    Zurliĭski, P

    1977-01-01

    The spread was studied of sheep and goat nematodirosis in the conditions prevailing in the district of Varna for the 1973-1975 period. Füleborn's method was employed with a total of 24,909 coprosamples taken from the animals as follows: sheep--12,690, weaned lambs--7370, lambs--3355, kids--782, and goats--712. Partial helminthologic postmortem examinations were carried out by the digestive method of 104 sheep, 33 goats, 142 weaned lambs, 96 lambs, and 35 kids. The percent of infected animals was determined on the basis of the coprostudies as follows; lambs--20.9 (per cent), kids--26.6, weaned lambs--54.9, sheep--22.6, goats--24.8. The necroscopic investigations revealed 32.3, 40, 64.8, 32.5, 39.4 per cent, respectively. The average parasite burden in number of helminths was 313 (lambs), 85 (kids), 1517 (weaned lambs), 586 (sheep), and goats--290. Greatest number of Nematodirus parasites was found in weaned lambs--16,000, followed by sheep--2260, lambs--1840, goats--800, and kids--240. As many as 100 per cent Nematodirus carriers were found on a sheep farm in the district. It was also found that the north plains of the district are less infected as against the south parts embracing the northern slopes of the Balkan mountain, and along the course of the Kamchia river. It was found that sheep on the private farm-holdings were more strongly infected than those on the cooperative farms. On the other hand, it was established that on farms where lambs and weaned lambs graze together with the adults the former prove more strongly infected than those that are on isolated grasslands.

  7. Induced pluripotent stem cells from goat fibroblasts.

    PubMed

    Song, Hui; Li, Hui; Huang, Mingrui; Xu, Dan; Gu, Chenghao; Wang, Ziyu; Dong, Fulu; Wang, Feng

    2013-12-01

    Embryonic stem cells (ESCs) are a powerful model for genetic engineering, studying developmental biology, and modeling disease. To date, ESCs have been established from the mouse (Evans and Kaufman, 1981, Nature 292:154-156), non-human primates (Thomson et al., , Proc Nat Acad Sci USA 92:7844-7848), humans (Thomson et al., 1998, Science 282:1145-1147), and rats (Buehr et al., , Cell 135:1287-1298); however, the derivation of ESCs from domesticated ungulates such as goats, sheep, cattle, and pigs have not been successful. Alternatively, induced pluripotent stem cells (iPSCs) can be generated by reprogramming somatic cells with several combinations of genes encoding transcription factors (OCT3/4, SOX2, KLF4, cMYC, LIN28, and NANOG). To date, iPSCs have been isolated from various species, but only limited information is available regarding goat iPSCs (Ren et al., 2011, Cell Res 21:849-853). The objectives of this study were to generate goat iPSCs from fetal goat primary ear fibroblasts using lentiviral transduction of four human transcription factors: OCT4, SOX2, KLF4, and cMYC. The goat iPSCs were successfully generated by co-culture with mitomycin C-treated mouse embryonic fibroblasts using medium supplemented with knockout serum replacement and human basic fibroblast growth factor. The goat iPSCs colonies are flat, compact, and closely resemble human iPSCs. They have a normal karyotype; stain positive for alkaline phosphatase, OCT4, and NANOG; express endogenous pluripotency genes (OCT4, SOX2, cMYC, and NANOG); and can spontaneously differentiate into three germ layers in vitro and in vivo.

  8. Isolation and characterization of orf viruses from Korean black goats.

    PubMed

    Oem, Jae-Ku; Chung, Joon-Yee; Kim, Yong-Joo; Lee, Kyoung-Ki; Kim, Seong-Hee; Jung, Byeong-Yeal; Hyun, Bang-Hun

    2013-01-01

    Five cases of orf virus infection in Korean black goats were diagnosed in our laboratory between 2010 and 2011. One orf virus (ORF/2011) was isolated from an ovine testis cell line (OA3.Ts) for use as a vaccine candidate. Sequences of the major envelope protein and orf virus interferon resistance genes were determined and compared with published reference sequences. Phylogenetic analyses revealed that orf viruses from Korean black goats were most closely related to an isolate (ORF/09/Korea) from dairy goats in Korea. This result indicates that the orf viruses might have been introduced from dairy goats into the Korean black goat population.

  9. The Hair Follicle: A Comparative Review of Canine Hair Follicle Anatomy and Physiology.

    PubMed

    Welle, Monika M; Wiener, Dominique J

    2016-06-01

    The hair follicle (HF) has a wide range of functions including thermoregulation, physical and immunological protection against external insults, sensory perception, social interactions, and camouflage. One of the most characteristic features of HFs is that they self-renew during hair cycle (HC) throughout the entire life of an individual to continuously produce new hair. HC disturbances are common in humans and comparable to some alopecic disorders in dogs. A normal HC is maintained by follicular stem cells (SCs), which are predominately found in an area known as the bulge. Due to similar morphological characteristics of the human and canine bulge area, the particularity of compound HFs in humans and dogs as well as similarities in follicular biomarker expression, the dog might be a promising model to study human HC and SC disorders. In this review, we give an overview of normal follicular anatomy, the HC, and follicular SCs and discuss the possible pathogenetic mechanisms of noninflammatory alopecia. PMID:27000375

  10. Thermoregulatory responses of goats in hot environments

    NASA Astrophysics Data System (ADS)

    Maia, Alex Sandro Campos; da Silva, Roberto Gomes; Nascimento, Sheila Tavares; Nascimento, Carolina Cardoso Nagib; Pedroza, Heloisa Paula; Domingos, Herica Girlane Tertulino

    2015-08-01

    Notwithstanding the solar radiation is recognized as a detrimental factor to the thermal balance and responses of animals on the range in tropical conditions, studies on the amount of thermal radiation absorbed by goats therein associated with data on their production and heat exchange are still lacking. Metabolic heat production and the heat exchange of goats in the sun and in the shade were measured simultaneously, aiming to observe its thermal equilibrium. The results showed that black goats absorb twice as much as the white goats under intense solar radiation (higher than 800 W m-2). This observation leads to a higher surface temperature of black goats, but it must not be seen as a disadvantage, because they increase their sensible heat flow in the coat-air interface, especially the convection heat flow at high wind speeds. In the shade, no difference between the coat colours was observed and both presented a lower absorption of heat and a lower sensible heat flow gain. When solar radiation levels increases from 300 to 1000 W m-2, we observed an increase of the heat losses through latent flow in both respiratory and cutaneous surface. Cutaneous evaporation was responsible for almost 90 % of the latent heat losses, independently of the coat colour. Goats decrease the metabolic heat production under solar radiation levels up to 800 W m-2, and increase in levels higher than this, because there is an increase of the respiratory rate and of the respiratory flow, but the fractions of consumed oxygen and produced carbon dioxide are maintained stable. The respiratory rate of black goats was higher than the white ones, under 300 W m-2 (55 and 45 resp min-1) and 1000 W m-2 (120 and 95 resp min-1, respectively). It was concluded that shade or any protection against solar radiation levels above 800 Wm-2 is critical to guarantee goat's thermal equilibrium. Strategies concerning the grazing period in accordance with the time of the day alone are not appropriate, because the

  11. Thermoregulatory responses of goats in hot environments.

    PubMed

    Maia, Alex Sandro Campos; da Silva, Roberto Gomes; Nascimento, Sheila Tavares; Nascimento, Carolina Cardoso Nagib; Pedroza, Heloisa Paula; Domingos, Herica Girlane Tertulino

    2015-08-01

    Notwithstanding the solar radiation is recognized as a detrimental factor to the thermal balance and responses of animals on the range in tropical conditions, studies on the amount of thermal radiation absorbed by goats therein associated with data on their production and heat exchange are still lacking. Metabolic heat production and the heat exchange of goats in the sun and in the shade were measured simultaneously, aiming to observe its thermal equilibrium. The results showed that black goats absorb twice as much as the white goats under intense solar radiation (higher than 800 W m(-2)). This observation leads to a higher surface temperature of black goats, but it must not be seen as a disadvantage, because they increase their sensible heat flow in the coat-air interface, especially the convection heat flow at high wind speeds. In the shade, no difference between the coat colours was observed and both presented a lower absorption of heat and a lower sensible heat flow gain. When solar radiation levels increases from 300 to 1000 W m(-2), we observed an increase of the heat losses through latent flow in both respiratory and cutaneous surface. Cutaneous evaporation was responsible for almost 90 % of the latent heat losses, independently of the coat colour. Goats decrease the metabolic heat production under solar radiation levels up to 800 W m(-2), and increase in levels higher than this, because there is an increase of the respiratory rate and of the respiratory flow, but the fractions of consumed oxygen and produced carbon dioxide are maintained stable. The respiratory rate of black goats was higher than the white ones, under 300 W m(-2) (55 and 45 resp min(-1)) and 1000 W m(-2) (120 and 95 resp min(-1), respectively). It was concluded that shade or any protection against solar radiation levels above 800 Wm(-2) is critical to guarantee goat's thermal equilibrium. Strategies concerning the grazing period in accordance with the time of the day alone are not

  12. Disposition of fenbendazole in the goat.

    PubMed

    Short, C R; Barker, S A; Hsieh, L C; Ou, S P; Davis, L E; Koritz, G; Neff-Davis, C A; Bevill, R F; Munsiff, I J; Sharma, G C

    1987-05-01

    The disposition of fenbendazole was studied in goats after oral or IV administration. Plasma concentration vs time profiles were determined for fenbendazole and all of its metabolites. The total excretion of the drug and its metabolites in urine and feces was also measured for 6 days. A biliary cannula was inserted in 1 goat to study the excretion of fenbendazole and its metabolites into the bile. Fenbendazole was converted to its sulfoxide (oxfendazole), and the sulfone, primary amine, and p-hydroxylated metabolites. The active metabolite, oxfendazole, appeared in plasma, but only trace amounts were found in feces or urine. The major excretory metabolite was p-hydroxyfenbendazole.

  13. Interspecific chimaerism between sheep and goat.

    PubMed

    Fehilly, C B; Willadsen, S M; Tucker, E M

    In rodents, chimaeric blastocysts produced by combining embryonic cells of two different species have been used in investigations of cell lineage and interaction during development (Mus musculus-Rattus norvegicus, M. musculus-Clethrionomys glareolus, M. musculus-Mus caroli). However, interspecific chimaerism also offers new approaches to the study of reproductive incompatibilities between species and may even allow such incompatibilities to be neutralized, thus improving the chances of successful hybridization and interspecific embryo transplantation. We report here the production of sheep-goat chimaeras by embryo manipulation and the use of interspecific chimaerism to allow successful interspecific embryo transplantation in sheep and goats. PMID:6694751

  14. Outpatient Follicle Monitoring: A Plea for Standardization in Ultrasound Based Follicle Monitoring and Data Transfer

    PubMed Central

    Murtinger, Maximilian; Zech, Mathias H.; Spitzer, Dietmar; Zech, Nicolas H.

    2014-01-01

    Background The complexity of assisted reproductive technology (ART) increased during the last decades. New scientific and medical findings as well as the statutory requirements for improving the safety and the outcome of ART were the main impetus for its development. While therapy planning is done and ART is used by the IVF centers, the medical support and monitoring of patients is conducted by referring gynecologists. Reported follicle measurements by the gynecologist allow the adoption of the therapy plan. Most notably, the crucial aspect is processing and interpretation of ultrasound scan (US). The results of the received US, the transfer of data between IVF center(s) and referred physician(s) as well as the subjective interpretation often culminate in interpretation and logistical problems. This might increase the error probability with considerable detriments for the patients and ART outcome. Methods The follicle monitoring was performed using Voluson I ultrasound system combined with SonoAVC® software. Results were communicated via DICOM language to DynaMed® software, a medical program for managing an IVF center with seamless integration of all processes needed for an accurate and precise workflow. Results In this study, no loss of data was detected. All data were integrated by DynaMed® software and were recallable in a fast and easy manner. Conclusion The broad usage of Voluson I ultrasound SonoAVC® software and communication of the results via Picture Archiving and Communication System (PACS) server between the IVF center and local gynecologist would provide more assistance for the patients and consequently the ART outcomes can be improved. PMID:24918083

  15. Candidate genes for the development of hair follicles in Hu sheep.

    PubMed

    Lv, X Y; Ni, R; Sun, W; Su, R; Musa, H H; Yin, J F; Wang, Q Z; Gao, W; Chen, L

    2016-01-01

    The aim of this study was to detect candidate genes for the development of hair follicles in the Hu sheep breed. Seven genes have been detected in large, medium, and small wave follicles of Hu sheep using gene chip technology. The histological features of the follicles of newborn Hu-lambs were combined with fluorescence quantitative PCR technology to detect the correlation between the expression of the seven genes and hair follicle development. Among the genes studied, matrix metalloproteinase 2 (MMP2), bone morphogenetic protein-7 (BMP7), and sideroflexin 1 (SFXN1) showed a significantly different pattern of expression in large, medium, and small wave follicles (P < 0.05). The expression of MMP2 had a significant positive correlation with secondary follicles in large waves (P < 0.05), while the expression of BMP7 had a significant correlation with primary follicle diameter in small wave follicles, and a highly significant positive correlation with the number of secondary follicles in the small waves (P < 0.01). The expression of SFXN1 was significantly and positively correlated with the diameters of small wave primary follicles; it also showed a highly significant positive correlation with secondary follicle diameters. Although other genes are associated with hair follicles, their expression in large, medium, and small wave follicles was not significant. We propose that BMP7, MMP2, and SFXN1 genes could be important candidate genes for use in breeding Hu lambs with early coat development. PMID:27525902

  16. Targeting to the hair follicles: current status and potential.

    PubMed

    Wosicka, Hanna; Cal, Krzysztof

    2010-02-01

    The pilosebaceous unit is a complex structure that undergoes a specific growth cycle and comprises a few important drug targeting sites. For example, drugs can be targeted to the bulge region with stem cells or to the sebaceous glands. Interest in pilosebaceous units is directed towards their utilization as reservoirs for localized therapy and also as a transport pathway for systemic drug delivery. Improved investigative methods, such as differential stripping, are being developed in order to determine follicular penetration. This article reviews relevant aspects of effective follicle-targeting formulations and delivery systems as well as the activity status of hair follicles, and variations in follicle size and distribution throughout various body regions. Each of these factors strongly affects follicular permeation. We provide examples of improved penetration of particle-based formulations and of a size-dependent manner of follicular penetration. Contradictions are also discussed, indicating the need for detailed future investigations. PMID:20060268

  17. Targeting to the hair follicles: current status and potential.

    PubMed

    Wosicka, Hanna; Cal, Krzysztof

    2010-02-01

    The pilosebaceous unit is a complex structure that undergoes a specific growth cycle and comprises a few important drug targeting sites. For example, drugs can be targeted to the bulge region with stem cells or to the sebaceous glands. Interest in pilosebaceous units is directed towards their utilization as reservoirs for localized therapy and also as a transport pathway for systemic drug delivery. Improved investigative methods, such as differential stripping, are being developed in order to determine follicular penetration. This article reviews relevant aspects of effective follicle-targeting formulations and delivery systems as well as the activity status of hair follicles, and variations in follicle size and distribution throughout various body regions. Each of these factors strongly affects follicular permeation. We provide examples of improved penetration of particle-based formulations and of a size-dependent manner of follicular penetration. Contradictions are also discussed, indicating the need for detailed future investigations.

  18. Communal goat production in Southern Africa: a review.

    PubMed

    Rumosa Gwaze, F; Chimonyo, M; Dzama, K

    2009-10-01

    Despite the fact that about 64% of goats in sub-Saharan Africa (SSA) are located in rural arid (38%) and semi-arid (26%) agro-ecological zones and that more than 90% of goats in these zones are indigenous, information on indigenous breeds is inadequate. This paper reviews the social and economic importance of goats to the communal farmer and assesses the potential of using goats in rural development in Southern Africa. Farmers in Southern Africa largely use the village goat management system. There are various goat breeds in Southern Africa, of which the Mashona, Matabele, Tswana, Nguni and the Landim are the dominant ones. It is, however, not clear if these breeds are distinct. Major constraints to goat production include high disease and parasite prevalence, low levels of management, limited forage availability and poor marketing management. Potential research areas that are required to ensure that goats are vehicles for rural development include evaluation of constraints to goat production, assessing the contribution of goats to household economies and food securities throughout the year, genetic and phenotypic characterisation of the indigenous breeds to identify appropriate strains and sustainable methods of goat improvement through either selection or crossbreeding.

  19. Reproductive outcomes of Alpine goats primed with progesterone and treated with human chorionic gonadotropin during the anestrus-to-estrus transition season.

    PubMed

    Alvarado-Espino, A S; Meza-Herrera, C A; Carrillo, E; González-Álvarez, V H; Guillen-Muñoz, J M; Ángel-García, O; Mellado, M; Véliz-Deras, F G

    2016-04-01

    This study aimed to determine the possible effects of a single injection of human chorionic gonadotropin (hCG) as a means for estrus induction in acyclic French-Alpine goats during the reproductive transition period at 25°N, 103°W. The potential effects of hCG upon ovarian function and reproductive performance of goats were also assessed. Multiparous acyclic French-Alpine goats (n = 39; 37.4 ± 8 .5 kg) were primed with 20mg progesterone (P4) 1 day prior to hCG administration. Thereafter, does were treated either with saline (hCG-0; n = 10), 50 (hCG-50; n = 9), 100 (hCG-100; n = 10), or 300 IU of hCG (hCG-300; n = 10). Ovarian structures and pregnancy were monitored by transrectal ultrasonography. In addition, after hCG application, goats were monitored twice daily (0800 and 1800 h) to detect estrus signs, with the use of aproned, sexually active bucks treated with testosterone. Goats were bred 12h after the onset of estrus. Two days after hCG administration, the number of large follicles was higher (P < 0.05) in the hCG-50 and hCG-300 groups (1.7 ± 0.1 and 1.8 ± 0.2, respectively) compared with the hCG-100 and hCG-0 groups (1.4 ± 0.2 and 1.1 ± 0.1, respectively). Although none of the hCG-0-goats depicted estrus, the estrus response from the hCG-50, hCG-100, and hCG-300 groups over the 7-d breeding period was 67%, 100%, and 90%, respectively (P > 0.05), being always accompanied by ovulation. Pregnancy rate (67, 100, and 70%), kidding rate (55%, 80%, and 70%), and litter size (1.6 ± 0.5, 1.5 ± 0.5, and 1.5 ± 0.5) for hCG-50, hCG-100, and hCG-300, respectively, did not differ among the hCG-treated does. Therefore, the combined use of P4-priming plus a 100-IU hCG injection is an effective protocol for inducing estrus in non-cycling Alpine goats during the anestrus-to-estrus transition period, which is of key importance for both goat producers and industrializers. PMID:26944772

  20. Genetic diversity in Swiss goat breeds based on microsatellite analysis.

    PubMed

    Saitbekova, N; Gaillard, C; Obexer-Ruff, G; Dolf, G

    1999-02-01

    Genetic diversity in eight Swiss goat breeds was estimated using PCR amplification of 20 bovine microsatellites on 20-40 unrelated animals per breed. In addition, the Creole breed from the Caribbean and samples of Ibex and Bezoar goat were included. A total of 352 animals were tested. The bovine microsatellites chosen amplified well in goat. The average heterozygosity within population was higher in domestic goat (0.51-0.58) than in Ibex (0.17) and Bezoar goat (0.19). Twenty-seven per cent of the genetic diversity in the total population could be attributed to differences between the populations. However, with the exclusion of Ibex from the total population, this proportion dropped to 17%. Principal component analysis showed that all Swiss goat breeds are closely related, whereas the Creole breed, Ibex and Bezoar goat are clearly distinct from all eight Swiss breeds.

  1. Two USA Ehrlichia spp. cause febrile illness in goats.

    PubMed

    Loftis, Amanda D; Levin, Michael L; Spurlock, J Paul

    2008-08-25

    Ehrlichia spp. are not currently recognized as a cause of illness in goats in the USA, but three Ehrlichia are enzootic in lone star ticks (Amblyomma americanum) in the eastern USA, and related bacteria in other countries cause illness in goats. We exposed naïve goats to Ehrlichia-infected Amblyomma and demonstrated that infection and clinical illness can be caused by two USA species, E. ewingii and the recently discovered Panola Mountain Ehrlichia sp. Clinical features in all five goats are described; ehrlichioses were associated with pyrexia, serous nasal discharge, inappetance, lethargy, decreased alkaline phosphatase, and, in most cases, neutropenia. Goats remained chronically infected for several months following exposure to ehrlichiae and transmitted the pathogens to uninfected ticks. In the eastern USA, undifferentiated febrile illness in goats might be caused by previously unrecognized ehrlichial infections, and pastures housing-infected goats could become infested with a large number of infected ticks. PMID:18328644

  2. Two USA Ehrlichia spp. cause febrile illness in goats.

    PubMed

    Loftis, Amanda D; Levin, Michael L; Spurlock, J Paul

    2008-08-25

    Ehrlichia spp. are not currently recognized as a cause of illness in goats in the USA, but three Ehrlichia are enzootic in lone star ticks (Amblyomma americanum) in the eastern USA, and related bacteria in other countries cause illness in goats. We exposed naïve goats to Ehrlichia-infected Amblyomma and demonstrated that infection and clinical illness can be caused by two USA species, E. ewingii and the recently discovered Panola Mountain Ehrlichia sp. Clinical features in all five goats are described; ehrlichioses were associated with pyrexia, serous nasal discharge, inappetance, lethargy, decreased alkaline phosphatase, and, in most cases, neutropenia. Goats remained chronically infected for several months following exposure to ehrlichiae and transmitted the pathogens to uninfected ticks. In the eastern USA, undifferentiated febrile illness in goats might be caused by previously unrecognized ehrlichial infections, and pastures housing-infected goats could become infested with a large number of infected ticks.

  3. Morphometry of human terminal and vellus hair follicles.

    PubMed

    Vogt, Annika; Hadam, Sabrina; Heiderhoff, Marc; Audring, Heike; Lademann, Juergen; Sterry, Wolfram; Blume-Peytavi, Ulrike

    2007-11-01

    Previous studies suggest that drug delivery systems based on particles can be used to deposit active compounds in hair follicles and to target hair follicle-associated cell populations. The development of application protocols is complicated by the fact that there is no information available on the size and the position of key target structures in the different hair follicle types and their intra- and interindividual variation. Therefore, we performed morphometric measurements on histological sections of human terminal (THF) and vellus hair follicles (VHF) from the scalp and the retroauricular region. With 3864 +/- 605 microm and 580 +/- 84 microm in THF compared to 646 +/- 140 microm and 225 +/- 34 microm in VHF, the total length and the length of the infundibulum differed significantly as determined by paired t-test (P < 0.0001). The same level of significance was observed for the position and the length of the bulge region. The thickness of the epithelial lining was lowest in VHF (45 +/- 14 microm at 100 microm from skin surface) compared to 65 +/- 20 microm at 150 microm in THF, while the thickness of the interfollicular epidermis ranged between 64 +/- 12 microm and 99 +/- 18 microm in VHF-bearing skin and 72 +/- 16 microm and 136 +/- 37 microm in THF-bearing skin. In addition, the diameter of the hair follicle opening was determined at 50 microm intervals from the skin surface. Our data suggest that hair follicle types in defined body regions represent rather homogenous groups and that particle-based drug delivery may be a feasible approach, also in larger numbers of individuals. We provide precise information on the size and the position of key target structures in VHF and THF.

  4. The cardiac biomarkers troponin I and CK-MB in nonpregnant and pregnant goats, goats with normal birth, goats with prolonged birth, and goats with pregnancy toxemia.

    PubMed

    Tharwat, M; Al-Sobayil, F; Al-Sobayil, K

    2012-10-15

    This study was designed to establish the reference range for the cardiac biomarkers cardiac troponin I (cTnI) and creatine kinase myocardial band (CK-MB) in nonpregnant and pregnant goats, goats with normal birth, goats with prolonged birth associated with dystocia, and goats with pregnancy toxemia. Fifty-seven does, categorized into three groups (G1 to G3), were used. These groups were comprised of 20 healthy does (G1), 19 does with prolonged birth (G2), and 18 does with pregnancy toxemia (G3). Six blood samples (T0 to T5) were collected from G1. The first blood sample (T0) was taken before insemination, the second (T1) at the first trimester, the third (T2) at the second trimester, the fourth (T3) at the last trimester, the fifth (T4) within 12 h of parturition, and the sixth blood sample (T5) was taken 10 days after parturition. A sample of blood was obtained from G2 and G3 upon admission to the hospital. At T0 to T3, no cTnI was detected in any of the 20 does in G1. At parturition (T4), seven of the 20 goats (35%) exhibited slightly elevated cTnI concentrations (range, 0.01 to 0.04 ng/mL). Ten days after parturition (T5), cTnI was not detected in any of the 20 goats. In 10 of the 19 goats (53%) with prolonged birth (G2), the cTnI was significantly elevated to a mean value of 0.094 ± 0.155 ng/mL, with a maximum value of 0.61 ng/mL. In 16 of the 18 goats (89%) with pregnancy toxemia (G3), the cTnI was significantly elevated to a value of 0.852 ± 1.472 ng/mL, with a maximum value of 5.219 ng/mL. Comparing the values of CK-MB in G1 (T0 to T5), G2 and G3 revealed nonsignificant differences. Only a slight elevation in the CK-MB levels in goats with prolonged birth (G2) was noted. We concluded that in healthy does, the cardiac biomarker cTnI is not elevated during normal pregnancy. The serum cTnI concentration may be elevated in a number of goats at normal vaginal or cesarean delivery. Finally, cTnI is significantly elevated in does with pregnancy toxemia and could

  5. Genetic Variation of Goat Interferon Regulatory Factor 3 Gene and Its Implication in Goat Evolution

    PubMed Central

    Shu, Liping; Zhang, Yesheng; Wang, Yangzi; Sanni, Timothy M.; Imumorin, Ikhide G.; Peters, Sunday O.; Zhang, Jiajin; Dong, Yang; Wang, Wen

    2016-01-01

    The immune systems are fundamentally vital for evolution and survival of species; as such, selection patterns in innate immune loci are of special interest in molecular evolutionary research. The interferon regulatory factor (IRF) gene family control many different aspects of the innate and adaptive immune responses in vertebrates. Among these, IRF3 is known to take active part in very many biological processes. We assembled and evaluated 1356 base pairs of the IRF3 gene coding region in domesticated goats from Africa (Nigeria, Ethiopia and South Africa) and Asia (Iran and China) and the wild goat (Capra aegagrus). Five segregating sites with θ value of 0.0009 for this gene demonstrated a low diversity across the goats’ populations. Fu and Li tests were significantly positive but Tajima’s D test was significantly negative, suggesting its deviation from neutrality. Neighbor joining tree of IRF3 gene in domesticated goats, wild goat and sheep showed that all domesticated goats have a closer relationship than with the wild goat and sheep. Maximum likelihood tree of the gene showed that different domesticated goats share a common ancestor and suggest single origin. Four unique haplotypes were observed across all the sequences, of which, one was particularly common to African goats (MOCH-K14-0425, Poitou and WAD). In assessing the evolution mode of the gene, we found that the codon model dN/dS ratio for all goats was greater than one. Phylogenetic Analysis by Maximum Likelihood (PAML) gave a ω0 (dN/dS) value of 0.067 with LnL value of -6900.3 for the first Model (M1) while ω2 = 1.667 in model M2 with LnL value of -6900.3 with positive selection inferred in 3 codon sites. Mechanistic empirical combination (MEC) model for evaluating adaptive selection pressure on particular codons also confirmed adaptive selection pressure in three codons (207, 358 and 408) in IRF3 gene. Positive diversifying selection inferred with recent evolutionary changes in domesticated goat

  6. Secretion of platelet-activating factor by periovulatory ovine follicles

    SciTech Connect

    Alexander, B.M.; Van Kirk, E.A.; Murdoch, W.J. )

    1990-01-01

    Secretion of platelet-activating factor (PAF) in vitro by ovine follicles and ovarian interstitium obtained at various times before, during and after the endogenous preovulatory surge of luteinizing hormone (LH) and ovulation was quantified by radioimmunoassay. Release of PAF by the preovulatory follicle increased within 2 h after initiation of the surge of LH. Capacity for secretion of PAF was greatest at the time of ovulation, then declined thereafter. Production of PAF by ovarian interstitium throughout the periovulatory period was relatively low and did not change with time. It appears that PAF could act as an intrafollicular mediator in the mechanisms of ovulation and(or) luteinization.

  7. Genetic relatedness between Ardi, Black Bedouin and Damascus goat breeds.

    PubMed

    Al-Atiyat, R M; Aljumaah, R S

    2014-06-18

    The present study aimed to analyze genetic relatedness and differentiation of common native goat populations in some countries of the Middle East. The populations were Ardi, Black Bedouin, and Damascus goats in the Kingdom of Saudi Arabia, Jordan, and Syria, respectively. Domesticated goats of the Middle East are mostly related to common ancestors, but there is limited molecular genetic evidence. Four microsatellite DNA markers were genotyped in 89 individuals of the three populations using an automated genetic analyzer. Ardi, Black Bedouin, and Damascus goats exhibited high average allele number and expected heterozygosity of 8.25, 9, and 7.25, and 0.750, 0.804, and 0.779, respectively. F-statistics for population differentiation showed 6.0% of total genetic variation, whereas 94.0% as differentiation between individuals within all populations. The least varied within populations was Ardi goats, then Damascus goats and finally Black Bedouin goats. Furthermore, the Damascus goat population was more differentiated from Black Bedouin goats than from Ardi goats. On the other hand, there was strong evidence of admixture between the majority of Ardi and Black Bedouin goat individuals but little with those of Damascus goats. Genetic distance between Ardi and Black Bedouin goats was the shortest, whereas it was the longest between Ardi and Damascus goats. The phylogenetic tree clearly revealed the expected degree of differentiation in the three populations. From a genetic conservation point of view, it is recommended to maintain the biodiversity of these distinct populations in case genetic migration of genetic resources and genetic conservation are absent.

  8. In vitro development of goat-sheep and goat-goat zona-free cloned embryos in different culture media.

    PubMed

    Khan, F A; Bhat, M H; Yaqoob, S H; Waheed, S M; Naykoo, N A; Athar, H; Khan, H M; Fazili, M R; Ganai, N A; Singla, S K; Shah, R A

    2014-02-01

    The gradual decline in the genetic diversity of farm animals has threatened their survival and risk of their extinction has increased many fold in the recent past. Endangered species could be rescued using interspecies embryo production. The objective of this study was to investigate the effect of three different culture media on the development of Handmade cloned intraspecies (goat-goat) and interspecies (goat-sheep) embryo reconstructs. Research vitro cleave media (RVCL) yielded higher cleavage and morula-blastocyst development in intraspecies and interspecies nuclear transfer groups compared with G1.G2 and modified synthetic oviductal fluid (mSOFaaci). Cleavage frequency of intraspecies cloned embryos in RVCL, mSOFaaci, and G1.G2 did not differ significantly (87.12%, 82.45%, and 92.52%, respectively). However, the morula/blastocyst frequency in RVCL was greater in mSOFaaci and G1.G2 (51.18% vs. 38.28% vs. 36.50%, respectively). Cleavage and morula/blastocyst frequency in interspecies cloned embryos was greater in RVCL than in mSOFaaci and G1.G2 (76.14% and 42.3% vs. 65.9% and 38.3% vs. 58.56% and 33.1%, respectively). Goat oocytes were parthenogenetically activated and cultured in RVCL, mSOFaaci, and G1.G2 and kept as control. Cleavage and morula/blastocyst frequency in this group was greater in RVCL than in mSOFaaci and G1.G2 (89.66% and 65.26% vs. 85.44% and 48.05% vs. 86.58% and 42.06%, respectively). Conclusively, the results suggest that not only can the interspecies embryos of goat be produced using sheep oocytes as donor cytoplast but also the percentages can be improved by using RVCL media for culturing of the embryos. PMID:24210966

  9. In vitro development of goat-sheep and goat-goat zona-free cloned embryos in different culture media.

    PubMed

    Khan, F A; Bhat, M H; Yaqoob, S H; Waheed, S M; Naykoo, N A; Athar, H; Khan, H M; Fazili, M R; Ganai, N A; Singla, S K; Shah, R A

    2014-02-01

    The gradual decline in the genetic diversity of farm animals has threatened their survival and risk of their extinction has increased many fold in the recent past. Endangered species could be rescued using interspecies embryo production. The objective of this study was to investigate the effect of three different culture media on the development of Handmade cloned intraspecies (goat-goat) and interspecies (goat-sheep) embryo reconstructs. Research vitro cleave media (RVCL) yielded higher cleavage and morula-blastocyst development in intraspecies and interspecies nuclear transfer groups compared with G1.G2 and modified synthetic oviductal fluid (mSOFaaci). Cleavage frequency of intraspecies cloned embryos in RVCL, mSOFaaci, and G1.G2 did not differ significantly (87.12%, 82.45%, and 92.52%, respectively). However, the morula/blastocyst frequency in RVCL was greater in mSOFaaci and G1.G2 (51.18% vs. 38.28% vs. 36.50%, respectively). Cleavage and morula/blastocyst frequency in interspecies cloned embryos was greater in RVCL than in mSOFaaci and G1.G2 (76.14% and 42.3% vs. 65.9% and 38.3% vs. 58.56% and 33.1%, respectively). Goat oocytes were parthenogenetically activated and cultured in RVCL, mSOFaaci, and G1.G2 and kept as control. Cleavage and morula/blastocyst frequency in this group was greater in RVCL than in mSOFaaci and G1.G2 (89.66% and 65.26% vs. 85.44% and 48.05% vs. 86.58% and 42.06%, respectively). Conclusively, the results suggest that not only can the interspecies embryos of goat be produced using sheep oocytes as donor cytoplast but also the percentages can be improved by using RVCL media for culturing of the embryos.

  10. 9 CFR 91.6 - Goats.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... exported if it is a scrapie-positive animal or an exposed animal, as defined in 9 CFR parts 54 and 79, or if it has ever been in an infected flock, source flock, or trace flock, as defined in 9 CFR parts 54...) Tuberculosis. All goats over 1 month of age shall be negative to a caudal intradermal tuberculin test using...

  11. 9 CFR 91.6 - Goats.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... exported if it is a scrapie-positive animal or an exposed animal, as defined in 9 CFR parts 54 and 79, or if it has ever been in an infected flock, source flock, or trace flock, as defined in 9 CFR parts 54...) Tuberculosis. All goats over 1 month of age shall be negative to a caudal intradermal tuberculin test using...

  12. 9 CFR 91.6 - Goats.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... exported if it is a scrapie-positive animal or an exposed animal, as defined in 9 CFR parts 54 and 79, or if it has ever been in an infected flock, source flock, or trace flock, as defined in 9 CFR parts 54...) Tuberculosis. All goats over 1 month of age shall be negative to a caudal intradermal tuberculin test using...

  13. Ectoparasites of goats in the UK.

    PubMed

    Cornall, Katherine; Wall, Richard

    2015-01-15

    The goat industry in the UK has expanded rapidly in recent years, but at present there is only a poor understanding of the prevalence of parasitic diseases in this farming system. Here, a questionnaire survey of 110 goat owners was used to address this issue. Problems with louse infestation in the previous 12 months were reported by 23% of owners and 19% reported mange. Chorioptic mange was the most common form, with 14 of 21 cases. Sarcoptic mage accounted for only 3 cases and demodex and psoroptic mange each made up 2 cases. Only 53 farmers (48%) said that they took preventative measures to protect their animals against ectoparasite infestation; 20 of these relied on macrocyclic lactones (MLs), the most common product specified was ivermectin. Therapeutic treatment was used by all respondents who said that they had experienced ectoparasites, and again ivermectin was the most common treatment. The use of fipronil was specified by 3 respondents, including one commercial meat producer. Four farmers said that they used antibiotics as an ectoparasiticide. This pattern of treatment for ectoparasites, with reliance on MLs, has implications for the inadvertent selection of resistance in endoparasites. The results suggest that ectoparasites are a major problem for many goat owners, both commercial and non-commercial, but that there is a need for better information for the goat producing community about the optimum approaches to parasite prevention and treatment. PMID:25465737

  14. Fatal melioidosis in goats in Bangkok, Thailand.

    PubMed

    Tonpitak, Walaiporn; Sornklien, Chulabha; Chawanit, Mongkol; Pavasutthipaisit, Suvarin; Wuthiekanun, Vanaporn; Hantrakun, Viriya; Amornchai, Premjit; Thaipadungpanit, Janjira; Day, Nicholas P J; Yingst, Samuel; Peacock, Sharon J; Limmathurotsakul, Direk

    2014-08-01

    Bangkok, Thailand, is a city considered to be at low risk for melioidosis. We describe 10 goats that died of melioidosis in Bangkok. Half of them were born and reared in the city. Multilocus sequence typing ruled out an outbreak. This finding challenges the assumption that melioidosis is rarely acquired in central Thailand.

  15. Viable transgenic goats derived from skin cells.

    PubMed

    Behboodi, Esmail; Memili, Erdogan; Melican, David T; Destrempes, Margaret M; Overton, Susan A; Williams, Jennifer L; Flanagan, Peter A; Butler, Robin E; Liem, Hetty; Chen, Li How; Meade, Harry M; Gavin, William G; Echelard, Yann

    2004-06-01

    The current study was undertaken to evaluate the possibility of expanding transgenic goat herds by means of somatic cell nuclear transfer (NT) using transgenic goat cells as nucleus donors. Skin cells from adult, transgenic goats were first synchronized at quiescent stage (G0) by serum starvation and then induced to exit G0 and proceed into G1. Oocytes collected from superovulated donors were enucleated, karyoplast-cytoplast couplets were constructed, and then fused and activated simultaneously by a single electrical pulse. Fused couplets were either co-cultured with oviductal cells in TCM-199 medium (in vitro culture) or transferred to intermediate recipient goat oviducts (in vivo culture) until final transfer. The resulting morulae and blastocysts were transferred to the final recipients. Pregnancies were confirmed by ultrasonography 25-30 days after embryo transfer. In vitro cultured NT embryos developed to morulae and blastocyst stages but did not produce any pregnancies while 30% (6/20) of the in vivo derived morulae and blastocysts produced pregnancies. Two of these pregnancies were resorbed early in gestation. Of the four recipients that maintained pregnancies to term, two delivered dead fetuses 2-3 days after their due dates, and two recipients gave birth to healthy kids at term. Fluorescence in situ hybridization (FISH) analysis confirmed that both kids were transgenic and had integration sites consistent with those observed in the adult cell line.

  16. Infiltrative Cutaneous Hemangiolipoma in a Goat

    PubMed Central

    Collier, Jessica R.; Byers, Stacey R.; Schaffer, Paula A.; Worley, Deanna R.; Ehrhart, E. J.; Duncan, Colleen G.; Grossman, Alicia N.; Holt, Timothy; Callan, Robert J.

    2013-01-01

    An approximately 4-year-old castrated male, Saanen cross goat presented to the Colorado State University Veterinary Teaching Hospital for evaluation and removal of a 22 cm × 22 cm, dark red, thickened, and crusted cutaneous lesion along the left ventrolateral thorax. An initial incisional biopsy performed approximately 8 weeks earlier was suspicious for cutaneous hemangiosarcoma. Surgical excision was deemed to be the most appropriate treatment option for this goat. A complete physical exam, complete blood count, and chemistry profile were performed and results were within normal limits. Thoracic radiographs and abdominal ultrasound were performed to rule out metastatic disease and comorbid conditions; no metastatic lesions or other abnormalities were observed. En bloc surgical excision of the affected skin was performed and the entire tissue was submitted for histopathology. A final diagnosis of cutaneous hemangiolipoma was reached upon extensive sectioning and histologic examination of the larger tissue specimen. The goat recovered well from surgery and has had no further complications up to 9 months postoperatively. To our knowledge, this is the first case report of a hemangiolipoma in a goat and surgical excision for such lesions appears to be a viable treatment method. PMID:23956926

  17. Culture and Co-Culture of Mouse Ovaries and Ovarian Follicles

    PubMed Central

    Morgan, Stephanie; Campbell, Lisa; Allison, Vivian; Murray, Alison; Spears, Norah

    2015-01-01

    The mammalian ovary is composed of ovarian follicles, each follicle consisting of a single oocyte surrounded by somatic granulosa cells, enclosed together within a basement membrane. A finite pool of follicles is laid down during embryonic development, when oocytes in meiotic arrest form a close association with flattened granulosa cells, forming primordial follicles. By or shortly after birth, mammalian ovaries contain their lifetime’s supply of primordial follicles, from which point onwards there is a steady release of follicles into the growing follicular pool. The ovary is particularly amenable to development in vitro, with follicles growing in a highly physiological manner in culture. This work describes the culture of whole neonatal ovaries containing primordial follicles, and the culture of individual ovarian follicles, a method which can support the development of follicles from an immature through to the preovulatory stage, after which their oocytes are able to undergo fertilization in vitro. The work outlined here uses culture systems to determine how the ovary is affected by exposure to external compounds. We also describe a co-culture system, which allows investigation of the interactions that occur between growing follicles and the non-growing pool of primordial follicles. PMID:25867892

  18. Cloning and functional analysis of goat SWEET1.

    PubMed

    Zhu, L Q; Bao, Z K; Hu, W W; Lin, J; Yang, Q; Yu, Q H

    2015-12-16

    SWEETs are a recently discovered class of sugar transporters that mediate glucose uptake in the intestine and mammary glands. Our objectives were to clone goat SWEET1 and conduct a functional analysis of its effect on glucose efflux in goat mammary gland epithelial cells. We cloned and sequenced the goat SWEET1 gene from goat mammary glands, then conducted an analysis of the structure of goat SWEET1, including a prediction of the transmembrane helices and potential N-glycosylation sites. To investigate the biological function of goat SWEET1, we also generated goat SWEET1-transfected goat mammary gland epithelial cells using the eukaryotic expression vector pcDNA3.1-gSWEET1. Goat SWEET1 overexpression can reduce glucose absorption in mammary gland epithelial cells with increasing expression of GLUT1, GLUT4, and GLUT12, which may be attributed to glucose efflux arising from the leading role played by goat SWEET1. This study will improve our understanding of the glucose balance in mammary glands and the level of glucose in milk.

  19. Use of microsatellite markers to assign goats to their breeds.

    PubMed

    Aljumaah, R S; Alobre, M M; Al-Atiyat, R M

    2015-08-07

    We investigated the potential of 17 microsatellite markers for assigning Saudi goat individuals to their breeds. Three local breeds, Bishi, Jabali, and Tohami were genotyped using these markers, and Somali goats were used as a reference breed. The majority of alleles were shared between the breeds, except for some that were specific to each breed. The Garza-Williamson index was lowest in the Bishi breed, indicating that a recent bottleneck event occurred. The overall results assigned the goat individuals (based on their genotypes) to the same breeds from which they were sampled, except in a few cases. The individuals' genotypes were sufficient to provide a clear distinction between the Somali goat breed and the others. In three factorial dimensions, the results of a correspondence analysis indicated that the total variation for the first and second factors was 48.85 and 31.43%, respectively. Consequently, Jabali, Bishi, and Tohami goats were in separate groups. The Jabali goat was closely related to the Bishi goat. Somali goats were distinguished from each other and from individuals of the other three goat breeds. The markers were successful in assigning individual goats to their breeds, based on the likelihood of a given individual's genotype.

  20. Potassium channel conductance as a control mechanism in hair follicles.

    PubMed

    Buhl, A E; Conrad, S J; Waldon, D J; Brunden, M N

    1993-07-01

    The opening of intracellular potassium channels is a common mechanism of action for a set of anti-hypertensive drugs that includes the hair-growth-inducing agent minoxidil. Recent work suggests potassium channel openers (PCOs) also influence hair growth. Correlative studies demonstrate that a series of PCOs including minoxidil, pinacidil, P-1075, an active pinacidil analog, RP-49,356, cromakalim, and nicorandil maintain hair growth in cultured vibrissa follicles. Studies using balding stumptail macaques verify that minoxidil, P-1075, and cromakalim but not RP-49,356 stimulate hair growth. The definition of potassium channels and documentation of drug effects on these channels is classically done using electrophysiologic techniques. Such studies require the identification and isolation of target cells. Both these are among the unsolved problems in the area of hair biology. Estimating K+ flux using 86Rb+ as a K+ tracer is an accepted method of assessing potassium channel conductance in other organ systems. Both pinacidil and RP-49,356 induce measurable Rb+ flux in isolated vibrissa follicles and a hair epithelial cell line whereas neither minoxidil nor minoxidil sulfate had measurable effects. Potassium channels have been studied successfully in other organ systems using specific pharmacologic blockers for the various channel subtypes. Blockers including glyburide, tetraethylammonium, and procaine failed to inhibit minoxidil stimulation of cultured follicles. The current explosion of knowledge on potassium channel biology, cloning of channels, and continued progress in hair biology promise to clarify the role of K+ ions in the control of hair follicles.

  1. Intravital imaging of hair follicle regeneration in the mouse

    PubMed Central

    Pineda, Cristiana M; Park, Sangbum; Mesa, Kailin R; Wolfel, Markus; Gonzalez, David G; Haberman, Ann M; Rompolas, Panteleimon; Greco, Valentina

    2015-01-01

    Hair follicles are mammalian skin organs that periodically and stereotypically regenerate from a small pool of stem cells. Hence, hair follicles are a widely studied model for stem cell biology and regeneration. This protocol describes the use of two-photon laser-scanning microscopy (TPLSM) to study hair regeneration within a living, uninjured mouse. TPLSM provides advantages over conventional approaches, including enabling time-resolved imaging of single hair follicle stem cells. Thus, it is possible to capture behaviors including apoptosis, proliferation and migration, and to revisit the same cells for in vivo lineage tracing. In addition, a wide range of fluorescent reporter mouse lines facilitates TPLSM in the skin. This protocol also describes TPLSM laser ablation, which can spatiotemporally manipulate specific cellular populations of the hair follicle or microenvironment to test their regenerative contributions. The preparation time is variable depending on the goals of the experiment, but it generally takes 30–60 min. Imaging time is dependent on the goals of the experiment. Together, these components of TPLSM can be used to develop a comprehensive understanding of hair regeneration during homeostasis and injury. PMID:26110716

  2. Heavy metal levels in goats from Notasulga, Alabama

    SciTech Connect

    Khan, A.T.; Diffay, B.C.; Forester, D.M.; Thompson, S.J.; Mielke, H.W.

    1994-12-31

    Goat meat farming is increasing in popularity in southeastern region of United States. In order to monitor environmental contamination of heavy metals in goat meat, samples of liver, kidney, and muscle were collected from 20 goats on a goat farm in Notasulga, Alabama. These samples were analyzed by Inductively Coupled Plasma Atomic Emission Spectroscopy. The copper concentration was significantly higher in livers than the concentration in kidneys and muscles. Lead, cadmium, and zinc levels did not show any significant differences between liver, kidney, and muscle samples. The concentrations of lead and copper in livers and cadmium in kidneys were significantly different in males when compared to females. However, in muscle, the concentrations of lead, cadmium, copper, and zinc showed no significant difference between male and female or between young and old goats. Further, the concentrations of lead in livers and cadmium in kidneys showed a significant difference between young and old goats.

  3. Paternal phylogeography and genetic diversity of East Asian goats.

    PubMed

    Waki, A; Sasazaki, S; Kobayashi, E; Mannen, H

    2015-06-01

    This study was a first analysis of paternal genetic diversity for extensive Asian domestic goats using SRY gene sequences. Sequencing comparison of the SRY 3'-untranslated region among 210 Asian goats revealed four haplotypes (Y1A, Y1B, Y2A and Y2B) derived from four variable sites including a novel substitution detected in this study. In Asian goats, the predominant haplotype was Y1A (62%) and second most common was Y2B (30%). Interestingly, the Y2B was a unique East Asian Y chromosomal variant, which differentiates eastern and western Eurasian goats. The SRY geographic distribution in Myanmar and Cambodia indicated predominant the haplotype Y1A in plains areas and a high frequency of Y2B in mountain areas. The results suggest recent genetic infiltration of modern breeds into South-East Asian goats and an ancestral SRY Y2B haplotype in Asian native goats. PMID:25917305

  4. Treatment of pieris ingestion in goats with intravenous lipid emulsion.

    PubMed

    Bischoff, Karyn; Smith, Mary C; Stump, Samuel

    2014-12-01

    Seven goats and one ram presented with clinical signs including regurgitation, obtundation, anorexia, apparent pain, and bloat. The animals had escaped from their barn, and it was discovered that they had ingested leaves of Pieris japonica, Japanese pieris, a grayanotoxin-containing plant. Animals were treated with antibiotics, calcium borogluconate, B vitamins, and activated charcoal within the first 24-h postexposure, which was followed by the recovery of the ram and two goats and the death of two goats. Approximately 36 h after Japanese pieris ingestion, one of the three remaining anorectic goats was dosed with intravenous lipid emulsion (ILE). This goat recovered within a few hours. The remaining two goats were given ILE the next day and appeared to recover, but one died a week later of aspiration pneumonia.

  5. Video tracking analysis of behavioral patterns during estrus in goats

    PubMed Central

    ENDO, Natsumi; RAHAYU, Larasati Puji; ARAKAWA, Toshiya; TANAKA, Tomomi

    2015-01-01

    Here, we report a new method for measuring behavioral patterns during estrus in goats based on video tracking analysis. Data were collected from cycling goats, which were in estrus (n = 8) or not in estrus (n = 8). An observation pen (2.5 m × 2.5 m) was set up in the corner of the female paddock with one side adjacent to a male paddock. The positions and movements of goats were tracked every 0.5 sec for 10 min by using a video tracking software, and the trajectory data were used for the analysis. There were no significant differences in the durations of standing and walking or the total length of movement. However, the number of approaches to a male and the duration of staying near the male were higher in goats in estrus than in goats not in estrus. The proposed evaluation method may be suitable for detailed monitoring of behavioral changes during estrus in goats. PMID:26560676

  6. Paternal phylogeography and genetic diversity of East Asian goats.

    PubMed

    Waki, A; Sasazaki, S; Kobayashi, E; Mannen, H

    2015-06-01

    This study was a first analysis of paternal genetic diversity for extensive Asian domestic goats using SRY gene sequences. Sequencing comparison of the SRY 3'-untranslated region among 210 Asian goats revealed four haplotypes (Y1A, Y1B, Y2A and Y2B) derived from four variable sites including a novel substitution detected in this study. In Asian goats, the predominant haplotype was Y1A (62%) and second most common was Y2B (30%). Interestingly, the Y2B was a unique East Asian Y chromosomal variant, which differentiates eastern and western Eurasian goats. The SRY geographic distribution in Myanmar and Cambodia indicated predominant the haplotype Y1A in plains areas and a high frequency of Y2B in mountain areas. The results suggest recent genetic infiltration of modern breeds into South-East Asian goats and an ancestral SRY Y2B haplotype in Asian native goats.

  7. The ovarian reserve of primordial follicles and the dynamic reserve of antral growing follicles: what is the link?

    PubMed

    Monniaux, Danielle; Clément, Frédérique; Dalbiès-Tran, Rozenn; Estienne, Anthony; Fabre, Stéphane; Mansanet, Camille; Monget, Philippe

    2014-04-01

    The growing follicles develop from a reserve of primordial follicles constituted early in life. From this pre-established reserve, a second ovarian reserve is formed, which consists of gonadotropin-responsive small antral growing follicles and is a dynamic reserve for ovulation. Its size, evaluated by direct antral follicular count or endocrine markers, determines the success of assisted reproductive technologies in humans and embryo production biotechnologies in animals. Strong evidence indicates that these two reserves are functionally related. The size of both reserves appears to be highly variable between individuals of similar age, but the equilibrium size of the dynamic reserve in adults seems to be specific to each individual. The dynamics of both follicular reserves appears to result from the fine tuning of regulations involving two main pathways, the phosphatase and tensin homolog (PTEN)/phosphatidylinositol-3 kinase (PI3K)/3-phosphoinositide-dependent protein kinase-1 (PDPK1)/v-akt murine thymoma viral oncogene homolog 1 (AKT1) and the bone morphogenetic protein (BMP)/anti-Müllerian hormone (AMH)/SMAD signaling pathways. Mutations in genes encoding the ligands, receptors, or signaling effectors of these pathways can accelerate or modulate the exhaustion rate of the ovarian reserves, causing premature ovarian insufficiency (POI) or increase in reproductive longevity, respectively. With female aging, the decline in primordial follicle numbers parallels the decrease in the size of the dynamic reserve of small antral follicles and the deterioration of oocyte quality. Recent progress in our knowledge of signaling pathways and their environmental and hormonal control during adult and fetal life opens new perspectives to improve the management of the ovarian reserves.

  8. Noninvasive Index of Cryorecovery and Growth Potential for Human Follicles In Vitro1

    PubMed Central

    Barrett, Susan L.; Shea, Lonnie D.; Woodruff, Teresa K.

    2010-01-01

    Cryopreservation of oocytes and embryos is commonly used to preserve fertility. However, women undergoing cancer treatment may not have the time or may not be good candidates for these options. Ovarian cortical tissue cryopreservation and subsequent tissue transplant has been proven successful yet inefficient in preserving larger secondary follicles, and is not recommended as a fertility preservation option for women with certain cancers. We evaluated cryopreservation of individual follicles as an alternative option in rodents, nonhuman primates, and human primates. Under optimal conditions, cryopreserved mouse secondary follicles were able to reestablish granulosa cell-oocyte interactions, which are essential for subsequent follicle growth. Individual secondary follicles survived cryopreservation, were able to be cultured in a three-dimensional alginate hydrogel matrix to the antral stage, and the enclosed oocytes were competent for fertilization. Using a vital imaging technique (pol-scope) employed in many fertility centers, we were able to bioassay the thawed, cultured follicles for the presence of transzonal connections between the somatic and germ cells. Perturbations in these linkages were shown to be reversed when follicles were cryopreserved under optimal freezing conditions. We applied the optimized cryopreservation protocol to isolated rhesus monkey and human secondary follicles, and using the birefringent bioassay, we were able to show good correlation between early follicle growth and healthy somatic cell-oocyte connections. Our results suggest that ovarian follicles can be cryopreserved, thawed, and analyzed noninvasively, making follicle preservation an additional option for young cancer patients. PMID:20200211

  9. Re-use of intravaginal progesterone devices associated with the Short-term Protocol for timed artificial insemination in goats.

    PubMed

    Vilariño, M; Rubianes, E; Menchaca, A

    2011-04-15

    Because intravaginal devices impregnated with 0.3 g of progesterone (i.e., CIDR-G) contain remaining hormone after their use in a Short-term Protocol (5 to 7 d of treatment), the reuse of these devices is proposed in goats. Two experiments were designed to study the effects of the reutilization of CIDR-G, establishing serum progesterone concentrations, follicular development, ovulatory response, and fertility. Experiment 1: Thirty dairy goats received a Short-term Protocol for 5 d using CIDR-G of first use (new devices, n = 10), second use (previously used for 5 d, n = 10), or third use (previously used twice for 5 d each time, n = 10). Goats were given (im) prostaglandin F(2α) (10 mg dinoprost) and eCG (300 IU) at device insertion and withdrawal, respectively. Serum progesterone concentrations induced by CIDR-G of first use were higher than CIDR-G of second or third use (P < 0.05); concentrations were consistently > 1 ng/mL in all females treated with reused devices. Estrus and ovulation were synchronized in 100% of goats (no differences among treatments). All females treated with new devices, but only 80% of females treated with re-used devices ovulated a new follicle that emerged after CIDR-G insertion (P = NS). Ovulation occurred between 60 and 70 h after device removal (no differences among groups). Experiment 2: In goats subjected to a Short-term Protocol followed by AI at 54 h after CIDR-G, pregnancy rates with CIDR-G of first, second, and third use were 75.3% (64/85), 67.4% (60/89), and 62.1% (54/87), respectively (devices of first versus third use, P < 0.05). In summary, intravaginal devices originally containing 0.3 g of progesterone appeared effective to synchronize estrus and ovulation after first, second and third use in the Short-term Protocol. Although the pregnancy rate with reused devices was acceptable (i.e., > 60%), it was significantly lower than that achieved with new devices and further studies to ensure adequate follicular turnover are

  10. Two Severe Cases of Tungiasis in Goat Kids in Uganda.

    PubMed

    Mutebi, Francis; Krücken, Jürgen; Mencke, Norbert; Feldmeier, Hermann; von Samson-Himmelstjerna, Georg; Waiswa, Charles

    2016-01-01

    Tungiasis ensues from the penetration and burrowing of female sand fleas (Tunga spp.; Siphonaptera: Tungidae) in the skin of mammals. There are few case reports of severe tungiasis in goats and in these cases the Tunga species were not in most cases clearly identified. Two cases of severe tungiasis caused by Tunga penetrans in goat kids from tungiasis-endemic rural Uganda are reported. These are the first severe cases of tungiasis in goats reported from outside South America. PMID:27012871

  11. Two Severe Cases of Tungiasis in Goat Kids in Uganda.

    PubMed

    Mutebi, Francis; Krücken, Jürgen; Mencke, Norbert; Feldmeier, Hermann; von Samson-Himmelstjerna, Georg; Waiswa, Charles

    2016-01-01

    Tungiasis ensues from the penetration and burrowing of female sand fleas (Tunga spp.; Siphonaptera: Tungidae) in the skin of mammals. There are few case reports of severe tungiasis in goats and in these cases the Tunga species were not in most cases clearly identified. Two cases of severe tungiasis caused by Tunga penetrans in goat kids from tungiasis-endemic rural Uganda are reported. These are the first severe cases of tungiasis in goats reported from outside South America.

  12. Two Severe Cases of Tungiasis in Goat Kids in Uganda

    PubMed Central

    Mutebi, Francis; Krücken, Jürgen; Mencke, Norbert; Feldmeier, Hermann; von Samson-Himmelstjerna, Georg; Waiswa, Charles

    2016-01-01

    Tungiasis ensues from the penetration and burrowing of female sand fleas (Tunga spp.; Siphonaptera: Tungidae) in the skin of mammals. There are few case reports of severe tungiasis in goats and in these cases the Tunga species were not in most cases clearly identified. Two cases of severe tungiasis caused by Tunga penetrans in goat kids from tungiasis-endemic rural Uganda are reported. These are the first severe cases of tungiasis in goats reported from outside South America. PMID:27012871

  13. Targeted expression of GFP in the hair follicle using ex vivo viral transduction.

    PubMed

    Hoffman, Robert M; Li, Lingna

    2008-01-01

    There are many cell types in the hair follicle, including hair matrix cells which form the hair shaft and stem cells which can initiate the hair shaft during early anagen, the growth phase of the hair cycle, as well as pluripotent stem cells that play a role in hair follicle growth but have the potential to differentiate to non-follicle cells such as neurons. These properties of the hair follicle are discussed. The various cell types of the hair follicle are potential targets for gene therapy. Gene delivery system for the hair follicle using viral vectors or liposomes for gene targeting to the various cell types in the hair follicle and the results obtained are also discussed [corrected]. PMID:19066571

  14. Oocyte-specific deletion of Pten causes premature activation of the primordial follicle pool.

    PubMed

    Reddy, Pradeep; Liu, Lian; Adhikari, Deepak; Jagarlamudi, Krishna; Rajareddy, Singareddy; Shen, Yan; Du, Chun; Tang, Wenli; Hämäläinen, Tuula; Peng, Stanford L; Lan, Zi-Jian; Cooney, Austin J; Huhtaniemi, Ilpo; Liu, Kui

    2008-02-01

    In the mammalian ovary, progressive activation of primordial follicles from the dormant pool serves as the source of fertilizable ova. Menopause, or the end of female reproductive life, occurs when the primordial follicle pool is exhausted. However, the molecular mechanisms underlying follicle activation are poorly understood. We provide genetic evidence that in mice lacking PTEN (phosphatase and tensin homolog deleted on chromosome 10) in oocytes, a major negative regulator of phosphatidylinositol 3-kinase (PI3K), the entire primordial follicle pool becomes activated. Subsequently, all primordial follicles become depleted in early adulthood, causing premature ovarian failure (POF). Our results show that the mammalian oocyte serves as the headquarters of programming of follicle activation and that the oocyte PTEN-PI3K pathway governs follicle activation through control of initiation of oocyte growth.

  15. Targeted expression of GFP in the hair follicle using ex vivo viral transduction.

    PubMed

    Hoffman, Robert M; Li, Lingna

    2008-03-17

    There are many cell types in the hair follicle, including hair matrix cells which form the hair shaft and stem cells which can initiate the hair shaft during early anagen, the growth phase of the hair cycle, as well as pluripotent stem cells that play a role in hair follicle growth but have the potential to differentiate to non-follicle cells such as neurons. These properties of the hair follicle are discussed. The various cell types of the hair follicle are potential targets for gene therapy. Gene delivery system for the hair follicle using viral vectors or liposomes for gene targeting to the various cell types in the hair follicle and the results obtained are also discussed [corrected].

  16. The hair follicle and its stem cells as drug delivery targets.

    PubMed

    Hoffman, Robert M

    2006-05-01

    The hair follicle is a skin appendage with a complex structure containing many cell types that produce highly specialised proteins. The hair follicle is in a continuous cycle: anagen is the hair growth phase, catagen the involution phase and telogen is the resting phase. The follicle offers many potential therapeutic targets. Hoffman and colleagues have pioneered hair-follicle-specific targeting using liposomes to deliver small and large molecules, including genes. They have also pioneered ex vivo hair-follicle targeting with continued expression of the introduced gene following transplantation. Recently, it has been discovered that hair follicle stem cells are highly pluripotent and can form neurons, glial cells and other cell types, and this has suggested that hair follicle stem cells may serve as gene therapy targets for regenerative medicine.

  17. SKPs derive from hair follicle precursors and exhibit properties of adult dermal stem cells.

    PubMed

    Biernaskie, Jeffrey; Paris, Maryline; Morozova, Olena; Fagan, B Matthew; Marra, Marco; Pevny, Larysa; Miller, Freda D

    2009-12-01

    Despite the remarkable regenerative capacity of mammalian skin, an adult dermal stem cell has not yet been identified. Here, we investigated whether skin-derived precursors (SKPs) might fulfill such a role. We show that SKPs derive from Sox2(+) hair follicle dermal cells and that these two cell populations are similar with regard to their transcriptome and functional properties. Both clonal SKPs and endogenous Sox2(+) cells induce hair morphogenesis, differentiate into dermal cell types, and home to a hair follicle niche upon transplantation. Moreover, hair follicle-derived SKPs self-renew, maintain their multipotency, and serially reconstitute hair follicles. Finally, grafting experiments show that follicle-associated dermal cells move out of their niche to contribute cells for dermal maintenance and wound-healing. Thus, SKPs derive from Sox2(+) follicle-associated dermal precursors and display functional properties predicted of a dermal stem cell, contributing to dermal maintenance, wound-healing, and hair follicle morphogenesis.

  18. Mitochondrial DNA diversity of Anatolian indigenous domestic goats.

    PubMed

    Akis, I; Oztabak, K; Mengi, A; Un, C

    2014-12-01

    Anatolia has been an important region for civilizations and agricultural revolution as a major domestication centre for livestock species. Goats (Capra hircus) were among the earliest domesticated animals in this region. In this study, genetic diversity of Anatolian goat breeds was characterized by comparison of mitochondrial DNA hypervariable region 1. A total of 295 individuals, including 99 Anatolian Black goats, 96 Angora goats and 100 Kilis goats, were used. Haplogroup A was found to be the dominant haplogroup in all three breeds. The highest haplogroup diversity, including haplogroups A, B2, C and G, was observed in the Anatolian Black breed. Haplogroup D was only observed in Kilis and Angora goats. Haplogroup G was found in Angora and Anatolian Black breeds. The Anatolian goat breeds had high genetic diversity values and a weak phylogeographical structure. The nucleotide diversity values were found to be higher than those in previously studied goat breeds. The fact that Anatolia is a domestication centre and its geographical position as a junction of trade routes may have caused the higher genetic diversity of Anatolian goat breeds.

  19. A Genetic Linkage Map of the Male Goat Genome

    PubMed Central

    Vaiman, D.; Schibler, L.; Bourgeois, F.; Oustry, A.; Amigues, Y.; Cribiu, E. P.

    1996-01-01

    This paper presents a first genetic linkage map of the goat genome. Primers derived from the flanking sequences of 612 bovine, ovine and goat microsatellite markers were gathered and tested for amplification with goat DNA under standardized PCR conditions. This screen made it possible to choose a set of 55 polymorphic markers that can be used in the three species and to define a panel of 223 microsatellites suitable for the goat. Twelve half-sib paternal goat families were then used to build a linkage map of the goat genome. The linkage analysis made it possible to construct a meiotic map covering 2300 cM, i.e., >80% of the total estimated length of the goat genome. Moreover, eight cosmids containing microsatellites were mapped by fluorescence in situ hybridization in goat and sheep. Together with 11 microsatellite-containing cosmids previously mapped in cattle (and supposing conservation of the banding pattern between this species and the goat) and data from the sheep map, these results made the orientation of 15 linkage groups possible. Furthermore, 12 coding sequences were mapped either genetically or physically, providing useful data for comparative mapping. PMID:8878693

  20. Goat paddock cryptoexplosion crater, Western Australia

    USGS Publications Warehouse

    Harms, J.E.; Milton, D.J.; Ferguson, J.; Gilbert, D.J.; Harris, W.K.; Goleby, B.

    1980-01-01

    Goat Paddock, a crater slightly over 5 km in diameter (18??20??? S, 126??40???E), lies at the north edge of the King Leopold Range/Mueller Range junction in the Kimberley district, Western Australia (Fig. 1). It was noted as a geological anomaly in 1964 during regional mapping by the Bureau of Mineral Resources, Geology and Geophysics and the Geological Survey of Western Australia. The possibility of its being a meteorite impact crater has been discussed1, although this suggestion was subsequently ignored2. Two holes were drilled by a mining corporation in 1972 to test whether kimberlite underlay the structure. Here we report the findings of five days of reconnaissance in August 1979 which established that Goat Paddock is a cryptoexplosion crater containing shocked rocks and an unusually well exposed set of structural features. ?? 1980 Nature Publishing Group.

  1. 9 CFR 91.6 - Goats.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... exported if it is a scrapie-positive animal or an exposed animal, as defined in 9 CFR parts 54 and 79, or if it has ever been in an infected flock, source flock, or trace flock, as defined in 9 CFR parts 54... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Goats. 91.6 Section 91.6 Animals...

  2. 9 CFR 91.6 - Goats.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... exported if it is a scrapie-positive animal or an exposed animal, as defined in 9 CFR parts 54 and 79, or if it has ever been in an infected flock, source flock, or trace flock, as defined in 9 CFR parts 54... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Goats. 91.6 Section 91.6 Animals...

  3. High-throughput sequencing analyses of XX genital ridges lacking FOXL2 reveal DMRT1 up-regulation before SOX9 expression during the sex-reversal process in goats.

    PubMed

    Elzaiat, Maëva; Jouneau, Luc; Thépot, Dominique; Klopp, Christophe; Allais-Bonnet, Aurélie; Cabau, Cédric; André, Marjolaine; Chaffaux, Stéphane; Cribiu, Edmond-Paul; Pailhoux, Eric; Pannetier, Maëlle

    2014-12-01

    FOXL2 loss of function in goats leads to the early transdifferentiation of ovaries into testes, then to the full sex reversal of XX homozygous mutants. By contrast, Foxl2 loss of function in mice induces an arrest of follicle formation after birth, followed by complete female sterility. In order to understand the molecular role of FOXL2 during ovarian differentiation in the goat species, putative FOXL2 target genes were determined at the earliest stage of gonadal sex-specific differentiation by comparing the mRNA profiles of XX gonads expressing the FOXL2 protein or not. Of these 163 deregulated genes, around two-thirds corresponded to testicular genes that were up-regulated when FOXL2 was absent, and only 19 represented female-associated genes, down-regulated in the absence of FOXL2. FOXL2 should therefore be viewed as an antitestis gene rather than as a female-promoting gene. In particular, the key testis-determining gene DMRT1 was found to be up-regulated ahead of SOX9, thus suggesting in goats that SOX9 primary up-regulation may require DMRT1. Overall, our results equated to FOXL2 being an antitestis gene, allowing us to propose an alternative model for the sex-determination process in goats that differs slightly from that demonstrated in mice.

  4. Bacteriocinogenic Bacteria Isolated from Raw Goat Milk and Goat Cheese Produced in the Center of México.

    PubMed

    Hernández-Saldaña, Oscar F; Valencia-Posadas, Mauricio; de la Fuente-Salcido, Norma M; Bideshi, Dennis K; Barboza-Corona, José E

    2016-09-01

    Currently, there are few reports on the isolation of microorganisms from goat milk and goat cheese that have antibacterial activity. In particular, there are no reports on the isolation of microorganisms with antibacterial activity from these products in central Mexico. Our objective was to isolate bacteria, from goat products, that synthesized antimicrobial peptides with activity against a variety of clinically significant bacteria. We isolated and identified Lactobacillus rhamnosus, L. plantarum, L. pentosus, L. helveticus and Enterococcus faecium from goat cheese, and Aquabacterium fontiphilum, Methylibium petroleiphilum, Piscinobacter aquaticus and Staphylococcus xylosus from goat milk. These bacteria isolated from goat cheese were able to inhibit Staphylococcus aureus, Bacillus cereus, Escherichia coli, Listeria monocytogenes, L. inoccua, Pseudomona aeruginosa, Shigella flexneri, Serratia marcescens, Enterobacter cloacae and Klebsiella pneumoniae. In addition, bacteria from goat milk showed inhibitory activity against B. cereus, L. lactis, E. coli, S. flexneri, E. cloacae and K. pneumonia; S. aureus, L. innocua, S. agalactiae and S. marcescens. The bacteriocins produced by these isolates were shown to be acid stable (pH 2-6) and thermotolerant (up to 100 °C), but were susceptible to proteinases. When screened by PCR for the presence of nisin, pediocin and enterocin A genes, none was found in isolates recovered from goat milk, and only the enterocin A gene was found in isolates from goat cheese.

  5. Bacteriocinogenic Bacteria Isolated from Raw Goat Milk and Goat Cheese Produced in the Center of México.

    PubMed

    Hernández-Saldaña, Oscar F; Valencia-Posadas, Mauricio; de la Fuente-Salcido, Norma M; Bideshi, Dennis K; Barboza-Corona, José E

    2016-09-01

    Currently, there are few reports on the isolation of microorganisms from goat milk and goat cheese that have antibacterial activity. In particular, there are no reports on the isolation of microorganisms with antibacterial activity from these products in central Mexico. Our objective was to isolate bacteria, from goat products, that synthesized antimicrobial peptides with activity against a variety of clinically significant bacteria. We isolated and identified Lactobacillus rhamnosus, L. plantarum, L. pentosus, L. helveticus and Enterococcus faecium from goat cheese, and Aquabacterium fontiphilum, Methylibium petroleiphilum, Piscinobacter aquaticus and Staphylococcus xylosus from goat milk. These bacteria isolated from goat cheese were able to inhibit Staphylococcus aureus, Bacillus cereus, Escherichia coli, Listeria monocytogenes, L. inoccua, Pseudomona aeruginosa, Shigella flexneri, Serratia marcescens, Enterobacter cloacae and Klebsiella pneumoniae. In addition, bacteria from goat milk showed inhibitory activity against B. cereus, L. lactis, E. coli, S. flexneri, E. cloacae and K. pneumonia; S. aureus, L. innocua, S. agalactiae and S. marcescens. The bacteriocins produced by these isolates were shown to be acid stable (pH 2-6) and thermotolerant (up to 100 °C), but were susceptible to proteinases. When screened by PCR for the presence of nisin, pediocin and enterocin A genes, none was found in isolates recovered from goat milk, and only the enterocin A gene was found in isolates from goat cheese. PMID:27407294

  6. Delivery and targeting of nanoparticles into hair follicles.

    PubMed

    Fang, Chia-Lang; Aljuffali, Ibrahim A; Li, Yi-Ching; Fang, Jia-You

    2014-01-01

    It has been demonstrated that nanoparticles used for follicular delivery provide some advantages over conventional pathways, including improved skin bioavailability, enhanced penetration depth, prolonged residence duration, fast transport into the skin and tissue targeting. This review describes recent developments using nanotechnology approaches for drug delivery into the follicles. Different types of nanosystems may be employed for management of follicular permeation, such as polymeric nanoparticles, metallic nanocrystals, liposomes, and lipid nanoparticles. This review systematically introduces the mechanisms of follicles for nanoparticulate penetration, highlighting the therapeutic potential of drug-loaded nanoparticles for treating skin diseases. Special attention is paid to the use of nanoparticles in treating appendage-related disorders, in particular, nanomedical strategies for treating alopecia, acne, and transcutaneous immunization. PMID:25375342

  7. Delivery and targeting of nanoparticles into hair follicles.

    PubMed

    Fang, Chia-Lang; Aljuffali, Ibrahim A; Li, Yi-Ching; Fang, Jia-You

    2014-01-01

    It has been demonstrated that nanoparticles used for follicular delivery provide some advantages over conventional pathways, including improved skin bioavailability, enhanced penetration depth, prolonged residence duration, fast transport into the skin and tissue targeting. This review describes recent developments using nanotechnology approaches for drug delivery into the follicles. Different types of nanosystems may be employed for management of follicular permeation, such as polymeric nanoparticles, metallic nanocrystals, liposomes, and lipid nanoparticles. This review systematically introduces the mechanisms of follicles for nanoparticulate penetration, highlighting the therapeutic potential of drug-loaded nanoparticles for treating skin diseases. Special attention is paid to the use of nanoparticles in treating appendage-related disorders, in particular, nanomedical strategies for treating alopecia, acne, and transcutaneous immunization.

  8. Follicle environment and quality of in vitro matured oocytes.

    PubMed

    Sirard, Marc-André

    2011-06-01

    In mammalian reproduction, the oocyte depends on the ovarian follicle for most of its growth. They form a bipolar partnership and the status of one will impact the functioning of the other. When oocytes are removed from their follicle by ovulation, they have normally completed all the steps required to begin their journey into the oviduct and drive the early embryonic development. When oocytes are removed from their follicle before natural ovulation, the process by which they acquire all the important components for their journey might not be completed and their ability to mature, fertilize or develop into embryos or to term might be compromised. Animal models have been useful to define the important steps required for the oocyte's growth phase, and in the mouse, when the oocyte has reached its full size, the program is ready. This is not the case in larger mammals where the completion of growth does not ensure that the oocyte is fully capable of undergoing all the steps to the embryo and to term. The final steps of oocyte preparation also involve a progressive condensation of the chromatin that may facilitate normal maturation but may also indirectly reduce the lifespan of the oocyte. In such a scenario, the oocyte would have an expiration date when fully competent. In humans, a number of indications may justify the aspiration of oocytes from unstimulated patients and the development of an in vitro maturation (IVM) process that would allow fertilization and subsequent development. This objective could be realized by a better understanding of the essential follicular contribution required before removing the oocyte. Therefore, this review will focus on the large animal models where IVM has been used and studied for more than 25 years. The status of the follicle at the time of oocyte recovery and the status of the oocyte's chromatin will be described in detail as they have a significant impact on the outcome.

  9. Maturation of ovarian follicles in the prepubertal gilt.

    PubMed

    Christenson, R K; Ford, J J; Redmer, D A

    1985-01-01

    The processes of follicle development and puberty are closely related, and both are associated with maturation of the hypothalamic-pituitary-ovarian axis. Prenatal development of the ovary is independent of gonadotrophic stimulation. Beyond 60 days of age (postnatally), tertiary follicles develop and gonadotrophins begin to influence ovarian follicular development. Negative feedback regulation of pituitary gonadotrophins by ovarian secretions develops between 60 and 100 days of age. In the prepubertal gilt, no consistent changes in peripheral FSH, oestrogen or progesterone concentrations have been identified which are associated with recruitment of the first set of preovulatory follicles. Whether LH secretion increases before this recruitment remains equivocal. Few details are available on how gonadotrophic hormones stimulate ovarian function in the prepubertal gilt. On the basis of a follicular maturation model that has been described for the rat, the actions of FSH, LH and oestrogens on follicular cell receptors and the regulation of aromatase activity seem paramount. Aromatization of androgens to oestrogens has been proposed as a central regulator for follicular maturation. In the prepubertal gilt, a selective increase in peripheral FSH concentration occurs on Day 1 after unilateral ovariectomy, followed by significant increases in ovarian venous concentrations of oestradiol and inhibin on Days 2 and 4 and compensatory growth as measured by follicular fluid volume on Days 2, 4 and 8. Administration of pig follicular fluid to the prepubertal gilt during and after unilateral ovariectomy suppresses compensatory ovarian hypertrophy by mechanisms yet to be determined. In pigs a number of intraovarian factors have been identified, but there is little information on how these factors regulate follicular recruitment and growth. The factor(s) that prevents ovarian follicles in the prepubertal gilt from progressing to ovulation after acquiring the ability to ovulate in

  10. Ultrastructure of the theca interna of ovarian follicles in sheep.

    PubMed

    O'Shea, J D; Cran, D G; Hay, M F; Moor, R M

    1978-03-13

    The theca interna of non-atretic ovarian follicles from 2.0 mm in diameter up to the stage shortly following ovulation was studied by light and electron microscopy. In follicles less than 3.0 mm in diameter, the theca interna consisted of about 8-12 layers of flattened cells, together with many capillaries and small bundles of collagen. Two main forms of cellular differentiation were seen. These were towards either fibroblast-like cells or presumed steroidogenic cells whose cytoplasm contained large amounts of predominantly smooth tubular endoplasmic reticulum, to which some ribosomes were attached. The majority of cells were of relatively undifferentiated or intermediate structure. In larger follicles up to the early stages of oestrus the theca interna cells became larger and less flattened, and cells rich in tubular endoplasmic reticulum became proportionately more numerous. By 18 h after the onset of oestrus the theca interna was oedematous, and many cells possessed pseudopodia. Many cells also contained numerous lipid droplets, but there were no signs of thecal cell degeneration or death. Shortly after ovulation the basal lamina of the membrana granulosa was incomplete, and it became more difficult to distinguish between theca and granulosa layers. Structural heterogeneity, with two major cell types and cells of intermediate structure, was present at all stages. It was concluded that: (1) the theca interna of 2.0-2.9 mm follicles contained many cells whose structure was compatible with a steroidogenic capacity; (2) changes in the differentiated thecal cells up to the early stages of oestrus were quantitative rather than qualitative, and suggestive of an increased steroidogenic capacity; (3) the accumulation of lipid in many cells of the theca interna by 18 h after the onset of oestrus probably reflected a reduction in steroidogenic activity; and (4) there was no evidence of any structural specialization to facilitate the transport of steroids from the theca

  11. Experimental pestivirus infections in pregnant goats.

    PubMed

    Løken, T; Bjerkås, I

    1991-08-01

    Fifty pregnant goats, inoculated intramuscularly at different gestational stages with a non-cytopathic ovine pestivirus or a cytopathic bovine pestivirus, all developed pestivirus-neutralizing antibodies within 5 weeks of inoculation. The incidence of reproductive failure was similar for the two agents. Parturition at term with only healthy kids occurred in 13 (26 per cent) of the goats. Viable kids were not born to any of the 17 goats inoculated at about day 40 of gestation. Three of the 17 delivered dead or weak kids, seven aborted and three of seven which were necropsied during pregnancy had markedly underdeveloped and autolysed or mummified fetuses in utero, while four were barren. When inoculated at around the 60th day of gestation, two of 18 animals gave birth to only healthy kids, 12 to dead and/or weak kids, two aborted and, at necropsy, a small, decomposed fetus was found in one goat while one other was barren. In this group, one kid was ataxic and seven others had body tremors characteristic of border disease. One of the latter kids was viable. Of 15 goats inoculated at around day 100 of gestation, 11 gave birth to healthy kids only, three to dead and/or weak kids and one aborted. In 23 progeny, histological changes in the central nervous system (CNS) consisted mainly of cerebral white matter necrosis, cerebellar dysplasia, hypercellular areas in white matter and lymphocytic perivascular cuffings. All seven weak-born kids with signs of border disease had CNS lesions, particularly cerebellar dysplasia and/or hypercellular areas. Non-cytopathic pestivirus was isolated from tissues from all eight progeny examined in the 40-day inoculation group, from tissues and/or serum from 10 of 23 progeny in the 60-day group, and from four of 24 in the 100-day group. Persistent infection was demonstrated in a healthy kid, in a viable shaker and in two other kids which appeared normal at birth. Examination of offspring before ingestion of colostrum revealed pestivirus

  12. Granulosa cells from bovine follicles activate different signal transduction pathways dependent on follicle health status and ability to convert androstenedione to estrogen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Since steroidogenesis is a critical component in the development of competent preovulatory follicles we hypothesized that granulosa cells from follicles of cows treated with normal levels of progesterone (CIDR) or with melengestrol acetate (MGA), which results in the development of persistent follic...

  13. Introduction of distillate rosemary leaves into the diet of the Murciano-Granadina goat: transfer of polyphenolic compounds to goats' milk and the plasma of suckling goat kids.

    PubMed

    Jordán, Maria José; Moñino, María Inmaculada; Martínez, Cristina; Lafuente, Arturo; Sotomayor, José Antonio

    2010-07-28

    The effect of the introduction of distilled rosemary leaves into the diet of the Murciano-Granadina goat on the polyphenolic profile of the goats' milk during the physiological stages of gestation and lactation was studied. The inclusion of rosemary leaves into the animal diet modified neither animal productivity (milk yield) nor milk quality. The following components were found in increased concentration (P < 0.05) in the goats' milk after the introduction of rosemary leaves into their diet: flavonoids hesperidin, naringin, and genkwanin; gallic acid; and phenolic diterpenes carnosol and carnosic acid. With regard to the transfer of polyphenols to the plasma of the suckling goat kid, a statistically significant increase (P < 0.05) in rosmarinic acid, carnosic acid, and carnosol concentrations was detected. From this point of view, distillate rosemary leaves can be proposed as an ingredient in ruminant feed because they both alter neither the yield nor the quality of Murciano-Granadina goats' milk and allow for an increased concentration of polyphenolic components in the goats' milk and in the plasma of the suckling goat kid.

  14. 7-Phloroeckol promotes hair growth on human follicles in vitro.

    PubMed

    Bak, Soon-Sun; Sung, Young Kwan; Kim, Se-Kwon

    2014-08-01

    7-Phloroeckol, phloroglucinol derivative isolated from marine brown algae, has anti-oxidative, anti-inflammatory responses and MMP inhibitory activities. In this study, we evaluated the hair growth-promoting effects of 7-phloroeckol in human hair follicles. To investigate cell viability of human dermal papilla cells (DPCs) and outer root sheath (ORS) cells in the presence or absence of 7-phloroeckol treatment, MTT assay was employed. Moreover, gene expression and protein concentration of insulin-like growth factor (IGF)-1 was measured by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. 7-Phloroeckol induced an increase in proliferation of DPCs and ORS cells. In addition, hair shaft growth was measured using the hair-follicle organ culture system. 7-Phloroeckol resulted in elongation of the hair shaft in cultured human hair follicles. 7-Phloroeckol induced an IGF-1 mRNA expression and protein concentration in DPCs and conditioned media, respectively. These results suggest that 7-phloroeckol promotes hair growth through stimulation of DPCs and ORS cells.

  15. Targeting gene expression to the wool follicle in transgenic sheep.

    PubMed

    Damak, S; Jay, N P; Barrell, G K; Bullock, D W

    1996-02-01

    To establish the feasibility of overexpressing foreign genes in the wool follicle, transgenic sheep were produced by pronuclear microinjection of a DNA construct consisting of a mouse ultrahigh-sulfur keratin promoter linked to the bacterial chloramphenicol acetyl transferase (CAT) gene. Four of 31 lambs born were transgenic. The overall efficiency of transgenesis was 1.1% of zygotes injected and transferred. Two transgenic rams were mated to nontransgenic ewes, and both transmitted the gene to their offspring in Mendelian fashion. CAT expression was found in the skin of one G0 ram and in 9 out of 26 transgenic G1 progeny. Two G1 lambs were sacrificed to study tissue specificity. Both had high levels of expression in skin but One had high expression in spleen and kidney with lower levels of expression in lung; the other had low expression in spleen, lung, and muscle. In situ hybridization demonstrated that transgene expression in the skin was confined to the keratogenous zone of the wool follicle cortex. Expression of CAT activity in skin was correlated with diet-induced or seasonal changes in the rate of wool growth. This keratin promoter appears useful for overexpressing factors in the wool follicle that might influence wool production or properties.

  16. Cementum matrix formation in vivo by cultured dental follicle cells.

    PubMed

    Handa, K; Saito, M; Yamauchi, M; Kiyono, T; Sato, S; Teranaka, T; Sampath Narayanan, A

    2002-11-01

    Dental follicle is the fibrous tissue that surrounds the developing tooth germ, and it is believed to contain progenitors for cementoblasts, periodontal ligament cells, and osteoblasts. In this study, we report the presence of cementoblast progenitors in cultures of bovine dental follicle cells and demonstrate their differentiation capacity. Bovine dental follicle cells (BDFC) obtained from tooth germs by collagenase digestion were compared with bovine alveolar bone osteoblasts (BAOB) and bovine periodontal ligament cells (BPDL) in vitro and in vivo. In culture, BDFC exhibited low levels of alkaline phosphatase activity and expressed mRNA for osteopontin (OP) and type I collagen (COLI), as well as low levels of osteocalcin (OC) mRNA. In contrast, cultured BAOB exhibited high alkaline phosphatase activity levels and expressed mRNA for OC, OP, COLI, and bone sialoprotein (BSP). To elucidate the differentiation capacity of BDFC in vivo, cells were transplanted into severe combined immunodeficiency (SCID) mice and analyzed after 4 weeks. Transplanted BDFC formed fibrous tissue and cementum-like matrix, which stained positive for anti-cementum attachment protein (CAP) monoclonal antibody (3G9), and expressed mRNA for OC, OP, COLI, and BSP. On the other hand, transplanted BAOB formed bone-like matrix, but were negative for anti-CAP monoclonal antibody. The BPDL transplants formed fibrous tissue that contained a few cells expressing CAP. These results indicate that cementoblast progenitors are present in BDFC, which can provide a useful model for investigating the molecular mechanisms of cementogenesis. PMID:12477575

  17. Signaling Involved in Hair Follicle Morphogenesis and Development

    PubMed Central

    Rishikaysh, Pisal; Dev, Kapil; Diaz, Daniel; Qureshi, Wasay Mohiuddin Shaikh; Filip, Stanislav; Mokry, Jaroslav

    2014-01-01

    Hair follicle morphogenesis depends on Wnt, Shh, Notch, BMP and other signaling pathways interplay between epithelial and mesenchymal cells. The Wnt pathway plays an essential role during hair follicle induction, Shh is involved in morphogenesis and late stage differentiation, Notch signaling determines stem cell fate while BMP is involved in cellular differentiation. The Wnt pathway is considered to be the master regulator during hair follicle morphogenesis. Wnt signaling proceeds through EDA/EDAR/NF-κB signaling. NF-κB regulates the Wnt pathway and acts as a signal mediator by upregulating the expression of Shh ligand. Signal crosstalk between epithelial and mesenchymal cells takes place mainly through primary cilia. Primary cilia formation is initiated with epithelial laminin-511 interaction with dermal β-1 integrin, which also upregulates expression of downstream effectors of Shh pathway in dermal lineage. PDGF signal transduction essential for crosstalk is mediated through epithelial PDGF-A and PDGFRα expressed on the primary cilia. Dermal Shh and PDGF signaling up-regulates dermal noggin expression; noggin is a potent inhibitor of BMP signaling which helps in counteracting BMP mediated β-catenin inhibition. This interplay of signaling between the epithelial and dermal lineage helps in epithelial Shh signal amplification. The dermal Wnt pathway helps in upregulation of epithelial Notch expression. Dysregulation of these pathways leads to certain abnormalities and in some cases even tumor outgrowth. PMID:24451143

  18. Follicle Loss and Apoptosis in Cyclophosphamide-Treated Mice: What’s the Matter?

    PubMed Central

    Chen, Xiu-Ying; Xia, He-Xia; Guan, Hai-Yun; Li, Bin; Zhang, Wei

    2016-01-01

    With increasing numbers of young female cancer survivors following chemotherapy, chemotherapy-induced fertility loss must be considered. Menstrual disorder and infertility are of particular concern in female cancer patients. We showed that treatment with the alkylating agent cyclophosphamide (CTX) could cause severe primordial follicle loss and growing follicle apoptosis, resulting in loss of ovarian reserve. SPF C57BL/6 female mice were treated with a single dose of 120 mg/kg of CTX or saline as a control, and both sides of ovaries were collected three or seven days after injection. Following CTX treatment, the ovaries were mostly composed of collapsed oocytes and presented marked cortical fibrosis and a reduced number of follicles, especially primordial follicles. The loss of primordial follicles was confirmed by primordial follicle counting, immunohistochemistry and Western blot detection of DDx4/MVH. Follicle apoptosis was tested by a TUNEL assay and the number of TUNEL-positive follicle cells increased, as expected, in CTX-treated mice. Furthermore, expression of APAF-1 and cleaved caspase-3 was also increased after CTX treatment. Analysis of the PI3K/Akt/mTOR signaling pathway showed that CTX increased phosphorylation of Akt, mTOR and downstream proteins without affecting total levels. These results demonstrated that the CTX treatment led to the hyperactivation of the PI3K/Akt/mTOR signaling pathway in ovaries which may be related to primordial follicle loss and growing follicle apoptosis. PMID:27248997

  19. Development of an automatic follicle isolation tool for ovarian ultrasonographic images

    NASA Astrophysics Data System (ADS)

    Sarty, Gordon E.; Sonka, Milan; Liang, Weidong; Pierson, Roger A.

    1997-04-01

    PURPOSE: To develop an automatic computer algorithm for isolating the follicles in ovarian ultrasonographic images. METHODS: A semi-automatic algorithm has been developed as the first step in the development of a totally automatic follicle isolation tool for use with ovarian ultrasonographic imaging. The algorithm is knowledge-based and depends upon the use of a priori information about the structure of the follicle. Graph searching techniques are used along with a method of assigning graph node costs that represent edge information combined with the a priori knowledge.Interactive identification of the follicle of interest is followed by, in some cases, a manual editing of an automatically defined interior boundary. After the interior boundary has been defined, the outer follicle wall border is found without further human intervention. RESULTS: Based on a test with 31 ultrasonographic images of women's ovaries made in vivo, the algorithm is able to locate the outer follicle wall to an rms accuracy of 0.59 mm +/- 0.28 mm in comparison with human expert manual boundary tracing. BREAKTHROUGHS: The isolation of ovarian follicles in ultrasonographic imaging has heretofore only been accomplished by manual tracing. Our semi-automated follicle border finding algorithm is, to the best of our knowledge, the first computerized method capable of finding the outer wall boundary of the follicle. CONCLUSIONS: The success of our semi-automatic follicle isolation algorithm clearly demonstrates the feasibility of a totally automatic tool that should have wide application in ultrasonographic studies of ovarian follicular dynamics.

  20. Oocyte Degeneration Associated with Follicle Cells in Female Mactra chinensis (Bivalvia: Mactridae)

    PubMed Central

    Kim, Sung Han; Chung, Ee-Yung; Lee, Ki-Young

    2014-01-01

    Ultrastructural studies of oocyte degeneration in the oocyte, and the functions of follicle cells during oocyte degeneration are described to clarify the reproductive mechanism on oocyte degeneration of Mactra chinensis using cytological methods. Commonly, the follicle cells are attached to the oocyte. Follicle cells play an important role in oocyte degeneration. In particular, the functions of follicle cells during oocyte degeneration are associated with phagocytosis and the intracellular digestion of products. In this study, morphologically similar degenerated phagosomes (various lysosomes), which were observed in the degenerated oocytes, appeared in the follicle cells. After the spawning of the oocytes, the follicle cells were involved in oocyte degeneration through phagocytosis by phagolysosomes. Therefore, it can be assumed that follicle cells reabsorb phagosomes from degenerated oocytes. In this study, the presence of lipid granules, which occurred from degenerating yolk granules, gradually increased in degenerating oocytes. The function of follicle cells can accumulate reserves of lipid granules and glycogen in the cytoplasm, which can be employed by the vitellogenic oocyte. Based on observations of follicle cells attached to degenerating oocytes after spawning, the follicle cells of this species are involved in the lysosomal induction of oocyte degeneration for the reabsorption of phagosomes (phagolysosomes) in the cytoplasm for nutrient storage, as seen in other bivalves. PMID:25949203

  1. Bisphenol A inhibits cultured mouse ovarian follicle growth partially via the aryl hydrocarbon receptor signaling pathway

    PubMed Central

    Ziv-Gal, Ayelet; Craig, Zelieann R.; Wang, Wei; Flaws, Jodi A.

    2013-01-01

    Bisphenol A (BPA) is an endocrine disruptor that inhibits growth of mouse ovarian follicles and disrupts steroidogenesis at a dose of 438 μM. However, the effects of lower doses of BPA and its mechanism of action in ovarian follicles are unknown. We hypothesized that low doses of BPA inhibit follicular growth and decrease estradiol levels through the aryl hydrocarbon receptor (AHR) pathway. Antral follicles from wild-type and Ahr knock-out (AhrKO) mice were cultured for 96 hours. Follicle diameters and estradiol levels then were compared in wild-type and AhrKO follicles ± BPA (0.004 - 438 μM). BPA inhibited follicle growth (110 - 438 μM) and decreased estradiol levels (43.8 - 438 μM) in wild-type and AhrKO follicles. However, at BPA 110 μM, inhibition of growth in AhrKO follicles was attenuated compared to wild-type follicles. These data suggest that BPA may inhibit follicle growth partially via the AHR pathway, whereas its effects on estradiol synthesis likely involve other mechanisms. PMID:23928317

  2. Bone morphogenetic protein 15 may promote follicle selection in the hen.

    PubMed

    Stephens, C S; Johnson, P A

    2016-09-01

    In the hen, optimal ovulation rate depends on selection of a single follicle into the pre-ovulatory hierarchy. Follicle selection is associated with increased oocyte growth and changes in gene expression in granulosa cells surrounding the oocyte, in preparation for ovulation. This study investigated the expression, function and regulation of bone morphogenetic protein-15 (BMP15) during follicle development in the hen. BMP15 mRNA expression was analyzed in the ooplasm and granulosa cells of 3mm follicles and was confirmed to be primarily in the ooplasm. BMP15 was detected by immunoblotting in 6 and 8mm follicles near the time of follicle selection. Expression of mRNA for BMP15 receptors (BMPR1B and BMPR2) in granulosa cells increased with follicle size, indicating that BMP15 may play an important role around follicle selection. The function of BMP15 was examined by culturing granulosa cells from 3-5mm and 6-8mm follicles with recombinant human BMP15 (rhBMP15). BMP15 increased expression of follicle stimulating hormone receptor (FSHR) mRNA and decreased anti-Müllerian hormone (AMH) mRNA and occludin (OCLN), factors associated with follicle maturation and growth in the hen. Hormonal regulation of BMP15 was assessed by whole follicle culture with estradiol (E2) which increased BMP15 mRNA expression. The distinct expression pattern of BMP15 and its receptors, coupled with the effects of BMP15 to increase FSHR mRNA and decrease AMH mRNA and OCLN mRNA and protein expression suggest that the oocyte may have a role in follicle selection in the chicken. PMID:27340039

  3. Bone morphogenetic protein 15 may promote follicle selection in the hen.

    PubMed

    Stephens, C S; Johnson, P A

    2016-09-01

    In the hen, optimal ovulation rate depends on selection of a single follicle into the pre-ovulatory hierarchy. Follicle selection is associated with increased oocyte growth and changes in gene expression in granulosa cells surrounding the oocyte, in preparation for ovulation. This study investigated the expression, function and regulation of bone morphogenetic protein-15 (BMP15) during follicle development in the hen. BMP15 mRNA expression was analyzed in the ooplasm and granulosa cells of 3mm follicles and was confirmed to be primarily in the ooplasm. BMP15 was detected by immunoblotting in 6 and 8mm follicles near the time of follicle selection. Expression of mRNA for BMP15 receptors (BMPR1B and BMPR2) in granulosa cells increased with follicle size, indicating that BMP15 may play an important role around follicle selection. The function of BMP15 was examined by culturing granulosa cells from 3-5mm and 6-8mm follicles with recombinant human BMP15 (rhBMP15). BMP15 increased expression of follicle stimulating hormone receptor (FSHR) mRNA and decreased anti-Müllerian hormone (AMH) mRNA and occludin (OCLN), factors associated with follicle maturation and growth in the hen. Hormonal regulation of BMP15 was assessed by whole follicle culture with estradiol (E2) which increased BMP15 mRNA expression. The distinct expression pattern of BMP15 and its receptors, coupled with the effects of BMP15 to increase FSHR mRNA and decrease AMH mRNA and OCLN mRNA and protein expression suggest that the oocyte may have a role in follicle selection in the chicken.

  4. Ultrastructural observations of previtellogenic ovarian follicles of the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii.

    PubMed

    Beyo, Reston S; Sreejith, Parameswaran; Divya, Lekha; Oommen, Oommen V; Akbarsha, Mohammad A

    2007-04-01

    The ultrastructural organization of the previtellogenic follicles of the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii, of the Western Ghats of India, were observed. Both species follow a similar seasonal reproductive pattern. The ovaries contain primordial follicles throughout the year with previtellogenic, vitellogenic, or postvitellogenic follicles, depending upon the reproductive status. The just-recruited primordial follicle includes an oocyte surrounded by a single layer of follicle and thecal cells. The differentiation of the theca into externa and interna layers, the follicle cells into dark and light variants, and the appearance of primordial yolk platelets and mitochondrial clouds in the ooplasm mark the transition of the primordial follicle into a previtellogenic follicle. During further development of the previtellogenic follicle the following changes occur: i) the theca loses distinction as externa and interna; ii) all the follicle cells become the dark variant and increase in the complexity of ultrastructural organization; iii) the nucleus of the oocyte transforms into the germinal vesicle and there is amplification of the nucleoli; iv) the primordial yolk platelets of the cortical cytoplasm of the oocyte increase in abundance; v) the mitochondrial clouds fragment and the mitochondria move away from the clouds, leaving behind the cementing matrix, which contains membrane-bound vesicles of various sizes, either empty or filled with a lipid material; vi) the perivitelline space appears first as troughs at the junctional points between the follicle cells and oocyte, which subsequently spread all around to become continuous; vii) macrovilli and microvilli develop from the follicle cells and oocyte, respectively; and viii) the precursor material of the vitelline envelop arrives at the perivitelline space. The sequential changes in the previtellogenic follicles of two species of caecilians are described.

  5. Ultrastructural observations of previtellogenic ovarian follicles of the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii.

    PubMed

    Beyo, Reston S; Sreejith, Parameswaran; Divya, Lekha; Oommen, Oommen V; Akbarsha, Mohammad A

    2007-04-01

    The ultrastructural organization of the previtellogenic follicles of the caecilians Ichthyophis tricolor and Gegeneophis ramaswamii, of the Western Ghats of India, were observed. Both species follow a similar seasonal reproductive pattern. The ovaries contain primordial follicles throughout the year with previtellogenic, vitellogenic, or postvitellogenic follicles, depending upon the reproductive status. The just-recruited primordial follicle includes an oocyte surrounded by a single layer of follicle and thecal cells. The differentiation of the theca into externa and interna layers, the follicle cells into dark and light variants, and the appearance of primordial yolk platelets and mitochondrial clouds in the ooplasm mark the transition of the primordial follicle into a previtellogenic follicle. During further development of the previtellogenic follicle the following changes occur: i) the theca loses distinction as externa and interna; ii) all the follicle cells become the dark variant and increase in the complexity of ultrastructural organization; iii) the nucleus of the oocyte transforms into the germinal vesicle and there is amplification of the nucleoli; iv) the primordial yolk platelets of the cortical cytoplasm of the oocyte increase in abundance; v) the mitochondrial clouds fragment and the mitochondria move away from the clouds, leaving behind the cementing matrix, which contains membrane-bound vesicles of various sizes, either empty or filled with a lipid material; vi) the perivitelline space appears first as troughs at the junctional points between the follicle cells and oocyte, which subsequently spread all around to become continuous; vii) macrovilli and microvilli develop from the follicle cells and oocyte, respectively; and viii) the precursor material of the vitelline envelop arrives at the perivitelline space. The sequential changes in the previtellogenic follicles of two species of caecilians are described. PMID:17345639

  6. Inhibition of phosphatase and tensin homologue (PTEN) in human ovary in vitro results in increased activation of primordial follicles but compromises development of growing follicles.

    PubMed

    McLaughlin, Marie; Kinnell, Hazel L; Anderson, Richard A; Telfer, Evelyn E

    2014-08-01

    In the mammalian ovary a small number of follicles are steadily recruited from the quiescent pool to undergo development. Follicle loss, maintenance and growth are strictly controlled by complex molecular interactions including the phosphoinositide 3-kinase (PI3K)-protein kinase B (Akt) signalling pathway. Stimulation of PI3K promotes phosphorylation of Akt resulting in follicle survival and activation of growth whereas this pathway is suppressed by the actions of the phosphatase and tensin homologue (PTEN). The aim of this study was to determine the effect of dipotassium bisperoxo(5-hydroxypyridine-2-carboxyl)oxovanadate (bpV), a reversible inhibitor of PTEN, on the activation, survival and development of human ovarian follicles in vitro. Biopsied ovarian tissue fragments were obtained from 17 women aged 23-46 years and exposed to 1 µM bpV(HOpic) (n = 146) or control medium (n = 128) for 24 h. Media were then replaced with control medium and all tissue incubated for a further 5 days. Ovarian tissue from each treatment group was fixed after the initial 24 h culture period and phosphorylated Akt was quantified by western blotting. After 6 days incubation all tissue fragments were inspected under light microscopy and any secondary follicles ≥100 µm isolated. Isolated follicles were cultured individually in control medium supplemented with 100 ng/ml recombinant human activin A. Tissue fragments without follicles suitable for isolation were fixed and processed for histological and immunohistochemical analysis. During 6 days culture, follicle activation occurred in tissue samples from both treatment groups but with significantly more follicles progressing to the secondary stage of development in the presence of 1 µM bpV(HOpic) compared with control (31 versus 16%; P < 0.05). Increased activation was associated with increased Akt phosphorylation and increased nuclear export of FOXO3. However isolated and cultured follicles that had been exposed to bpV(HOpic) showed

  7. Chromatin and cytoplasmic characteristics of equine oocytes recovered by transvaginal ultrasound-guided follicle aspiration are influenced by the developmental stage of their follicle of origin.

    PubMed

    Vernunft, Andreas; Alm, Hannelore; Tuchscherer, Armin; Kanitz, Wilhelm; Hinrichs, Katrin; Torner, Helmut

    2013-07-01

    Dynamic follicular changes occur during the equine estrus cycle, but little is known about their impact on the properties of recovered oocytes. The aim of this study was to characterize the cytoplasmic and chromatin status of equine oocytes in relation to the time of recovery during the follicle wave. Transvaginal ultrasound-guided follicle aspiration was performed two times in relation to the follicle wave: estrus-subordinate, from the subordinate follicles of mares in estrus, 24 hours after human chorionic gonadotropin stimulation of a dominant preovulatory follicle, and new-wave, from the follicles of the subsequent induced follicular wave, at the time of dominant follicle divergence (largest follicle 23 mm diameter). A total of 1011 follicles were aspirated. The oocyte recovery rate in the new-wave group was significantly lower than that for the estrus-subordinate group (12% vs. 26%, respectively); this was associated with a significantly higher proportion of oocytes with compact cumuli (44% vs. 27%, respectively). Estradiol concentrations were markedly higher in follicular fluid from new-wave follicles (885.6 ± 123.2 ng/mL vs. 54.3 ± 18.9 ng/mL, for estrus-subordinate; P < 0.001), indicating greater viability. Aspiration group did not affect glucose-6-phosphate dehydrogenase activity in recovered oocytes. Fibrillar (more juvenile) chromatin was more prevalent in new-wave oocytes, whereas estrus-subordinate oocytes showed more condensed chromatin or resumption of meiosis (P < 0.05). Mitochondrial activity was higher in oocytes with expanded cumuli in the new-wave group, but not in the estrus-subordinate group. In conclusion, our results clearly showed that the time of aspiration in relation to the follicle wave is associated with significant differences in follicle status and oocyte characteristics: new-wave oocytes were from a more viable follicle population and had more juvenile chromatin and cytoplasmic characteristics, whereas estrus-subordinate oocytes

  8. Inhibition of phosphatase and tensin homologue (PTEN) in human ovary in vitro results in increased activation of primordial follicles but compromises development of growing follicles

    PubMed Central

    McLaughlin, Marie; Kinnell, Hazel L.; Anderson, Richard A.; Telfer, Evelyn E.

    2014-01-01

    In the mammalian ovary a small number of follicles are steadily recruited from the quiescent pool to undergo development. Follicle loss, maintenance and growth are strictly controlled by complex molecular interactions including the phosphoinositide 3-kinase (PI3K)-protein kinase B (Akt) signalling pathway. Stimulation of PI3K promotes phosphorylation of Akt resulting in follicle survival and activation of growth whereas this pathway is suppressed by the actions of the phosphatase and tensin homologue (PTEN). The aim of this study was to determine the effect of dipotassium bisperoxo(5-hydroxypyridine-2-carboxyl)oxovanadate (bpV), a reversible inhibitor of PTEN, on the activation, survival and development of human ovarian follicles in vitro. Biopsied ovarian tissue fragments were obtained from 17 women aged 23–46 years and exposed to 1 µM bpV(HOpic) (n = 146) or control medium (n = 128) for 24 h. Media were then replaced with control medium and all tissue incubated for a further 5 days. Ovarian tissue from each treatment group was fixed after the initial 24 h culture period and phosphorylated Akt was quantified by western blotting. After 6 days incubation all tissue fragments were inspected under light microscopy and any secondary follicles ≥100 µm isolated. Isolated follicles were cultured individually in control medium supplemented with 100 ng/ml recombinant human activin A. Tissue fragments without follicles suitable for isolation were fixed and processed for histological and immunohistochemical analysis. During 6 days culture, follicle activation occurred in tissue samples from both treatment groups but with significantly more follicles progressing to the secondary stage of development in the presence of 1 µM bpV(HOpic) compared with control (31 versus 16%; P < 0.05). Increased activation was associated with increased Akt phosphorylation and increased nuclear export of FOXO3. However isolated and cultured follicles that had been exposed to bp

  9. Follicle Diameter and Systemic Hormone Interrelationships during Induction of Follicle Collapse with Intrafollicular Prostaglandin E2 and F2α in Mares.

    PubMed

    Martínez-Boví, R; Cuervo-Arango, J

    2016-04-01

    The objectives were to determine: (i) whether intrafollicular administration of PGE2 and PGF2α to mares would hasten follicle collapse and (ii) the differences in reproductive hormone characteristics in mares with spontaneous and prostaglandin-induced follicle collapses. Six mares were followed for two oestrous cycles each: when the mares reached a follicle diameter of 30-35 mm and showed mild-to-moderate endometrial oedema, mares were administered a single 0.5 ml dose containing 500 μg PGE2 and 125 μg PGF2α (treatment cycle) or a placebo (0.5 ml of water for injection; control cycle) into the preovulatory follicle (Hour 0). Blood samples were collected, and serial ultrasound examinations were performed until follicle collapse. Treated mares showed follicle collapse significantly earlier (20.0 ± 5.9 h) than the control mares (72.0 ± 10.7 h). The LH, progesterone, total oestrogens and oestradiol concentrations did not differ between groups; however, the progesterone concentration increased more between 48 and 72 h after follicle injection in the treatment compared to the control cycles (P < 0.05). In conclusion, intrafollicular treatment with PGE2 and PGF2α hastened follicle collapse in mares without the simultaneous use of an inductor of ovulation; despite the early induction of follicle collapse, the profiles of LH and oestradiol were not altered. This study provides information on the role of prostaglandins (PGs) in the process of follicle wall rupture and collapse and suggests that this may happen even before the beginning of the sharp rise in circulating LH at the final stage of the ovulatory surge.

  10. Proteomic analysis of mare follicular fluid during late follicle development

    PubMed Central

    2011-01-01

    Background Follicular fluid accumulates into the antrum of follicle from the early stage of follicle development. Studies on its components may contribute to a better understanding of the mechanisms underlying follicular development and oocyte quality. With this objective, we performed a proteomic analysis of mare follicular fluid. First, we hypothesized that proteins in follicular fluid may differ from those in the serum, and also may change during follicle development. Second, we used four different approaches of Immunodepletion and one enrichment method, in order to overcome the masking effect of high-abundance proteins present in the follicular fluid, and to identify those present in lower abundance. Finally, we compared our results with previous studies performed in mono-ovulant (human) and poly-ovulant (porcine and canine) species in an attempt to identify common and/or species-specific proteins. Methods Follicular fluid samples were collected from ovaries at three different stages of follicle development (early dominant, late dominant and preovulatory). Blood samples were also collected at each time. The proteomic analysis was carried out on crude, depleted and enriched follicular fluid by 2D-PAGE, 1D-PAGE and mass spectrometry. Results Total of 459 protein spots were visualized by 2D-PAGE of crude mare follicular fluid, with no difference among the three physiological stages. Thirty proteins were observed as differentially expressed between serum and follicular fluid. Enrichment method was found to be the most powerful method for detection and identification of low-abundance proteins from follicular fluid. Actually, we were able to identify 18 proteins in the crude follicular fluid, and as many as 113 in the enriched follicular fluid. Inhibins and a few other proteins involved in reproduction could only be identified after enrichment of follicular fluid, demonstrating the power of the method used. The comparison of proteins found in mare follicular fluid

  11. The GOAT Effect's Impact upon Educational R and D.

    ERIC Educational Resources Information Center

    Kean, Michael H.; McNamara, Thomas C.

    1979-01-01

    The "Goodbye To All That" (GOAT) Effect is introduced as a special research and evaluation "outcome" effect characterizing decision making unduly influenced by abandoning "write-off" tendencies. The "gradual refinement" approach offers an antidote to the GOAT Effect because it does not use the systems analysis approach. (Author/RL)

  12. ADAPTmap: International coordinated data resource for improving goat production effiency

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Goats provide vital food and economic security, particularly in developing countries. We created a database that is a nexus for all performance, type, geographic information system (GIS), production environment, and genome information on goats. This resource provides a platform for meta-analysis tha...

  13. [Veterinary aspects of the raising of mountain goats (Oreamnos americanus)].

    PubMed

    Seidel, B

    1991-03-01

    An empirical report outlines veterinary problems in Rocky Mountain Goats kept in climatic conditions of Central Europe. Described are therapeutic findings in the treatment of infections, parasitosis, disorders of the extremities, disturbances of reproduction, and injuries as well as haematological findings and experiences made during immobilization of Rocky Mountain Goats.

  14. Research update: finishing lambs and meat goat kids on pasture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditional sheep (Ovis aries), hair sheep and meat goat (Capra hircus) industries are growing rapidly in the Appalachian Region to help produce meats for ethnic markets. This niche market offers an economic opportunity for owners of small farms. Control of gastrointestinal (GI) parasites in goats...

  15. Ghrelin O-acyltransferase (GOAT) and energy metabolism.

    PubMed

    Li, Ziru; Mulholland, Michael; Zhang, Weizhen

    2016-03-01

    Ghrelin O-acyltransferase (GOAT), a member of MBOATs family, is essential for octanoylation of ghrelin, which is required for active ghrelin to bind with and activate its receptor. GOAT is expressed mainly in the stomach, pancreas and hypothalamus. Levels of GOAT are altered by energy status. GOAT contains 11 transmembrane helices and one reentrant loop. Its invariant residue His-338 and conserved Asn-307 are located in the endoplasmic reticulum lumen and cytosol respectively. GOAT contributes to the regulation of food intake and energy expenditure, as well as glucose and lipids homeostasis. Deletion of GOAT blocks the acylation of ghrelin leading to subsequent impairment in energy homeostasis and survival when mice are challenged with high energy diet or severe caloric restriction. GO-CoA-Tat, a peptide GOAT inhibitor, attenuates acyl-ghrelin production and prevents weight gain induced by a medium-chain triglycerides-rich high fat diet. Further, GO-CoA-Tat increases glucose- induced insulin secretion. Overall, inhibition of GOAT is a novel strategy for treatment of obesity and related metabolic disorders. PMID:26732975

  16. Milk production of West African Dwarf goats in the Gambia.

    PubMed

    Jaitner, J; Njie, M; Corr, N; Dempfle, L

    2006-04-01

    Goats are important in the low-input systems of West Africa and their main importance lies in their role for income and saving. In addition, it is known that milk offtake for home consumption is also important. In order to obtain information about the real importance of milk offtake, a recording scheme was operated in 27 villages in the Central River Division of The Gambia from July 1998 until January 2000. Detailed information was obtained from about 1500 kiddings. In the recording scheme, any sheep being milked as well as the goats of the International Trypanotolerance Centre nucleus flock were also recorded. In the villages, 36% of all lactations were used for milk offtake, but the fraction milked was lower for the first two lactations. The average length of lactation was 127 days and the average daily milk offtake was 0.18 L. Goats are milked once a day and the residual milk is left for the kids. Milking starts about one week after parturition and stops when the goat becomes pregnant or the kid(s) die or the goat is drying off. The repeatability of the 90-day milk offtake was 0.24 +/- 0.09. Sixty-five percent of goat owner were women and a large fraction of goat owners also owned cattle. Goat milk was used exclusively for home consumption. It is concluded that in breeding and extension work more attention should be given to aspects of milk production.

  17. The resolution of rayless goldenrod (Isocoma pluriflora) poisoning in goats

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rayless goldenrod (Isocoma pluriflora) occasionally poisons livestock causing myocardial and skeletal muscle degeneration and necrosis. The objectives of this study were to describe the resolution of the clinical and pathologic changes of rayless goldenrod poisoning in goats. Eight goats were gava...

  18. Dairy goat demography and Q fever infection dynamics

    PubMed Central

    2013-01-01

    Between 2007 and 2009, the largest human Q fever epidemic ever described occurred in the Netherlands. The source was traced back to dairy goat farms, where abortion storms had been observed since 2005. Since one putative cause of these abortion storms is the intensive husbandry systems in which the goats are kept, the objective of this study was to assess whether these could be explained by herd size, reproductive pattern and other demographic aspects of Dutch dairy goat herds alone. We adapted an existing, fully parameterized simulation model for Q fever transmission in French dairy cattle herds to represent the demographics typical for Dutch dairy goat herds. The original model represents the infection dynamics in a herd of 50 dairy cows after introduction of a single infected animal; the adapted model has 770 dairy goats. For a full comparison, herds of 770 cows and 50 goats were also modeled. The effects of herd size and goat versus cattle demographics on the probability of and time to extinction of the infection, environmental bacterial load and abortion rate were studied by simulation. The abortion storms could not be fully explained by demographics alone. Adequate data were lacking at the moment to attribute the difference to characteristics of the pathogen, host, within-herd environment, or a combination thereof. The probability of extinction was higher in goat herds than in cattle herds of the same size. The environmental contamination was highest within cattle herds, which may be taken into account when enlarging cattle farming systems. PMID:23621908

  19. Sarcocystis oreamni n. sp. from the mountain goat (oreamnos americanus)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Numerous species of Sarcocystis have been reported from wild ruminants but none has been named from the Rocky Mountain goat (Oreamnos americanus). Mature sarcocysts were found in frozen muscle samples of 3 of 7 mountain goats from Alaska, USA. Two morphological types of sarcocysts were found; 1 had ...

  20. Stimulation of regressing subordinate follicles of wave 2 with a gonadotropin product in heifers.

    PubMed

    Ginther, O J; Siddiqui, M A R; Baldrighi, J M; Hoffman, M M

    2016-04-01

    The recovery of regressing wave-2 subordinate follicles was studied by treating heifers with a gonadotropin product that had about 84% and 16% of follicle-stimulating hormone and luteinizing hormone activity, respectively. A treated group (n = 8) received a single dose of 50 mg (2.5 mL) of the gonadotropin product, and a control group (n = 8) received 2.5 mL of saline vehicle. The group assignment of heifers was not known to the ultrasonographer who tracked the follicles and measured follicle diameters. Follicle measurements began on the day of expected follicle deviation in wave 2 (largest follicle closest to 8.5 mm), and treatment (hour 0) was given on Day 13.4 ± 0.2 (Day 0 = ovulation) when the dominant follicles of waves 1 and 2 were 14.1 ± 0.3 mm and 10.7 ± 0.1 mm, respectively. Subordinate follicles of wave 2 that had regressed to a 3-mm category (3.0-3.9 mm) or 4-mm category by hour 0 decreased in diameter for at least 48 h before hour 0, whereas follicles that were in the 5-mm or 6-mm categories at hour 0 did not change significantly in diameter during the previous 48 h. About 55% of the follicles that had regressed to the 3-mm and 4-mm categories at hour 0% and 78% of the follicles in the 5-mm and 6-mm categories increased in diameter after gonadotropin treatment, whereas follicles in the control group continued to decrease (regress) in diameter. The follicles for each of the 4 diameter categories were greater (P < 0.05) in diameter 9 h after treatment in the treated group than in the control group. The dominant follicle of wave 1 and the largest subordinate follicle of wave 2 in the treated group also increased in diameter so that diameter was greater (P < 0.05) than in the controls at hour 9. The results demonstrated that subordinate follicles of wave 2 that had decreased in diameter (regressed) for at least 48 h retained the capability to recover as indicated by a diameter increase when exposed to a gonadotropin product. PMID:26773367

  1. The placenta shed from goats with classical scrapie is infectious to goat kids and lambs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Classical scrapie is a natural prion disease of sheep in which the immediate postpartum period and, in particular, the placenta have long been known to play key roles in natural horizontal transmission. Goats, too, are a natural host of classical scrapie and are frequently raised with sheep; but the...

  2. Inter- and intraspecific placentae in sheep, goats and sheep-goat chimaeras.

    PubMed

    MacLaren, L A; Anderson, G B; BonDurant, R H; Edmondson, A J

    1992-04-01

    These studies compared inter- and intraspecific placentae during implantation and at full-term in sheep, goats and interspecific sheep-goat chimaeras. Histological sections prepared from intra- and interspecific day-26, 32 and 36 placentae in ewes and does indicated an altered ability of the trophoblast to invade the maternal caruncle in interspecific pregnancies. Two sheep-in-goat pregnancies were less, and two goat-in-sheep pregnancies were more, invasive than homologous pregnancies. Caprine pregnancies in chimaeras generally terminated before timed samples could be obtained, but biopsy samples collected at laparotomy between days 42 and 48 demonstrated both normal and abnormal placentation in chimaeras after breedings to rams. In six of 11 full-term fetal placentae from ovine pregnancies in chimaeras, there was abnormal retention of maternal caruncular tissue to the extent that macroscopic lesions were visible on the surface of the fetal cotyledons. Histological observations indicated that proliferation of maternal septa and hyalinization of maternal vessels had occurred at the expense of the fetal villi. Overall, the results suggested that the physiological events that regulate implantation are different in the two species, despite anatomical similarities between the ovine and caprine placenta. The caprine conceptus is likely to be rejected in the ovine or chimaeric uterus because of its over-invasiveness in the early stages of implantation, whereas the ovine conceptus can survive in the potentially chimaeric uterus. PMID:1602061

  3. Molecular phylogeography and genetic diversity of East Asian goats.

    PubMed

    Lin, B Z; Odahara, S; Ishida, M; Kato, T; Sasazaki, S; Nozawa, K; Mannen, H

    2013-02-01

    The domestic goat is one of the most important livestock species, but its origins and genetic diversity still remain uncertain. Multiple highly divergent maternal lineages of goat have been reported in previous studies. Although one of the mitochondrial DNA lineages, lineage B, was detected only in eastern and southern Asia, the geographic distribution of these lineages was previously unclear. Here, we examine the genetic diversity and phylogeographic structure of Asian goats by mitochondrial DNA sequences and morphological characteristics. The analyses of a total of 1661 Asian goats from 12 countries revealed a high frequency of lineage B in Southeast Asia. The frequency of this lineage tended to be higher in mountain areas than in plain areas in Southeast Asian countries, and there was a significant correlation between its frequency and morphological traits. The results suggest an original predominance of lineage B in Southeast Asia and the recent infiltration of lineage A into Southeast Asian goats. PMID:22524237

  4. Epidemiological Observations on Cryptosporidiosis in Diarrheic Goat Kids in Greece.

    PubMed

    Giadinis, Nektarios D; Papadopoulos, Elias; Lafi, Shawkat Q; Papanikolopoulou, Vasiliki; Karanikola, Sofia; Diakou, Anastasia; Vergidis, Vergos; Xiao, Lihua; Ioannidou, Evi; Karatzias, Harilaos

    2015-01-01

    This study aimed at investigating the occurrence of Cryptosporidium spp. in diarrheic goat kids in Greece and the risk factors associated with cryptosporidiosis. Altogether, 292 diarrheic 4-15-day-old goat kids from 54 dairy goat herds of Northern Greece were examined. Oocysts of Cryptosporidium spp. were detected in 223 of 292 (76.4%) goat kids and the intensity of infection was scored as "high" in 142 samples, "moderate" in 45 samples, and "low" in 36 samples. Larger herds (>200 animals) had higher infection rates than smaller ones, although this difference was not statistically significant. Significantly higher infection rates were observed in herds during late kidding season (1 January to 30 April) compared to the early one (1 September to 31 December). These results suggest that cryptosporidiosis is very common in diarrheic goat kids in Greece, especially in large herds during the late parturition season.

  5. Molecular phylogeography and genetic diversity of East Asian goats.

    PubMed

    Lin, B Z; Odahara, S; Ishida, M; Kato, T; Sasazaki, S; Nozawa, K; Mannen, H

    2013-02-01

    The domestic goat is one of the most important livestock species, but its origins and genetic diversity still remain uncertain. Multiple highly divergent maternal lineages of goat have been reported in previous studies. Although one of the mitochondrial DNA lineages, lineage B, was detected only in eastern and southern Asia, the geographic distribution of these lineages was previously unclear. Here, we examine the genetic diversity and phylogeographic structure of Asian goats by mitochondrial DNA sequences and morphological characteristics. The analyses of a total of 1661 Asian goats from 12 countries revealed a high frequency of lineage B in Southeast Asia. The frequency of this lineage tended to be higher in mountain areas than in plain areas in Southeast Asian countries, and there was a significant correlation between its frequency and morphological traits. The results suggest an original predominance of lineage B in Southeast Asia and the recent infiltration of lineage A into Southeast Asian goats.

  6. Epidemiological Observations on Cryptosporidiosis in Diarrheic Goat Kids in Greece

    PubMed Central

    Giadinis, Nektarios D.; Papadopoulos, Elias; Lafi, Shawkat Q.; Papanikolopoulou, Vasiliki; Karanikola, Sofia; Diakou, Anastasia; Vergidis, Vergos; Xiao, Lihua; Ioannidou, Evi; Karatzias, Harilaos

    2015-01-01

    This study aimed at investigating the occurrence of Cryptosporidium spp. in diarrheic goat kids in Greece and the risk factors associated with cryptosporidiosis. Altogether, 292 diarrheic 4–15-day-old goat kids from 54 dairy goat herds of Northern Greece were examined. Oocysts of Cryptosporidium spp. were detected in 223 of 292 (76.4%) goat kids and the intensity of infection was scored as “high” in 142 samples, “moderate” in 45 samples, and “low” in 36 samples. Larger herds (>200 animals) had higher infection rates than smaller ones, although this difference was not statistically significant. Significantly higher infection rates were observed in herds during late kidding season (1 January to 30 April) compared to the early one (1 September to 31 December). These results suggest that cryptosporidiosis is very common in diarrheic goat kids in Greece, especially in large herds during the late parturition season. PMID:26819799

  7. Cytogenetic analysis of experimental interspecies goat-sheep chimera.

    PubMed

    Jaszczak, K; Członkowska, M; Guszkiewicz, A; Parada, R

    1991-01-01

    Chromosomal analysis was carried out on blood lymphocytes, skin fibroblasts, and germinal cells of an interspecies goat-sheep chimera. This chimera was produced by aggregation of blastomeres of goat and sheep embryos. A cell chimerism 54,XX/60,XY was found in blood lymphocytes and skin fibroblasts. At birth the percentage of lymphocytes with karyotype 54,XX (sheep) amounted to 80% and with karyotype 60,XY (goat) to 20%. With age the percentage of lymphocytes with chromosome complement 54,XX increased, so that at 18 months it was 94% sheep and 6% goat. At the same age, in skin fibroblasts the percentage of cells with goat karyotype reached 25%. Analysis of germinal cells showed in spermatogonia the presence of only karyotype 60,XY and in primary spermatocytes of 29 autosomal bivalents and the sex bivalent XY. PMID:2061596

  8. Epidemiological Observations on Cryptosporidiosis in Diarrheic Goat Kids in Greece.

    PubMed

    Giadinis, Nektarios D; Papadopoulos, Elias; Lafi, Shawkat Q; Papanikolopoulou, Vasiliki; Karanikola, Sofia; Diakou, Anastasia; Vergidis, Vergos; Xiao, Lihua; Ioannidou, Evi; Karatzias, Harilaos

    2015-01-01

    This study aimed at investigating the occurrence of Cryptosporidium spp. in diarrheic goat kids in Greece and the risk factors associated with cryptosporidiosis. Altogether, 292 diarrheic 4-15-day-old goat kids from 54 dairy goat herds of Northern Greece were examined. Oocysts of Cryptosporidium spp. were detected in 223 of 292 (76.4%) goat kids and the intensity of infection was scored as "high" in 142 samples, "moderate" in 45 samples, and "low" in 36 samples. Larger herds (>200 animals) had higher infection rates than smaller ones, although this difference was not statistically significant. Significantly higher infection rates were observed in herds during late kidding season (1 January to 30 April) compared to the early one (1 September to 31 December). These results suggest that cryptosporidiosis is very common in diarrheic goat kids in Greece, especially in large herds during the late parturition season. PMID:26819799

  9. Goat milk acceptance and promotion methods in Japan: The questionnaire survey to middle class households.

    PubMed

    Ozawa, Takeyuki; Mukuda, Kumiko; Fujita, Masaru; Nishitani, Jiro

    2009-04-01

    A consumer questionnaire conducted with the purpose of ascertaining the acceptability of goat milk and related products in Japan was carried out on 345 guarantees of Nippon Veterinary and Life Science University in December 2006. 275 effective responses (79%) representing middle class urban households were returned. The results revealed that (1) 30% of respondents have experienced drinking goat milk and only 10% are aware of the current retail situation of goat milk and related products; (2) over 70% of goat milk drinkers raised goats by hand at some point in their past and their first experience drinking goat milk was in infancy; (3) those with experience in drinking goat milk expressed a vague evaluation and minimal understanding of drinking goat milk; (4) respondents who were inexperienced goat milk drinkers expressed a strong desire to taste and a weak desire to purchase goat milk; (5) respondents expressed low recognition regarding retailed goat milk products, but those who had already purchased goat milk products expressed a high evaluation and strong desire to purchase these products again; and (6) recognition of goat milk characteristics is low, but those with high recognition also rate goat milk highly. Goats are perceived as being 'mild and familiar.' It is necessary for those who manage goat husbandry to present goat milk and related product tasting opportunities to consumers. The key point is to make the functional differences between cow and goat milk clear and present the advantages of goat milk at the fore of this promotion. Goat milk should not be promoted merely as a drink that is similar to cow milk, but must be positioned as a functional drink or health food in order to expand the Japanese goat milk market.

  10. Goat milk acceptance and promotion methods in Japan: The questionnaire survey to middle class households.

    PubMed

    Ozawa, Takeyuki; Mukuda, Kumiko; Fujita, Masaru; Nishitani, Jiro

    2009-04-01

    A consumer questionnaire conducted with the purpose of ascertaining the acceptability of goat milk and related products in Japan was carried out on 345 guarantees of Nippon Veterinary and Life Science University in December 2006. 275 effective responses (79%) representing middle class urban households were returned. The results revealed that (1) 30% of respondents have experienced drinking goat milk and only 10% are aware of the current retail situation of goat milk and related products; (2) over 70% of goat milk drinkers raised goats by hand at some point in their past and their first experience drinking goat milk was in infancy; (3) those with experience in drinking goat milk expressed a vague evaluation and minimal understanding of drinking goat milk; (4) respondents who were inexperienced goat milk drinkers expressed a strong desire to taste and a weak desire to purchase goat milk; (5) respondents expressed low recognition regarding retailed goat milk products, but those who had already purchased goat milk products expressed a high evaluation and strong desire to purchase these products again; and (6) recognition of goat milk characteristics is low, but those with high recognition also rate goat milk highly. Goats are perceived as being 'mild and familiar.' It is necessary for those who manage goat husbandry to present goat milk and related product tasting opportunities to consumers. The key point is to make the functional differences between cow and goat milk clear and present the advantages of goat milk at the fore of this promotion. Goat milk should not be promoted merely as a drink that is similar to cow milk, but must be positioned as a functional drink or health food in order to expand the Japanese goat milk market. PMID:20163593

  11. Phagocytosis of sperm by follicle cells of the carnivorous sponge Asbestopluma occidentalis (Porifera, Demospongiae).

    PubMed

    Riesgo, Ana

    2010-06-01

    During spermatogenesis of the carnivorous sponge Asbestopluma occidentalis, follicle cells that lined the spermatocysts phagocytosed unreleased mature sperm. Such follicle cells are part of the complex envelope that limits spermatocysts of A. occidentalis, which is also comprised of a collagen layer, a thick layer of intertwined cells, and spicules. Follicle cells showed vesicles containing single phagocytosed spermatozoa within their cytoplasm. Additionally, lipids and other inclusions were observed within the cytoplasm of follicle cells. It is likely that follicle cells recapture nutrients by phagocytosing spermatozoa and use them to form lipids and other inclusions. Such sperm phagocytosis is usually performed in higher invertebrates and vertebrates by Sertoli cells that are located in the testis wall. While Sertoli cells develop a wide range of functions such as creating a blood-testis barrier, providing crucial factors to ensure correct progression of spermatogenesis, and phagocytosis of aberrant, degenerating, and unreleased sperm cells, sponge follicle cells may only display phagocytotic activity on spermatogenic cells.

  12. Canonical and non-canonical Wnt signaling control the regeneration of amputated rodent vibrissae follicles.

    PubMed

    Yuan, Yan-Ping; Huang, Keng; Xu, Yan-Min; Chen, Xian-Cai; Li, Hai-Hong; Cai, Bo-Zhi; Liu, Yang; Zhang, Huan; Li, Yu; Lin, Chang-Min

    2016-02-01

    Although mammals are notoriously poor at regeneration compared with many lower-order species, the hair follicle, particular to mammals, is capable of regeneration following partial amputation. The detailed internal mechanism of this phenomenon is still unclear. Development and regrowth of the hair follicle depends on dermal-epidermal interaction within the hair follicle. Previous studies have shown that Wnt/β-catenin, Shh, Bmp, PDGF, TGF and Notch signals all take part in the development and growth of the hair follicle, and the Wnt/β-catenin signaling additionally plays an indispensable role in hair follicle morphogenesis and regrowth. In this study, we investigated the localization, as well as, protein levels of Wnt/β-catenin signaling molecules during amputated whisker follicle regeneration. PMID:26742765

  13. 9 CFR 79.2 - Identification of sheep and goats in interstate commerce.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Identification of sheep and goats in... SCRAPIE IN SHEEP AND GOATS § 79.2 Identification of sheep and goats in interstate commerce. (a) No sheep or goat that is required to be individually identified or premises identified by § 79.3 may be...

  14. 9 CFR 51.28 - Moving goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Moving goats, sheep, and horses to be... DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.28 Moving goats, sheep, and horses to be destroyed. Goats, sheep, and horses to be destroyed because of brucellosis must...

  15. 9 CFR 51.27 - Identification of goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Identification of goats, sheep, and... DISEASES ANIMALS DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.27 Identification of goats, sheep, and horses to be destroyed. The claimant must ensure that any goats, sheep,...

  16. 9 CFR 79.2 - Identification of sheep and goats in interstate commerce.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Identification of sheep and goats in... SCRAPIE IN SHEEP AND GOATS § 79.2 Identification of sheep and goats in interstate commerce. (a) No sheep or goat that is required to be individually identified or premises identified by § 79.3 may be...

  17. 9 CFR 51.28 - Moving goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Moving goats, sheep, and horses to be... DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.28 Moving goats, sheep, and horses to be destroyed. Goats, sheep, and horses to be destroyed because of brucellosis must...

  18. 9 CFR 51.27 - Identification of goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Identification of goats, sheep, and... DISEASES ANIMALS DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.27 Identification of goats, sheep, and horses to be destroyed. The claimant must ensure that any goats, sheep,...

  19. 9 CFR 51.27 - Identification of goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Identification of goats, sheep, and... DISEASES ANIMALS DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.27 Identification of goats, sheep, and horses to be destroyed. The claimant must ensure that any goats, sheep,...

  20. 9 CFR 51.27 - Identification of goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Identification of goats, sheep, and... DISEASES ANIMALS DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.27 Identification of goats, sheep, and horses to be destroyed. The claimant must ensure that any goats, sheep,...

  1. 9 CFR 51.28 - Moving goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Moving goats, sheep, and horses to be... DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.28 Moving goats, sheep, and horses to be destroyed. Goats, sheep, and horses to be destroyed because of brucellosis must...

  2. 9 CFR 51.28 - Moving goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Moving goats, sheep, and horses to be... DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.28 Moving goats, sheep, and horses to be destroyed. Goats, sheep, and horses to be destroyed because of brucellosis must...

  3. 9 CFR 51.28 - Moving goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Moving goats, sheep, and horses to be... DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.28 Moving goats, sheep, and horses to be destroyed. Goats, sheep, and horses to be destroyed because of brucellosis must...

  4. Major proteins of the goat milk fat globule membrane.

    PubMed

    Cebo, C; Caillat, H; Bouvier, F; Martin, P

    2010-03-01

    Fat is present in milk as droplets of triglycerides surrounded by a complex membrane derived from the mammary epithelial cell called milk fat globule membrane (MFGM). Although numerous studies have been published on human or bovine MFGM proteins, to date few studies exist on MFGM proteins from goat milk. The objective of this study was thus to investigate the protein composition of the goat MFGM. Milk fat globule membrane proteins from goat milk were separated by 6% and 10% sodium dodecyl sulfate-PAGE and were Coomassie or periodic acid-Schiff stained. Most of MFGM proteins [mucin-1, fatty acid synthase, xanthine oxidase, butyrophilin, lactadherin (MFG EGF-8, MFG-E8), and adipophilin] already described in cow milk were identified in goat milk using peptide mass fingerprinting. In addition, lectin staining provided a preliminary characterization of carbohydrate structures occurring on MFGM proteins from goat milk depending on alpha(S1)-casein genotype and lactation stage. We provide here first evidence of the presence of O-glycans on fatty acid synthase and xanthine oxidase from goat milk. A prominent difference between the cow and the goat species was demonstrated for lactadherin. Indeed, whereas 2 polypeptide chains were easily identified by peptide mass fingerprinting matrix-assisted laser desorption/ionization-time of flight analysis within bovine MFGM proteins, lactadherin from goat milk consisted of a single polypeptide chain. Another striking observation was the presence of caseins associated with MFGM preparations from goat milk, whereas virtually no caseins were found in MFGM extracts from bovine milk. Taken together, these observations strongly support the existence of a singular secretion mode previously hypothesized in the goat.

  5. Peripheral plasma levels of progesterone in pregnant goats and in pregnant goats treated with prostaglandin F2a.

    PubMed

    Bosu, W T; Serna Garibay, J A; Barker, C A

    1979-02-01

    Prostaglandin or prostaglandin analogues have been shown to be luteolytic in the pregnant goat. In this study the temporal changes in the plasma concentrations of progesterone during pregnancy and after administration of PGF2a to pregnant goats are described. PGF2a administration to pregnant goats at 30 and 65 days after breeding induced abortion within 34 to 75 hours. These abortions were accompanied by estrus and profuse muco-hemorrhagic discharges. When PGF2a was administered to pregnant goats 140 or 142 days after breeding, premature parturition occurred within 42 to 76 hours. Live kids were delivered in all cases. The plasma levels of progesterone in all pregnant goats showed dramatic decreases within 24 hours after the prostaglandin injections and continued to decrease gradually until abortions or premature parturition. Thereafter, the progesterone levels remained low for several days. PMID:16725398

  6. Isolation and Culture of Neural Crest Stem Cells from Human Hair Follicles.

    PubMed

    Yang, Ruifeng; Xu, Xiaowei

    2016-01-01

    The hair follicle undergoes lifelong cycling and growth. Previous studies have been focused on epithelial stem cells in the hair follicles. Neural crest stem cells (NCSCs) are pluripotent cells that can persist in adult tissues. We have previously demonstrated that human NCSCs can be isolated from hair follicles. Here, we present a protocol to isolate NCSCs from human hair follicles based on their specific surface-marker expression of CD271/HNK1 or CD271/CD49D (alpha4 integrin). NCSCs can be expanded in the culture as neural spheres or attached cells.

  7. Fibrin-Mediated Delivery of an Ovarian Follicle Pool in a Mouse Model of Infertility

    PubMed Central

    Smith, Rachel M.; Shikanov, Ariella; Kniazeva, Ekaterina; Ramadurai, Deepa; Woodruff, Teresa K.

    2014-01-01

    The cryopreservation and autotransplantation of ovarian tissue is emerging as a powerful approach for preserving fertility. However, for cancer patients, it may not be possible to transplant ovarian tissue due to the risk of re-seeding disease. We investigated strategies for transplantation of individually isolated follicles to minimize the risk of re-introducing cancer cells present within the vasculature of ovarian stroma. Procedures for large-scale isolation of early-stage follicles and their encapsulation into fibrin hydrogels were developed. For in vivo validation studies, mice were ovariectomized and transplanted with encapsulated follicles into the ovarian bursa. A substantial increase in the number of secondary follicles was observed in the graft at 9 days after transplantation, and antral follicles by day 21, demonstrating primordial follicle recruitment into the growing pool. Initially, elevated follicle-stimulating hormone levels declined substantially by day 21, indicating feedback from the graft; presence of corpora lutea showed the graft's capability of restoring hormone cyclicity. Taken together, the transplanted follicles were able to engraft, mature, and restore ovarian function in an infertile mouse. This biomaterial may, thus, provide a platform for follicle transplantation with a low risk of cancer contamination and for developing strategies that preserve fertility for women facing a cancer diagnosis. PMID:24802617

  8. Activin B promotes initiation and development of hair follicles in mice.

    PubMed

    Jia, Qin; Zhang, Min; Kong, Yanan; Chen, Shixuan; Chen, Yinghua; Wang, Xueer; Zhang, Lei; Lang, Weiya; Zhang, Lu; Zhang, Lin

    2013-01-01

    Activin B has been reported to promote the regeneration of hair follicles during wound healing. However, its role in the development and life cycle of hair follicles has not been elucidated. In our study, the effect of activin B on mouse hair follicles of cultured and neonatal mouse skin was investigated. In these models, PBS or activin B (5, 10 or 50 ng/ml) was applied, and hair follicle development was monitored. Hair follicle initiation and development was examined using hematoxylin and eosin staining, alkaline phosphatase activity staining, Oil Red O+ staining, and the detection of TdT-mediated dUTP-biotin nick end-labeling cell apoptosis. Activin B was found to efficiently induce the initiation of hair follicles in the skin of both cultured and neonatal mice and to promote the development of hair follicles in neonatal mouse skin. Moreover, activin-B-treated hair follicles were observed to enter the anagen stage from the telogen stage and to remain in the anagen stage. These results demonstrate that activin B promotes the initiation and development of hair follicles in mice.

  9. Syndecan-1 is strongly expressed in the anagen hair follicle outer root sheath and in the dermal papilla but expression diminishes with involution of the hair follicle.

    PubMed

    Bayer-Garner, Ilene B; Sanderson, Ralph D; Smoller, Bruce R

    2002-12-01

    Syndecan-1 is the prototypic member of a family of heparan sulfate-bearing cell surface proteoglycans that function in adhesion, cell-extracellular matrix interactions, migration, and proliferation. During embryogenesis, syndecan-1 expression in the epithelium is downregulated when the epithelium gives rise to motile mesenchymal cells, whereas mesenchymal syndecan-1 expression is upregulated during organ formation. In aggressive basal cell carcinomas, syndecan-1 expression is evident in the stroma. Some neoplastic cells induce stroma to meet needs for growth, and it may be the mesenchymal cells that produce and shed syndecan-1 into the stroma. The physiologic mechanism by which the hair follicle undergoes its cyclic process of involution and formation of a new active hair follicle is not well understood. Sixty scalp biopsies and a large scalp resection were evaluated for syndecan-1 expression within hair follicles in the growing (anagen), involuting (catagen), and resting (telogen) phases. Strong syndecan-1 immunoreactivity was evident in the outer root sheath (ORS) of the anagen hair follicle, but this expression diminished in intensity with the involution and resting stages in the hair follicle cycle. The diminution of syndecan-1 immunoreactivity in the ORS of involuting and resting hair follicles may be a result of terminal keratinocyte differentiation. Syndecan-1 was also present in the dermal papilla of the anagen hair follicle, where it may promote growth factor-mediated cell signaling that induces and maintains growth of the hair shaft and the inner root sheath.

  10. Cardiac troponin I in healthy newborn goat kids and in goat kids with cardiac nutritional muscular dystrophy.

    PubMed

    Tharwat, Mohamed; Al-Sobayil, Fahd; El-Sayed, Mehana

    2013-12-01

    This study was designed to establish serum cardiac troponin I (cTnI) concentrations in healthy newborn goat kids and in those with cardiac nutritional muscular dystrophy (NMD). Thirty-five single full-term newborn goat kids (20 males and 15 females; age: 6.1 ± 3.5 h; weight 3.4 ± 0.68 kg), together with their respective mothers (Group 1; G1) were enrolled consecutively. Thirty-one goat kids (age: 9.5 ± 4.3 days) with NMD, together with 20 control goat kids (age: 7.8 ± 4.3 days) were also included in this study (Group 2; G2). Blood samples were collected from G1 within 12 h of birth and from G2 on admission. Serum samples were collected and analysed for cTnI. In G1, the mean serum concentration of cTnI in goat kids was 0.290 ± 0.37 ng/mL, with no statistically significant difference between male and female kids (P = 0.61). The mean cTnI concentration in the does was 0.017 ± 0.04, ng/mL. Serum values of cTnI in the goat kids and in their respective mothers differed significantly (P = 0.0001). In G2, the mean cTnI concentration was 0.02 ± 0.05 ng/mL in the control and 11.18 ± 20.07 ng/mL in the diseased goat kids, with a statistically significant difference between diseased and control goat kids (P = 0.017). Serum concentrations of cTnI are higher in goat kids than in their respective mothers. In conclusion, the cTnI assay appears to be a sensitive and specific marker for myocardial injury in goat kids.

  11. Changes in the ovarian dynamics and endocrine profiles in goats treated with a progesterone antagonist during the early luteal phase of the estrous cycle.

    PubMed

    Suganuma, Chiho; Kuroiwa, Takenobu; Tanaka, Tomomi; Kamomae, Hideo

    2007-10-01

    The aim of the present study was to determine the physiological role of endogenous progesterone in the regulation of ovarian dynamics, gonadotropin and progesterone secretion during the early luteal phase in the goat. Cycling Shiba goats received subcutaneously a vehicle (control group, n=5) or 50 mg of RU486 (RU486 group, n=4) daily from 1 to 7 days after ovulation (day 0) determined by transrectal ultrasonography. Ovarian dynamics were monitored by the ultrasonography and blood samples were collected daily until the subsequent ovulation for analysis of progesterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion. Blood samples were also collected at 10 min intervals for 6 h on day 3 and day 7 for the analysis of pulsatile patterns of LH and FSH. The LH pulse frequency was significantly (P<0.05) higher in the RU486 group than in the control group on day 7 (4.8+/-1.1 pulses/6 h versus 1.2+/-0.4 pulses/6 h). The shape of the FSH pulses was unclear on day 3 and day 7 in both groups and the overall means of FSH concentration for 6 h on day 3 and day 7 were not significantly different between the RU486 and the control groups. The pattern of daily FSH concentrations showed a wave-like fluctuation in both groups. There was no significant difference in the inter-peak intervals of the wave-like pattern of daily FSH secretion between the RU486 and the control groups (4.1+/-0.6 days versus 4.5+/-0.6 days). The maximum diameter of the largest follicle that grew from day 1 to day 7 in the RU486 group tended to be greater than that in control goats (6.4+/-0.8 mm versus 5.0+/-0.8 mm, P=0.050), whereas no significant difference was detected in the size of the corpus luteum and progesterone concentrations between the control and RU486 groups on almost all days during the treatment period. These results indicate that the rise of the progesterone concentration suppresses the pulsatile LH secretion and follicular growth, whereas progesterone has no

  12. Structural changes occurring during atresia in sheep ovarian follicles.

    PubMed

    Hay, M R; Cran, D G; Moor, R M

    1976-07-01

    The structural changes that characterize primary, secondary and tertiary atresia in sheep Graafian follicles have been studied by means of histological, histochemical and ultrastructural techniques. In primary atresia vacuoles representing swollen endoplasmic reticulum are prominent along the antral border together with disorganized granulosa cells containing pyknotic nuclei. Phagocytic cells, which increase in number as atresia progresses, were seen within the membrana granulosa and are considered to be transformed granulosa cells. Even in follicles classified as nonatretic, a few antral vacuoles and occasional pyknotic nuclei are present. During secondary atresia there is a large increase in the number of cells with pyknotic nuclei; many of these nuclei had been extruded and had fused to form the characteristic Feulgen-positive atretic bodies found along the edge of the antral cavity. These bodies usually have a diameter of up to 15 mum but occasionally reached as much as 400 mum. A second area of degeneration is frequently present in the membrana granulosa, two or three cell layers from the basal lamina, and it is at this level that exfoliation of granulosa cells occurs in tertiary atresia. In contrast to the membrana granulosa, there are during secondary atresia, only slight indications of degeneration in the cumulus. In tertiary atresia the membrana granulosa is highly disorganized; the atretic bodies are often fewer in number than at earlier stages. The basal lamina remains essentially intact. It is at this stage that the first clear signs of degeneration occur in the theca interna. Despite some disintegration of the cumulus, the integrity of the oocyte is maintained and its nucleus remains vesicular. Changes in the thecal microcirculation may plan a key role in atresia: adjacent to the basal lamina of non-atretic follicles, there is a well-developed capillary network which is significantly reduced as atresia progresses. PMID:991198

  13. PHYSICOCHEMICAL CHARACTERIZATION OF PITUITARY FOLLICLE-STIMULATING HORMONE

    PubMed Central

    Li, Choh Hao; Pedersen, Kai O.

    1952-01-01

    The physiochemical characteristics of the follicle-stimulating hormone (FSH) from whole sheep pituitary glands have been studied. The hormone behaves as a single protein in electrophoresis, diffusion, and ultracentrifugation. It has an isoelectric point at pH 4.5 and a molecular weight of 67,000 and contains 1.23 per cent hexose and 1.51 per cent hexosamine. The amino acid composition has also been determined in large part. The stability of the hormone to acid and heat has been investigated. PMID:14898040

  14. Multiple teeth in a single dentigerous cyst follicle: A perplexity

    PubMed Central

    Agrawal, Mamta; Raghavendra, Pramod D. S.; Singh, Bhawana; Agrawal, Neha

    2011-01-01

    Dentigerous cysts are always associated with an embedded or unerupted tooth; involvement of more than one permanent tooth in a single cyst follicle is exceedingly rare and only a few such cases can be found in the literature. The cysts reported in the literature involving multiple teeth typically consist of mesiodens or odontomas. Here we report a rare case of dentigerous cyst associated with two permanent teeth causing displacement of maxillary canine into the osteo meatal complex and maxillary premolar to lateral nasal wall. The possible etiopathogenesis of such cases is also discussed here. PMID:23483109

  15. Are cattle, sheep, and goats endangered species?

    PubMed

    Taberlet, P; Valentini, A; Rezaei, H R; Naderi, S; Pompanon, F; Negrini, R; Ajmone-Marsan, P

    2008-01-01

    For about 10 000 years, farmers have been managing cattle, sheep, and goats in a sustainable way, leading to animals that are well adapted to the local conditions. About 200 years ago, the situation started to change dramatically, with the rise of the concept of breed. All animals from the same breed began to be selected for the same phenotypic characteristics, and reproduction among breeds was seriously reduced. This corresponded to a strong fragmentation of the initial populations. A few decades ago, the selection pressures were increased again in order to further improve productivity, without enough emphasis on the preservation of the overall genetic diversity. The efficiency of modern selection methods successfully increased the production, but with a dramatic loss of genetic variability. Many industrial breeds now suffer from inbreeding, with effective population sizes falling below 50. With the development of these industrial breeds came economic pressure on farmers to abandon their traditional breeds, and many of these have recently become extinct as a result. This means that genetic resources in cattle, sheep, and goats are highly endangered, particularly in developed countries. It is therefore important to take measures that promote a sustainable management of these genetic resources; first, by in situ preservation of endangered breeds; second, by using selection programmes to restore the genetic diversity of industrial breeds; and finally, by protecting the wild relatives that might provide useful genetic resources. PMID:17927711

  16. Conservation genetics of cattle, sheep, and goats.

    PubMed

    Taberlet, Pierre; Coissac, Eric; Pansu, Johan; Pompanon, François

    2011-03-01

    Cattle, sheep and goats were domesticated about 10,000 years ago, spread out of the domestication centers in Europe, Asia, and Africa during the next few thousands years, and gave many populations locally adapted. After a very long period of soft selection, the situation changed dramatically 200 years ago with the emergence of the breed concept. The selection pressure strongly increased, and the reproduction among breeds was seriously reduced, leading to the fragmentation of the initial gene pool. More recently, the selection pressure was increased again via the use of artificial insemination, leading to a few industrial breeds with very high performances, but with low effective population sizes. Beside this performance improvement of industrial breeds, genetic resources are being lost, because of the replacement of traditional breeds by high performance industrial breeds at the worldwide level, and because of the loss of genetic diversity in these industrial breeds. Many breeds are already extinct, and genetic resources in cattle, sheep, and goats are thus highly endangered, particularly in developed countries. The recent development of next generation sequencing technologies opens new avenues for properly characterizing the genetic resources, not only in the very diverse domestic breeds, but also in their wild relatives. Based on sound genetic characterization, urgent conservation measures must be taken to avoid an irremediable loss of farm animal genetic resources, integrating economical, sociological, and political parameters. PMID:21377620

  17. Microbiological quality of goat's milk obtained under different production systems.

    PubMed

    Kyozaire, J K; Veary, C M; Petzer, I M; Donkin, E F

    2005-06-01

    In order to determine the safety of milk produced by smallholder dairy goat farms, a farm-based research study was conducted on commercial dairy goat farms to compare the microbiological quality of milk produced using 3 different types of dairy goat production systems (intensive, semi-intensive and extensive). A survey of dairy goat farms in and around Pretoria carried out by means of a questionnaire revealed that most of the smallholder dairy goat farms surveyed used an extensive type of production system. The method of milking varied with the type of production system, i.e. machine milking; bucket system machine milking and hand-milking, respectively. Udder half milk samples (n=270) were analysed, of which 31.1% were infected with bacteria. The lowest intra-mammary infection was found amongst goats in the herd under the extensive system (13.3%), compared with 43.3% and 36.7% infection rates under the intensive and semi-intensive production systems, respectively. Staphylococcus intermedius (coagulase positive), Staphylococcus epidermidis and Staphylococcus simulans (both coagulase negative), were the most common cause of intramammary infection with a prevalence of 85.7% of the infected udder halves. The remaining 14.3% of the infection was due to Staphylococcus aureus. Bacteriology of bulk milk samples on the other hand, showed that raw milk obtained by the bucket system milking machine had the lowest total bacterial count (16,450 colony forming units (CFU)/ml) compared to that by pipeline milking machine (36,300 CFU/ml) or hand-milking (48,000 CFU/ml). No significant relationship was found between the somatic cell counts (SCC) and presence of bacterial infection in goat milk In comparison with the herds under the other 2 production systems, it was shown that dairy goat farming under the extensive production system, where hand-milking was used, can be adequate for the production of safe raw goat milk.

  18. Atresia revisited: two basic patterns of atresia of bovine antral follicles.

    PubMed

    Irving-Rodgers, H F; van Wezel, I L; Mussard, M L; Kinder, J E; Rodgers, R J

    2001-11-01

    Our observations of bovine follicles indicated that the original histological classifications of atresia were inaccurate. A detailed histological, ultrastructural and immunohistochemical study of antral follicles from bovine ovaries collected from an abattoir and from animals whose large follicles had been monitored by ultrasonography was conducted to investigate this further. Nidogen and CD68 were immunolocalized to observe the follicular basal lamina and macrophages, respectively. In randomly collected ovaries, approximately one quarter of all antral follicles were undergoing antral atresia, as designated in this study. Antral atresia was characterized by early destruction of the layers of the membrana granulosa closest to the antrum, whereas the most basal cells remained intact. Numerous pyknotic nuclei were observed in the most antral layers and in the antrum close to the membrana granulosa. This is the classic description of atretic follicles and was observed at all sizes of follicle development and almost universally in large follicles (> 5 mm in diameter), including dominant follicles. Basal atretic follicles, as designated in this study, were almost as prevalent as the antral atretic follicles, and were characterized by initial destruction of the most basal layer of granulosa cells, whereas the cells in the most antral layers remained associated with each other and were predominantly healthy. Pyknotic nuclei and the nuclei of dying basal cells budded into apoptotic bodies were observed rarely. The basal lamina of basal atretic follicles was often breached by macrophages, which were phagocytosing dying basal granulosa cells. The theca was characterized by an increased deposition of collagen, and the cells were orientated randomly, rather than lying parallel to the membrana granulosa as in healthy follicles. Basal atresia occurred in small (< 5 mm in diameter) follicles only. Importantly, these basal atretic follicles were originally identified incorrectly in

  19. Lymphocyte Depletion in Ileal Peyer’s Patch Follicles in Lambs Infected with Eimeria ovinoidalis

    PubMed Central

    Aleksandersen, Mona; Lie, Kai-Inge; Gjerde, Bjørn; Landsverk, Thor

    2002-01-01

    A total of 14 lambs were experimentally infected with Eimeria ovinoidalis in two separate experiments in two consecutive years. Nine lambs served as uninoculated controls. Material was collected from the ileum 2 weeks after infection in eight lambs and 3 weeks after infection in six lambs. Lambs examined 2 weeks after infection had normal follicles. After three weeks, the follicle-associated epithelium covering the lymphoid follicles of the ileal Peyer’s patches showed fusions with adjacent absorptive epithelium, focal hyperplasia, and occasionally necrosis. Macrogametes, microgamonts, and oocysts were often found in the follicle-associated epithelium and the dome region. Various degrees of lymphocyte depletion were present in the ileal lymphoid follicles in all six infected lambs 3 weeks after infection, and four lambs had decreased follicle size. Reduced staining for leukocyte common antigen (CD45), B-cell markers, and the proliferation marker Ki-67 was present in these lambs. Application of the terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling method for apoptotic cells revealed decreased staining in the ileal lymphoid follicles 3 weeks after infection. A marker of follicular dendritic cells, 5′- nucleotidase, showed increased reactivity, probably due to condensation of reticular cells following loss of follicle lymphocytes. Reduced staining for carbonic anhydrase in the follicle-associated epithelium and the domes was present in all six lambs examined 3 weeks after infection, indicating decreased production of carbonic anhydrase-reactive 50-nm particles and a decreased lymphoproliferative stimulus. In conclusion, the present study shows that severe E. ovinoidalis infection in lambs causes lesions of the follicle-associated epithelium and may result in lymphocyte depletion and atrophy of the ileal Peyer’s patch follicles. PMID:11777834

  20. A Follicle Rupture Assay Reveals an Essential Role for Follicular Adrenergic Signaling in Drosophila Ovulation.

    PubMed

    Deady, Lylah D; Sun, Jianjun

    2015-10-01

    Ovulation is essential for the propagation of the species and involves a proteolytic degradation of the follicle wall for the release of the fertilizable oocyte. However, the precise mechanisms for regulating these proteolytic events are largely unknown. Work from our lab and others have shown that there are several parallels between Drosophila and mammalian ovulation at both the cellular and molecular levels. During ovulation in Drosophila, posterior follicle cells surrounding a mature oocyte are selectively degraded and the residual follicle cells remain in the ovary to form a corpus luteum after follicle rupture. Like in mammals, this rupturing process also depends on matrix metalloproteinase 2 (Mmp2) activity localized at the posterior end of mature follicles, where oocytes exit. In the present study, we show that Mmp2 activity is regulated by the octopaminergic signaling in mature follicle cells. Exogenous octopamine (OA; equivalent to norepinephrine, NE) is sufficient to induce follicle rupture when isolated mature follicles are cultured ex vivo, in the absence of the oviduct or ovarian muscle sheath. Knocking down the alpha-like adrenergic receptor Oamb (Octoampine receptor in mushroom bodies) in mature follicle cells prevents OA-induced follicle rupture ex vivo and ovulation in vivo. We also show that follicular OA-Oamb signaling induces Mmp2 enzymatic activation but not Mmp2 protein expression, likely via intracellular Ca2+ as the second messenger. Our work develops a novel ex vivo follicle rupture assay and demonstrates the role for follicular adrenergic signaling in Mmp2 activation and ovulation in Drosophila, which is likely conserved in other species.

  1. Hypoxia-mediated carbohydrate metabolism and transport promote early-stage murine follicle growth and survival.

    PubMed

    Makanji, Yogeshwar; Tagler, David; Pahnke, Jennifer; Shea, Lonnie D; Woodruff, Teresa K

    2014-04-15

    Oxygen tension is critical for follicle growth and metabolism, especially for early-stage follicles, where vascularity is limited. Its role and underlying mechanism in the in vitro activation and maturation of immature to ovulatory follicles is largely unknown. In this study, early secondary (110 μm) murine follicles were isolated and encapsulated in alginate hydrogels to replicate the in vivo environment of the growing/maturing follicle. Encapsulated follicles were cultured for 8 days at either 2.5 or 20% O2. Survival (2.6-fold) and growth (1.2-fold) were significantly higher for follicles cultured at 2.5% compared with 20% O2. Using a mouse hypoxia-signaling pathway qRT-PCR array and GeneGo Metacore analysis, we found that direct target genes of the hypoxia-activated HIF1-complex were significantly upregulated in follicles cultured for 8 days at 2.5% compared with 20% O2, including the carbohydrate transport and metabolism genes Slc2a3, Vegfa, Slc2a1, Edn1, Pgk1, Ldha, and Hmox1. Other upregulated genes included carbohydrate transporters (Slc2a1, Slc2a3, and Slc16a3) and enzymes essential for glycolysis (Pgk1, Hmox1, Hk2, Gpi1, Pfkl, Pfkp, Aldoa, Gapdh, Pgam1, Eno1, Pkm2, and Ldha). For follicles cultured at 2.5% O2, a 7.2-fold upregulation of Vegfa correlated to an 18-fold increase in VEGFA levels, and a 3.2-fold upregulation of Ldha correlated to a 4.8-fold increase in lactate levels. Both VEGFA and lactate levels were significantly higher in follicles cultured at 2.5% compared with 20% O2. Therefore, enhanced hypoxia-mediated glycolysis is essential for growth and survival of early secondary follicles and provides vital insights into improving in vitro culture conditions.

  2. Epidemiological survey of helminths of goats in southern Punjab, Pakistan.

    PubMed

    Ayaz, Muhammad Mazhar; Raza, Muhammad Asif; Murtaza, Saeed; Akhtar, Saleem

    2013-03-01

    The main purpose of this study was to investigate the prevalence of helminths of goats such as Fasciola hepatica, Haemonchus contortus, Paramphistomum cervi, Oesophagostomum columbian, Cotylophoron cotylophorum, Monezia expansa, Oestertagia oestertagi and Oestertagia circumcincta. The overall prevalence of all species of helminthes was 52% in goat. The study was designed to investigate the factors of helminths prevalence on the basis of sex and age of goat with the help of Chi-square. All the results obtained were non-significant due to some factors which directly affects the prevalence of helminths.

  3. Endocrine and ovarian responses associated with the first-wave dominant follicle in cattle.

    PubMed

    Badinga, L; Driancourt, M A; Savio, J D; Wolfenson, D; Drost, M; De La Sota, R L; Thatcher, W W

    1992-11-01

    To examine endocrine and biochemical differences between dominant and subordinate follicles and how the dominant follicle affects the hypothalamic-pituitary-ovarian axis in Holstein cows, the ovary bearing the dominant follicle was unilaterally removed on Day 5 (n = 8), 8 (n = 8), or 12 (n = 8) of synchronized estrous cycles. Follicular development was followed daily by ultrasonography from the day of detected estrus (Day 0) until 5 days after ovariectomy. Aromatase activity and steroid concentrations in first-wave dominant and subordinate follicles were measured. Intact dominant and subordinate follicles were cultured in 4 ml Minimum Essential Medium supplemented with 100 microCi 3H-leucine to evaluate de novo protein synthesis. Five days after unilateral ovariectomy, cows were resynchronized and the experiment was repeated. Follicular growth was characterized by the development of single large dominant follicles, which was associated with suppression of other follicles. Concentrations of estradiol-17 beta (E2) in follicular fluid and aromatase activity of follicular walls were higher in dominant follicles (438.9 +/- 45.5 ng/ml; 875.4 +/- 68.2 pg E2/follicle) compared to subordinate follicles (40.6 +/- 69.4 ng/ml; 99.4 +/- 104.2 pg E2/follicle). Aromatase activity in first-wave dominant follicles was higher at Days 5 (1147.1 +/- 118.1 pg E2/follicle) and 8 (1028.2 +/- 118.1 pg E2/follicle) compared to Day 12 (450.7 +/- 118.1 pg E2/follicle). Concentrations of E2 and androstenedione in first-wave dominant follicles were higher at Day 5 (983.2 +/- 78.2 and 89.5 +/- 15.7 ng/ml) compared to Days 8 (225.1 +/- 78.6 and 5.9 +/- 14.8 ng/ml) and 12 (108.5 +/- 78.6 and 13.0 +/- 14.8 ng/ml). Concentrations of progesterone in subordinate follicles increased linearly between Days 5 and 12 of the estrous cycle. Plasma concentrations of FSH increased from 17.9 +/- 1.4 to 32.5 +/- 1.4 ng/ml between 0 and 32 h following unilateral removal of the ovary with the first-wave dominant

  4. Triggering of drug release of particles in hair follicles.

    PubMed

    Mak, Wing Cheung; Patzelt, Alexa; Richter, Heike; Renneberg, Reinhardt; Lai, Kwok Kei; Rühl, Eckart; Sterry, Wolfram; Lademann, Jürgen

    2012-06-28

    Particulate drug delivery via hair follicles represents a promising concept, although requirements are high. This process must be realized at the desired depth and at the appropriate time, due to the fact that the particles themselves are not able to overcome the follicular skin barrier. In the present study, a novel triggering concept for the release of a model drug from the delivering particles is presented based on the application of two different particle types of the same size, where one particle type is the drug carrier, and the second one is loaded with a protease. The latter particle type is supposed to interact with the drug-carrying particles to trigger the drug release. A mixture of both particles was applied onto porcine skin samples, followed by follicular analysis. As a control, the particles were applied unaided without protease, whereas one skin area remained untreated. The investigations revealed that the protease was able to release the model drug from the delivering particles in significant depths within the hair follicle (866±62nm). Additionally, an uptake of the model drug in the sebaceous gland was observed after release providing a promising novel approach for the development of treatment strategies for different skin diseases. PMID:22516090

  5. Hyperproliferative Hepatocellular Alterations after Intraportal Transplantation of Thyroid Follicles

    PubMed Central

    Dombrowski, Frank; Klotz, Luisa; Hacker, Hans Jörg; Li, Yanhua; Klingmüller, Dietrich; Brix, Klaudia; Herzog, Volker; Bannasch, Peter

    2000-01-01

    The thyroid hormone 3,5,3′-triiodo-l-thyronine (T3) is a strong direct hepatocyte mitogen in vivo. The effects of T3 resemble those of peroxisome proliferators, which are known to induce hepatocellular tumors in rats. With the aim of studying long-term local effects of thyroid hormones on liver parenchyma, small pieces of thyroid tissue were transplanted via the portal veins into the livers of thyroidectomized male Lewis rats. At 1 week, 3 weeks, 3 months, and 18 months after transplantation, the transplants were found to proliferate, to synthesize thyroglobulin, and to release thyroxine and T3. At 3 and 18 months after transplantation, the hepatocytes of the liver acini downstream of the transplanted follicles showed an increase in cytoplasmic basophilia, a loss of glycogen, an enlargement and hyperchromasia of their nuclei, and a strong increase in cell turnover compared with unaltered liver acini. The altered hepatocytes exhibited an increase in the activities of glucose-6-phosphate dehydrogenase, glucose-6-phosphatase, malic enzyme, mitochondrial glycerol-3-phosphate dehydrogenase, cytochrome-c-oxidase, and acid phosphatase; the activities of glycogen synthase and glycogen phosphorylase were strongly decreased. The hepatocytic alterations downstream of the transplanted follicles could be explained by effects of T3. On the other hand, they resembled alterations characteristic of amphophilic preneoplastic liver foci observed in different models of hepatocarcinogenesis. PMID:10623658

  6. Palmitoylation Regulates Epidermal Homeostasis and Hair Follicle Differentiation

    PubMed Central

    Fukata, Yuko; Tsutsumi, Ryouhei; Fukata, Masaki; Keighren, Margaret; Porter, Rebecca M.; McKie, Lisa; Smyth, Ian; Jackson, Ian J.

    2009-01-01

    Palmitoylation is a key post-translational modification mediated by a family of DHHC-containing palmitoyl acyl-transferases (PATs). Unlike other lipid modifications, palmitoylation is reversible and thus often regulates dynamic protein interactions. We find that the mouse hair loss mutant, depilated, (dep) is due to a single amino acid deletion in the PAT, Zdhhc21, resulting in protein mislocalization and loss of palmitoylation activity. We examined expression of Zdhhc21 protein in skin and find it restricted to specific hair lineages. Loss of Zdhhc21 function results in delayed hair shaft differentiation, at the site of expression of the gene, but also leads to hyperplasia of the interfollicular epidermis (IFE) and sebaceous glands, distant from the expression site. The specific delay in follicle differentiation is associated with attenuated anagen propagation and is reflected by decreased levels of Lef1, nuclear β-catenin, and Foxn1 in hair shaft progenitors. In the thickened basal compartment of mutant IFE, phospho-ERK and cell proliferation are increased, suggesting increased signaling through EGFR or integrin-related receptors, with a parallel reduction in expression of the key differentiation factor Gata3. We show that the Src-family kinase, Fyn, involved in keratinocyte differentiation, is a direct palmitoylation target of Zdhhc21 and is mislocalized in mutant follicles. This study is the first to demonstrate a key role for palmitoylation in regulating developmental signals in mammalian tissue homeostasis. PMID:19956733

  7. Soft matrix supports osteogenic differentiation of human dental follicle cells

    SciTech Connect

    Viale-Bouroncle, Sandra; Voellner, Florian; Moehl, Christoph; Kuepper, Kevin; Brockhoff, Gero; Reichert, Torsten E.; Schmalz, Gottfried; Morsczeck, Christian

    2011-07-08

    Highlights: {yields} Rigid stiffness supports osteogenic differentiation in mesenchymal stem cells (MSCs). {yields} Our study examined stiffness and differentiation of dental follicle cells (DFCs). {yields} Soft ECMs have a superior capacity to support the osteogenic differentiation of DFCs. {yields} DFCs and MSCs react contrarily to soft and rigid surface stiffness. -- Abstract: The differentiation of stem cells can be directed by the grade of stiffness of the developed tissue cells. For example a rigid extracellular matrix supports the osteogenic differentiation in bone marrow derived mesenchymal stem cells (MSCs). However, less is known about the relation of extracellular matrix stiffness and cell differentiation of ectomesenchymal dental precursor cells. Our study examined for the first time the influence of the surface stiffness on the proliferation and osteogenic differentiation of human dental follicle cells (DFCs). Cell proliferation of DFCs was only slightly decreased on cell culture surfaces with a bone-like stiffness. The osteogenic differentiation in DFCs could only be initiated with a dexamethasone based differentiation medium after using varying stiffness. Here, the softest surface improved the induction of osteogenic differentiation in comparison to that with the highest stiffness. In conclusion, different to bone marrow derived MSCs, soft ECMs have a superior capacity to support the osteogenic differentiation of DFCs.

  8. Storage of bovine isolated follicles: a new alternative way to improve the recovery rate of viable embryos from ovarian follicles of slaughtered cows.

    PubMed

    Pavlok, A; Cech, S; Kubelka, M; Lopatárová, M; Holý, L; Jindra, M

    2006-11-01

    The vitality of bovine oocytes stored in isolated follicles was examined. The aim of this work was to prolong the time of in vitro manipulation of oocytes before their maturation and develop a new alternative of oocyte "capacitation" to improve the quality of in vitro produced embryos. Follicles were dissected from the ovaries of slaughtered cows; subsequently, follicles were divided according to their diameter into three categories (2-3, 3-4 and 4-6 mm), and stored at 17-18 degrees C for 24 or 48 h in a modified tissue culture medium-199 (TCM-199) with reduced pH. After that time, the cumulus-oocyte complexes (COCs) were isolated, matured, fertilized, and embryos cultured in vitro for a total of 9 days. The percentage of total blastocysts, and hatched blastocysts developed from oocytes, initially kept ("capacitated") for 24h at 17-18 degrees C, within follicles of 3-6mm size categories, were significantly higher than that oocytes of the control [of control oocytes] (44.9 and 30.3% versus 36.2 and 20.4%, respectively). The oocytes of follicles stored for 48 h at 17-18 degrees C already had decreased developmental capacity. Interesting data were obtained when COCs of the 3-4 and 4-6 categories were additionally divided into two subgroups according to their presumed developmental history (originating from the supposed growing "fit" in contrast to the supposed regressing "unfit" follicles). The higher improvement in the rate of hatched blastocysts from 24h stored oocytes was observed only in the subgroup originated from "fit" COCs (15.3 versus 25.0%, and 20.0 versus 34.4%, in the 3-4 and 4-6mm categories, respectively). The transfer of 26 blastocysts (developed of follicles kept for 24h at 17-18 degrees C) to 26 recipient heifers resulted in 18 pregnancies. Storage of follicles at 17-18 degrees C in vitro resulted not only in recovery of higher numbers of blastocysts of better quality but also facilitated the safe transport of follicles for a long distance. The

  9. Methoxychlor and its metabolites inhibit growth and induce atresia of baboon antral follicles.

    PubMed

    Gupta, Rupesh K; Aberdeen, Graham; Babus, Janice K; Albrecht, Eugene D; Flaws, Jodi A

    2007-08-01

    Methoxychlor (MXC), an organochlorine pesticide, inhibits growth and induces atresia of antral follicles in rodents. MXC metabolites, mono-OH MXC (mono-OH) and bis-OH MXC (HPTE), are thought to be more toxic than the parent compound. Although studies have examined effects of MXC in rodents, few studies have evaluated the effects of MXC in primates. Therefore, the present study tested the hypothesis that MXC, mono-OH, and HPTE inhibit growth and induce atresia of baboon antral follicles. To test this hypothesis, antral follicles were isolated from adult baboon ovaries and cultured with vehicle (dimethylsulfoxide; DMSO), MXC (1-100 micro g/ml), mono-OH (0.1-10 micro g/ml), or HPTE (0.1-10 micro g/ml) for 96 hr. Growth was monitored at 24 hr intervals. After culture, follicles were processed for histological evaluation of atresia. MXC, mono-OH, and HPTE significantly inhibited follicular growth and increased atresia compared to DMSO. Moreover, the adverse effects of MXC and its metabolites on growth and atresia in baboon antral follicles were observed at lower (100-fold) doses than those causing similar effects in rodents. These data suggest that MXC and its metabolites inhibit growth and induce atresia of baboon antral follicles, and that primate follicles are more sensitive to MXC than rodent follicles.

  10. Quantitative bioluminescence imaging of transgene expression in intact porcine antral follicles in vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The porcine oocyte maturation in vivo occurs within the ovarian follicle and is regulated by the interactions between oocytes and surrounding follicular components, including theca, granulosa, and cumulus cells, and follicular fluid. Therefore, the antral follicle is an essential microenvironment fo...

  11. The Utility of Stage-specific Mid-to-late Drosophila Follicle Isolation

    PubMed Central

    Spracklen, Andrew J.; Tootle, Tina L.

    2013-01-01

    Drosophila oogenesis or follicle development has been widely used to advance the understanding of complex developmental and cell biologic processes. This methods paper describes how to isolate mid-to-late stage follicles (Stage 10B-14) and utilize them to provide new insights into the molecular and morphologic events occurring during tight windows of developmental time. Isolated follicles can be used for a variety of experimental techniques, including in vitro development assays, live imaging, mRNA expression analysis and western blot analysis of proteins. Follicles at Stage 10B (S10B) or later will complete development in culture; this allows one to combine genetic or pharmacologic perturbations with in vitro development to define the effects of such manipulations on the processes occurring during specific periods of development. Additionally, because these follicles develop in culture, they are ideally suited for live imaging studies, which often reveal new mechanisms that mediate morphological events. Isolated follicles can also be used for molecular analyses. For example, changes in gene expression that result from genetic perturbations can be defined for specific developmental windows. Additionally, protein level, stability, and/or posttranslational modification state during a particular stage of follicle development can be examined through western blot analyses. Thus, stage-specific isolation of Drosophila follicles provides a rich source of information into widely conserved processes of development and morphogenesis. PMID:24326735

  12. Topical liposome targeting of dyes, melanins, genes, and proteins selectively to hair follicles.

    PubMed

    Hoffman, R M

    1998-01-01

    For therapeutic and cosmetic modification of hair, we have developed a hair-follicle-selective macromolecule and small molecule targeting system with topical application of phosphatidylcholine-based liposomes. Liposome-entrapped melanins, proteins, genes, and small-molecules have been selectively targeted to the hair follicle and hair shafts of mice. Liposomal delivery of these molecules is time dependent. Negligible amounts of delivered molecules enter the dermis, epidermis, or bloodstream thereby demonstrating selective follicle delivery. Naked molecules are trapped in the stratum corneum and are unable to enter the follicle. The potential of the hair-follicle liposome delivery system for therapeutic use for hair disease as well as for cosmesis has been demonstrated in 3-dimensional histoculture of hair-growing skin and mouse in vivo models. Topical liposome selective delivery to hair follicles has demonstrated the ability to color hair with melanin, the delivery of the active lac-Z gene to hair matrix cells and delivery of proteins as well. Liposome-targeting of molecules to hair follicles has also been achieved in human scalp in histoculture. Liposomes thus have high potential in selective hair follicle targeting of large and small molecules, including genes, opening the field of gene therapy and other molecular therapy of the hair process to restore hair growth, physiologically restore or alter hair pigment, and to prevent or accelerate hair loss.

  13. Characterization and Small RNA Content of Extracellular Vesicles in Follicular Fluid of Developing Bovine Antral Follicles

    PubMed Central

    Navakanitworakul, Raphatphorn; Hung, Wei-Ting; Gunewardena, Sumedha; Davis, John S.; Chotigeat, Wilaiwan; Christenson, Lane K.

    2016-01-01

    Exosomes and microvesicles (i.e., extracellular vesicles: EVs) have been identified within ovarian follicular fluid and recent evidence suggests that EVs are able to elicit profound effects on ovarian cell function. While existence of miRNA within EVs has been reported, whether EV size and concentration as well as their cargos (i.e., proteins and RNA) change during antral follicle growth remains unknown. Extracellular vesicles isolated from follicular fluid of small, medium and large bovine follicles were similar in size, while concentration of EVs decreased progressively as follicle size increased. Electron microscopy indicated a highly purified population of the lipid bilayer enclosed vesicles that were enriched in exosome biomarkers including CD81 and Alix. Small RNA sequencing identified a large number of known and novel miRNAs that changed in the EVs of different size follicles. Ingenuity Pathway Analysis (IPA) indicated that miRNA abundant in small follicle EV preparations were associated with cell proliferation pathways, while those miRNA abundant in large follicle preparations were related to inflammatory response pathways. These studies are the first to demonstrate that EVs change in their levels and makeup during antral follicle development and point to the potential for a unique vesicle-mediated cell-to-cell communication network within the ovarian follicle. PMID:27158133

  14. Effect of helium-neon laser irradiation on hair follicle growth cycle of Swiss albino mice.

    PubMed

    Shukla, S; Sahu, K; Verma, Y; Rao, K D; Dube, A; Gupta, P K

    2010-01-01

    We report the results of a study carried out to investigate the effect of helium-neon (He-Ne) laser (632.8 nm) irradiation on the hair follicle growth cycle of testosterone-treated and untreated mice. Both histology and optical coherence tomography (OCT) were used for the measurement of hair follicle length and the relative percentage of hair follicles in different growth phases. A positive correlation (R = 0.96) was observed for the lengths of hair follicles measured by both methods. Further, the ratios of the lengths of hair follicles in the anagen and catagen phases obtained by both methods were nearly the same. However, the length of the hair follicles measured by both methods differed by a factor of 1.6, with histology showing smaller lengths. He-Ne laser irradiation (at approximately 1 J/cm(2)) of the skin of both the control and the testosterone-treated mice was observed to lead to a significant increase (p < 0.05) in % anagen, indicating stimulation of hair growth. The study also demonstrates that OCT can be used to monitor the hair follicle growth cycle, and thus hair follicle disorders or treatment efficacy during alopecia. PMID:20016249

  15. Proximate analysis of two breeds of goat meat (chevon) and assessment of perception on goat consumption

    NASA Astrophysics Data System (ADS)

    Baharuddin, Azan Azuwan; Abdullah, Aminah

    2015-09-01

    Goat Meat (chevon) has been used as a source of protein and its demand for consumption is increasing yearly in Malaysia. The objective of the research was to determine proximate composition namely moisture, protein, fat and ash level in chevon from different type of breed i.e. Boer and Katjang. The goat breed proximate analyses were compared with sheep meat (mutton). The results for goat breeds were showing that the percentage of moisture, fat, protein and ash were of 73.06 - 74.99, 2.76 - 2.94, 20.81 - 22.47 and 0.97 - 1.21 respectively. Meanwhile the concentration of moisture, fat, protein and ash for mutton were 73.52, 5.06, 22.50 and 1.17 respectively. The pilot study on perception survey indicated that the highest respondent percentage on consumption of chevon was once in a month. Based on the survey, Boer was the most known breed compared to the other breed. Majority of respondents reported that chevon was fairly expensive meat but easier to obtain in the survey area.

  16. Sphingosine-1-phosphate and ceramide are associated with health and atresia of bovine ovarian antral follicles.

    PubMed

    Hernández-Coronado, C G; Guzmán, A; Espinosa-Cervantes, R; Romano, M C; Verde-Calvo, J R; Rosales-Torres, A M

    2015-02-01

    The follicle destiny towards ovulation or atresia is multi-factorial in nature and involves outcries, paracrine and endocrine factors that promote cell proliferation and survival (development) or unchain apoptosis as part of the atresia process. In several types of cells, sphingosine-1-phospate (S1P) promotes cellular proliferation and survival, whereas ceramide (CER) triggers cell death, and the S1P/CER ratio may determine the fate of the cell. The aim of present study was to quantify S1P and CER concentrations and their ratio in bovine antral follicles of 8 to 17 mm classified as healthy and atretic antral follicles. Follicles were dissected from cow ovaries collected from a local abattoir. The theca cell layer, the granulosa cells and follicular fluid were separated, and 17β-estradiol (E2) and progesterone (P4) concentrations were measured in the follicular fluid by radioimmunoassay. Based on the E2/P4 ratio, the follicles were classified as healthy (2.2±0.3) or atretic (0.2±0.3). In both follicular compartments (granulosa and theca cell layer), sphingolipids were extracted and S1P and CER concentrations were quantified by HPLC (XTerra RP18; 5 µm, 3.0×150 mm column). Results showed that in both follicular compartments, S1P concentrations were higher in healthy antral follicles than in atretic antral follicles (P<0.05). The concentration of CER in the granulosa cells was higher in atretic antral follicles than in healthy antral follicles, but no differences were observed in the theca cell layer. The S1P/CER ratio in both follicular compartments was also higher in healthy antral follicles. Interestingly, in these follicles, there was a 45-fold greater concentration of S1P than CER in the granulosa cells (P<0.05), whereas in the theca cell layer, S1P had only a 14-fold greater concentration than CER when compared with atretic antral follicles. These results suggest that S1P plays a role in follicle health, increasing cellular proliferation and survival. In

  17. Sphingosine-1-phosphate and ceramide are associated with health and atresia of bovine ovarian antral follicles.

    PubMed

    Hernández-Coronado, C G; Guzmán, A; Espinosa-Cervantes, R; Romano, M C; Verde-Calvo, J R; Rosales-Torres, A M

    2015-02-01

    The follicle destiny towards ovulation or atresia is multi-factorial in nature and involves outcries, paracrine and endocrine factors that promote cell proliferation and survival (development) or unchain apoptosis as part of the atresia process. In several types of cells, sphingosine-1-phospate (S1P) promotes cellular proliferation and survival, whereas ceramide (CER) triggers cell death, and the S1P/CER ratio may determine the fate of the cell. The aim of present study was to quantify S1P and CER concentrations and their ratio in bovine antral follicles of 8 to 17 mm classified as healthy and atretic antral follicles. Follicles were dissected from cow ovaries collected from a local abattoir. The theca cell layer, the granulosa cells and follicular fluid were separated, and 17β-estradiol (E2) and progesterone (P4) concentrations were measured in the follicular fluid by radioimmunoassay. Based on the E2/P4 ratio, the follicles were classified as healthy (2.2±0.3) or atretic (0.2±0.3). In both follicular compartments (granulosa and theca cell layer), sphingolipids were extracted and S1P and CER concentrations were quantified by HPLC (XTerra RP18; 5 µm, 3.0×150 mm column). Results showed that in both follicular compartments, S1P concentrations were higher in healthy antral follicles than in atretic antral follicles (P<0.05). The concentration of CER in the granulosa cells was higher in atretic antral follicles than in healthy antral follicles, but no differences were observed in the theca cell layer. The S1P/CER ratio in both follicular compartments was also higher in healthy antral follicles. Interestingly, in these follicles, there was a 45-fold greater concentration of S1P than CER in the granulosa cells (P<0.05), whereas in the theca cell layer, S1P had only a 14-fold greater concentration than CER when compared with atretic antral follicles. These results suggest that S1P plays a role in follicle health, increasing cellular proliferation and survival. In

  18. Goats as an osteopenic animal model.

    PubMed

    Leung, K S; Siu, W S; Cheung, N M; Lui, P Y; Chow, D H; James, A; Qin, L

    2001-12-01

    A large osteopenic animal model that resembles human osteoporotic changes is essential for osteoporosis research. This study aimed at establishing a large osteopenic animal model in goats. Twenty-five Chinese mountain goats were used in which they were either ovariectomized (OVX) and fed with a low-calcium diet (n = 16) or sham-operated (SHAM; n = 9). Monthly photodensitometric analysis on proximal tibial metaphysis and calcaneus was performed. Two iliac crest biopsy specimens obtained before and 6 months after OVX were used for bone mineral density (BMD) measurement with peripheral quantitative computed tomography (pQCT). Lumbar vertebrae (L2 and L7), humeral heads, and calcanei were collected for BMD measurement after euthanasia. The humeral heads and calcanei were used in biomechanical indentation test. BMD measurement showed a significant 25.0% (p = 0.006) decrease in BMD of the iliac crest biopsy specimens 6 months after OVX. It also was statistically significant when compared with the SHAM (p = 0.028). BMD at L2, L7, calcaneus, and humeral head reduced by 24-33% (p ranged from 0.001 to 0.011) when compared with the SHAM. Photodensitometry showed a continuous decrease in bone density after OVX. There were significant decreases of 18.9% in proximal tibial metaphysis (p = 0.003) and 21.8% in calcaneus (p = 0.023) in the OVX group 6 months postoperatively. Indentation test on the humeral head and calcaneus showed a significant decrease 52% (p = 0.006) and 54% (p = 0.001), respectively, in energy required for displacement of 3 mm in the OVX group compared with the SHAM group. The decreases correlated significantly to the decrease in BMD of the corresponding specimens (r2 = 0.439 and 0.581; p < 0.001 for both). In conclusion, this study showed that OVX plus a low-calcium diet could induce significant osteopenia and deterioration of mechanical properties of the cancellous bone in goats.

  19. Anti-angiogenic VEGFA164B isoform mRNA is more abundant in E2-inactive, atretic follicles while expression of angiogenic VEGFA isoforms is greater in granulosa cells from developing bovine follicles prior to the LH surge

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Vascular endothelial growth factor A (VEGFA) is expressed by granulosa cells of the follicle and if its actions are blocked, ovulation and antral follicle development is inhibited. However, the role of anti-angiogenic VEGFA isoforms in bovine dominant follicle development, especially prior to and a...

  20. Bald scalp in men with androgenetic alopecia retains hair follicle stem cells but lacks CD200-rich and CD34-positive hair follicle progenitor cells

    PubMed Central

    Garza, Luis A.; Yang, Chao-Chun; Zhao, Tailun; Blatt, Hanz B.; Lee, Michelle; He, Helen; Stanton, David C.; Carrasco, Lee; Spiegel, Jeffrey H.; Tobias, John W.; Cotsarelis, George

    2011-01-01

    Androgenetic alopecia (AGA), also known as common baldness, is characterized by a marked decrease in hair follicle size, which could be related to the loss of hair follicle stem or progenitor cells. To test this hypothesis, we analyzed bald and non-bald scalp from AGA individuals for the presence of hair follicle stem and progenitor cells. Cells expressing cytokeratin15 (KRT15), CD200, CD34, and integrin, α6 (ITGA6) were quantitated via flow cytometry. High levels of KRT15 expression correlated with stem cell properties of small cell size and quiescence. These KRT15hi stem cells were maintained in bald scalp samples. However, CD200hiITGA6hi and CD34hi cell populations — which both possessed a progenitor phenotype, in that they localized closely to the stem cell–rich bulge area but were larger and more proliferative than the KRT15hi stem cells — were markedly diminished. In functional assays, analogous CD200hiItga6hi cells from murine hair follicles were multipotent and generated new hair follicles in skin reconstitution assays. These findings support the notion that a defect in conversion of hair follicle stem cells to progenitor cells plays a role in the pathogenesis of AGA. PMID:21206086

  1. Observations on Mycoplasma mycoides subsp. mycoides infection in Saanen goats.

    PubMed

    Bar-Moshe, B; Rapapport, E

    1981-07-01

    An epizootic in white Saanen goats, caused by Mycoplasma mycoides subsp. mycoides is described. Twenty-five flocks totalling approximately 4,500 animals were involved. The disease was characterized by a high, transient temperature, general malaise and mastitis in the lactating does, and a keratoconjunctivitis, arthritis, mycoplasmaemia and death among the kids. In one goat flock there was a precipitous change in the character of the disease, from a predominantly mastitis syndrome to a fulminating pleuropneumonia. In another goat flock, twin kids were born with an advanced purulent, proliferative arthritis, suggesting early congenital infection. In yet another infected flock there were cases of subcutaneous abscesses from which both M. mycoides and Corynebacterium pyogenes were cultured. M. mycoides was also isolated from synovial fluid and the parenchymal organs of an Ibex mountain goat that died of a purulent polyarthritis. Experimental infection in kids caused a diffuse cellulitis at the site of inoculation, a high fever, polyarthritis and death.

  2. Oligosaccharides in goat milk: structure, health effects and isolation.

    PubMed

    Kiskini, A; Difilippo, E

    2013-11-03

    Oligosaccharides have been widely recognized for their prebiotic and anti-infective properties. Among the different types of mammalian milk, the one of humans is the richest source of naturally derived oligosaccharides. However, their use as a basis for functional foods is hampered, due to their structural complexity, which in turn makes their re-synthesis extremely difficult. Thus, oligosaccharides from other sources have to be used. In this sense, goat milk constitutes a very appealing candidate, as it contains the highest amount of oligosaccharides among domestic animals, while goat milk oligosaccharides show significant similarities to human milk oligosaccharides from a structural point of view. Studies on goat milk oligosaccharides are scant, and more data is required in order to provide solid clinical evidence of their beneficial effects on humans. The aim of this review is to collect and present the main research findings on goat milk oligosaccharides structure, health effects and isolation.

  3. Complete mitochondrial genome of Nanjiang Yellow goat (Capra hircus).

    PubMed

    Li, Haijun; Meng, Xiangren; Zhang, Hao; Duan, Xiaoyue; Niu, Lili; Wang, Linjie; Li, Li; Zhang, Hongping; Wu, Hongda; Zhong, Tao

    2016-01-01

    Nanjiang Yellow goat (Capra hircus) is the first cultured mutton breed in China. In this study, the complete mitochondrial genome sequence of Nanjiang Yellow goat has been identified for the first time. The total length of the mitochondrial genome was 16,639 bp, with the base composition of 33.54% A, 26.05% C, 13.11% G and 27.30% T. It contained 37 genes (22 transfer RNA genes, 2 ribosomal RNA genes, and 13 protein-coding genes) and a major non-coding control region (D-loop). Most of the genes have ATG initiation codons, whereas ND2, ND3 and ND5 start with ATA. The complete mitochondrial genome sequence of Nanjiang Yellow goat provides an important data set for further estimation on the phylogeographic structure of domestic goats.

  4. Ultrasonic biparietal diameter of second trimester Pygmy goat fetuses.

    PubMed

    Reichle, J K; Haibel, G K

    1991-04-01

    Four does pregnant with seven Pygmy goat fetuses were scanned transabdominally from Day 36 to Day 102 of gestation using a real-time ultrasound 5 MHz linear-array scanhead. Fetal biparietal diameter (BPD) was measured on symmetrical frozen images using internal electronic calipers. The relationship between gestational age (GA) in days and BPD in millimeters in Pygmy goat fetuses is expressed as: [GA = 23.2 + 2.08 BPD]. This equation can assign GA to Pygmy goat fetuses of unknown conception dates and allow confinement of does near kidding for induced or observed parturition. Mean kid birthweight was 2.0 + 0.45 kg SEM. These observations are consistent with relationships which have been determined for Toggenburg, Nubian and Angora goat fetuses.

  5. Immobilization of mountain goats with xylazine and reversal with idazoxan.

    PubMed

    Haviernick, M; Côté, S D; Festa-Bianchet, M

    1998-04-01

    Mountain goats (Oreamnos americanus) were captured in traps and immobilized with xylazine, later reversed with idazoxan. One hundred and forty-one goats were immobilized, 94 with a single injection and 47 with multiple injections. Dosage (mg/kg of body weight) of xylazine received, induction time, and recovery time after handling did not differ among sex-age classes. Increasing the dosage did not shorten induction time. The first injection of xylazine in multiple-injection captures was lower than the dose given in single-injection captures, suggesting that insufficient initial doses of xylazine made multiple injections necessary. Xylazine is an effective drug for immobilization of mountain goats captured in traps, at dosages of about 4.9 mg/kg. The dosage of xylazine required to immobilize mountain goats is higher than that reported for bighorn sheep and white-tailed deer.

  6. Sheep-goat hybrid born under natural conditions.

    PubMed

    Mine; Kedikilwe; Ndebele; Nsoso

    2000-07-01

    The paper reports a 5 year old male sheep-goat interspecific hybrid born under natural conditions. The hybrid was castrated at 10 months of age. Karyotype analysis confirmed that the animal was a male and a hybrid with 57 chromosomes, an intermediate number between sheep (2n=54) and goat (2n=60). Morphological characteristics of the animal were compared with those of castrated male Tswana goats of the same age. Data on castrated male Tswana sheep of the same age are not available. The hybrid at 5 years of age weighed 93kg compared to 53.73+/-13.83kg, the average weight of castrated Tswana male goats of the same age. The paper also raises a question of whether the animal could be of commercial interest in meat production in Botswana. PMID:10818315

  7. In-vitro and in-vivo study of dye diffusion into the human skin and hair follicles

    NASA Astrophysics Data System (ADS)

    Genina, Elina A.; Bashkatov, Alexey N.; Sinichkin, Yurii P.; Kochubey, Vyacheslav I.; Lakodina, Nina A.; Perpelitzina, Olga A.; Altshuler, Gregory B.; Tuchin, Valery V.

    2000-11-01

    We present experimental results on in vitro and in vivo investigation of dye diffusion into the human skin and hair follicles. It was shown that dyeing as a method of enhancement of the absorption coefficient of hair follicle tissue components can be used for selective photodestruction of hair follicle and surrounding tissues. Strength and depth of hair follicle dyeing inside the skin were determined for various dyes.

  8. Renal mechanisms of calcium homeostasis in sheep and goats.

    PubMed

    Herm, G; Muscher-Banse, A S; Breves, G; Schröder, B; Wilkens, M R

    2015-04-01

    In small ruminants, the renal excretion of calcium (Ca) and phosphate (Pi) is not modulated in response to dietary Ca restriction. Although this lack of adaptation was observed in both sheep and goats, differences in renal function between these species cannot be excluded. Recent studies demonstrated that compared with sheep, goats have a greater ability to compensate for challenges to Ca homeostasis, probably due to a more pronounced increase in calcitriol production. Therefore, the aim of the present study was to examine the effect of 1) dietary Ca restriction, 2) administration of calcitriol, and 3) lactation on Ca and Pi transport mechanisms and receptors as well as enzymes involved in vitamin D metabolism in renal tissues of sheep and goats. Whereas RNA expression of renal transient receptor potential vanilloid channel type 5 was unaffected by changes in dietary Ca content, a significant stimulation was observed with administration of calcitriol in both sheep (P < 0.001) and goats (P < 0.01). Calbindin-D28K was downregulated during dietary Ca restriction in goats (P < 0.05). Expression of the sodium/Ca exchanger type 1 was decreased by low Ca intake in sheep (P < 0.05) and upregulated by calcitriol treatment in goats (P < 0.05). A significant reduction in RNA expression of the cytosolic and the basolateral Ca transporting proteins was also demonstrated for lactating goats in comparison to dried-off animals. Species differences were found for vitamin D receptor expression, which was stimulated by calcitriol treatment in sheep (P < 0.01) but not in goats. As expected, expression of 1α-hydroxylase was upregulated by dietary Ca restriction (P < 0.001; P < 0.05) and inhibited by exogenous calcitriol (P < 001; P < 0.05) in both sheep and goats. However, whereas 24-hydroxylase expression was stimulated to the same extent by calcitriol treatment in sheep, irrespective of the diet (P < 0.001), a modulatory effect of dietary Ca supply on 24-hydroxylase induction was

  9. Renal mechanisms of calcium homeostasis in sheep and goats.

    PubMed

    Herm, G; Muscher-Banse, A S; Breves, G; Schröder, B; Wilkens, M R

    2015-04-01

    In small ruminants, the renal excretion of calcium (Ca) and phosphate (Pi) is not modulated in response to dietary Ca restriction. Although this lack of adaptation was observed in both sheep and goats, differences in renal function between these species cannot be excluded. Recent studies demonstrated that compared with sheep, goats have a greater ability to compensate for challenges to Ca homeostasis, probably due to a more pronounced increase in calcitriol production. Therefore, the aim of the present study was to examine the effect of 1) dietary Ca restriction, 2) administration of calcitriol, and 3) lactation on Ca and Pi transport mechanisms and receptors as well as enzymes involved in vitamin D metabolism in renal tissues of sheep and goats. Whereas RNA expression of renal transient receptor potential vanilloid channel type 5 was unaffected by changes in dietary Ca content, a significant stimulation was observed with administration of calcitriol in both sheep (P < 0.001) and goats (P < 0.01). Calbindin-D28K was downregulated during dietary Ca restriction in goats (P < 0.05). Expression of the sodium/Ca exchanger type 1 was decreased by low Ca intake in sheep (P < 0.05) and upregulated by calcitriol treatment in goats (P < 0.05). A significant reduction in RNA expression of the cytosolic and the basolateral Ca transporting proteins was also demonstrated for lactating goats in comparison to dried-off animals. Species differences were found for vitamin D receptor expression, which was stimulated by calcitriol treatment in sheep (P < 0.01) but not in goats. As expected, expression of 1α-hydroxylase was upregulated by dietary Ca restriction (P < 0.001; P < 0.05) and inhibited by exogenous calcitriol (P < 001; P < 0.05) in both sheep and goats. However, whereas 24-hydroxylase expression was stimulated to the same extent by calcitriol treatment in sheep, irrespective of the diet (P < 0.001), a modulatory effect of dietary Ca supply on 24-hydroxylase induction was

  10. Increased sensitivity of estrogen receptor alpha overexpressing antral follicles to methoxychlor and its metabolites.

    PubMed

    Paulose, Tessie; Hernández-Ochoa, Isabel; Basavarajappa, Mallikarjuna S; Peretz, Jackye; Flaws, Jodi A

    2011-04-01

    Methoxychlor (MXC), an organochlorine pesticide, and its metabolites, mono-hydroxy MXC (MOH) and bis-hydroxy MXC (HPTE) are known ovarian toxicants and can cause inhibition of antral follicle growth. Since these chemicals bind to estrogen receptor alpha (ESR1), we hypothesized that ovaries overexpressing ESR1 (ESR1 OE) would be more susceptible to toxicity induced by MXC and its metabolites because the chemicals can bind to more ESR1 in the antral follicles. We cultured antral follicles from controls and ESR1 OE mouse ovaries with either the vehicle dimethylsulfoxide (DMSO), MXC, MOH, or HPTE. The data show that at 96 h, the cultured antral follicles from ESR1 OE antral follicles are more susceptible to toxicity induced by MXC, MOH, and HPTE because low doses of these chemicals cause follicle growth inhibition in ESR1 OE mice but not in control mice. On comparing gene expression levels of nuclear receptors in the cultured antral follicles of ESR1 OE and control follicles, we found differential messenger RNA (mRNA) expression of Esr1, estrogen receptor beta (Esr2), androgen receptor (Ar), progesterone receptor (Pr), and aryl hydrocarbon receptor (Ahr) between the genotypes. We also analyzed mRNA levels of Cyp3a41a, the enzyme metabolizing MOH and HPTE, in the cultured follicles and found that Cyp3a41a was significantly lower in DMSO-treated ESR1 OE follicles compared with controls. In ESR1 OE livers, we found that Cyp3a41a levels were significantly lower compared with control livers. Collectively, these data suggest that MXC and its metabolites cause differential gene expression in ESR1 OE mice compared with controls. The results also suggest that the increased sensitivity of ESR1 OE mouse ovaries to toxicity induced by MXC and its metabolites is due to low clearance of the metabolites by the liver and ovary.

  11. Wnt/β-catenin signaling in dermal condensates is required for hair follicle formation

    PubMed Central

    Tsai, Su-Yi; Sennett, Rachel; Rezza, Amélie; Clavel, Carlos; Grisanti, Laura; Zemla, Roland; Najam, Sara; Rendl, Michael

    2014-01-01

    Broad dermal Wnt signaling is required for patterned induction of hair follicle placodes and subsequent Wnt signaling in placode stem cells is essential for induction of dermal condensates, cell clusters of precursors for the hair follicle dermal papilla (DP). Progression of hair follicle formation then requires coordinated signal exchange between dermal condensates and placode stem cells. However, it remains unknown whether continued Wnt signaling in DP precursor cells plays a role in this process, largely due to the long-standing inability to specifically target dermal condensates for gene ablation. Here we use the Tbx18Cre knockin mouse line to ablate the Wnt-responsive transcription factor β-catenin specifically in these cells at E14.5 during the first wave of guard hair follicle formation. In the absence of β-catenin, canonical Wnt signaling is effectively abolished in these cells. Sox2+ dermal condensates initiate normally, however by E16.5 guard hair follicle numbers are strongly reduced and by E18.5 most whiskers and guard hair follicles are absent, suggesting that active Wnt signaling in dermal condensates is important for hair follicle formation to proceed after induction. To explore the molecular mechanisms by which Wnt signaling in dermal condensates regulates hair follicle formation, we analyze genome-wide the gene expression changes in embryonic β-catenin null DP precursor cells. We find altered expression of several signaling pathway genes, including Fgfs and Activin, both previously implicated in hair follicle formation. In summary, these data reveal a functional role of Wnt signaling in DP precursors for embryonic hair follicle formation and identify Fgf and Activin signaling as potential effectors of Wnt signaling-regulated events. PMID:24309208

  12. In Vitro Growth and Steroidogenesis of Dog Follicles as Influenced by the Physical and Hormonal Microenvironment

    PubMed Central

    Songsasen, N; Woodruff, TK; Wildt, DE

    2011-01-01

    The present study examined the influences of physical and hormonal microenvironment on in vitro growth and steroidogenesis of dog follicles. Follicles were enzymatically-isolated and individually-encapsulated in 0.5% (w/v; n = 17) or 1.5% (n = 10) alginate and cultured with 0.5 IU/ml equine chorionic gonadotropin for 192 h. In a separate experiment, follicles were encapsulated in 0.5% alginate and cultured with 0 (n = 22), 1 (n= 23), 10 (n = 20) or 100 (n = 21) µg/ml FSH for 240 h. Follicle diameter and steroid production were assessed every 48 h in both studies. Follicles encapsulated in the 0.5% alginate grew faster (P < 0.05) than those cultured in the 1.5% concentration. Oestradiol (E2) and progesterone (P4) increased consistently (P < 0.05) over time, and follicles in the 1.5% alginate produced more (P < 0.05) P4 than those in the 0.5% solution. Follicles cultured in the highest FSH concentration (100 µg/ml) increased 100% in size after 240 h compared to 50 to 70% in lower dosages. E2 concentration remained unchanged over time (P > 0.05) across FSH dosages. However, P4 increased (P < 0.05) as culture progressed and with increasing FSH concentration. Results demonstrate that dog follicles cultured in alginate retain structural integrity, grow in size and are hormonally active. Lower alginate and increasing FSH concentrations promote in vitro follicle growth. However, the absence of an E2 rise in follicles cultured in FSH alone suggests the need for luteinizing hormone supplementation to support thecal cell differentiation and granulosa cell function. PMID:21502334

  13. Gene expression in mouse ovarian follicle development in vivo versus an ex vivo alginate culture system.

    PubMed

    Parrish, Elizabeth M; Siletz, Anaar; Xu, Min; Woodruff, Teresa K; Shea, Lonnie D

    2011-08-01

    Ovarian follicle maturation results from a complex interplay of endocrine, paracrine, and direct cell-cell interactions. This study compared the dynamic expression of key developmental genes during folliculogenesis in vivo and during in vitro culture in a 3D alginate hydrogel system. Candidate gene expression profiles were measured within mouse two-layered secondary follicles, multi-layered secondary follicles, and cumulus-oocyte complexes (COCs). The expression of 20 genes involved in endocrine communication, growth signaling, and oocyte development was investigated by real-time PCR. Gene product levels were compared between i) follicles of similar stage and ii) COCs derived either in vivo or by in vitro culture. For follicles cultured for 4 days, the expression pattern and the expression level of 12 genes were the same in vivo and in vitro. Some endocrine (cytochrome P450, family 19, subfamily A, polypeptide 1 (Cyp19a1) and inhibin βA subunit (Inhba)) and growth-related genes (bone morphogenetic protein 15 (Bmp15), kit ligand (Kitl), and transforming growth factor β receptor 2 (Tgfbr2)) were downregulated relative to in vivo follicles. For COCs obtained from cultured follicles, endocrine-related genes (inhibin α-subunit (Inha) and Inhba) had increased expression relative to in vivo counterparts, whereas growth-related genes (Bmp15, growth differentiation factor 9, and kit oncogene (Kit)) and zona pellucida genes were decreased. However, most of the oocyte-specific genes (e.g. factor in the germline α (Figla), jagged 1 (Jag1), and Nlrp5 (Mater)) were expressed in vitro at the same level and with the same pattern as in vivo-derived follicles. These studies establish the similarities and differences between in vivo and in vitro cultured follicles, guiding the creation of environments that maximize follicle development and oocyte quality. PMID:21610168

  14. Genistein exposure inhibits growth and alters steroidogenesis in adult mouse antral follicles.

    PubMed

    Patel, Shreya; Peretz, Jackye; Pan, Yuan-Xiang; Helferich, William G; Flaws, Jodi A

    2016-02-15

    Genistein is a naturally occurring isoflavone phytoestrogen commonly found in plant products such as soybeans, lentils, and chickpeas. Genistein, like other phytoestrogens, has the potential to mimic, enhance, or impair the estradiol biosynthesis pathway, thereby potentially altering ovarian follicle growth. Previous studies have inconsistently indicated that genistein exposure may alter granulosa cell proliferation and hormone production, but no studies have examined the effects of genistein on intact antral follicles. Thus, this study was designed to test the hypothesis that genistein exposure inhibits follicle growth and steroidogenesis in intact antral follicles. To test this hypothesis, antral follicles isolated from CD-1 mice were cultured with vehicle (dimethyl sulfoxide; DMSO) or genistein (6.0 and 36μM) for 18-96h. Every 24h, follicle diameters were measured to assess growth. At the end of each culture period, the media were pooled to measure hormone levels, and the cultured follicles were collected to measure expression of cell cycle regulators and steroidogenic enzymes. The results indicate that genistein (36μM) inhibits growth of mouse antral follicles. Additionally, genistein (6.0 and 36μM) increases progesterone, testosterone, and dehydroepiandrosterone (DHEA) levels, but decreases estrone and estradiol levels. The results also indicate that genistein alters the expression of steroidogenic enzymes at 24, 72 and 96h, and the expression of cell cycle regulators at 18h. These data indicate that genistein exposure inhibits antral follicle growth by inhibiting the cell cycle, alters sex steroid hormone levels, and dysregulates steroidogenic enzymes in cultured mouse antral follicles. PMID:26792615

  15. Wnt/β-catenin signaling in dermal condensates is required for hair follicle formation.

    PubMed

    Tsai, Su-Yi; Sennett, Rachel; Rezza, Amélie; Clavel, Carlos; Grisanti, Laura; Zemla, Roland; Najam, Sara; Rendl, Michael

    2014-01-15

    Broad dermal Wnt signaling is required for patterned induction of hair follicle placodes and subsequent Wnt signaling in placode stem cells is essential for induction of dermal condensates, cell clusters of precursors for the hair follicle dermal papilla (DP). Progression of hair follicle formation then requires coordinated signal exchange between dermal condensates and placode stem cells. However, it remains unknown whether continued Wnt signaling in DP precursor cells plays a role in this process, largely due to the long-standing inability to specifically target dermal condensates for gene ablation. Here we use the Tbx18(Cre) knockin mouse line to ablate the Wnt-responsive transcription factor β-catenin specifically in these cells at E14.5 during the first wave of guard hair follicle formation. In the absence of β-catenin, canonical Wnt signaling is effectively abolished in these cells. Sox2(+) dermal condensates initiate normally; however by E16.5 guard hair follicle numbers are strongly reduced and by E18.5 most whiskers and guard hair follicles are absent, suggesting that active Wnt signaling in dermal condensates is important for hair follicle formation to proceed after induction. To explore the molecular mechanisms by which Wnt signaling in dermal condensates regulates hair follicle formation, we analyze genome-wide the gene expression changes in embryonic β-catenin null DP precursor cells. We find altered expression of several signaling pathway genes, including Fgfs and Activin, both previously implicated in hair follicle formation. In summary, these data reveal a functional role of Wnt signaling in DP precursors for embryonic hair follicle formation and identify Fgf and Activin signaling as potential effectors of Wnt signaling-regulated events.

  16. Spatial epidemiology of Toxoplasma gondii infection in goats in Serbia.

    PubMed

    Djokic, Vitomir; Klun, Ivana; Musella, Vincenzo; Rinaldi, Laura; Cringoli, Giuseppe; Sotiraki, Smaragda; Djurkovic-Djakovic, Olgica

    2014-05-01

    A major risk factor for Toxoplasma gondii infection is consumption of undercooked meat. Increasing demand for goat meat is likely to promote the role of this animal for human toxoplasmosis. As there are virtually no data on toxoplasmosis in goats in Serbia, we undertook a cross-sectional serological study, including prediction modelling using geographical information systems (GIS). Sera from 431 goats reared in 143 households/farms throughout Serbia, sampled between January 2010 and September 2011, were examined for T. gondii antibodies by a modified agglutination test. Seroprevalence was 73.3% at the individual level and 84.6% at the farm level. Risk factor analysis showed above two-fold higher risk of infection for goats used for all purposes compared to dairy goats (P = 0.012), almost seven-fold higher risk for goats kept as sole species versus those kept with other animals (P = 0.001) and a two-fold lower risk for goats introduced from outside the farm compared to those raised on the farm (P = 0.027). Moreover, households/farms located in centre-eastern Serbia were found to be less often infected than those in northern Serbia (P = 0.004). The risk factor analysis was fully supported by spatial analysis based on a GIS database containing data on origin, serology, land cover, elevation, meteorology and a spatial prediction map based on kriging analysis, which showed western Serbia as the area most likely for finding goats positive for T. gondii and centre-eastern Serbia as the least likely. In addition, rainfall favoured seropositivity, whereas temperature, humidity and elevation did not. PMID:24893025

  17. Monitoring of clinical signs in goats with transmissible spongiform encephalopathies

    PubMed Central

    2010-01-01

    Background As there is limited information about the clinical signs of BSE and scrapie in goats, studies were conducted to describe the clinical progression of scrapie and BSE in goats and to evaluate a short clinical protocol for its use in detecting scrapie-affected goats in two herds with previously confirmed scrapie cases. Clinical assessments were carried out in five goats intracerebrally infected with the BSE agent as well as five reported scrapie suspects and 346 goats subject to cull from the two herds, 24 of which were retained for further monitoring. The brain and selected lymphoid tissue were examined by postmortem tests for disease confirmation. Results The sensitivity and specificity of the short clinical protocol in detecting a scrapie case in the scrapie-affected herds was 3.9% and 99.6%, respectively, based on the presence of tremor, positive scratch test, extensive hair loss, ataxia and absent menace response. All BSE- and scrapie-affected goats displayed abnormalities in sensation (over-reactivity to external stimuli, startle responses, pruritus, absent menace response) and movement (ataxia, tremor, postural deficits) at an advanced clinical stage but the first detectable sign associated with scrapie or BSE could vary between animals. Signs of pruritus were not always present despite similar prion protein genotypes. Clinical signs of scrapie were also displayed by two scrapie cases that presented with detectable disease-associated prion protein only in lymphoid tissues. Conclusions BSE and scrapie may present as pruritic and non-pruritic forms in goats. Signs assessed for the clinical diagnosis of scrapie or BSE in goats should include postural and gait abnormalities, pruritus and visual impairment. However, many scrapie cases will be missed if detection is solely based on the display of clinical signs. PrPd accumulation in the brain appeared to be related to the severity of clinical disease but not to the display of individual neurological signs

  18. Hyperimmune goat serum for amyotrophic lateral sclerosis.

    PubMed

    Mackenzie, R; Kiernan, M; McKenzie, D; Youl, B D

    2006-12-01

    The authors report a patient with amyotrophic lateral sclerosis (ALS) who showed a lessening of deterioration in respiratory muscle strength during treatment with hyperimmune goat serum (HGS) (Aimspro). Respiratory function tests (RFTs) were measured by established protocols, and all measurements were expressed as a percentage of normal predicted values. The rate of decline was calculated by linear regression analysis. Respiratory muscle strength decline was less during 13 months of treatment with HGS (mean 1.3% per month, range 0.8-1.7%) compared to the preceding 13 months (mean 2.3% per month, range 1.2-3.1%), while a greater decline would be expected with disease progression. Comparison with similarly affected patients in the literature suggest that a decline of 4-5% per month of predicted values may be expected during the treatment phase.

  19. Advanced assisted reproduction technologies (ART) in goats.

    PubMed

    Baldassarre, H; Karatzas, C N

    2004-07-01

    Assisted reproduction technologies (ART) are reviewed with special emphasis on goat genetic improvement programs. Estrous synchronization and artificial insemination are the most commonly used ART worldwide because of their simplicity and excellent cost/benefit, especially when proven sires are used. Multiple ovulation and embryo transfer (MOET) has not become widely used due to its unpredictability. In vitro embryo production using oocytes collected by laparoscopy from valuable donors has the potential to improve the results obtained from MOET and expand its applications (for example, using prepubertal donors). However, the costs and inefficiencies of the system might restrict its use to special situations. Finally, transgenesis and cloning are expected to have a significant impact on the future genetic improvement of livestock. However, because of low efficiencies and high costs, their present use is restricted to applications with high returns such as the production of recombinant proteins of pharmaceutical and biomedical interest.

  20. Toxicity of Commiphora myrrha to goats.

    PubMed

    Omer, S A; Adam, S E

    1999-10-01

    An investigation was made of Commiphora myrrha used in traditional medicine for the treatment of various ailments. Twelve 6-mo-old male Nubian goat kids were assigned to 4 groups: undosed controls, C myrrha-dosed at 0.25 g plant resin/kg/d, C myrrha-dosed at 1 g resin/kg/d and C myrrha dosed at 5 g plant resin/kg/d. Results of hepatorenal function tests were correlated with clinical and pathological changes. The use of 1 or 5 g plant resin/kg/d caused grinding of teeth, salivation, soft feces, inappetence, jaundice, dyspnea, ataxia and recumbency. Death occurred between 5 and 16 d. Enterohepatonephrotoxicity was accompanied by anemia, leucopenia, increases in serum ALP activity and concentrations of bilirubin, cholesterol, triglycerides and creatinine, and decreases in total protein and albumin. The oral dose of 0.25 g plant resin/kg/d was not toxic.

  1. The advantages of hair follicle pluripotent stem cells over embryonic stem cells and induced pluripotent stem cells for regenerative medicine.

    PubMed

    Amoh, Yasuyuki; Katsuoka, Kensei; Hoffman, Robert M

    2010-12-01

    Multipotent adult stem cells have many potential therapeutic applications. Our recent findings suggest that hair follicles are a promising source of easily accessible multipotent stem cells. Stem cells in the hair follicle area express the neural stem cell marker nestin, suggesting that hair-follicle stem cells and neural stem cells have common features. Nestin-expressing hair follicle stem cells can form neurons and other cell types, and thus adult hair follicle stem cells could have important therapeutic applications, particularly for neurologic diseases. Transplanted hair follicle stem cells promote the functional recovery of injured peripheral nerve and spinal cord. Recent findings suggest that direct transplantation of hair-follicle stem cells without culture can promote nerve repair, which makes them potentially clinically practical. Human hair follicle stem cells as well as mouse hair follicle stem cells promote nerve repair and can be applied to test the hypothesis that human hair follicle stem cells can provide a readily available source of neurologically therapeutic stem cells. The use of hair follicle stem cells for nerve regeneration overcomes critical problems of embryonic stem cells or induced pluripotent stem cells in that the hair follicle stem cells are multipotent, readily accessible, non-oncogenic, and are not associated with ethical issues.

  2. Focusing on the goat casein complex.

    PubMed

    Caroli, A; Chiatti, F; Chessa, S; Rignanese, D; Bolla, P; Pagnacco, G

    2006-08-01

    The analysis of casein polymorphisms in goat species is rather difficult, because of a large number of mutations at each locus, and the tight linkage involving the 4 casein genes. Three goat breeds from Northern Italy, Orobica, Verzasca, and Frisa, were analyzed at the casein complex by milk isoelectrofocusing and analyses at the DNA level to identify the majority of all known polymorphisms. The casein gene structure of the 3 local breeds at alpha(S1)-casein (CSN1S1), beta-casein (CSN2), alpha(S2)-casein (CSN1S2), and kappa-casein (CSN3) was compared with that of Camosciata, a more widely distributed breed. A new allele was identified and characterized at CSN2 gene, which seemed to be specific to the Frisa breed. It was named CSN2*E, and was characterized by a transversion TCT --> TAT responsible for the amino acid exchange Ser(166) --> Tyr(166) in the mature protein. The casein haplotype structure is highly different among breeds. A total of 26 haplotypes showed a frequency higher than 0.01 in at least 1 of the 4 breeds considered, with 12, 3, 5, and 19 haplotypes in Frisa, Orobica, Verzasca, and Camosciata breeds, respectively. Only 13 haplotypes occurred at a frequency higher than 0.05 in at least 1 breed. With the molecular knowledge of each locus, the ancestral haplotype coding for CSN1S1*B, CSN2*A, CSN1S2*A, and CSN3*B protein variants can be postulated. A protein evolutionary model considering the whole casein haplotype is proposed.

  3. Phylogenetic analysis of Sicilian goats reveals a new mtDNA lineage.

    PubMed

    Sardina, M T; Ballester, M; Marmi, J; Finocchiaro, R; van Kaam, J B C H M; Portolano, B; Folch, J M

    2006-08-01

    The mitochondrial hypervariable region 1 (HVR1) sequence of 67 goats belonging to the Girgentana, Maltese and Derivata di Siria breeds was partially sequenced in order to present the first phylogenetic characterization of Sicilian goat breeds. These sequences were compared with published sequences of Indian and Pakistani domestic goats and wild goats. Mitochondrial lineage A was observed in most of the Sicilian goats. However, three Girgentana haplotypes were highly divergent from the Capra hircus clade, indicating that a new mtDNA lineage in domestic goats was found.

  4. A serologic survey of Mycoplasma putrefaciens infection in goats.

    PubMed

    Abegunde, T O; Adler, H E; Farver, T B; DaMassa, A J

    1981-10-01

    The prevalence of Mycoplasma putrefaciens infection in goat populations in Mendocino and Sonoma counties of northern California was studied, using the plate and tube agglutination tests. On a county basis, Mendocino had a higher antibody prevalence (13%) than Sonoma (10%). The overall antibody prevalence among the 377 goat serum samples tested was 11%. There was no statistical evidence to show any significant difference in prevalence on the basis of herd size. Of the common goat breeds in California, the American La Mancha had the lowest prevalence (4.7%), the Toggenberg, highest (10.8%). Angora goats shipped from Texas showed a much higher prevalence (67%) than any of the California breeds. The age-specific risk calculations indicate that all age groups were more susceptible to M putrefaciens than 4-year-old goats, with the lowest prevalence of 3.8%. The highest prevalence (21.3%) was observed in the Angora goats. Males had a lower prevalence (10.7%) than females (16.1%). A flock of sheep included in the survey showed a prevalence of 15%. PMID:7325445

  5. Carnivore fecal chemicals suppress feeding by Alpine goats (Capra hircus).

    PubMed

    Weldon, P J; Graham, D P; Mears, L P

    1993-12-01

    The efficacy of carnivore and ungulate fecal chemicals in suppressing the feeding behavior of Alpine goats (Capra hircus) was examined. In the first four experiments, goats were offered food covered with paper strips treated with fecal extracts of the Bengal tiger, Siberian tiger, African lion, and brown bear, respectively; food covered with solvent-treated and untreated (plain) papers served as controls in each experiment. Goats made fewer head entries into, and ate less food from, buckets containing fecal extracts. In the fifth experiment, goats were offered food covered with paper strips treated with fecal extracts of the puma, Dorcas gazelle, white-bearded gnu, and conspecifics; food covered with solvent-treated and plain papers again served as controls. The amounts of food consumed from buckets containing puma, gazelle, gnu, and solvent treatments were statistically indistinguishable, but less food was consumed from them than from buckets containing the goat-scented or plain papers. No significant differences among treatments were detected with respect to head entries. Field experiments are needed on the use of predator-derived chemicals to reduce damage by goats to vegetation.

  6. Microsatellite-based phylogeny of Indian domestic goats

    PubMed Central

    Rout, Pramod K; Joshi, Manjunath B; Mandal, Ajoy; Laloe, D; Singh, Lalji; Thangaraj, Kumarasamy

    2008-01-01

    Background The domestic goat is one of the important livestock species of India. In the present study we assess genetic diversity of Indian goats using 17 microsatellite markers. Breeds were sampled from their natural habitat, covering different agroclimatic zones. Results The mean number of alleles per locus (NA) ranged from 8.1 in Barbari to 9.7 in Jakhrana goats. The mean expected heterozygosity (He) ranged from 0.739 in Barbari to 0.783 in Jakhrana goats. Deviations from Hardy-Weinberg Equilibrium (HWE) were statistically significant (P < 0.05) for 5 loci breed combinations. The DA measure of genetic distance between pairs of breeds indicated that the lowest distance was between Marwari and Sirohi (0.135). The highest distance was between Pashmina and Black Bengal. An analysis of molecular variance indicated that 6.59% of variance exists among the Indian goat breeds. Both a phylogenetic tree and Principal Component Analysis showed the distribution of breeds in two major clusters with respect to their geographic distribution. Conclusion Our study concludes that Indian goat populations can be classified into distinct genetic groups or breeds based on the microsatellites as well as mtDNA information. PMID:18226239

  7. [The seaweed Sargassum (Sargassaceae) as tropical alternative for goats' feeding].

    PubMed

    Casas-Valdez, M; Hernández-Contreras, H; Marín-Alvarez, A; Aguila-Ramírez, R N; Hernández-Guerrero, C J; Sánchez-Rodríguez, I; Carrillo-Domínguez, S

    2006-03-01

    The seaweed Sargassum (Sargassaceae) as tropical alternative for goats' feeding. The nutritive value of seaweed (Sargassum spp.) was studied in Baja California Sur, Mexico. Twenty female Nubian goats (43-weeks old) were randomly distributed into two groups of 10 goats each and were housed in individual pens. One group was fed with a control diet and the other with a diet supplemented with 25% of Sargassum spp. Feed and water intake were recorded daily and individually for 60 days. The weight of each goat was recorded every 15 days. The nutritional content of Sargassum spp. was 89% dry mater, 8% crude protein, 31% ash, 2% ether extract, and 39% carbohydrates. Fiber fractions, minerals, vitamins, fatty acids, and antinutritional factors were also determined. There were no significant differences in body weight (8.6 kg control and 9 kg experimental), feed intake (1.3 kg control and 1.6 kg experimental), and feed conversion rate (11.1 control and 12.6 experimental). Water consumption was greater in the goats that ate the Sargassum diet (5.3 1). From these results, Sargassum spp. can be considered as an alternative feedstuff for goats.

  8. Comparison of the Fecal Microbiota in Feral and Domestic Goats

    PubMed Central

    De Jesús-Laboy, Kassandra M.; Godoy-Vitorino, Filipa; Piceno, Yvette M.; Tom, Lauren M.; Pantoja-Feliciano, Ida G.; Rivera-Rivera, Michelle J.; Andersen, Gary L.; Domínguez-Bello, María G.

    2011-01-01

    Animals have co-evolved with mutualistic microbial communities, known as the microbiota, which are essential for organ development and function. We hypothesize that modern animal husbandry practices exert an impact on the intestinal microbiota. In this study, we compared the structure of the fecal microbiota between feral and domestic goats using the G2 PhyloChip and assessed the presence of five tetracycline resistance genes [tet(M), tet(S), tet(O), tet(Q) and tet(W)] by PCR. Feces were collected from 10 goats: 5 domestic from a farm in the main island of Puerto Rico and 5 feral from the remote dry island of Mona. There were 42 bacterial phyla from 153 families detected in the goats’ feces. A total of 84 PhyloChip-OTUs were different in the fecal microbiota of feral and domestic goat. Both feral and domestic goats carried antibiotic resistance genes tet(O) and tet(W), but domestic goats additionally carried tet(Q). Diet, host genetics and antibiotic exposure are likely determinant factors in shaping the intestinal microbiota and may explain the differences observed between feral and domestic goats fecal microbiota. PMID:24704840

  9. Carnivore fecal chemicals suppress feeding by Alpine goats (Capra hircus).

    PubMed

    Weldon, P J; Graham, D P; Mears, L P

    1993-12-01

    The efficacy of carnivore and ungulate fecal chemicals in suppressing the feeding behavior of Alpine goats (Capra hircus) was examined. In the first four experiments, goats were offered food covered with paper strips treated with fecal extracts of the Bengal tiger, Siberian tiger, African lion, and brown bear, respectively; food covered with solvent-treated and untreated (plain) papers served as controls in each experiment. Goats made fewer head entries into, and ate less food from, buckets containing fecal extracts. In the fifth experiment, goats were offered food covered with paper strips treated with fecal extracts of the puma, Dorcas gazelle, white-bearded gnu, and conspecifics; food covered with solvent-treated and plain papers again served as controls. The amounts of food consumed from buckets containing puma, gazelle, gnu, and solvent treatments were statistically indistinguishable, but less food was consumed from them than from buckets containing the goat-scented or plain papers. No significant differences among treatments were detected with respect to head entries. Field experiments are needed on the use of predator-derived chemicals to reduce damage by goats to vegetation. PMID:24248787

  10. The timing of oestrus, the preovulatory LH surge and ovulation in Blanca Andaluza goats synchronised by intravaginal progestagen sponge treatment is modified by season but not by body condition score.

    PubMed

    Zarazaga, L A; Gatica, M C; Gallego-Calvo, L; Celi, I; Guzmán, J L

    2014-05-01

    The aim of this study was to determine whether a seasonal pattern of reproductive events is followed after synchronisation by intravaginal progestagen sponge treatment in female Blanca Andaluza goats, and whether the timing of these events is affected by body condition score (BCS). During seasonal anoestrus (March), and again during the breeding season (November), the same 32 does were distributed into four subgroups according to their BCS: ≤2.25, =2.50, =2.75, and ≥3.00 (n=8 in all cases). They were then synchronised using a commercial intravaginal sponge treatment. Every 4h over the 72h following sponge removal, oestrous activity, the LH concentration and each doe's number of follicles were followed by transrectal ultrasonography. The does synchronised during seasonal anoestrus produced more follicles than those synchronised during the breeding season (P<0.01). The time elapsed between sponge removal and the onset of oestrus, the LH surge and time of ovulation, was also shorter in these does (P<0.001). The BCS only modified the number of follicles present in the ovary just before ovulation; this number was significantly lower in the =2.50 BCS subgroup than in the other subgroups (P<0.05). The present results show that the time to ovulation, and all events around it, are modified by the season in which Blanca Andaluza does are synchronised, but not by BCS.

  11. Seroprevalence and risk factors of Q fever in goats on commercial dairy goat farms in the Netherlands, 2009-2010

    PubMed Central

    2011-01-01

    Background The aim of this study was to estimate the seroprevalence of Coxiella burnetii in dairy goat farms in the Netherlands and to identify risk factors for farm and goat seropositivity before mandatory vaccination started. We approached 334 eligible farms with more than 100 goats for serum sampling and a farm questionnaire. Per farm, median 21 goats were sampled. A farm was considered positive when at least one goat tested ELISA positive. Results In total, 2,828 goat serum samples from 123 farms were available. Farm prevalence was 43.1% (95%CI: 34.3%-51.8%). Overall goat seroprevalence was 21.4% (95%CI: 19.9%-22.9%) and among the 53 positive farms 46.6% (95%CI: 43.8%-49.3%). Multivariable logistic regression analysis included 96 farms and showed that farm location within 8 kilometres proximity from a bulk milk PCR positive farm, location in a municipality with high cattle density (≥ 100 cattle per square kilometre), controlling nuisance animals through covering airspaces, presence of cats or dogs in the goat stable, straw imported from abroad or unknown origin and a herd size above 800 goats were independent risk factors associated with Q fever on farm level. At animal level almost identical risk factors were found, with use of windbreak curtain and artificial insemination as additional risk factors. Conclusion In 2009-2010, the seroprevalence in dairy goats in the Netherlands increased on animal and farm level compared to a previous study in 2008. Risk factors suggest spread from relatively closely located bulk milk-infected small ruminant farms, next to introduction and spread from companion animals, imported straw and use of artificial insemination. In-depth studies investigating the role of artificial insemination and bedding material are needed, while simultaneously general biosecurity measures should be updated, such as avoiding companion animals and vermin entering the stables, next to advice on farm stable constructions on how to prevent introduction

  12. Effect of copulation on estrus duration and ovulation time in goats.

    PubMed

    Romano, Juan E; Alkar, Abdalhamid; Amstalden, Marcel

    2016-01-15

    The objective of the present study was to evaluate the effect of copulation on estrus duration and ovulation in goats. During the fall season, 14 multiparous Boer does were estrous synchronized with controlled internal drug release (300 mg), maintained in the vagina for 7 days, and received 50 μg of intramuscular GnRH device insertion and 5 mg of natural intramuscular PGF2α at device removal. The does were randomly divided into two equal groups: a treatment group (TRE; n = 7) and a control group (CON; n = 7). The TRE group received two copulas by fertile bucks within the first 4 hours of estrus onset, and the CON group received only mounts by the same males equipped with canvas aprons. Estrus detection was performed every 12 hours after controlled internal drug release removal within the first 24 hours and then every 4 hours for 5 days. Estrus was defined when a doe accepted mounting by the bucks equipped with canvas aprons. Each doe in estrus got the first transrectal ultrasonography at 24 hours after estrus onset and then every 4 hours until all the preovulatory follicles ovulated. Estrus onset for the TRE and CON groups was 40.3 ± 17.4 (mean ± standard deviation) and 43.3 ± 12.2 hours (P = 0.72), respectively. Estrus duration for the same groups was 28.6 ± 5.4 and 36.7 ± 5.3 hours (P = 0.02), respectively. The mean ovulation time for the TRE and CON groups was 31.4 ± 2.2 and 35.7 ± 3.7 hours (P = 0.04), respectively. The proportion of ovulations that occurred after the end of estrus in the TRE group was higher than in the CON group (86% vs. 33%, respectively; P = 0.05). The number of ovulations for the TRE group was 2.1 ± 0.7; for the CON group, there were 2.2 ± 0.5 ovulations (P = 0.92). It was concluded that copulation by a buck at the beginning of estrus reduced estrus duration and hastened the ovulation time in Boer goats.

  13. Efficacy of albendazole against nematode parasites isolated from a goat farm in Ethiopia: relationship between dose and efficacy in goats.

    PubMed

    Eguale, Tadesse; Chaka, Hassen; Gizaw, Daniel

    2009-10-01

    A suspected case of albendazole resistance in a goat farm of Hawassa University was examined using faecal egg count reduction test (FECRT), controlled anthelmintic efficacy test and egg hatch assay (EHA) to verify the development of resistance and/or the need for higher doses of the drug in goats than in sheep. The experiment was conducted in 12 sheep (2 groups: treatment versus control) and 24 goats (4 groups: 3 treatments versus control, n = 6; per group) following artificial infection with infective larvae of Haemonchus contortus and Oesophagostomum columbianum. The first group of sheep and goats were treated orally with albendazole at the dose rate of 3.8 mg/kg body weight (i.e. manufacturer's recommended dose for sheep) while the second group of sheep and the fourth group of goats were left untreated. The second and the third group of goats were treated with albendazole at 5.7 and 7.6 mg/kg respectively. The FECRT showed an efficacy of albendazole in goats to be 65.5, 81.4 and 84.1% at the dose rate of 3.8, 5.7 and 7.6 mg/kg body weight respectively while in sheep it was 62% at the dose rate of 3.8 mg/kg. Increasing the dose to 1.5 the sheep recommended dose induced minor improvement of efficacy in goats; however the efficacy was almost the same at 1.5 and twice the dose recommended for sheep. Worm counts at day 15 post-treatment revealed that H. contortus has developed resistance to albendazole. EHA results also supported these findings. On the other hand, O. columbianum was 100% susceptible at all dose levels tested.

  14. Wnt10b promotes differentiation of mouse hair follicle melanocytes.

    PubMed

    Ye, Jixing; Yang, Tian; Guo, Haiying; Tang, Yinhong; Deng, Fang; Li, Yuhong; Xing, Yizhan; Yang, Li; Yang, Ke

    2013-01-01

    Previous research has revealed that Wnt10b activates canonical Wnt signaling, which is integral to melanocyte differentiation in hair follicles (HFs). However, the function of Wnt10b in HF melanocytes remains poorly understood. We determined using Dct-LacZ transgenic mice that Wnt10b is mainly expressed near and within melanocytes of the hair bulbs during the anagen stage of the hair cycle. We also found that Wnt10b promotes an increase in melanocyte maturation and pigmentation in the hair bulbs of the mouse HF. To further explore the potential functions of Wnt10b in mouse HF melanocytes, we infected iMC23 cells with Ad-Wnt10b to overexpress Wnt10b. We demonstrated that Wnt10b promotes the differentiation of melanocytes by activating canonical Wnt signaling in melanocytes.

  15. Morphology of preovulatory bovine follicles as related to oocyte maturation.

    PubMed

    de Loos, F A; Bevers, M M; Dieleman, S J; Kruip, T A

    1991-03-01

    Thirty-three preovulatory bovine oocytes and their follicles were collected during the period of final maturation in normally cyclic cows. Cell density of the membrana granulosa, mitotic index of the membrana granulosa, and the occurrence of eosinophilic granulocytes around the basal membrane as well as the maturational stage of the oocyte were determined. Cell density decreased during the period of final maturation. Mitotic indices also decreased after an initial high level in the first hours of the final maturation. Eosinophilic granulocytes were only seen during the last hours of final maturation. The maturational stages of the oocytes were related to distinct maturational stages of the follicular wall as determined by morphological characteristics. We propose a scoring system for the maturity of the follicular wall based on cell density, presence of mitotic figures and the presence of eosinophilic granulocytes outside the vascular compartment. PMID:16726922

  16. Androgen actions on the human hair follicle: perspectives.

    PubMed

    Inui, Shigeki; Itami, Satoshi

    2013-03-01

    Androgens stimulate beard growth but suppress hair growth in androgenetic alopecia (AGA). This condition is known as 'androgen paradox'. Human pilosebaceous units possess enough enzymes to form the active androgens testosterone and dihydrotestosterone. In hair follicles, 5α-reductase type 1 and 2, androgen receptors (AR) and AR coactivators can regulate androgen sensitivity of dermal papillae (DP). To regulate hair growth, androgens stimulate production of IGF-1 as positive mediators from beard DP cells and of TGF-β1, TGF-β2, dickkopf1 and IL-6 as negative mediators from balding DP cells. In addition, androgens enhance inducible nitric oxide synthase from occipital DP cells and stem cell factor for positive regulation of hair growth in beard and negative regulation of balding DP cells. Moreover, AGA involves crosstalk between androgen and Wnt/β-catenin signalling. Finally, recent data on susceptibility genes have provided us with the impetus to investigate the molecular pathogenesis of AGA. PMID:23016593

  17. Haemonchotolerance in West African Dwarf goats: contribution to sustainable, anthelmintics-free helminth control in traditionally managed Nigerian dwarf goats.

    PubMed

    Chiejina, Samuel N; Behnke, Jerzy M; Fakae, Barineme B

    2015-01-01

    West African Dwarf (WAD) goats are extremely important in the rural village economy of West Africa, but still little is known about their biology, ecology and capacity to cope with gastrointestinal nematode (GIN) infections. Here, we summarise the history of this breed and explain its economic importance in rural West Africa. We review recent work showing that Nigerian WAD goats are highly trypanotolerant and resist infections with Haemonchus contortus more effectively than other breeds of domestic goat (haemonchotolerance). We believe that haemonchotolerance is largely responsible for the generally low level GIN infections and absence of clinical haemonchosis in WADs under field conditions, and has contributed to the relatively successful and sustainable, anthelmintics-free, small-scale system of goat husbandry in Nigeria's humid zone, and is immunologically based and genetically controlled. If haemonchotolerance can be shown to be genetically controlled, it should be possible to exploit the underlying genes to improve GIN resistance among productive fibre and milk producing breeds of goats, most of which are highly susceptible to nematode infections. Genetic resistance to GIN and trypanosome infections would obviate the need for expensive chemotherapy, mostly unaffordable to small-holder farmers in Africa, and a significant cost of goat husbandry in more developed countries. Either introgression of resistance alleles into susceptible breeds by conventional breeding, or transgenesis could be used to develop novel parasite-resistant, but highly productive breeds, or to improve the resistance of existing breeds, benefitting the local West African rural economy as well as global caprine livestock agriculture.

  18. Male goat vocalizations stimulate the estrous behavior and LH secretion in anestrous goats that have been previously exposed to bucks.

    PubMed

    Delgadillo, José Alberto; Vielma, Jesús; Hernandez, Horacio; Flores, José Alfredo; Duarte, Gerardo; Fernández, Ilda Graciela; Keller, Matthieu; Gelez, Hélène

    2012-09-01

    We investigated whether live vocalizations emitted by bucks interacting with anestrous females stimulate secretion of LH, estrous behavior and ovulation in anestrous goats. In experiment 1, bucks rendered sexually active by exposure to long days followed by natural photoperiod were exposed in a light-proof-building to five anestrous females. Buck vocalizations were reproduced through a microphone-amplifier-loudspeaker system to an open pen where one group of goats (n=6) was exposed for 10 days to these live vocalizations. Another group of females (n=6) was isolated from males and vocalizations. The proportion of goats displaying estrous behavior was significantly higher in females exposed to buck vocalizations than in females isolated from males. The proportion of goats that ovulated did not differ between the 2 groups (exposed to males versus isolated). In experiment 2, female goats that either had previous contact with males (n=7), or no previous contact with males (n=7) were exposed to live buck vocalizations, reproduced as described in experiment 1, for 5 days. The number and amplitude of LH pulses did not differ between groups before exposition to buck vocalizations. Five days of exposure to male vocalizations significantly increased LH pulsatility only in females that had previous contact with males, while LH pulse amplitude was not modified. We concluded that live buck vocalizations can stimulate estrous behavior and LH secretion in goats if they have had previous contact with bucks.

  19. Targeting the exogenous htPAm gene on goat somatic cell beta-casein locus for transgenic goat production.

    PubMed

    Shen, Wei; Lan, Guocheng; Yang, Xueyi; Li, Lan; Min, Lingjiang; Yang, Zhengtian; Tian, Liyuan; Wu, Xiaojie; Sun, Yujiang; Chen, Hong; Tan, Jinghe; Deng, Jixian; Pan, Qingjie

    2007-04-01

    Combining gene targeting of animal somatic cells with nuclear transfer technique has provided a powerful method to produce transgenic animal mammary gland bioreactor. The objective of this study is to make an efficient and reproducible gene targeting in goat fetal fibroblasts by inserting the exogenous htPAm cDNA into the beta-casein locus with liposomes or electroporation so that htPAm protein might be produced in gene-targeted goat mammary gland. By gene-targeting technique, the exogenous htPAm gene was inserted to milk goat beta-casein gene sequences. Fetal fibroblasts were isolated from Day 35 fetuses of Guanzhong milk goats, and transfected with linear gene-targeting vector pGBC4htPAm using Lipefectamin-2000 and electoporation, respectively. Forty-eight gene-targeted cell colonies with homologous recombination were obtained, and three cell colonies were verified by DNA sequence analysis within the homologous recombination region. Using gene-targeted cell lines as donor cells for nuclear transfer, a total of 600 reconstructed embryos had been obtained, and 146 developed cloned embryos were transferred to 16 recipient goats, and finally three goats showed pregnancy at Day 90.

  20. Multifactorial regulation of prostaglandin synthesis in preovulatory goldfish ovarian follicles.

    PubMed

    Kellner, R G; Van der Kraak, G

    1992-04-01

    Goldfish preovulatory ovarian follicles (prior to germinal vesicle breakdown) were utilized for studies investigating the actions of activators of different signal transduction pathways on prostaglandin (PG) production. The protein kinase C (PKC) activators phorbol 12-myristate 13-acetate (PMA; 100-400 nM), 1-oleoyl-2-acetylglycerol (5 and 25 micrograms/ml), and 1,2-dioctanoylglycerol (10 and 50 micrograms/ml) stimulated PGE production; the inactive phorbol 4 alpha-phorbol didecanoate, which does not activate PKC, had no effect. Calcium ionophore A23187 (0.25-4.0 microM) stimulated PGE production and acted in a synergistic manner with activators of PKC. Although produced in lower amounts than PGE, PGF was stimulated by PMA and A23187. The direct activator of phospholipase A2, melittin (0.1-1.0 microM), stimulated a dose-related increase in PGE production, whereas chloroquine (100 microM), a putative inhibitor of phospholipase A2, blocked basal and PMA + A23187-stimulated PGE production. Several drugs known to elevate intracellular levels of cAMP including the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0.1-1.0 mM), forskolin (10 microM), and dibutyryl cAMP (dbcAMP; 5 mM) attenuate PMA + A23187-stimulated PGE production. Melittin-stimulated production of PGE was inhibited by dbcAMP, suggesting that the action of cAMP was distal to the activation of phospholipase A2. In summary, these studies demonstrate that activation of PKC and elevation of intracellular calcium levels stimulate PG production, in part, through activation of phospholipase A2. The adenylate cyclase/cAMP signalling pathway is inhibitory to PG production by goldfish ovarian follicles. PMID:1315582

  1. Analysis of polymorphisms in milk proteins from cloned and sexually reproduced goats.

    PubMed

    Xing, H; Shao, B; Gu, Y Y; Yuan, Y G; Zhang, T; Zang, J; Cheng, Y

    2015-12-08

    This study evaluates the relationship between the genotype and milk protein components in goats. Milk samples were collected from cloned goats and normal white goats during different postpartum (or abortion) phases. Two cloned goats, originated from the same somatic line of goat mammary gland epithelial cells, and three sexually reproduced normal white goats with no genetic relationships were used as the control. The goats were phylogenetically analyzed by polymerase chain reaction-restriction fragment length polymorphism. The milk protein components were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results indicated that despite the genetic fingerprints being identical, the milk protein composition differed between the two cloned goats. The casein content of cloned goat C-50 was significantly higher than that of cloned goat C-4. Conversely, although the genetic fingerprints of the normal white goats N-1, N-2, and N-3 were not identical, the milk protein profiles did not differ significantly in their milk samples (obtained on postpartum day 15, 20, 25, 30, and 150). These results indicated an association between milk protein phenotypes and genetic polymorphisms, epigenetic regulation, and/or non-chromosomal factors. This study extends the knowledge of goat milk protein polymorphisms, and provides new strategies for the breeding of high milk-yielding goats.

  2. Analysis of polymorphisms in milk proteins from cloned and sexually reproduced goats.

    PubMed

    Xing, H; Shao, B; Gu, Y Y; Yuan, Y G; Zhang, T; Zang, J; Cheng, Y

    2015-01-01

    This study evaluates the relationship between the genotype and milk protein components in goats. Milk samples were collected from cloned goats and normal white goats during different postpartum (or abortion) phases. Two cloned goats, originated from the same somatic line of goat mammary gland epithelial cells, and three sexually reproduced normal white goats with no genetic relationships were used as the control. The goats were phylogenetically analyzed by polymerase chain reaction-restriction fragment length polymorphism. The milk protein components were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results indicated that despite the genetic fingerprints being identical, the milk protein composition differed between the two cloned goats. The casein content of cloned goat C-50 was significantly higher than that of cloned goat C-4. Conversely, although the genetic fingerprints of the normal white goats N-1, N-2, and N-3 were not identical, the milk protein profiles did not differ significantly in their milk samples (obtained on postpartum day 15, 20, 25, 30, and 150). These results indicated an association between milk protein phenotypes and genetic polymorphisms, epigenetic regulation, and/or non-chromosomal factors. This study extends the knowledge of goat milk protein polymorphisms, and provides new strategies for the breeding of high milk-yielding goats. PMID:26662412

  3. Follicle Detection on the USG Images to Support Determination of Polycystic Ovary Syndrome

    NASA Astrophysics Data System (ADS)

    Adiwijaya; Purnama, B.; Hasyim, A.; Septiani, M. D.; Wisesty, U. N.; Astuti, W.

    2015-06-01

    Polycystic Ovary Syndrome(PCOS) is the most common endocrine disorders affected to female in their reproductive cycle. This has gained the attention from married couple which affected by infertility. One of the diagnostic criteria considereded by the doctor is analysing manually the ovary USG image to detect the number and size of ovary's follicle. This analysis may affect low varibilites, reproducibility, and efficiency. To overcome this problems. automatic scheme is suggested to detect the follicle on USG image in supporting PCOS diagnosis. The first scheme is determining the initial homogeneous region which will be segmented into real follicle form The next scheme is selecting the appropriate regions to follicle criteria. then measuring the segmented region attribute as the follicle. The measurement remains the number and size that aimed at categorizing the image into the PCOS or non-PCOS. The method used is region growing which includes region-based and seed-based. To measure the follicle diameter. there will be the different method including stereology and euclidean distance. The most optimum system plan to detect PCO is by using region growing and by using euclidean distance on quantification of follicle.

  4. Primordial Follicle Transplantation within Designer Biomaterial Grafts Produce Live Births in a Mouse Infertility Model

    PubMed Central

    Kniazeva, E.; Hardy, A. N.; Boukaidi, S. A.; Woodruff, T. K.; Jeruss, J. S.; Shea, L. D.

    2015-01-01

    The gonadotoxic effects of chemotherapy and radiation may result in premature ovarian failure in premenopausal oncology patients. Although autotransplantation of ovarian tissue has led to successful live births, reintroduction of latent malignant cells inducing relapse is a significant concern. In this report, we investigated the design of biomaterial grafts for transplantation of isolated ovarian follicles as a means to preserve fertility. Primordial and primary ovarian follicles from young female mice were extracted and encapsulated into biomaterials for subsequent transplantation into adult mice. Among the formulations tested, aggregated follicles encapsulated within fibrin had enhanced survival and integration with the host tissue following transplantation relative to the fibrin-alginate and fibrin-collagen composites. All mice transplanted with fibrin-encapsulated follicles resumed cycling, and live births were achieved only for follicles transplanted within VEGF-loaded fibrin beads. The extent to which these procedures reduce the presence of metastatic breast cancer cells among the isolated follicles was evaluated, with significantly reduced numbers of cancer cells present relative to intact ovaries. This ability to obtain live births by transplanting isolated primordial and primary follicles, while also reducing the risk of re-seeding disease relative to ovarian tissue transplantation, may ultimately provide a means to preserve fertility in premenopausal oncology patients. PMID:26633657

  5. Context based algorithmic framework for identifying and classifying embedded images of follicle units

    NASA Astrophysics Data System (ADS)

    Rahman, Md. Mahbubur; Iyengar, S. S.; Zeng, Wei; Hernandez, Frank; Nusbaum, Bernard P.; Rose, Paul

    2014-03-01

    Medical image processing has been very emerging research areas in recent days. These types of images are naturally so noisy. To count the target objects is never easy. But the proper treatment depends on the accuracy of the successful locating and counting of the desired objects in an image. Some research work can do this type of segmentation of images, but they include so many constraints on the input images that these solutions cannot be applied in a generalized way to most of the images. Even a slight variation in nature of an input image can lead to a major incorrectness of the result. So we developed a generalized way to count a very noisy part of human body, the hair follicle on the scalp. The objective of this research is to count the number of hair follicle groups and the number of follicles into each group in a microscopic image of human scalp. The follicles are nonstandard in shape i.e. they do not maintain any standard shape like rectangle, oval, circle etc. Moreover the follicles are overlapping with one another in many cases. So it is hard to separate them. Here we will present a technique to count the number of follicle group as well as number of follicles in each group. We also applied well-known techniques to cluster the objects detected and a method to generate a neighboring connected graph in order to calculate the inter follicular distances.

  6. Profiling of SOD isoenzymes in compartments of the developing bovine antral follicle

    PubMed Central

    Combelles, Catherine M.H.; Holick, Emily A.; Paolella, Louis J.; Walker, David C.; Wu, Qiaqia

    2011-01-01

    The antral follicle constitutes a complex and regulated ovarian microenvironment that influences oocyte quality. Oxidative stress is a cellular state that may play a role during folliculogenesis and oogenesis, although direct supporting evidence is currently lacking. We thus evaluated the expression of the three isoforms (SOD1, 2, 3) of the enzymatic antioxidant superoxide dismutase in all of the cellular (granulosa cells, cumulus cells, oocytes) and extracellular (follicular fluid) compartments of the follicle. Comparisons were performed in bovine ovaries across progressive stages of antral follicular development. Follicular fluid possessed increased amounts of SOD1, 2, 3 in small when compared to large antral follicles; concomitantly, total SOD activity was highest in follicular fluids from smaller diameter follicles. SOD1, 2, and 3 proteins were expressed in granulosa cells without any fluctuations with follicle sizes. All three SOD isoforms were present but distributed differently in oocytes from small, medium, or large antral follicles. Cumulus cells expressed high levels of SOD3, some SOD2, but no detectable SOD1. Our studies provide a temporal and spatial expression profile of the three SOD isoforms in the different compartments of the developing bovine antral follicles. These results lay the ground for future investigations into the potential regulation and roles of antioxidants during folliculogenesis and oogenesis. PMID:20197373

  7. Combination of infrared thermography and reflectance spectroscopy for precise classification of hair follicle stage

    NASA Astrophysics Data System (ADS)

    Wang, Jianru; Guan, Yue; Liu, Caihua; Zhu, Dan

    2015-03-01

    Hair follicles enjoy continual cycle of anagen, catagen and telogen all life. They not only provide a unique opportunity to study the physiological mechanism of organ regeneration, but also benefit to guide the treatment of organ repair in regenerative medicine. Usually, the histological examination as a gold standard has been applied to determine the stage of hair follicle cycle, but noninvasive classification of hair cycle in vivo remains unsolved. In this study, the thermal infrared imager was applied to measure the temperature change of mouse dorsal skin with hair follicle cycle, and the change of diffuse reflectance was monitored by the optical fiber spectrometer. Histological examination was used to verify the hair follicle stages. The results indicated that the skin temperature increased at the beginning of anagen. After having stayed a high value for several days, the temperature began to decrease. At the same time, the skin diffuse reflectance decreased until the end of this period. Then the temperature increased gradually after slightly decreased when the hair follicle entered into catagen stage, and the diffuse reflectance increased at this time. In telogen, both the temperature and the diffuse reflectance went back to a steady state all the time. Sub-stages of hair follicle cycle could be distinguished based on the joint curves. This study provided a new method to noninvasively recognize the hair follicle stage, and should be valuable for the basic and therapeutic investigations on hair regeneration.

  8. High follicle density does not decrease sweat gland density in Huacaya alpacas.

    PubMed

    Moore, K E; Maloney, S K; Blache, D

    2015-01-01

    When exposed to high ambient temperatures, mammals lose heat evaporatively by either sweating from glands in the skin or by respiratory panting. Like other camelids, alpacas are thought to evaporate more water by sweating than panting, despite a thick fleece, unlike sheep which mostly pant in response to heat stress. Alpacas were brought to Australia to develop an alternative fibre industry to sheep wool. In Australia, alpacas can be exposed to ambient temperatures higher than in their native South America. As a young industry there is a great deal of variation in the quality and quantity of the fleece produced in the national flock. There is selection pressure towards animals with finer and denser fleeces. Because the fibre from secondary follicles is finer than that from primary follicles, selecting for finer fibres might alter the ratio of primary and secondary follicles. In turn the selection might alter sweat gland density because the sweat glands are associated with the primary follicle. Skin biopsy and fibre samples were obtained from the mid-section of 33 Huacaya alpacas and the skin sections were processed into horizontal sections at the sebaceous gland level. Total, primary, and secondary follicles and the number of sweat gland ducts were quantified. Fibre samples from each alpaca were further analysed for mean fibre diameter. The finer-fibred animals had a higher total follicle density (P<0.001) and more sweat glands (P<0.001) than the thicker-fibred animals. The fibre diameter and total follicle density were negatively correlated (R(2)=0.56, P<0.001). Given that the finer-fibred animals had higher follicle density and more sweat glands than animals with thicker fibres, we conclude that alpacas with high follicle density should not be limited for potential sweating ability.

  9. Methoxychlor inhibits growth of antral follicles by altering cell cycle regulators.

    PubMed

    Gupta, Rupesh K; Meachum, Sharon; Hernández-Ochoa, Isabel; Peretz, Jackye; Yao, Humphrey H; Flaws, Jodi A

    2009-10-01

    Methoxychlor (MXC) reduces fertility in female rodents, decreases antral follicle numbers, and increases atresia through oxidative stress pathways. MXC also inhibits antral follicle growth in vitro. The mechanism by which MXC inhibits growth of follicles is unknown. The growth of follicles is controlled, in part, by cell cycle regulators. Thus, we tested the hypothesis that MXC inhibits follicle growth by reducing the levels of selected cell cycle regulators. Further, we tested whether co-treatment with an antioxidant, N-acetyl cysteine (NAC), prevents the MXC-induced reduction in cell cycle regulators. For in vivo studies, adult cycling CD-1 mice were dosed with MXC or vehicle for 20 days. Treated ovaries were subjected to immunohistochemistry for proliferating cell nuclear antigen (PCNA) staining. For in vitro studies, antral follicles isolated from adult cycling CD-1 mouse ovaries were cultured with vehicle, MXC, and/or NAC for 48, 72 and 96 h. Levels of cyclin D2 (Ccnd2) and cyclin dependent kinase 4 (Cdk4) were measured using in vivo and in vitro samples. The results indicate that MXC decreased PCNA staining, and Ccnd2 and Cdk4 levels compared to controls. NAC co-treatment restored follicle growth and expression of Ccnd2 and Cdk4. Collectively, these data indicate that MXC exposure reduces the levels of Ccnd2 and Cdk4 in follicles, and that protection from oxidative stress restores Ccnd2 and Cdk4 levels. Therefore, MXC-induced oxidative stress may decrease the levels of cell cycle regulators, which in turn, results in inhibition of the growth of antral follicles.

  10. Magnetic resonance imaging for the study of ovarian follicles in the mouse.

    PubMed

    Stephenson, A P; Tyler, D J; Carr, C A; Williams, S A

    2012-10-01

    Additional tools to analyze follicle development would be highly advantageous because current methods require sacrifice of animals at specific times and time-consuming sectioning of tissues for histologic analysis. Magnetic resonance imaging (MRI) may provide a less involved, faster and more cost-effective method to analyze follicles in whole ovaries. Fixed ovaries were collected at different stages of the estrus cycle and after stimulation with gonadotrophins (24 and 48 h post pregnant mares serum (PMSG), and 10 and 24 h post human chorionic gonadotrophin (hCG)) with or without administration of the contrast agent gadodiamide. The MR images were generated using a vertical-bore, 11.7 Tesla MR system. Analysis of the MR images revealed large antral follicles in fixed ovaries with the oocyte and cumulus mass identifiable within preovulatory follicles. The use of gadodiamide had no impact on the quality of MR images obtained. The fixed ovaries were paraffin embedded, sectioned, and hematoxylin stained. Follicles were counted using the MR images and the histology sections. Preovulatory follicle numbers determined using MR images were comparable to those using histology; however counts of smaller follicles were inconsistent. MRI of gonadotrophin-stimulated ovaries in situ did not reveal discernable ovarian structures. Therefore, MRI is a useful tool for studying whole fixed ovaries leaving the ovary intact for additional analyses or for selection of samples based on morphology. The MRI is also useful for identifying preovulatory follicles, although analysis of smaller follicles is not possible, and thus the potential exists for cyst analysis in mouse models of polycystic ovarian syndrome (PCOS).

  11. Epidermal stem cells and skin tissue engineering in hair follicle regeneration.

    PubMed

    Balañá, María Eugenia; Charreau, Hernán Eduardo; Leirós, Gustavo José

    2015-05-26

    The reconstitution of a fully organized and functional hair follicle from dissociated cells propagated under defined tissue culture conditions is a challenge still pending in tissue engineering. The loss of hair follicles caused by injuries or pathologies such as alopecia not only affects the patients' psychological well-being, but also endangers certain inherent functions of the skin. It is then of great interest to find different strategies aiming to regenerate or neogenerate the hair follicle under conditions proper of an adult individual. Based upon current knowledge on the epithelial and dermal cells and their interactions during the embryonic hair generation and adult hair cycling, many researchers have tried to obtain mature hair follicles using different strategies and approaches depending on the causes of hair loss. This review summarizes current advances in the different experimental strategies to regenerate or neogenerate hair follicles, with emphasis on those involving neogenesis of hair follicles in adult individuals using isolated cells and tissue engineering. Most of these experiments were performed using rodent cells, particularly from embryonic or newborn origin. However, no successful strategy to generate human hair follicles from adult cells has yet been reported. This review identifies several issues that should be considered to achieve this objective. Perhaps the most important challenge is to provide three-dimensional culture conditions mimicking the structure of living tissue. Improving culture conditions that allow the expansion of specific cells while protecting their inductive properties, as well as methods for selecting populations of epithelial stem cells, should give us the necessary tools to overcome the difficulties that constrain human hair follicle neogenesis. An analysis of patent trends shows that the number of patent applications aimed at hair follicle regeneration and neogenesis has been increasing during the last decade. This

  12. Methoxychlor inhibits growth of antral follicles by altering cell cycle regulators

    SciTech Connect

    Gupta, Rupesh K. Meachum, Sharon Hernandez-Ochoa, Isabel Peretz, Jackye Yao, Humphrey H. Flaws, Jodi A.

    2009-10-01

    Methoxychlor (MXC) reduces fertility in female rodents, decreases antral follicle numbers, and increases atresia through oxidative stress pathways. MXC also inhibits antral follicle growth in vitro. The mechanism by which MXC inhibits growth of follicles is unknown. The growth of follicles is controlled, in part, by cell cycle regulators. Thus, we tested the hypothesis that MXC inhibits follicle growth by reducing the levels of selected cell cycle regulators. Further, we tested whether co-treatment with an antioxidant, N-acetyl cysteine (NAC), prevents the MXC-induced reduction in cell cycle regulators. For in vivo studies, adult cycling CD-1 mice were dosed with MXC or vehicle for 20 days. Treated ovaries were subjected to immunohistochemistry for proliferating cell nuclear antigen (PCNA) staining. For in vitro studies, antral follicles isolated from adult cycling CD-1 mouse ovaries were cultured with vehicle, MXC, and/or NAC for 48, 72 and 96 h. Levels of cyclin D2 (Ccnd2) and cyclin dependent kinase 4 (Cdk4) were measured using in vivo and in vitro samples. The results indicate that MXC decreased PCNA staining, and Ccnd2 and Cdk4 levels compared to controls. NAC co-treatment restored follicle growth and expression of Ccnd2 and Cdk4. Collectively, these data indicate that MXC exposure reduces the levels of Ccnd2 and Cdk4 in follicles, and that protection from oxidative stress restores Ccnd2 and Cdk4 levels. Therefore, MXC-induced oxidative stress may decrease the levels of cell cycle regulators, which in turn, results in inhibition of the growth of antral follicles.

  13. Transcriptome Profiling of the Theca Interna in Transition from Small to Large Antral Ovarian Follicles

    PubMed Central

    Hatzirodos, Nicholas; Hummitzsch, Katja; Irving-Rodgers, Helen F.; Rodgers, Raymond J.

    2014-01-01

    The theca interna layer of the ovarian follicle forms during the antral stage of follicle development and lies adjacent to and directly outside the follicular basal lamina. It supplies androgens and communicates with the granulosa cells and the oocyte by extracellular signaling. To better understand developmental changes in the theca interna, we undertook transcriptome profiling of the theca interna from small (3–5 mm, n = 10) and large (9–12 mm, n = 5) healthy antral bovine follicles, representing a calculated >7-fold increase in the amount of thecal tissue. Principal Component Analysis and hierarchical classification of the signal intensity plots for the arrays showed no clustering of the theca interna samples into groups depending on follicle size or subcategories of small follicles. From the over 23,000 probe sets analysed, only 76 were differentially expressed between large and small healthy follicles. Some of the differentially expressed genes were associated with processes such as myoblast differentiation, protein ubiquitination, nitric oxide and transforming growth factor β signaling. The most significant pathway affected from our analyses was found to be Wnt signaling, which was suppressed in large follicles via down-regulation of WNT2B and up-regulation of the inhibitor FRZB. These changes in the transcriptional profile could have been due to changes in cellular function or alternatively since the theca interna is composed of a number of different cell types it could have been due to any systematic change in the volume density of any particular cell type. However, our study suggests that the transcriptional profile of the theca interna is relatively stable during antral follicle development unlike that of granulosa cells observed previously. Thus both the cellular composition and cellular behavior of the theca interna and its contribution to follicular development appear to be relatively constant throughout the follicle growth phase examined

  14. Nestin-expressing hair follicle-accessible pluripotent stem cells for nerve and spinal cord repair.

    PubMed

    Hoffman, Robert M

    2014-01-01

    Nestin-expressing stem cells of the hair follicle, discovered by our laboratory, have been shown to be able to form neurons and other nonfollicle cell types. We have shown that the nestin-expressing stem cells from the hair follicle can effect the repair of peripheral nerve and spinal cord injury. The hair follicle stem cells differentiate into neuronal and glial cells after transplantation to the injured peripheral nerve and spinal cord, and enhance injury repair and locomotor recovery. We have termed these cells hair follicle-accessible pluripotent (HAP) stem cells. When the excised hair follicle with its nerve stump was placed in Gelfoam 3D histoculture, HAP stem cells grew and extended the hair follicle nerve which consisted of βIII-tubulin-positive fibers with F-actin expression at the tip. These findings indicate that βIII-tubulin-positive fibers elongating from the whisker follicle sensory nerve stump were growing axons. The growing whisker sensory nerve was highly enriched in HAP stem cells, which appeared to play a major role in its elongation and interaction with other nerves in 3D Gelfoam histoculture, including the sciatic nerve, the trigeminal nerve, and the trigeminal nerve ganglion. Our results suggest that a major function of the HAP stem cells in the hair follicle is for growth of the follicle sensory nerve. HAP stem cells have critical advantages over embryonic stem cells and induced pluripotent stem cells in that they are highly accessible, require no genetic manipulation, are nontumorigenic, and do not present ethical issues for regenerative medicine.

  15. Epidermal stem cells and skin tissue engineering in hair follicle regeneration

    PubMed Central

    Balañá, María Eugenia; Charreau, Hernán Eduardo; Leirós, Gustavo José

    2015-01-01

    The reconstitution of a fully organized and functional hair follicle from dissociated cells propagated under defined tissue culture conditions is a challenge still pending in tissue engineering. The loss of hair follicles caused by injuries or pathologies such as alopecia not only affects the patients’ psychological well-being, but also endangers certain inherent functions of the skin. It is then of great interest to find different strategies aiming to regenerate or neogenerate the hair follicle under conditions proper of an adult individual. Based upon current knowledge on the epithelial and dermal cells and their interactions during the embryonic hair generation and adult hair cycling, many researchers have tried to obtain mature hair follicles using different strategies and approaches depending on the causes of hair loss. This review summarizes current advances in the different experimental strategies to regenerate or neogenerate hair follicles, with emphasis on those involving neogenesis of hair follicles in adult individuals using isolated cells and tissue engineering. Most of these experiments were performed using rodent cells, particularly from embryonic or newborn origin. However, no successful strategy to generate human hair follicles from adult cells has yet been reported. This review identifies several issues that should be considered to achieve this objective. Perhaps the most important challenge is to provide three-dimensional culture conditions mimicking the structure of living tissue. Improving culture conditions that allow the expansion of specific cells while protecting their inductive properties, as well as methods for selecting populations of epithelial stem cells, should give us the necessary tools to overcome the difficulties that constrain human hair follicle neogenesis. An analysis of patent trends shows that the number of patent applications aimed at hair follicle regeneration and neogenesis has been increasing during the last decade. This

  16. Epidermal stem cells and skin tissue engineering in hair follicle regeneration.

    PubMed

    Balañá, María Eugenia; Charreau, Hernán Eduardo; Leirós, Gustavo José

    2015-05-26

    The reconstitution of a fully organized and functional hair follicle from dissociated cells propagated under defined tissue culture conditions is a challenge still pending in tissue engineering. The loss of hair follicles caused by injuries or pathologies such as alopecia not only affects the patients' psychological well-being, but also endangers certain inherent functions of the skin. It is then of great interest to find different strategies aiming to regenerate or neogenerate the hair follicle under conditions proper of an adult individual. Based upon current knowledge on the epithelial and dermal cells and their interactions during the embryonic hair generation and adult hair cycling, many researchers have tried to obtain mature hair follicles using different strategies and approaches depending on the causes of hair loss. This review summarizes current advances in the different experimental strategies to regenerate or neogenerate hair follicles, with emphasis on those involving neogenesis of hair follicles in adult individuals using isolated cells and tissue engineering. Most of these experiments were performed using rodent cells, particularly from embryonic or newborn origin. However, no successful strategy to generate human hair follicles from adult cells has yet been reported. This review identifies several issues that should be considered to achieve this objective. Perhaps the most important challenge is to provide three-dimensional culture conditions mimicking the structure of living tissue. Improving culture conditions that allow the expansion of specific cells while protecting their inductive properties, as well as methods for selecting populations of epithelial stem cells, should give us the necessary tools to overcome the difficulties that constrain human hair follicle neogenesis. An analysis of patent trends shows that the number of patent applications aimed at hair follicle regeneration and neogenesis has been increasing during the last decade. This

  17. Shh expression is required for embryonic hair follicle but not mammary gland development.

    PubMed

    Michno, Kinga; Boras-Granic, Kata; Mill, Pleasantine; Hui, C C; Hamel, Paul A

    2003-12-01

    The embryonic mammary gland and hair follicle are both derived from the ventral ectoderm, and their development depends on a number of common fundamental developmental pathways. While the Hedgehog (Hh) signaling pathway is required for hair follicle morphogenesis, the role of this pathway during embryonic mammary gland development remains undetermined. We demonstrate here that, unlike the hair follicle, both Shh and Ihh are expressed in the developing embryonic mouse mammary rudiment as early as E12.5. In Shh(-/-) embryos, hair follicle development becomes arrested at an early stage, while the mammary rudiment, which continues to express Ihh, develops in a manner indistinguishable from that of wild-type littermates. The five pairs of mammary buds in Shh(-/-) female embryos exhibit normal branching morphogenesis at E16.5, forming a rudimentary ductal structure identical to wild-type embryonic mammary glands. We further demonstrate that loss of Hh signaling causes altered cyclin D1 expression in the embryonic dermal mesenchyme. Specifically, cyclin D1 is expressed at E14.5 principally in the condensed mesenchymal cells of the presumptive hair follicles and in both mesenchymal and epithelial cells of the mammary rudiments in wild-type and Shh-deficient embryos. By E18.5, robust cyclin D1 expression is maintained in mammary rudiments of both wild-type and Shh-deficient embryos. In hair follicles of wild-type embryos by E18.5, cyclin D1 expression switches to follicular epithelial cells. In contrast, strong cyclin D1 expression is observed principally in the mesenchymal cells of arrested hair follicles in Shh(-/-) embryos at E18.5. These data reveal that, despite the common embryonic origin of hair follicles and mammary glands, distinct patterns of Hh-family expression occur in these two tissues. Furthermore, these data suggest that cyclin D1 expression in the embryonic hair follicle is mediated by both Hh-independent and Hh-dependent mechanisms.

  18. Histopathological and Radiographic Analysis of Dental Follicle of Impacted Teeth Using Modified Gallego’s Stain

    PubMed Central

    Satheesan, Evie; Tamgadge, Avinash; Bhalerao, Sudhir; Periera, Treville

    2016-01-01

    Introduction In the WHO classification of odontogenic tumours, hard tissue formation has been considered as a sub-classification however, this parameter has not been much explored in dental follicle in literature. Epithelial-mesenchymal interactions play an important role in odontogenesis and its associated pathologies; therefore research on dental follicle should also include mesenchymal components along with epithelial components. Additionally, special stains to identify the nature of such depositions in dental follicle have been less explored. Modified Gallego’s stain is such an example which has not been tried in odontogenic lesions which makes this study unique. Aim Aim of this study was to study histopathological variations in dental follicle, the nature of calcification and depositions using Modified Gallego’s stain and to correlate histological features of dental follicle with pericoronal width radiographically. Materials and Methods A prospective histological study of the dental follicles of 50 impacted teeth was carried out to microscopically evaluate the dental follicular tissues for pathological changes, and to correlate it with pericoronal radiolucency. Impacted teeth with pericoronal radiographic width less than 3mm were included in the study and symptomatic teeth were excluded. Further Modified Gallego stain was used to differentiate the nature of hard tissue formation in dental follicle tissues. Results Dental follicle histologically showed pathological changes resembling dentigerous cyst, ameloblastoma, odontogenic fibroma (Simple and WHO Type), clear cell odontogenic tumour, neurofibroma, neurilemmoma and mucoepidermoid carcinoma. Conclusion The dental follicle surrounding an impacted tooth has the potential to differentiate into a wide variety of tissue types, and thus shows the potential for cyst and tumour development which was observed in this study in most of the specimens with normal follicular width radiographically. PMID:27437341

  19. Traditional goat husbandry may substantially contribute to human toxoplasmosis exposure.

    PubMed

    Paştiu, Anamaria I; Ajzenberg, Daniel; Györke, Adriana; Şuteu, Ovidiu; Balea, Anamaria; Rosenthal, Benjamin M; Kalmár, Zsuzsa; Domşa, Cristian; Cozma, Vasile

    2015-02-01

    Raising goats in settings that are highly contaminated with oocysts of Toxoplasma gondii may contribute significantly to human exposure to this zoonotic parasite. Increasing consumption of young goats in countries where goats are frequently reared in backyards that are also homes to cats (the definitive host of this parasite) elevates such concern. To date, there has been little attention to either the prevalence or genotypic characteristics of T. gondii isolates in young ruminant food animals in Europe. Here, we estimated the prevalence of T. gondii goat-kids raised in backyards and slaughtered for human consumption during Easter. We collected 181 paired samples of serum and diaphragm. Serum samples were analyzed by enzyme-linked immunosorbent assay for antibodies against T. gondii , and muscle tissues were analyzed by polymerase chain reaction to detect T. gondii DNA. Thirty-two diaphragm samples were also bioassayed in mice, and the isolates were genotyped using microsatellite markers. The overall seroprevalence of T. gondii infection in goat-kids was 33.1% (60/181; 95% confidence interval [CI] 26.3-40.5%), and T. gondii DNA was found in 6.1% (11/181; 95% CI 3.1-10.6) of the diaphragm samples. We isolated the parasite from 2 of 32 goat-kids, and the T. gondii strains belonged to genotype II. The results showed that 1/3 of 3-mo-old goats may be infected with T. gondii, and their consumption during Easter (as barbecue) may seriously compromise food safety as a result.

  20. Variation in the prion protein sequence in Dutch goat breeds.

    PubMed

    Windig, J J; Hoving, R A H; Priem, J; Bossers, A; van Keulen, L J M; Langeveld, J P M

    2016-10-01

    Scrapie is a neurodegenerative disease occurring in goats and sheep. Several haplotypes of the prion protein increase resistance to scrapie infection and may be used in selective breeding to help eradicate scrapie. In this study, frequencies of the allelic variants of the PrP gene are determined for six goat breeds in the Netherlands. Overall frequencies in Dutch goats were determined from 768 brain tissue samples in 2005, 766 in 2008 and 300 in 2012, derived from random sampling for the national scrapie surveillance without knowledge of the breed. Breed specific frequencies were determined in the winter 2013/2014 by sampling 300 breeding animals from the main breeders of the different breeds. Detailed analysis of the scrapie-resistant K222 haplotype was carried out in 2014 for 220 Dutch Toggenburger goats and in 2015 for 942 goats from the Saanen derived White Goat breed. Nine haplotypes were identified in the Dutch breeds. Frequencies for non-wild type haplotypes were generally low. Exception was the K222 haplotype in the Dutch Toggenburger (29%) and the S146 haplotype in the Nubian and Boer breeds (respectively 7 and 31%). The frequency of the K222 haplotype in the Toggenburger was higher than for any other breed reported in literature, while for the White Goat breed it was with 3.1% similar to frequencies of other Saanen or Saanen derived breeds. Further evidence was found for the existence of two M142 haplotypes, M142 /S240 and M142 /P240 . Breeds vary in haplotype frequencies but frequencies of resistant genotypes are generally low and consequently selective breeding for scrapie resistance can only be slow but will benefit from animals identified in this study. The unexpectedly high frequency of the K222 haplotype in the Dutch Toggenburger underlines the need for conservation of rare breeds in order to conserve genetic diversity rare or absent in other breeds. PMID:26991480

  1. The prevalence of lymphoid follicles in Helicobacter pylori associated gastritis in patients with ulcers and non-ulcer dyspepsia.

    PubMed Central

    Zaitoun, A M

    1995-01-01

    AIMS--To determine the prevalence of lymphoid follicles in Helicobacter pylori positive and negative gastritis in antral and body type gastric mucosa in patients with non-ulcer dyspepsia (NUD), duodenal ulcer, or gastric ulcer; to correlate follicle presence with patient age; to evaluate the correlation between the prevalence of lymphoid follicles and active and inactive gastritis and its severity; and to assess the positive predictive value of lymphoid follicle prevalence with respect to H pylori infection. METHODS--Gastric biopsy specimens, graded according to the Sydney system, from 337 patients were studied. RESULTS--Lymphoid follicles occurred more often in antral mucosa (78%) than in body type mucosa (41%) and were observed in 85% of patients with H pylori positive gastritis. There was no significant difference between NUD and gastric and duodenal ulcer disease with regard to the presence of lymphoid follicles. The positive predictive value of the presence of lymphoid follicles in H pylori infection was 96%. Lymphoid follicles were more commonly observed in patients aged between 10 and 29 years. Lymphoid follicles were more frequently found in pangastritis of all subtypes than in antral gastritis and also in active gastritis than in inactive gastritis. The presence of lymphoid follicles correlated strongly with the degree and severity of gastritis. CONCLUSION--Lymphoid follicles are a constant morphological feature of H pylori associated gastritis. Images PMID:7615851

  2. Wnt7b is an important intrinsic regulator of hair follicle stem cell homeostasis and hair follicle cycling.

    PubMed

    Kandyba, Eve; Kobielak, Krzysztof

    2014-04-01

    The hair follicle (HF) is an exceptional mini-organ to study the mechanisms which regulate HF morphogenesis, cycling, hair follicle stem cell (hfSCs) homeostasis, and progeny differentiation. During morphogenesis, Wnt signaling is well-characterized in the initiation of HF patterning but less is known about which particular Wnt ligands are required and whether individual Wnt ligands act in an indispensable or redundant manner during postnatal hfSCs anagen onset and HF cycle progression. Previously, we described the function of the bone morphogenetic protein (BMP) signaling target gene WNT7a in intrinsic regulation of hfSCs homeostasis in vivo. Here, we investigated the role of Wnt7b, which was also intrinsically upregulated in hfSCs during physiological and precocious anagen after BMP inhibition in vivo. We demonstrated Wnt7b to be a direct target of canonical BMP signaling in hfSCs and using Wnt7b conditional gene targeting during HF morphogenesis revealed disrupted HF cycling including a shorter anagen, premature catagen onset with overall shorter hair production, and diminished HF differentiation marker expression. Additionally, we observed that postnatal ablation of Wnt7b resulted in delayed HF activation, affecting both the hair germ and bulge hfSCs but still maintaining a two-step sequence of HF stimulation. Interestingly, Wnt7b cKO hfSCs participated in reformation of the new HF bulge, but with slower self-renewal. These findings demonstrate the importance of intrinsic Wnt7b expression in hfSCs regulation and normal HF cycling and surprisingly reveal a nonredundant role for Wnt7b in the control of HF anagen length and catagen entry which was not compensated by other Wnt ligands. PMID:24222445

  3. Wnt7b is an important intrinsic regulator of hair follicle stem cell homeostasis and hair follicle cycling.

    PubMed

    Kandyba, Eve; Kobielak, Krzysztof

    2014-04-01

    The hair follicle (HF) is an exceptional mini-organ to study the mechanisms which regulate HF morphogenesis, cycling, hair follicle stem cell (hfSCs) homeostasis, and progeny differentiation. During morphogenesis, Wnt signaling is well-characterized in the initiation of HF patterning but less is known about which particular Wnt ligands are required and whether individual Wnt ligands act in an indispensable or redundant manner during postnatal hfSCs anagen onset and HF cycle progression. Previously, we described the function of the bone morphogenetic protein (BMP) signaling target gene WNT7a in intrinsic regulation of hfSCs homeostasis in vivo. Here, we investigated the role of Wnt7b, which was also intrinsically upregulated in hfSCs during physiological and precocious anagen after BMP inhibition in vivo. We demonstrated Wnt7b to be a direct target of canonical BMP signaling in hfSCs and using Wnt7b conditional gene targeting during HF morphogenesis revealed disrupted HF cycling including a shorter anagen, premature catagen onset with overall shorter hair production, and diminished HF differentiation marker expression. Additionally, we observed that postnatal ablation of Wnt7b resulted in delayed HF activation, affecting both the hair germ and bulge hfSCs but still maintaining a two-step sequence of HF stimulation. Interestingly, Wnt7b cKO hfSCs participated in reformation of the new HF bulge, but with slower self-renewal. These findings demonstrate the importance of intrinsic Wnt7b expression in hfSCs regulation and normal HF cycling and surprisingly reveal a nonredundant role for Wnt7b in the control of HF anagen length and catagen entry which was not compensated by other Wnt ligands.

  4. Di (2-ethylhexyl) phthalate inhibits growth of mouse ovarian antral follicles through an oxidative stress pathway

    SciTech Connect

    Wang, Wei Craig, Zelieann R. Basavarajappa, Mallikarjuna S. Gupta, Rupesh K. Flaws, Jodi A.

    2012-01-15

    Di (2-ethylhexyl) phthalate (DEHP) is a plasticizer that has been shown to inhibit growth of mouse antral follicles, however, little is known about the mechanisms by which DEHP does so. Oxidative stress has been linked to follicle growth inhibition as well as phthalate-induced toxicity in non-ovarian tissues. Thus, we hypothesized that DEHP causes oxidative stress and that this leads to inhibition of the growth of antral follicles. To test this hypothesis, antral follicles isolated from CD-1 mice (age 31–35 days) were cultured with vehicle control (dimethylsulfoxide [DMSO]) or DEHP (1–100 μg/ml) ± N-acetyl cysteine (NAC, an antioxidant at 0.25–1 mM). During culture, follicles were measured daily. At the end of culture, follicles were collected and processed for in vitro reactive oxygen species (ROS) assays to measure the presence of free radicals or for measurement of the expression and activity of various key antioxidant enzymes: Cu/Zn superoxide dismutase (SOD1), glutathione peroxidase (GPX) and catalase (CAT). The results indicate that DEHP inhibits the growth of follicles compared to DMSO control and that NAC (0.25–1 mM) blocks the ability of DEHP to inhibit follicle growth. Furthermore, DEHP (10 μg/ml) significantly increases ROS levels and reduces the expression and activity of SOD1 compared to DMSO controls, whereas NAC (0.5 mM) rescues the effects of DEHP on ROS levels and SOD1. However, the expression and activity of GPX and CAT were not affected by DEHP treatment. Collectively, these data suggest that DEHP inhibits follicle growth by inducing production of ROS and by decreasing the expression and activity of SOD1. -- Highlights: ► DEHP inhibits growth and increases reactive oxygen species in ovarian antral follicles in vitro. ► NAC rescues the effects of DEHP on the growth and reactive oxygen species levels in follicles. ► DEHP decreases the expression and activity of Cu/Zn superoxide dismutase, which can be rescued by NAC, in antral

  5. Isolation of Mouse Hair Follicle Bulge Stem Cells and Their Functional Analysis in a Reconstitution Assay.

    PubMed

    Zheng, Ying; Hsieh, Jen-Chih; Escandon, Julia; Cotsarelis, George

    2016-01-01

    The hair follicle (HF) is a dynamic structure readily accessible within the skin, and contains various pools of stem cells that have a broad regenerative potential during normal homeostasis and in response to injury. Recent discoveries demonstrating the multipotent capabilities of hair follicle stem cells and the easy access to skin tissue make the HF an attractive source for isolating stem cells and their subsequent application in tissue engineering and regenerative medicine. Here, we describe the isolation and purification of hair follicle bulge stem cells from mouse skin, and hair reconstitution assays that allows the functional analysis of multipotent stem cells. PMID:27431247

  6. Insulin-like growth factor binding proteins in follicular fluid from morphologically distinct healthy and atretic bovine antral follicles.

    PubMed

    Irving-Rodgers, H F; Catanzariti, K D; Master, M; Grant, P A; Owens, P C; Rodgers, R J

    2003-01-01

    In bovine follicles 2-5 mm in diameter, two morphologically distinct types of healthy follicles and two types of atretic follicles have been described recently. Healthy follicles either have columnar basal granulosa cells with follicular basal lamina composed of many layers or 'loops' or they have rounded basal cells with a conventional single-layered, aligned follicular basal lamina. In atretic follicles, cell death either commences at the basal layer and progresses to the antrum (basal atresia) with macrophage penetration of the membrana granulosa or death progresses from the antrum in a basal direction (antral atresia). Little is known about how these different phenotypes develop. To determine whether insulin-like growth factor binding protein (IGFBP) levels in follicular fluid differ between these different types of follicles, we measured IGFBP levels in fluids from these follicles. A total of 61 follicles were assessed by light microscopy and characterized by morphological analysis as either healthy, with columnar or rounded basal granulosa cells, or as undergoing antral or basal atresia. The IGFBP concentration in the follicular fluid of individual follicles from the four groups (n = 12-20 per group) was identified by Western ligand blots using (125)I-insulin-like growth factor (IGF)-II as a probe. Insulin-like growth factor binding proteins 2, 3 (44 and 40 kDa), 4 (glycosylated and non-glycosylated) and 5 were observed. The levels (per volume of fluid) of IGFBPs 2, 4 and 5 were greater in atretic follicles than in healthy follicles. However, there were no statistical differences in levels of each IGFBP between either the two types of healthy follicle or between the two types of atretic follicles. Thus, IGFBP levels are not related to the different types of healthy or atretic follicles. PMID:12921699

  7. Transplantation of goat bone marrow stromal cells to the degenerating intervertebral disc in a goat disc-injury model

    PubMed Central

    Zhang, Yejia; Drapeau, Susan; An, Howard S.; Thonar, Eugene J-M.A.; Anderson, D. Greg

    2010-01-01

    Study Design In vivo randomized controlled study in the goat intervertebral disc (IVD) injury model. Objective To define the effects of allogeneic bone marrow-derived stromal cell injected into the degenerating goat IVDs. Summary of Background Data Transplantation of bone marrow stromal cells to the degenerating disc has been suggested as a means to correct the biologic incompetence of the disc. However, large animal models with IVDs similar in shape and size to those of humans are needed to define the efficacy and safety of this approach. Methods Goat IVD degeneration was induced by stabbing with a #15 blade. One month after disc injury, the injured discs were randomly selected to receive goat bone marrow-derived stromal cell (suspended in hydrogel), saline (control), or hydrogel (control) injections. Three and 6 months after stem cell transplantation, goats were euthanized and the IVD were examined for biochemical content and tissue morphology. MR images at 3- and 6-month time points were also examined. Results The goat large animal model shows early degenerative changes following disc injury. Degenerating IVDs injected with bone marrow stromal cells showed significantly increased proteoglycan (PG) accumulation within their nucleus pulposus (NP) region. However, collagen content, MRI grade and histology did not show statistically significant differences between the cell-treated and control IVDs. Conclusions Following transplantation of bone marrow stromal cells, NP tissue contained more PG than control discs. Although this result was promising, the rate and severity of degeneration in this goat disc injury were modest, suggesting that a more severe injury and a larger sample size is indicated for future studies to better define the utility of cell therapies in this model. PMID:20890267

  8. Goat Meat Does Not Cause Increased Blood Pressure

    PubMed Central

    Sunagawa, Katsunori; Kishi, Tetsuya; Nagai, Ayako; Matsumura, Yuka; Nagamine, Itsuki; Uechi, Shuntoku

    2014-01-01

    While there are persistent rumors that the consumption of goat meat dishes increases blood pressure, there is no scientific evidence to support this. Two experiments were conducted to clarify whether or not blood pressure increases in conjunction with the consumption of goat meat dishes. In experiment 1, 24 Dahl/Iwai rats (15 weeks old, body weight 309.3±11.1 g) were evenly separated into 4 groups. The control group (CP) was fed a diet containing 20% chicken and 0.3% salt on a dry matter basis. The goat meat group (GM) was fed a diet containing 20% goat meat and 0.3% salt. The goat meat/salt group (GS) was fed a diet containing 20% goat meant and 3% to 4% salt. The Okinawan mugwort (Artemisia Princeps Pampan)/salt group (GY) was fed a diet containing 20% goat meat, 3% to 4% salt and 5% of freeze-dried mugwort powder. The experiment 1 ran for a period of 14 weeks during which time the blood pressure of the animals was recorded. The GS, and GY groups consumed significantly more water (p<0.01) than the CP and GM groups despite the fact that their diet consumption levels were similar. The body weight of animals in the CP, GM, and GS groups was similar while the animals in the GY group were significantly smaller (p<0.01). The blood pressure in the GM group was virtually the same as the CP group throughout the course of the experiment. In contrast, while the blood pressure of the animals in the GS and GY group from 15 to 19 weeks old was the same as the CP group, their blood pressures were significantly higher (p<0.01) after 20 weeks of age. The GY group tended to have lower blood pressure than the GS group. In experiment 2, in order to clarify whether or not the increase in blood pressure in the GS group and the GY group in experiment 1 was caused by an excessive intake of salt, the effects on blood pressure of a reduction of salt in diet were investigated. When amount of salt in the diet of the GS and GY group was reduced from 4% to 0.3%, the animal’s blood pressure

  9. Induction of parturition with aglepristone in the Majorera goat.

    PubMed

    Batista, M; Reyes, R; Santana, M; Alamo, D; Vilar, J; González, F; Cabrera, F; Gracia, A

    2011-10-01

    This study assessed the efficacy of aglepristone at inducing parturition in pregnant goats. Six experimental groups were defined: group A-5 (n = 12), group A-3.3 (n = 12), group A-2.5 (n = 12) and group A-1.5 (n = 12) in which goats were injected SC once with 5.0, 3.3, 2.5 and 1.5 mg of aglepristone per kg body weight of goat, respectively, group L (n = 11), which was treated IM with 3.75 mg of luprostiol; and group Ct (n = 11), which was injected SC with 1 ml of saline solution. Different parameters associated with parturition were thereafter investigated. In addition, plasma progesterone concentrations were defined after treatments till parturition. Aglepristone effectively induced parturition in all of the goats. In the A-5, A-3.3 and A-2.5 groups, the time to parturition was around 30-34 h, and the majority of goats (97.2%, 35/36) started kidding between 25 and 40 h after the aglepristone injection. However, the goats in group A-1.5 showed a significantly (p < 0.01) higher time to parturition (mean: 46.8 h). Overall, the incidence of dystocia registered in aglepristone-induced goats (20.8%, 10/48) and luprostiol-induced goats was not different from that observed after a spontaneous parturition. The percentage of live kids was very similar between A-5, A-3.3, A.2.5 and L groups (95.7, 95.3, 95.0 and 96.3%, respectively) but was higher that observed in the control (83.4%) and A-1.5 (81.2%) groups. In addition, no maternal mortality was registered in any groups. No changes in plasma progesterone were observed during the first 24 h after treatment, and high plasma progesterone concentrations were present at kidding (6.7, 5.5, 4.5 and 3.6 ng/ml for groups A-5, A-3.3, A-2.5 and A-1.5, respectively), confirming that aglepristone does not induce parturition via luteolysis. This study demonstrates that aglepristone can be used to induce parturition in goats with satisfactory efficacy, inducing pregnancy termination without direct or immediate modifications of luteal

  10. 9 CFR 51.22 - Payment to owners for goats, sheep, and horses destroyed.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Payment to owners for goats, sheep... DISEASES ANIMALS DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.22 Payment to owners for goats, sheep, and horses destroyed. (a) The Administrator may authorize the payment of...

  11. 9 CFR 51.22 - Payment to owners for goats, sheep, and horses destroyed.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Payment to owners for goats, sheep... DISEASES ANIMALS DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.22 Payment to owners for goats, sheep, and horses destroyed. (a) The Administrator may authorize the payment of...

  12. New cryptic karyotypic differences between cattle (Bos taurus) and goat (Capra hircus).

    PubMed

    De Lorenzi, Lisa; Planas, Jordi; Rossi, Elena; Malagutti, Luca; Parma, Pietro

    2015-06-01

    Cattle (Bos taurus) and goat (Capra hircus) belong to the Bovidae family, and they share a common ancestor 19.7-21.5 Ma ago (MYA). The Bovidae family apparently experienced a rapid species radiation in the middle Miocene. The present day cattle and goat possess the same diploid chromosome number (2n = 60) and structurally similar autosomes, except that a small subcentromeric portion of cattle chromosome nine has been translocated to goat chromosome 14. In this study, we adopted a new strategy that involves the use of bioinformatics approach to detect unknown cryptic chromosome divergences between cattle and goat using and subsequent validation using the fluorescence in situ hybridization (FISH) of bacterial artificial chromosome clones. We identified two hypothetical discrepancies between the cattle and goat genome assemblies: an inversion in the goat chromosome 13 and a transposition in the goat chromosome 6. The FISH technique allowed clear validation of the existence of a new 7.4 Mb chromosomal inversion in the goat chromosome 13. Regarding the transposition in the goat chromosome six, FISH analyses revealed that the cattle and goat genomes shared the same organization, with the assembly of the goat genome being the correct one. Moreover, we defined, for the first time, the size and orientation of the translocated fragment involved in the evolutionary translocation between cattle chromosomes 9 and goat chromosome 14. Our results suggest that bioinformatics represents an efficient method for detecting cryptic chromosome divergences among species.

  13. Molecular cloning, polymorphisms, and expression analysis of the RERG gene in indigenous Chinese goats.

    PubMed

    Sui, M X; Wang, H H; Wang, Z W

    2015-01-01

    The current study aimed to investigate the coding sequence, polymorphisms, and expression of the RERG gene in indigenous Chinese goats. cDNA of RERG, obtained through reverse transcription PCR was analyzed using bioinformatic techniques. Polymorphisms in the exon regions of the RERG gene were identified and their associations with growth traits in three varieties of indigenous Chinese goats were investigated. Expression of the RERG gene in three goat breeds of the same age was detected using real-time quantitative PCR. The results revealed that the cDNA of RERG, which contained a complete open reading frame of 20-620 bp, was 629 bp in length. The associated accession numbers in GenBank are JN672576, JQ917222, and JN580309 for the QianBei Ma goat, the GuiZhou white goat, and the GuiZhou black goat, respectively. Four consistent SNP sites were found in the exon regions of the RERG gene for the three goat breeds. mRNA expression of the RERG gene differed between different tissues in adult goats of same age. The highest expression was observed in lung and spleen tissues, while the lowest expression was recorded in thymus gland tissue. In addition, the expression of the RERG gene in the muscle of Guizhou white goat, GuiZhou black goat, and QianBei Ma goat decreased sequentially. Our results lay the foundations for further investigation into the role of the RERG gene in goat growth traits. PMID:26634455

  14. 9 CFR 93.428 - Sheep and goats and wild ruminants from Mexico.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Sheep and goats and wild ruminants... PRODUCTS; REQUIREMENTS FOR MEANS OF CONVEYANCE AND SHIPPING CONTAINERS Ruminants Mexico 10 § 93.428 Sheep and goats and wild ruminants from Mexico. (a) Sheep and goats intended for importation from...

  15. 9 CFR 93.428 - Sheep and goats and wild ruminants from Mexico.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Sheep and goats and wild ruminants... PRODUCTS; REQUIREMENTS FOR MEANS OF CONVEYANCE AND SHIPPING CONTAINERS Ruminants Mexico 10 § 93.428 Sheep and goats and wild ruminants from Mexico. (a) Sheep and goats intended for importation from...

  16. 9 CFR 93.428 - Sheep and goats and wild ruminants from Mexico.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Sheep and goats and wild ruminants... PRODUCTS; REQUIREMENTS FOR MEANS OF CONVEYANCE AND SHIPPING CONTAINERS Ruminants Mexico 10 § 93.428 Sheep and goats and wild ruminants from Mexico. (a) Sheep and goats intended for importation from...

  17. 9 CFR 93.428 - Sheep and goats and wild ruminants from Mexico.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Sheep and goats and wild ruminants... PRODUCTS; REQUIREMENTS FOR MEANS OF CONVEYANCE AND SHIPPING CONTAINERS Ruminants Mexico 10 § 93.428 Sheep and goats and wild ruminants from Mexico. (a) Sheep and goats intended for importation from...

  18. 9 CFR 93.428 - Sheep and goats and wild ruminants from Mexico.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Sheep and goats and wild ruminants... PRODUCTS; REQUIREMENTS FOR MEANS OF CONVEYANCE AND SHIPPING CONTAINERS Ruminants Mexico 10 § 93.428 Sheep and goats and wild ruminants from Mexico. (a) Sheep and goats intended for importation from...

  19. 9 CFR 51.27 - Identification of goats, sheep, and horses to be destroyed.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... horses to be destroyed. 51.27 Section 51.27 Animals and Animal Products ANIMAL AND PLANT HEALTH... DISEASES ANIMALS DESTROYED BECAUSE OF BRUCELLOSIS Indemnity for Sheep, Goats, and Horses § 51.27 Identification of goats, sheep, and horses to be destroyed. The claimant must ensure that any goats, sheep,...

  20. Comparative Response of the West African Dwarf Goats to Experimental Infections with Red Sokoto and West African Dwarf Goat Isolates of Haemonchus contortus.

    PubMed

    Ngongeh, Lucas Atehmengo; Onyeabor, Amaechi

    2015-01-01

    Response of the West African Dwarf (WAD) goats to two different isolates of Haemonchus contortus, the Red Sokoto (RS) goat isolate (RSHc) and the WAD goat isolate (WADHc) (isolated from WAD goats), was studied by experimental infections of 4-6-month-old male WAD goat kids. Group 1 and Group 2 goats were each infected with 4500 infective larvae (L3) of RSHc and WADHc, respectively. Group 3 animals served as uninfected control. Prepatent period (PPP), faecal egg counts (FEC), worm burden (WB), body weight (BW), packed cell volume (PCV), and body condition score (BCS) were determined. WAD goats infected with RSHc isolate and the ones infected with WADHc isolate had mean PPP of 19.63 ± 0.26 and 19.50 ± 0.19, respectively. Goats infected with WADHc isolate had significantly higher FEC (P = 0.004) and WB (P = 0.001). BW were significantly higher (P = 0.004) both in the controls and in Group 2 goats infected with WADHc isolate than in Group 1 goats infected with the RSHc isolate. BCS of animals in both infected groups dropped significantly (P = 0.001). There was a significant drop in PCV (P = 0.004) of both infected groups in comparison. Both isolates of H. contortus were pathogenic to the host. PMID:26697224

  1. Comparative Response of the West African Dwarf Goats to Experimental Infections with Red Sokoto and West African Dwarf Goat Isolates of Haemonchus contortus.

    PubMed

    Ngongeh, Lucas Atehmengo; Onyeabor, Amaechi

    2015-01-01

    Response of the West African Dwarf (WAD) goats to two different isolates of Haemonchus contortus, the Red Sokoto (RS) goat isolate (RSHc) and the WAD goat isolate (WADHc) (isolated from WAD goats), was studied by experimental infections of 4-6-month-old male WAD goat kids. Group 1 and Group 2 goats were each infected with 4500 infective larvae (L3) of RSHc and WADHc, respectively. Group 3 animals served as uninfected control. Prepatent period (PPP), faecal egg counts (FEC), worm burden (WB), body weight (BW), packed cell volume (PCV), and body condition score (BCS) were determined. WAD goats infected with RSHc isolate and the ones infected with WADHc isolate had mean PPP of 19.63 ± 0.26 and 19.50 ± 0.19, respectively. Goats infected with WADHc isolate had significantly higher FEC (P = 0.004) and WB (P = 0.001). BW were significantly higher (P = 0.004) both in the controls and in Group 2 goats infected with WADHc isolate than in Group 1 goats infected with the RSHc isolate. BCS of animals in both infected groups dropped significantly (P = 0.001). There was a significant drop in PCV (P = 0.004) of both infected groups in comparison. Both isolates of H. contortus were pathogenic to the host.

  2. Comparative Response of the West African Dwarf Goats to Experimental Infections with Red Sokoto and West African Dwarf Goat Isolates of Haemonchus contortus

    PubMed Central

    Ngongeh, Lucas Atehmengo; Onyeabor, Amaechi

    2015-01-01

    Response of the West African Dwarf (WAD) goats to two different isolates of Haemonchus contortus, the Red Sokoto (RS) goat isolate (RSHc) and the WAD goat isolate (WADHc) (isolated from WAD goats), was studied by experimental infections of 4–6-month-old male WAD goat kids. Group 1 and Group 2 goats were each infected with 4500 infective larvae (L3) of RSHc and WADHc, respectively. Group 3 animals served as uninfected control. Prepatent period (PPP), faecal egg counts (FEC), worm burden (WB), body weight (BW), packed cell volume (PCV), and body condition score (BCS) were determined. WAD goats infected with RSHc isolate and the ones infected with WADHc isolate had mean PPP of 19.63 ± 0.26 and 19.50 ± 0.19, respectively. Goats infected with WADHc isolate had significantly higher FEC (P = 0.004) and WB (P = 0.001). BW were significantly higher (P = 0.004) both in the controls and in Group 2 goats infected with WADHc isolate than in Group 1 goats infected with the RSHc isolate. BCS of animals in both infected groups dropped significantly (P = 0.001). There was a significant drop in PCV (P = 0.004) of both infected groups in comparison. Both isolates of H. contortus were pathogenic to the host. PMID:26697224

  3. Susceptibility of goats and calves after experimental inoculation or contact exposure to a Canadian strain of Mycoplasma mycoides subsp. mycoides isolated from a goat.

    PubMed Central

    Rosendal, S

    1983-01-01

    Transmissibility of Mycoplasma mycoides subsp. mycoides infection from experimentally inoculated goats to other goats and calves was studied. Eight goats and six calves were housed in an 18 m2 room. Six of the goats were inoculated endobronchially with strain D44 isolated from a natural case of polyarthritis in Ontario. These six goats died within a week of Mycoplasma septicemia. The two contact goats or the six calves never showed signs of disease and M. mycoides subsp. mycoides was not recovered from these animals. The contact goats and four calves were killed 25 days after exposure. They were all seronegative, M. mycoides subsp. mycoides was not recovered at necropsy and none had pathomorphological changes attributable to this Mycoplasma. The two remaining calves were inoculated endobronchially with 10(9) CFU of strain D44 and observed for 20 days. They never showed signs of disease and did not have significant lesions at necropsy. Both developed a significant serological response to M. mycoides subsp. mycoides, although this organism was not recovered during the experimental period or at necropsy. This study did not provide evidence for transmission of M. mycoides subsp. mycoides from endobronchially inoculated goats to contact goats or calves and endobronchially inoculated calves did not develop pneumonia. This would suggest that the infection of the goat population in Canada with this pathogen would not be a significant threat to the cattle population. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. PMID:6365296

  4. Immunological and biochemical studies of fascioliasis in goats and cattle

    SciTech Connect

    Reddington, J.J.

    1985-01-01

    Using the goat as a susceptible host and cattle as a resistant species to Fasciola hepatica infections, the humoral response of these animals to the surface of the newly excysted juvenile (NEJ) fluke was examined. Tegumental proteins of the NEJ were labeled with /sup 125/I by lactoperoxidase and analyzed after immunoprecipitation using a double antibody system. In addition, a comparison was made between the infected sera's capacity to immunoprecipitate surface antigens and their in vitro cytotoxic activity against the NEJ. In both goats and cattle the levels of NEJ surface antigens precipitated increased during the first 4 weeks PI. The peak immunoprecipitation of NEJ surface antigens by cattle sera (58%) was significantly higher than that of infected goat sera (33%). Immunoprecipitation of the available radiolabeled NEJ surface proteins by the infected cattle sera remained consistently higher than goat sera until the 16th week PI. The cytotoxic effects of these same caprine sera on NEJs in vitro was limited, while the cytotoxicity of the infected bovine sera closely approximated the sera's ability to precipitate NEJ surface antigens. There was also a qualitative difference between the species in their recognition of /sup 35/S and /sup 125/I radiolabeled NEJ surface antigens. Uninfected goat or cattle sera failed to precipitate any /sup 125/I or /sup 35/S-labeled surface proteins.

  5. Microbiological Assessment of Raw Goat Milk Collected from Sardinian Herds

    PubMed Central

    Carusillo, Francesca; Rosu, Valentina; Fancello, Cipriana; Pirino, Tonino; Bandino, Ennio

    2014-01-01

    With Regulation EC 853/04, the European Parliament and the Council laid down general rules for food business operators regarding the hygiene of foodstuffs. In particular, the regulation established ≤1.500.000 cfu/mL as the maximum-tolerated value for total bacterial count in raw goat milk. Moreover, in order to enhance the hygiene of dairy farms, the Sardinia Region has funded the F measure programme which provides rewards for farmers showing improvements in herd management and animal welfare practices. This work aimed to evaluate the microbiological quality of raw goat milk samples collected during the F measure enforcement. A total of 536 raw goat samples, collected from dairy farms in the Sardinian province of Nuoro and Ogliastra, were analised for total bacterial count at 30°C. Results showed that total bacterial count was ≤1.500.000 ufc/mL in 456 (85.1%) raw milk goat samples, most of them (80.2%) with a total bacterial contamination <500.000 cfu/mL. This study confirms the hygienic good quality of raw goat milk collected from Sardinian dairy farms. PMID:27800332

  6. Interspecific transmission of small ruminant lentiviruses from goats to sheep.

    PubMed

    Souza, Thiago S de; Pinheiro, Raymundo R; Costa, Joselito N; Lima, Carla C V de; Andrioli, Alice; Azevedo, Dalva A A de; Santos, Vanderlan W S dos; Araújo, Juscilânia F; Sousa, Ana Lídia M de; Pinheiro, Danielle N S; Fernandes, Flora M C; Costa Neto, Antonio O

    2015-01-01

    This study was conducted in order to evaluate the transmission of caprine lentivirus to sheep using different experimental groups. The first one (colostrum group) was formed by nine lambs receiving colostrum from goats positive for small ruminant lentiviruses (SRLV). The second group (milk group) was established by nine lambs that received milk of these goats. Third was a control group, consisting of lambs that suckled colostrum and milk of negative mothers. Another experimental group (contact group) was formed by eight adult sheep, confined with two naturally infected goats. The groups were monitored by immunoblotting (IB), enzyme-linked immunosorbent assay (ELISA), agar gel immunodiffusion (AGID) and nested polymerase chain reaction (nPCR). All lambs that suckled colostrum and milk of infected goats and six sheep of the contact group had positive results in the nPCR, although seroconversion was detected only in three of the exposed animals, with no clinical lentiviruses manifestation, in 720 days of observation. There was a close relationship between viral sequences obtained from infected animals and the prototype CAEV-Cork. Thus, it was concluded that SRLV can be transmitted from goats to sheep, however, the degree of adaptation of the virus strain to the host species probably interferes with the infection persistence and seroconversion rate. PMID:26413072

  7. Kappa casein gene polymorphism in local Tunisian goats.

    PubMed

    Jemmali, B; Ben Gara, A; Selmi, H; Ammari, Z; Bouheni, C; Ben Larbi, M; Hammami, M; Amraoui, M; Kamoun, M; Rouissi, H; Rekik, B

    2013-12-15

    The genetic polymorphism of the goat Kappa casein was investigated in Tunisian goats. Blood samples were collected from local goat breeds. Samples of genomic DNA were obtained from leukocytes of 175 dairy goats and regions of interest in the gene were amplified by Polymerase Chain Reaction (PCR) and then evaluated in agarose gel. For a better characterization of the single nucleotide polymorphism, a PCR-Restriction Fragment Length Polymorphism was performed employing the endonuclease DNA amplification using 459 bp primers. The PCR products of primers (459 bp) digested by restriction enzyme Alw44I produced two fragments of 459 and 381 bp. The Kappa casein allelic variants in tested animals revealed different genotypes, two of them were homozygous: AA or BB, AC or BC and CC. Genotypic frequencies were 12.5, 60.5 and 27% for AA or BB, C