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1

Cryopreservation of hamster oocytes: effects of vitrification or freezing on human sperm penetration of zona-free hamster oocytes.  

PubMed

Three experiments were conducted for evaluation of the efficacy of conventional freezing or vitrification of hamster oocytes for use in a human sperm penetration assay (hSPA). In experiment 1, oocytes were cryopreserved and evaluated for survival on the basis of morphologic criteria. Survival of vitrified oocytes and that of frozen oocytes were not different, whereas all cryopreserved groups had lower survival than noncryopreserved controls. In experiment 2, oocytes were conventionally frozen or vitrified and evaluated in an hSPA. Vitrified oocytes had a lower frequency of sperm penetration than frozen oocytes, and all cryopreserved groups had lower penetration rates than untreated controls. In experiment 3, oocytes were exposed to the cryoprotectant used to vitrify (VS1) or freeze (DMSO) but not cooled prior to evaluation in an hSPA. Exposure to DMSO but not VS1 reduced hSPA values. It is concluded from these experiments that while all cryopreserved oocytes do not survive, at current stages of development conventionally frozen oocytes perform better than vitrified oocytes in the hSPA and losses associated with conventional freezing procedures may be related to cryoprotectant exposure, whereas vitrification losses are more probably due to events associated with rapid cooling and/or warming of the oocytes. PMID:3732535

Critser, J K; Arneson, B W; Aaker, D V; Ball, G D

1986-08-01

2

An alternative medicine study of herbal effects on the penetration of zona-free hamster oocytes and the integrity of sperm deoxyribonucleic acid  

Microsoft Academic Search

Objective: To analyze the effects of certain herbs on sperm DNA and on the fertilization process.Design: Prospective comparative study.Setting: Clinical and academic research environment.Patient(s): Donor sperm specimens.Intervention(s): Zona-free hamster oocytes were incubated for 1 hour in saw palmetto (Serenoa repens), echinacea purpura, ginkgo biloba, St. John’s wort (Hypericum perforatum), or control medium before sperm-oocyte interaction. The DNA of herb-treated sperm

Richard R. Ondrizek; Philip J. Chan; William C. Patton; Alan King

1999-01-01

3

Ejaculate-hormonal traits in the leopard cat (Felis bengalensis) and sperm function as measured by in vitro penetration of zona-free hamster ova and zona-intact domestic cat oocytes  

Microsoft Academic Search

Electroejaculate traits and cir- culating follicle-stimulating hormone (FSH), lutein- izing hormone (LhH), and testosterone concentra- tions were analyzed in adult leopard cats {Fells bengalensis), a rare felid species indigenous to east Asia. The ability of leopard cat sperm to bind and penetrate zona-free hamster ova and zona- intact domestic cat oocytes in vitro was examined as a means of testing

Jogayle Howard; David E. Wildt

1990-01-01

4

Comparative study of extra and intrafollicular hamster oocyte maturation  

E-print Network

of intra- and extra-follicular maturation of hamster oocytes checked by in vitro fertilization showed many possibilities for in vitro studies on fertilization and on their further development. However of gonadotropins. The ability of hamster sperm to be capacitated in vitro and to fertilize ovulated oocytes (Barros

Paris-Sud XI, Université de

5

Thermostability of sperm nuclei assessed by microinjection into hamster oocytes  

EPA Science Inventory

Nuclei isolated from spermatozoa of various species (golden hamster, mouse, human, rooster, and the fish tilapia) were heated at 60 degrees-125 degrees C for 20-120 min and then microinjected into hamster oocytes to determine whether they could decondense and develop into pronucl...

6

THERMOSTABILITY OF SPERM NUCLEI ASSESSED BY MICROINJECTION INTO HAMSTER OOCYTES  

EPA Science Inventory

Nuclei isolated from spermatozoa of various species (golden hamster, mouse, human, rooster, and the fish tilapia) were heated at 60 degrees - 125 degrees for 20-120 min and then microinjected into hamster oocytes to determine whether they could decondense and develop into pronucl...

7

Role of the follicle in zona pellucida maturation of hamster oocyte  

E-print Network

Role of the follicle in zona pellucida maturation of hamster oocyte Jacqueline MANDELBAUM Michèle. Hamster oocytes matured in vitro are not fer- tilizable (Mandelbaum and Plachot, 1977). When immature). In this report, we re-investigate hamster oocyte maturation, comparing extra- and intrafollicular maturations

Paris-Sud XI, Université de

8

Ultrastructural study of the interactions and fusion of ram spermatozoa with zona-free hamster oocytes  

E-print Network

Ultrastructural study of the interactions and fusion of ram spermatozoa with zona-free hamster, Czechoslovakia. Summary. The interaction of preincubated ram sperm with zona-free hamster oocytes was studied undertaken ultrastructural observations of the interactions of zona- free hamster oocytes with ram sperm

Paris-Sud XI, Université de

9

Changes of spontaneous parthenogenetic activation and development potential of golden hamster oocytes during the aging process.  

PubMed

The golden hamster is an excellent animal experimental model for oocyte research. The hamster oocytes are very useful in clinical examination of human spermatozoan activity. Non-fertile oocytes can lead to time-dependent processes of aging, which will affect the results of human spermatozoa examination. As a consequence there is a need to investigate the aging and anti-aging processes of golden hamster oocytes. In order to study the aging processes and parthenogenetic activation of golden hamster oocytes, in vivo oocytes, oocytes cultured with or without cumulus cells, and oocytes treated with Trichostatin A (TSA) or caffeine were collected and investigated. We found that: (1) spontaneous parthenogenetic activation, developmental potential (cleavage rate), and zona pellucida (ZP) hardening undergo age-dependent changes in in vivo, in vitro, and after TSA or caffeine treatment; (2) in vivo, oocytes became spontaneously parthenogenetic 25 h post-hCG treatment; (3) in vitro, cumulus cells did not significantly increase the parthenogenetic activation rate of cultured hamster oocytes; and (4) TSA or caffeine could delay spontaneous oocyte parthenogenetic activation and the aging processes by at least 5h, but also accelerated the hardening of the ZP. These results define the conditions for the aging and anti-aging processes in golden hamster oocytes. TSA and caffeine play roles in controlling spontaneous activation, which could facilitate the storage and use of golden hamster oocytes for studying processes relevant to human reproduction. PMID:25480399

Jiang, Han; Wang, Ce; Guan, Jiyu; Wang, Lingyan; Li, Ziyi

2015-01-01

10

Preservation of hamster oocytes to assay the fertilizing capacity of human spermatozoa.  

PubMed

Between 70 and 80% of zona-intact hamster ova survived freezing after slow cooling (approximately 0.3 degrees C/min) to -80 degrees C in Medium PB1 containing 1.5 or 2.0 M-DMSO before transfer to -196 degrees C. After slow warming (approximately 8 degrees C/min), there was no difference in survival if the DMSO was diluted out by a slow stepwise or a rapid single addition of medium. When slow cooling was terminated at -40 degrees C by direct transfer to -196 degrees C, up to 75% of the ova survived rapid warming (approximately 500 degrees C/min) and rapid dilution if the medium contained 2.0 M-DMSO. The survival rates were calculated on the basis of the number of thawed ova which retained their normal morphological appearance after a 1 h incubation before removal of the zona pellucida with trypsin. All of these ova were penetrated after incubation with mouse spermatozoa, indicating that the freezing procedure per se does not adversely affect the penetration of frozen-thawed hamster ova by heterologous spermatozoa. There was no difference in the penetration rate of human spermatozoa into frozen (34%) or fresh (42%) oocytes when a Hepes-buffered Tyrode solution containing 30 mg BSA/ml and 2.0 M-DMSO was used as the freezing medium. However, fewer ova frozen in Medium PB1 containing 4 mg BSA/ml and 2.0 M-DMSO were penetrated by human spermatozoa (18%) compared with freshly collected ova (38%). Zona-free ova did not survive the freezing procedure as well as zona-intact ova. The survival of hamster oocytes stored at -196 degrees C offers a convenient means of supplying and transporting these ova for the assessment of the fertilizing capacity of human and other heterologous spermatozoa. PMID:7120180

Quinn, P; Barros, C; Whittingham, D G

1982-09-01

11

Age-Associated Metabolic and Morphologic Changes in Mitochondria of Individual Mouse and Hamster Oocytes  

PubMed Central

Background In human oocytes, as in other mammalian ova, there is a significant variation in the pregnancy potential, with approximately 20% of oocyte-sperm meetings resulting in pregnancies. This frequency of successful fertilization decreases as the oocytes age. This low proportion of fruitful couplings appears to be influenced by changes in mitochondrial structure and function. In this study, we have examined mitochondrial biogenesis in both hamster (Mesocricetus auratus) and mouse (Mus musculus) ova as models for understanding the effects of aging on mitochondrial structure and energy production within the mammalian oocyte. Methodology/Principal Findings Individual metaphase II oocytes from a total of 25 young and old mice and hamsters were collected from ovarian follicles after hormone stimulation and prepared for biochemical or structural analysis. Adenosine triphosphate levels and mitochondrial DNA number were determined within individual oocytes from young and old animals. In aged hamsters, oocyte adenosine triphosphate levels and mitochondrial DNA molecules were reduced 35.4% and 51.8%, respectively. Reductions of 38.4% and 44% in adenosine triphosphate and mitochondrial genomes, respectively, were also seen in aged mouse oocytes. Transmission electron microscopic (TEM) analysis showed that aged rodent oocytes had significant alterations in mitochondrial and cytoplasmic lamellae structure. Conclusions/Significance In both mice and hamsters, decreased adenosine triphosphate in aged oocytes is correlated with a similar decrease in mtDNA molecules and number of mitochondria. Mitochondria in mice and hamsters undergo significant morphological change with aging including mitochondrial vacuolization, cristae alterations, and changes in cytoplasmic lamellae. PMID:23741435

Simsek-Duran, Fatma; Li, Fang; Ford, Wentia; Swanson, R. James; Jones, Howard W.; Castora, Frank J.

2013-01-01

12

DNA (DEOXYRIBONUCLEIC ACID) SYNTHESIS FOLLOWING MICROINJECTION OF HETEROLOGOUS SPERM AND SOMATIC CELL NUCLEI INTO HAMSTER OOCYTES  

EPA Science Inventory

The authors have investigated the ability of the hamster oocyte to initiate DNA synthesis in nuclei differing in basic protein content. DNA synthesis was studied by autoradiography in oocytes that had been incubated in 3H-thymidine after being parthenogenetically activated by sha...

13

Human sperm chromosome analysis after subzonal sperm insemination of hamster oocytes  

SciTech Connect

Sperm microinjection techniques, subzonal sperm insemination (SUZI) and intracytoplasmic sperm injection (ICSI), have achieved a wide spread clinical application for the treatment of male infertility. To date, only one study has focused on sperm karyotypes after microinjection. Martin et al. reported a very high incidence of abnormal human sperm complements after ICSI into hamster oocytes. In the present study, are reported the first human sperm karyotypes after SUZI of hamster oocytes. Spermatozoa from two control donors were treated by calcium ionophore A23187 and injected under the zona of hamster eggs. The microinjected eggs were then cultured for cytogenetic analysis of the pronuclei. Out of 47 analyzed sperm chromosome metaphases, 5 (10.6%) were abnormal, 4 (8.5%) were hypohaploid and 1 (2.1%) had a structural abnormality. The sex ratio was not significantly different from the expected 1:1 ratio. Rates of chromosomal abnormalities in microinjected spermatozoa were similar to those observed in spermatozoa inseminated with zona free eggs, suggesting that SUZI procedure per se does not increase sperm chromosomal abnormalities.

Cozzi, J. [Medical School of Grenoble (France)

1994-09-01

14

The rates of premature chromosome condensation and embryo development after injection of irradiated sperms into hamster oocytes  

PubMed Central

Background: Irradiation is one of the major causes of induced sperm DNA damage. Various studies suggested a relation between sperm DNA damage and fertilization rate after intra-cytoplasmic sperm injection (ICSI). Objective: In this study, fertilization rate and premature chromosome condensation (PCC) formation after ICSI of hamster oocytes with irradiated sperms from normal and oligosperm individuals was investigated. Materials and Methods: Human sperms were classified according to counts to normal and oligosperm. Ten samples were used for each group. Golden hamster oocytes were retrieved after super ovulation by PMSG and HCG injection. From retrieved oocytes, 468 were in metaphase II. Control and 4 Gy gamma irradiated sperms were then injected into oocytes. After pronuclei formation in injected oocytes and formation of 8 cells embryos, slides were prepared using Tarkowskie's standard air-drying technique. The frequency of embryos and PCC were analyzed using 1000× microscope after staining in 5% Giemsa. Results: The extent of embryo development in oocytes injected by irradiated sperms was lower than those injected by non-irradiated sperms (p=0.0001). The frequency of PCC in failed fertilized oocytes was significantly higher in oligosperms (46%) compared with normal ones (0%), but there was no significant difference between irradiated and non-irradiated samples in each group (p=0.12). Conclusion: The results showed that irradiation of sperms might influence the fertilization outcome possibly due to sperm DNA damage. One possible cause of precluding oocytes from fertilization in oligosperm individuals might be the formation of PCC. PMID:24639771

Moghbelinejad, Sahar; Mozdarani, Hossein; Rezaeian, Zahra

2013-01-01

15

CARBENDAZIM (MBC) DISRUPTS OOCYTE SPINDLE FUNCTION AND INDUCES ANEUPLOIDY IN HAMSTERS EXPOSED DURING FERTILIZATION (MEIOSIS II)  

EPA Science Inventory

Peri-fertilization exposure to Carbendazim (MBC; a microtubule poison) induces infertility and early pregnancy loss (EPL) in hamsters. resently, both in vivo and in vitro techniques were employed to characterize the effects of MBC on cellular aspects of fertilization in hamsters....

16

Recombinant Hamster Oviductin Is Biologically Active and Exerts Positive Effects on Sperm Functions and Sperm-Oocyte Binding  

PubMed Central

Studies carried out in several mammalian species suggest that oviductin, also known as oviduct-specific glycoprotein or OVGP1, plays a key role in sperm capacitation, fertilization, and development of early embryos. In the present study, we used recombinant DNA technology to produce, for the first time, recombinant hamster OVGP1 (rHamOVGP1) in human embryonic kidney 293 (HEK293) cells. rHamOVGP1 secreted in the culture medium was purified by affinity chromatography. The resulting protein migrated as a poly-dispersed band of 160-350 kDa on SDS-PAGE corresponding to the molecular mass of the native HamOVGP1. Subsequent mass spectrometric analysis of the purified rHamOVGP1 confirmed its identity as HamOVGP1. Immunocytochemistry demonstrated binding of rHamOVGP1 to the mid-piece and head of hamster sperm and to the zona pellucida (ZP) of ovarian oocytes. In vitro functional experiments showed that addition of rHamOVGP1 in the capacitation medium further enhanced tyrosine phosphorylation of two sperm proteins of approximately 75 kDa and 83 kDa in a time-dependent manner. After 3 hours of incubation in the presence of rHamOVGP1, a significant increase in acrosome reaction was measured. Pretreatment of either sperm or oocyte with 20 ?g/ml of rHamOVGP1 prior to sperm-egg binding assay significantly increased the number of sperm bound to the ZP. Addition of rHamOVGP1 in the medium during sperm-egg binding with either oocyte or sperm pretreated with rHamOVGP1 also saw an increase in the number of sperm bound to ZP. In all experimental conditions, the effect of rHamOVGP1 on sperm-oocyte binding was negated by the addition of monoclonal anti-HamOVGP1 antibody. The successful production and purification of a biologically active rHamOVGP1 will allow further exploration of the function of this glycoprotein in reproductive function. PMID:25849110

Yang, Xiaojing; Zhao, Yuewen; Yang, Xiaolong; Kan, Frederick W. K.

2015-01-01

17

Recombinant hamster oviductin is biologically active and exerts positive effects on sperm functions and sperm-oocyte binding.  

PubMed

Studies carried out in several mammalian species suggest that oviductin, also known as oviduct-specific glycoprotein or OVGP1, plays a key role in sperm capacitation, fertilization, and development of early embryos. In the present study, we used recombinant DNA technology to produce, for the first time, recombinant hamster OVGP1 (rHamOVGP1) in human embryonic kidney 293 (HEK293) cells. rHamOVGP1 secreted in the culture medium was purified by affinity chromatography. The resulting protein migrated as a poly-dispersed band of 160-350 kDa on SDS-PAGE corresponding to the molecular mass of the native HamOVGP1. Subsequent mass spectrometric analysis of the purified rHamOVGP1 confirmed its identity as HamOVGP1. Immunocytochemistry demonstrated binding of rHamOVGP1 to the mid-piece and head of hamster sperm and to the zona pellucida (ZP) of ovarian oocytes. In vitro functional experiments showed that addition of rHamOVGP1 in the capacitation medium further enhanced tyrosine phosphorylation of two sperm proteins of approximately 75 kDa and 83 kDa in a time-dependent manner. After 3 hours of incubation in the presence of rHamOVGP1, a significant increase in acrosome reaction was measured. Pretreatment of either sperm or oocyte with 20 ?g/ml of rHamOVGP1 prior to sperm-egg binding assay significantly increased the number of sperm bound to the ZP. Addition of rHamOVGP1 in the medium during sperm-egg binding with either oocyte or sperm pretreated with rHamOVGP1 also saw an increase in the number of sperm bound to ZP. In all experimental conditions, the effect of rHamOVGP1 on sperm-oocyte binding was negated by the addition of monoclonal anti-HamOVGP1 antibody. The successful production and purification of a biologically active rHamOVGP1 will allow further exploration of the function of this glycoprotein in reproductive function. PMID:25849110

Yang, Xiaojing; Zhao, Yuewen; Yang, Xiaolong; Kan, Frederick W K

2015-01-01

18

Hamsters  

NSDL National Science Digital Library

Scientists believe that most behaviors, from fighting to mating, are controlled by brain chemistry. In this Science Update, you'll hear how researchers are using hamsters to understand the roots of aggression.

Science Update

2000-03-15

19

Effect of fertility regulating agents on motility and zona-free hamster egg penetration by spermatozoa of bonnet monkey.  

PubMed

Administration of STS-557 (17 alpha-cyanomethyl-17 beta-hydroxyestra 4,9(10)-dien-3-one; 12 mg/monkey daily) for 4 weeks either alone or in combination with 20 Aet-1 (testosterone-trans-4-n-butyl cyclohexyl carboxylate; code CDB 1781; 40 mg/monkey single administration) had no significant effect on motility and zona free hamster egg penetration by spermatozoa of bonnet monkey, but continuation of the treatment for 12 weeks reduced (in one monkey treated with STS-557) or abolished (one treated with STS-557 and two with STS-557 + 20 Aet-1) the motility as well as zona-free hamster egg penetration (by spermatozoa of all treated monkeys). Motility and the ability to penetrate zona-free hamster egg returned to normalcy after 10 weeks of withdrawal of treatments. Active immunization of monkeys with ovine FSH (4 weeks after booster) had no adverse effect on motility of spermatozoa but none of the zona-free hamster eggs was fertilized. The correlation between motility and the capacity to penetrate the zona-free hamster eggs by monkey spermatozoa varies with the treatment. Such correlation was apparent in monkeys treated with STS-557 but not in monkeys immunized with ovine FSH. PMID:1293043

Sharma, R K; Das, R P

1992-11-01

20

Serial study of the effect of radiotherapy on semen parameters, hamster egg penetration rates, and lymphocyte chromosome abnormalities  

SciTech Connect

This study was designed to assess the long-term effects of radiotherapy (RT) on male fertility and the induction of lymphocyte and sperm chromosome abnormalities. This preliminary report provides information on 11 cancer patients (mainly seminomas) treated by RT (testicular dose, 44 to 499 rads). All 11 men were studied pre-RT and at intervals post-RT. The pre-RT semen profile varied considerably, but, in general, the profile was poor with a mean sperm concentration of 19.4 x 10/sup 6/ ml and a mean hamster egg penetration rate of 5%. One month after RT, the sperm concentration decreased and hamster egg penetration was 0% in all men. At 3 and 12 months post-RT, all but two patients were azoospermic. By 24 months post-RT, 9 of 11 patients had regained sperm production and 5 had sperm capable of hamster egg penetration. The three men who have been studied 36 months post-RT had a mean sperm concentration of 45.3 x 10/sup 6/ ml, and all had positive hamster egg penetration tests, although two of the three men had very low penetration rates (2% and 4%). Lymphocyte chromosome analysis demonstrated a striking frequency of chromosome abnormalities post-RT which decreased with time (pre-RT, 0%; 1 month, 42.4%; 3 months, 24.7%; 12 months, 13.8%; 24 months, 11.2%; and 36 months, 10.0%). Thus, it appears that sperm production starts to recover 2 to 3 years after RT when the frequency of lymphocyte chromosome abnormalities has decreased, but the sperm may not be fully functional at this time, as evidenced by poor rates of hamster egg penetration. Future studies of sperm chromosome analysis in these men will determine whether this impairment of the sperm is associated with meiotic chromosome abnormalities.

Martin, R.H.; Barnes, M.; Arthur, K.; Ringrose, T.; Douglas, G.

1984-02-01

21

IMPORTANCE OF GLUTATHIONE IN THE ACQUISITION AND MAINTENANCE OF SPERM NUCLEAR DECONDENSING ACTIVITY IN MATURING HAMSTER OOCYTES  

EPA Science Inventory

Sperm nuclear decondensing activity in mammalian oocytes is dependent upon the maturational state of the oocyte. It is maximal in mature, metaphase II oocytes and minimal or absent in immature germinal vesicle (GV) and fertilized pronuclear oocytes. Previous studies suggested tha...

22

Pattern and frequency of nondisjunction in oocytes from the Djungarian hamster are determined by the stage of first meiotic spindle inhibition.  

PubMed

In order to study the mechanisms of nondisjunction at meiosis I in oocytes gonadotropin-stimulated Djungarian hamsters were treated at two stages [4.5 and 6 h post human chorionic gonadotropin (HCG)] during the preovulatory period with 1000 mg/kg Carbendazim (MBC). The compound, known to bind fast but reversibly to mammalian tubulin, was chosen to investigate whether the stage at which spindle function is inhibited affects the pattern of nondisjunction. Ovulated oocytes were cytologically prepared and scored for hyperhaploidy, diploidy and presegregation. Application at an early spindle phase, 4.5 h post HCG, to females stimulated with a low gonadotropin dose [3 IU pregnant mares serum (PMS); 2 IU HCG] caused a high frequency of nondisjunction (40.6%) with a more or less nonspecific pattern of malsegregated bivalents. Treatment at a late stage of spindle function (6 h post HCG) resulted in a less frequent (22.5%) but highly preferential malsegregation of those A-D group bivalents thought earlier to be late segregators. On the other hand, oocytes from females primed with a high (10 IU PMS and HCG) gonadotropin dose, a treatment assumed to delay meiosis by approximately 1.5 h, responded to MBC treatment at the late stage (6 h) with a nonspecific pattern and a high frequency (71.2%) of nondisjunction. The latter result is comparable to that in which MBC was given at the early stage (4.5 h) and after a low gonadotropin dose. The high nondisjunction response additionally indicates that spindles in hypergonadotropic stimulated oocytes are more susceptible and/or that the concentration of the inhibitor is higher in such oocytes. Only few oocytes with presegregation (3.1%; 0.0%; 1.7%) and few diploid oocytes (3.3%; 1.5%; 3.2%) with complete inhibition of meiosis I were observed. We conclude, that in Djungarian hamsters (1) the segregation of bivalents at meiosis I is asynchronous with the large A-D bivalents segregating last, (2) the phase in which spindle function is inhibited determines the pattern of nondisjunction, and (3) the resumption of meiosis I - from dictyotene to metaphase II - does not follow a rigidly timed programme but depends on the conditions of follicular maturation. PMID:3219919

Hummler, E; Hansmann, I

1988-11-01

23

Proteins of the accessory sex glands associated with the oocyte-penetrating capacity of cauda epididymal sperm from holstein bulls of documented fertility  

Microsoft Academic Search

We previously reported that ac- cessory sex gland fluid (AGF) from high fertility (HF) bulls influenced the oocyte-penetrating capacity of cauda epididymal sperm from low fertility (LF) bulls, based on in vitro fertilization (IVF) assays. The present study determined if AGF proteins were associated with these effects. Nineteen IVF assays with 12 bulls were grouped as follows. Group I (n

Arlindo A. Moura; David A. Chapman; Gary J. Killian

2007-01-01

24

Identification of a sperm penetration factor in the oviduct of the golden hamster.  

PubMed

Previously, we found oviductal eggs to be significantly more penetrable and fertilizable in vitro than ovulated eggs collected from the ovarian bursa, while bursal eggs were comparable to mature (unovulated) follicular eggs. Incubation of follicular eggs with a soluble eluate of oviductal egg cumulus complexes (COF) increased sperm penetration: the activity was macromolecular, was destroyed at 56 degrees C, and was produced in the oviduct. We now report purification of this oviductal factor that enhances penetration of follicular eggs and have identified it as oviductin (OVN). Oviducts, 1-1.5 h post-LH from eCG-primed females, were homogenized and the cytosolic fraction was chromatographed on a Helix pomatia lectin affinity column; specific proteins were eluted with 0.2 M N-acetyl-D-galactosamine. Fractions were monitored by dot-blot assay using as the primary antibody monoclonal antibody (mAb) 1C4 against OVN. Proteins were resolved by one-dimensional SDS-gel electrophoresis, followed by electrotransfer and immunostaining of Western blots. OVN fractions were indexed to COF by quantitative dot-blot assay, and activity was bioassayed by penetration of follicular eggs within 1 h of coincubation with precapacitated sperm +/- factors: COF and BSA (high and low controls, respectively) and fractions from the lectin-isolated peak. The mean penetration rates for three isolations were 17 +/- 4.0a, 51.7 +/- 5.0b, and 49 +/- 2.7b% for BSA, COF, and column fractions, respectively (p < or = 0.05). Purified OVN bound to follicular zonae during culture. Acrosome-intact sperm heads bound OVN during 30 min of incubation both before (t = 0 h) and after capacitation (t = 5.5 h) (visualized by indirect immunofluorescence).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7711179

Boatman, D E; Magnoni, G E

1995-01-01

25

USE OF THE FUNGICIDE CARBENDAZIM AS A MODEL COMPOUND TO DETERMINE THE IMPACT OF ACUTE CHEMICAL EXPOSURE DURING OOCYTE MATURATION AND FERTILIZATION ON PREGNANCY OUTCOME IN THE HAMSTER  

EPA Science Inventory

Here we use a hamster animal model to identify early pregnancy loss due to an acute chemical exposure to the female during the perifertilization interval. The fungicide carbendazim (methyl 1H-benzimidazole-2-carbamate), a microtubule poison with antimitotic activity, was selected...

26

Disposition and metabolic profiling of the penetration enhancer Azone. I. In vitro studies: Urinary profiles of hamster, rat, monkey, and man  

SciTech Connect

Chain-labeled {sup 14}C-Azone was intravenously administered to hamster, monkey, and rat, to compare its metabolic profile with that obtained previously in humans after dermal application. Azone-derived radioactivity was excreted predominantly in the urine for both hamster and monkey, which is similar to the disposition in humans. Metabolic profiling in urine revealed extensive systemic metabolism to occur in all species studied. The main fraction of the metabolites was most polar in man, followed by rat, monkey, and hamster. Traces of the parent compound were detectable only in hamster urine. Although some of the polar major human metabolites were also present in rat urine, the animals were unsuitable for collecting metabolites of Azone observed in humans. In rats, complete cleavage of the dodecyl side chain was ruled out by administering Azone that had been labeled at two distinct positions of the molecule. Additionally, oral administration of Azone to rats resulted in the same metabolic profile as intravenous administration, indicating that gastrointestinal metabolism does not occur or is similar to systemic metabolism.

Wiechers, J.W.; Drenth, B.F.; Adolfsen, F.A.; Prins, L.; de Zeeuw, R.A. (Groningen Centre for Drug Research (Netherlands))

1990-05-01

27

Cryopreservation of starfish oocytes.  

PubMed

Research from many laboratories over the past several decades indicates that invertebrate oocytes and eggs are extraordinarily difficult to freeze. Since starfish oocytes, eggs, and embryos are an important cell and developmental biology model system, there is great interest to cryopreserve these cells. Previous starfish oocyte cryopreservation studies using slow cooling protocols revealed that these cells are highly sensitive to osmotic stress and form intracellular ice at very high sub-zero temperatures, suggesting that common freezing methodologies may not prove useful. We report here that a short exposure to 1.5 M Me2SO/1 M trehalose in hypotonic salt solution followed by ultra-rapid cooling to cryogenic temperatures allows starfish oocytes to be cryopreserved with the average survival rate of 34% when normalized to control oocytes that were exposed to CPA, but not frozen. On average, 51% of the oocytes in 77% of the batches of frozen oocytes underwent meiotic maturation in response to the starfish maturation hormone, 1-methyladenine. In one experiment, eggs developing from thawed oocytes were capable of being fertilized and two developed into embryos. These data suggests that successful cryopreservation of starfish oocytes is possible, but will need further refinement to increase the numbers of fully competent embryos. PMID:15710368

Hamarato?lu, Fisun; Ero?lu, Ali; Toner, Mehmet; Sadler, Kirsten C

2005-02-01

28

Reduction of polyspermic penetration using biomimetic microfluidic technology during in vitro fertilization  

E-print Network

of spermatozoa past the oocytes similar to the pattern in the oviduct. In vitro fertilization of porcine oocytes are similar. Here we demonstrate that the biomimetic microchannel in vitro fertilization system can reduce in vitro production efficiency. Introduction Polyspermic penetration of oocytes fertilized in vitro remains

Beebe, David J.

29

Maturation, fertilization and complete development of porcine oocytes matured under different systems  

Microsoft Academic Search

This study was designed 1) to determine the effectiveness of 2 in vitro\\u000a maturation systems commonly employed to produce nuclear and\\u000a cytoplasmicly mature pig oocytes, 2) to assess the effects of boar,\\u000a sperm concentration and maturation system on oocyte penetrability and\\u000a male pronucleus formation and 3) to determine the ability of the in\\u000a vitro matured oocytes to be fertilized in

P. Coy; S. Ruiz; R. Romar; I. Campos; J. Gadea

1999-01-01

30

Molecular and Cellular Mechanisms of Sperm-Oocyte Interactions Opinions Relative to in Vitro Fertilization (IVF)  

PubMed Central

One of the biggest prerequisites for pregnancy is the fertilization step, where a human haploid spermatozoon interacts and penetrates one haploid oocyte in order to produce the diploid zygote. Although fertilization is defined by the presence of two pronuclei and the extraction of the second polar body the process itself requires preparation of both gametes for fertilization to take place at a specific time. These preparations include a number of consecutive biochemical and molecular events with the help of specific molecules and with the consequential interaction between the two gametes. These events take place at three different levels and in a precise order, where the moving spermatozoon penetrates (a) the outer vestments of the oocyte, known as the cumulus cell layer; (b) the zona pellucida (ZP); where exocytosis of the acrosome contents take place and (c) direct interaction of the spermatozoon with the plasma membrane of the oocyte, which involves a firm adhesion of the head of the spermatozoon with the oocyte plasma membrane that culminates with the fusion of both sperm and oocyte membranes (Part I). After the above interactions, a cascade of molecular signal transductions is initiated which results in oocyte activation. Soon after the entry of the first spermatozoon into the oocyte and oocyte activation, the oocyte’s coat (the ZP) and the oocyte’s plasma membrane seem to change quickly in order to initiate a fast block to a second spermatozoon (Part II). Sometimes, two spermatozoa fuse with one oocyte, an incidence of 1%–2%, resulting in polyploid fetuses that account for up to 10%–20% of spontaneously aborted human conceptuses. The present review aims to focus on the first part of the human sperm and oocyte interactions, emphasizing the latest molecular and cellular mechanisms controlling this process. PMID:25054321

Anifandis, George; Messini, Christina; Dafopoulos, Konstantinos; Sotiriou, Sotiris; Messinis, Ioannis

2014-01-01

31

Oocyte ageing and epigenetics  

PubMed Central

It has become a current social trend for women to delay childbearing. However, the quality of oocytes from older females is compromised and the pregnancy rate of older women is lower. With the increased rate of delayed childbearing, it is becoming more and more crucial to understand the mechanisms underlying the compromised quality of oocytes from older women, including mitochondrial dysfunctions, aneuploidy and epigenetic changes. Establishing proper epigenetic modifications during oogenesis and early embryo development is an important aspect in reproduction. The reprogramming process may be influenced by external and internal factors that result in improper epigenetic changes in germ cells. Furthermore, germ cell epigenetic changes might be inherited by the next generations. In this review, we briefly summarise the effects of ageing on oocyte quality. We focus on discussing the relationship between ageing and epigenetic modifications, highlighting the epigenetic changes in oocytes from advanced-age females and in post-ovulatory aged oocytes as well as the possible underlying mechanisms. PMID:25391845

Ge, Zhao-Jia; Schatten, Heide; Zhang, Cui-Lian; Sun, Qing-Yuan

2015-01-01

32

Oocyte ageing and epigenetics.  

PubMed

It has become a current social trend for women to delay childbearing. However, the quality of oocytes from older females is compromised and the pregnancy rate of older women is lower. With the increased rate of delayed childbearing, it is becoming more and more crucial to understand the mechanisms underlying the compromised quality of oocytes from older women, including mitochondrial dysfunctions, aneuploidy and epigenetic changes. Establishing proper epigenetic modifications during oogenesis and early embryo development is an important aspect in reproduction. The reprogramming process may be influenced by external and internal factors that result in improper epigenetic changes in germ cells. Furthermore, germ cell epigenetic changes might be inherited by the next generations. In this review, we briefly summarise the effects of ageing on oocyte quality. We focus on discussing the relationship between ageing and epigenetic modifications, highlighting the epigenetic changes in oocytes from advanced-age females and in post-ovulatory aged oocytes as well as the possible underlying mechanisms. PMID:25391845

Ge, Zhao-Jia; Schatten, Heide; Zhang, Cui-Lian; Sun, Qing-Yuan

2015-03-01

33

TIMING OF HAMSTER SPERM NUCLEAR DECONDENSATION AND MALE PRONUCLEUS FORMATION IS RELATED TO SPERM NUCLEAR DISULFIDE BOND CONTENT  

EPA Science Inventory

The relationship between the timing of both sperm nuclear decondensation and male pronucleus formation in the oocyte and the relative level of disulfide bonds within the sperm nucleus was evaluated. Four types of hamster sperm nuclei, in which the extent of S-S bonding differed, ...

34

Curiosity in the hamster  

Microsoft Academic Search

Running time for repeated alley runs by hamsters to end boxes containing (a) nothing, (b) a constant set of objects, or (c) a changing set of objects was in the order a > b > c: thus, novelty appeared reinforcing. Length of intersession interval (2, 24, 48 hr.) affected only Condition b: thus, \\

Gerald E. Schneider; Charles G. Gross

1965-01-01

35

Intracytoplasmic sperm injection improves in vitro embryo production from poor quality bovine oocytes.  

PubMed

The objective was to use subzonal sperm injection (SUZI) to understand sperm penetration patterns and to use intracytoplasmic sperm injection (ICSI) to improve production of bovine embryos using poor quality gametes. In experiment 1, poor versus good quality oocytes were fertilized with sperm from two bulls, A and B, with poor and good sperm vigor, respectively. The blastocyst rate was higher for good versus poor quality oocytes (23.3% vs. 11.1%, P < 0.05), regardless of the bull used. There was no significant difference in blastocyst rate for bull A (low vigor) regardless of oocyte quality, and for bull B (high vigor), blastocyst rate was better for good versus poor quality oocytes (25.7% vs. 9.2%, P < 0.05). In experiment 2, poor quality oocytes were subjected to SUZI. The oocyte penetration rate was lower for bull A than for bull B (29.6% vs. 53.8%, P < 0.05), when SUZI was performed within 1 hour after sperm processing. However, when SUZI was performed 2 to 3 hours after sperm processing, penetrating capacity was similar between bulls, but for bull B, penetrating capacity significantly decreased after 3 hours of sperm processing. In an attempt to overcome sperm penetrating disorders, poor and good quality oocytes were subjected to ICSI (experiment 3). Irrespective of the bull or of the oocyte quality grade, there were no differences in cleavage or blastocyst rates. Both bulls had distinct IVF embryo production rates, which we inferred were because of particular individual sperm characteristics. In conclusion, ICSI was an effective means to achieve in vitro production of bovine embryos with gametes of variable quality. PMID:23312719

Ohlweiler, L U; Brum, D S; Leivas, F G; Moyses, A B; Ramos, R S; Klein, N; Mezzalira, J C; Mezzalira, A

2013-03-15

36

Fertilization Ability of Porcine Oocytes Reconstructed from Ooplasmic Fragments Produced and Characterized after Serial Centrifugations  

PubMed Central

Abstract Mitochondria are reported to be critical in in vitro maturation of oocytes and subsequent embryo development after fertilization, but their contribution for fertilization has not been investigated in detail. In the present study, we investigate the contribution of mitochondria to fertilization using reconstructed porcine oocytes by fusion of ooplasmic fragments produced by serial centrifugations (centri-fusion). Firstly, we evaluated the characteristics of ooplasmic fragments. Three types of fragments were obtained by centrifugation of porcine oocytes matured in vitro for 46 h: brownish (B), transparent (T) and large (L) fragments containing both B and T parts in a fragment. The production efficiencies of these types of fragments were 71.7, 91.0 and 17.8 fragments/100 oocytes, respectively. In experiments, L fragments were excluded because they contained both brownish and transparent components that were apparently intermediate between B and T fragments. Observations by confocal microscopy after staining with MitoTracker Red CMXRos® and transmission electron microscopy revealed highly condensed active mitochondria in B fragments in contrast to T fragments that contained only sparse organelles. We reconstructed oocytes by fusion of a karyoplast and two cytoplasts from B and T fragments (B and T oocytes, respectively). The B oocytes showed higher sperm penetration (95.8%) and male pronuclear formation rates (94.2%) by in vitro fertilization than T oocytes (66.7% and 50.0%, respectively). These results suggest that the active mitochondria in oocytes may be related to their ability for fertilization. PMID:23965685

Viet LINH, Nguyen; KIKUCHI, Kazuhiro; NAKAI, Michiko; TANIHARA, Fuminori; NOGUCHI, Junko; KANEKO, Hiroyuki; DANG-NGUYEN, Thanh Quang; MEN, Nguyen Thi; VAN HANH, Nguyen; SOMFAI, Tamas; NGUYEN, Bui Xuan; NAGAI, Takashi; MANABE, Noboru

2013-01-01

37

Oocyte cryopreservation in oncological patients.  

PubMed

The use of chemotherapy and radiotherapy in oncological patients may reduce their reproductive potential. Sperm cryopreservation has been already used in men affected by neoplastic disease. Oocyte cryopreservation might be an important solution for these patients at risk of losing ovarian function. A program of oocyte cryopreservation for oncological patients is also present in our center. From June 1996 to January 2000, 18 patients awaiting chemotherapy and radiotherapy for neoplastic disease were included in our oocyte cryopreservation program. Our experience documents that oocyte storage may be a concrete and pragmatic alternative for oncological patients. The duration of oocyte storage does not seem to interfere with oocyte survival as pregnancies occurred even after several years of gamete cryopreservation in liquid nitrogen. PMID:15041124

Porcu, Eleonora; Fabbri, Raffaella; Damiano, Giuseppe; Fratto, Rosita; Giunchi, Susanna; Venturoli, Stefano

2004-04-01

38

The control of reactive oxygen species influences porcine oocyte in vitro maturation.  

PubMed

The aim of this study was to examine the effect of varying intracellular reactive oxygen species (ROS) levels during oocyte in vitro maturation with enzymatic ROS production systems (xanthine + xanthine oxidase or xanthine + xanthine oxidase + catalase), scavenger systems (catalase or superoxide dismutase + catalase) or cysteine on porcine oocyte maturation. Oocyte ROS levels showed an increase when H2 O2 or O2 ?(-) production systems were added to the culture medium (p < 0.05). On the other hand, the presence of ROS scavengers in the maturation medium did not modify oocyte ROS levels compared with the control after 48 h of maturation, but the addition of cysteine induced a decrease in oocyte ROS levels (p < 0.05). The ROS production systems used in this work did not modified the percentage of oocyte nuclear maturation, but increased the decondensation of sperm head (p < 0.05) and decreased the pronuclear formation (p < 0.05). In turn, the addition of O2 ?(-) and H2 O2 scavenging systems during in vitro maturation did not modify the percentage of oocytes reaching metaphase II nor the oocytes with decondensed sperm head or pronuclei after fertilization. However, both parameters increased in the presence of cysteine (p < 0.05). The exogenous generation of O2 ?(-) and H2 O2 during oocyte in vitro maturation would not affect nuclear maturation or later sperm penetration, but most of the spermatozoa cannot progress to form the pronuclei after fusion with the oocyte. The decrease in endogenous ROS levels by the addition of cysteine would improve pronuclear formation after sperm penetration. PMID:25522082

Alvarez, G M; Morado, S A; Soto, M P; Dalvit, G C; Cetica, P D

2015-04-01

39

Photoperiodic Control of Hamster Testis  

Microsoft Academic Search

The response of the testes of juvenile and adult hamsters to various photoperiods was examined. The testes of juvenile animals reached maturity regardless of the light cycle on which the animals were raised. However, the testes of adult hamsters required at least 12.5 hours of light per day to maintain spermatogenesis and prevent degeneration. This is one of the few

Suzanne Gaston; Michael Menaker

1967-01-01

40

Fertilization failure of frozen mouse oocytes is not due to premature cortical granule release.  

PubMed

The proportion of mouse oocytes that were fertilized in vitro after storage at -196 degrees C in the presence of 1.5 M dimethylsulfoxide (DMSO) was significantly lower than in unfrozen controls (39% vs. 81%). Sperm failed to penetrate the zona pellucida of approximately 80% of the frozen oocytes that remained unfertilized. Removal of the zona restored fertilization to control levels, indicating that changes induced in the zona during freezing and/or thawing were the primary cause of fertilization failure. Sperm-oocyte fusion, sperm nucleus decondensation, and the resumption of meiosis in frozen oocytes appeared to be delayed but subsequently fertilization progressed normally. No evidence was found to suggest zona modification by the premature release of fucosyl-rich glycoconjugates of cortical granule origin onto the surface of the plasma membrane of frozen oocytes stained immediately after thawing with fluorescently labeled Ulex europaeus lectin. Only a few frozen (less than 5%) and control (less than 3%) oocytes that failed to fertilize in vitro had fucosylated molecules on the plasma membrane. Prolonged exposure of fertilized oocytes to DMSO at 4 degrees C did not alter the pattern of lectin binding. In conclusion, fertilization is inhibited in frozen-thawed oocytes by as yet undefined modifications to the zona pellucida which do not involve the premature release of cortical granules. PMID:1391317

Wood, M J; Whittingham, D G; Lee, S H

1992-06-01

41

Oocyte destruction is activated during viral infection.  

PubMed

Viral infection has been associated with a starvation-like state in Drosophila melanogaster. Because starvation and inhibiting TOR kinase activity in vivo result in blocked oocyte production, we hypothesized that viral infection would also result in compromised oogenesis. Wild-type flies were injected with flock house virus (FHV) and survival and embryo production were monitored. Infected flies had a dose-responsive loss of fecundity that corresponded to a global reduction in Akt/TOR signaling. Highly penetrant egg chamber destruction mid-way through oogenesis was noted and FHV coat protein was detected within developing egg chambers. As seen with in vivo TOR inhibition, oogenesis was partially rescued in loss of function discs large and merlin mutants. As expected, mutants in genes known to be involved in virus internalization and trafficking [Clathrin heavy chain (chc) and synaptotagmin] survive longer during infection. However, oogenesis was rescued only in chc mutants. This suggests that viral response mechanisms that control fly survival and egg chamber survival are separable. The genetic and signaling requirements for oocyte destruction delineated here represent a novel host-virus interaction with implications for the control of both fly and virus populations. PMID:22173880

Thomson, Travis C; Schneemann, Anette; Johnson, Joshua

2012-06-01

42

Microinjection of Follicle-Enclosed Mouse Oocytes  

NASA Astrophysics Data System (ADS)

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.

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

43

Microinjection of follicle-enclosed mouse oocytes  

PubMed Central

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

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

2011-01-01

44

Vitamin A deprivation in hamsters  

Microsoft Academic Search

Summary  The effects of vitamin A deprivation on the tracheal epithelium of young hamsters were investigated. Colchicine was administered\\u000a 6 h prior to death to induce metaphase arrest, thus making it possible to quantify the mitotic rates of basal cells and secretory\\u000a (mucous) cells in the epithelium. Blood samples were taken from all hamsters, and liver samples from some, in order

Judy M. Strum; Patricia S. Latham; Martin L. Schmidt; Elizabeth M. McDowell

1986-01-01

45

Gynogenetic activation of porcine oocytes.  

PubMed

The possibility of fertilization without male contribution to the embryonic genome was investigated in pig oocytes. Mature oocytes were co-incubated with sperm, and in an attempt to prevent the incorporation of the sperm head into the ooplasm, the actin polymerization inhibitor cytochalasin B was added to the fertilization medium. We found that perturbing actin filament integrity did not affect the pattern of the sperm-induced Ca(2+) signal or the process of cortical granule exocytosis, and it did not alter the percentage of activated oocytes compared to the control (oocytes fertilized in the absence of the inhibitor). However, over 20% of the cytochalasin B-treated oocytes formed only a single pronucleus after fertilization, indicating that the inhibitor blocked sperm head incorporation at least in some oocytes. In most cases, cytochalasin B also prevented the integration of the male chromosomes into the embryonic genome as determined by the absence of the SRY gene in the embryonic blastomeres or by the frequency of embryos showing green fluorescence after sperm from a GFP-transgenic boar was used for fertilization. Finally, the percentage of embryos that developed beyond the four-cell stage and the total number of nuclei in the resultant blastocysts were higher when oocytes reconstructed by nuclear transfer were activated by fertilization in the presence of cytochalasin B compared to the control group, where activation was induced by electroporation. These results suggest that fertilization in the presence of cytochalasin B can induce oocyte activation while it also prevents integration of the male genome into the embryo. This method has the potential to be used as an alternative to inducing embryonic development after nuclear transfer. PMID:24661186

Lee, Kiho; Wang, Chunmin; Spate, Lee; Murphy, Clifton N; Prather, Randall S; Machaty, Zoltan

2014-04-01

46

Technical aspects of oocyte cryopreservation  

Microsoft Academic Search

Since the successful development in the mouse, the oocyte cryopreservation has been applied with varying success to a number of different species including the human. The recently reported successes in terms of pregnancies obtained by human oocyte cryopreservation are encouraging. Several studies typically reported different rates of survival (20–80%), fertilization (30–60%) and cleavage (32–100%). This variability of results throws some

R Fabbri; E Porcu; T Marsella; M. R Primavera; G Rocchetta; P. M Ciotti; O Magrini; R Seracchioli; S Venturoli; C Flamigni

2000-01-01

47

Effect of DMSO and DMBA hamster pouch carcinogenesis  

SciTech Connect

The penetration of mucosal surfaces by chemical carcinogens is required for tumor induction. The effectiveness of dimethyl sulfoxide (DMSO) as a carrier for carcinogen is controversial. The purpose of this study was to determine what effect DMSO would have on the 9,10-dimethyl- 1,2-benzanthracene (DMBA)-induced carcinogenesis in the hamster cheek pouch. Thirty Syrian golden hamsters were divided into two groups: the control group received a topical application of 0.5% DMBA in mineral oil three times per week for 16 weeks, while the experimental group received a topical application of DMSO previous to each DMBA application. At autopsy, both groups had developed tumors, the tumor ratio of control to experimental was 3.5:1.9 and the average size of tumors was 2.2 to 1.9 mm sq. The results suggest that DMSO interfered with the usual DMBA induction mechanism.

Rivera-Hidalgo, F.; Miller, E.G.; Binnie, W.H.

1987-01-01

48

Nanoliter droplet vitrification for oocyte cryopreservation  

PubMed Central

Aim Oocyte cryopreservation remains largely experimental, with live birth rates of only 2–4% per thawed oocyte. In this study, we present a nanoliter droplet technology for oocyte vitrification. Materials & methods An ejector-based droplet vitrification system was designed to continuously cryopreserve oocytes in nanoliter droplets. Oocyte survival rates, morphologies and parthenogenetic development after each vitrification step were assessed in comparison with fresh oocytes. Results Oocytes were retrieved after cryoprotectant agent loading/unloading, and nanoliter droplet encapsulation showed comparable survival rates to fresh oocytes after 24 h in culture. Also, oocytes recovered after vitrification/thawing showed similar morphologies to those of fresh oocytes. Additionally, the rate of oocyte parthenogenetic activation after nanoliter droplet encapsulation was comparable with that observed for fresh oocytes. This nanoliter droplet technology enables the vitrification of oocytes at higher cooling and warming rates using lower cryoprotectant agent levels (i.e., 1.4 M ethylene glycol, 1.1 M dimethyl sulfoxide and 1 M sucrose), thus making it a potential technology to improve oocyte cryopreservation outcomes. PMID:22188180

Zhang, Xiaohui; Khimji, Imran; Shao, Lei; Safaee, Hooman; Desai, Khanjan; Keles, Hasan Onur; Gurkan, Umut Atakan; Kayaalp, Emre; Nureddin, Aida; Anchan, Raymond M; Maas, Richard L; Demirci, Utkan

2011-01-01

49

Oocyte maturation: converting the zebrafish oocyte to the fertilizable egg.  

PubMed

The process of oogenesis culminates in steroid-induced oocyte maturation to produce the fertilizable egg. A quintessential biological entity, the egg is central to the production of new individuals. The result of egg fertilization by a sperm cell is the production of the mother of all stem cells (i.e. the zygote). Furthermore, the egg cytoplasm is the only one known to support reprogramming a transplanted nucleus to give rise to an individual (i.e. animal cloning). Zebrafish oocyte maturation is a complex event encompassing a number of cellular changes including germinal vesicle migration (GVM) and dissolution or breakdown (GVD), ooplasmic clearing (OC) with correlated yolk protein changes (YP), development of osmoregulation (OR) in fresh water, the formation of the future embryonic pole, the blastodisc (BF) and activatibility (AC) or cortical maturation. In zebrafish, and many other teleosts, 17alpha, 20beta-dihydroxy-4-pregnen-3-one (17alpha, 20beta-DP) has been shown to be the normal inducer of oocyte maturation. A 17alpha, 20beta-DP membrane-resident receptor mediates oocyte maturation via non-genomic mechanisms that are beginning to be understood. This paper will highlight some of the cellular markers resulting from the signaling initiated by 17alpha, 20beta-DP. By describing these markers, it is hoped that workers in the field will have additional tools to help further elucidate the signaling events of oocyte maturation. PMID:19027744

Lessman, Charles A

2009-03-01

50

Cryopreservation of Human Oocytes and Ovarian Tissue  

Microsoft Academic Search

Oocyte cryopreservation has the potential to be an important adjunct to assisted reproductive technologies and bypasses some\\u000a ethical, moral, and religious dilemmas posed by human embryo cryopreservation. The success of human oocyte cryopreservation\\u000a depends on morphological and biophysical factors that could influence oocyte survival after thawing. Among the morphological\\u000a factors, the maturity, quality, size of the oocyte, the presence or

Raffaella Fabbri

2006-01-01

51

Penetrating trauma.  

PubMed

Pneumothorax occurs when air enters the pleural space. Currently there is increasing incidence of road traffic accidents, increasing awareness of healthcare leading to more advanced diagnostic procedures, and increasing number of admissions in intensive care units are responsible for traumatic (non iatrogenic and iatrogenic) pneumothorax. Pneumothorax has a clinical spectrum from asymptomatic patient to life-threatening situations. Diagnosis is usually made by clinical examination and imaging techniques. In our current work we focus on the treatment of penetrating trauma. PMID:25337403

Kuhajda, Ivan; Zarogoulidis, Konstantinos; Kougioumtzi, Ioanna; Huang, Haidong; Li, Qiang; Dryllis, Georgios; Kioumis, Ioannis; Pitsiou, Georgia; Machairiotis, Nikolaos; Katsikogiannis, Nikolaos; Papaiwannou, Antonis; Lampaki, Sofia; Zaric, Bojan; Branislav, Perin; Dervelegas, Konstantinos; Porpodis, Konstantinos; Zarogoulidis, Paul

2014-10-01

52

Penetrating trauma  

PubMed Central

Pneumothorax occurs when air enters the pleural space. Currently there is increasing incidence of road traffic accidents, increasing awareness of healthcare leading to more advanced diagnostic procedures, and increasing number of admissions in intensive care units are responsible for traumatic (non iatrogenic and iatrogenic) pneumothorax. Pneumothorax has a clinical spectrum from asymptomatic patient to life-threatening situations. Diagnosis is usually made by clinical examination and imaging techniques. In our current work we focus on the treatment of penetrating trauma. PMID:25337403

Kuhajda, Ivan; Zarogoulidis, Konstantinos; Kougioumtzi, Ioanna; Huang, Haidong; Li, Qiang; Dryllis, Georgios; Kioumis, Ioannis; Pitsiou, Georgia; Machairiotis, Nikolaos; Katsikogiannis, Nikolaos; Papaiwannou, Antonis; Lampaki, Sofia; Zaric, Bojan; Branislav, Perin; Dervelegas, Konstantinos; Porpodis, Konstantinos

2014-01-01

53

Original article Chromosome analysis of horse oocytes  

E-print Network

Original article Chromosome analysis of horse oocytes cultured in vitro* WA King1 M Desjardins2 KP%) and fixed for chromosome analysis. To determine the time required for nuclear maturation, oocytes were fixed of culture and MII was reached by 24 h. To examine the chromosome features, an additional 113 oocyte- cumulus

Paris-Sud XI, Université de

54

Blastocyst development after intergeneric nuclear transfer of mountain bongo antelope somatic cells into bovine oocytes.  

PubMed

Intergeneric embryos were constructed by nuclear transfer using Mountain Bongo antelope somatic cells fused with enucleated bovine oocytes and their subsequent development in vitro was investigated. After two to six passages, starved or non-starved skin fibroblast cells were used as donor nuclei. In vitro matured bovine oocytes were enucleated by squeezing the first polar body and surrounding cytoplasm through a slit in the zona pellucida. After injection of a somatic cell into the perivitelline space, couplets were fused electrically and activated chemically, then subjected to different embryo culture treatments. Serum starvation had no effect on the frequency of cleavage to two cells or on development to the blastocyst stage in either sequential hamster embryo culture medium (HECM)-6/TCM-199 + serum or HECM-9/TC-199 + serum, or modified synthetic oviduct fluid (mSOF) culture medium. When couplets from non-starved donor nuclei were cultured, the frequency of cleavage (66 +/- 8% vs. 44 +/- 5%), development to >/=9 cells (46 +/- 6% vs. 24 +/- 4%), and formation of blastocysts (24 +/- 5% vs. 11 +/- 2%) were all significantly higher (p < 0.05) in the HECM-6 medium than in mSOF medium. In conclusion, bovine oocytes can support blastocyst development after intergeneric fusion with bongo fibroblasts. This technique could potentially be used as an alternative to using scarce bongo oocytes in attempts to propagate these endangered animals. PMID:12713698

Lee, Byeongchun; Wirtu, Gemechu G; Damiani, Philip; Pope, Earle; Dresser, Betsy L; Hwang, Woosuk; Bavister, Barry D

2003-01-01

55

Mouse oocytes fertilised by ICSI during in vitro maturation retain the ability to be activated after refertilisation in metaphase II and can generate Ca2+ oscillations  

PubMed Central

Background At fertilisation, mammalian oocytes are activated by oscillations of intracellular Ca2+ ([Ca2+]i). Phospholipase C?, which is introduced by fertilising spermatozoon, triggers [Ca2+]i oscillations through the generation of inositol 1,4,5-triphosphate (IP3), which causes Ca2+ release by binding to IP3 receptors located on the endoplasmic reticulum (ER) of the oocyte. Ability to respond to this activating stimulus develops during meiotic maturation of the oocyte. Here we examine how the development of this ability is perturbed when a single spermatozoon is introduced into the oocyte prematurely, i.e. during oocyte maturation. Results Mouse oocytes during maturation in vitro were fertilised by ICSI (intracytoplasmic sperm injection) 1 – 4 h after germinal vesicle break-down (GVBD) and were subsequently cultured until they reached metaphase II (MII) stage. At MII stage they were fertilised in vitro for the second time (refertilisation). We observed that refertilised oocytes underwent activation with similar frequency as control oocytes, which also went through maturation in vitro, but were fertilised only once at MII stage (87% and 93%, respectively). Refertilised MII oocytes were able to develop [Ca2+]i oscillations in response to penetration by spermatozoa. We found however, that they generated a lower number of transients than control oocytes. We also showed that the oocytes, which were fertilised during maturation had a similar level of MPF activity as control oocytes, which were not subjected to ICSI during maturation, but had reduced level of IP3 receptors. Conclusion Mouse oocytes, which were experimentally fertilised during maturation retain the ability to generate repetitive [Ca2+]i transients, and to be activated after completion of maturation. PMID:17584490

J?drusik, Agnieszka; Ajduk, Anna; Pomorski, Pawe?; Maleszewski, Marek

2007-01-01

56

Embryological, clinical and ultrastructural study of human oocytes presenting indented zona pellucida.  

PubMed

Human oocyte dysmorphisms attain a large proportion of retrieved oocytes from assisted reproductive technology (ART) treatment cycles. Extracytoplasmic defects involve abnormal morphology of the zona pellucida (ZP), perivitelline space and first polar body. The aim of the present study was to describe a novel dysmorphism affecting the ZP, indented ZP. We also evaluated the clinical, embryological and ultrastructural features of these cases. We evaluated all ART treatment cycles during 7 consecutive years and found 13 treatment cycles (six patients) with all oocytes presenting an indented ZP. In addition, these oocytes presented total or partial absence of the perivitelline space, absence of resistance to ZP and oolemma penetration during microinjection, and low ooplasm viscosity during aspiration. This novel described dysmorphism was recurrent and attained all oocytes in three cases that had more than one treatment cycle. When compared with controls, data showed significant low oocyte maturity (42% versus 81.6%) and high cycle cancellation (30.8% versus 8.5%) rates, normal degeneration (3.4% versus 6.3%) and fertilization rates (69% versus 69.5%), and low pregnancy (15.4% versus 33.3%) and live-birth delivery (7.7% versus 27.7%) rates per cycle. Ultrastructure analysis revealed a zona pellucida structure with large empty electrolucent regions, an outer ZP layer with an indented surface with protuberances and a thick inner ZP that obliterated the perivitelline space. There was evidence of exocytosis of ZP material by the oocyte. In conclusion, oocytes with this novel described dysmorphism (indented ZP) are associated with low maturity, pregnancy and live-birth delivery rates. PMID:23992046

Sousa, M; Teixeira da Silva, J; Silva, J; Cunha, M; Viana, P; Oliveira, E; Sá, R; Soares, C; Oliveira, C; Barros, A

2015-02-01

57

Recent Progress in Cryopreservation of Bovine Oocytes  

PubMed Central

Principle of oocyte cryoinjury is first overviewed and then research history of cryopreservation using bovine oocytes is summarized for the last two decades with a few special references to recent progresses. Various types of cryodevices have been developed to accelerate the cooling rate and applied to the oocytes from large domestic species enriched with cytoplasmic lipid droplets. Two recent approaches include the qualitative improvement of IVM oocytes prior to the vitrification and the short-term recovery culture of vitrified-warmed oocytes prior to the subsequent IVF. Supplementation of L-carnitine to IVM medium of bovine oocytes has been reported to reduce the amount of cytoplasmic lipid droplets and improve the cryotolerance of the oocytes, but it is still controversial whether the positive effect of L-carnitine is reproducible. Incidence of multiple aster formation, a possible cause for low developmental potential of vitrified-warmed bovine oocytes, was inhibited by a short-term culture of the postwarm oocytes in the presence of Rho-associated coiled-coil kinase (ROCK) inhibitor. Use of an antioxidant ?-tocopherol, instead of the ROCK inhibitor, also supported the revivability of the postwarm bovine oocytes. Further improvements of the vitrification procedure, combined with pre- and postvitrification chemical treatment, would overcome the high sensitivity of bovine oocytes to cryopreservation. PMID:24738063

Hochi, Shinichi

2014-01-01

58

Mammalian oocyte growth and development in vitro.  

PubMed

This paper is a review of the current status of technology for mammalian oocyte growth and development in vitro. It compares and contrasts the characteristics of the various culture systems that have been devised for the culture of either isolated preantral follicles or the oocyte-granulosa cell complexes form preantral follicles. The advantages and disadvantages of these various systems are discussed. Endpoints for the evaluation of oocyte development in vitro, including oocyte maturation and embryogenesis, are described. Considerations for the improvement of the culture systems are also presented. These include discussions of the possible effects of apoptosis and inappropriate differentiation of oocyte-associated granulosa cells on oocyte development. Finally, the potential applications of the technology for oocyte growth and development in vitro are discussed. For example, studies of oocyte development in vitro could help to identify specific molecules produced during oocyte development that are essential for normal early embryogenesis and perhaps recognize defects leading to infertility or abnormalities in embryonic development. Moreover, the culture systems may provide the methods necessary to enlarge the populations of valuable agricultural, pharmaceutical product-producing, and endangered animals, and to rescue the oocytes of women about to undergo clinical procedures that place oocytes at risk. PMID:9115726

Eppig, J J; O'Brien, M; Wigglesworth, K

1996-06-01

59

Sperm capacitation in the domestic cat (Felis catus) and leopard cat (Felis bengalensis) as studied with a salt-stored zona pellucida penetration assay  

Microsoft Academic Search

The ability of domestic cat or leop- ard cat spermatozoa to penetrate zonae pellucidae (ZP) of salt-stored, domestic cat oocytes was examined as an assay for sperm capacitation. Ovarian oocytes were recov- ered after ovariectomy and matured in vitro for 18-36 h. Following removal of cumulus cells, the oocytes were used fresh, or stored (4°C, 0.5-24 weeks) in a HEPES-buffered

J. C. Andrews; J. G. Howard; B. D. Bavister; D. E. Wildt

1992-01-01

60

How is plasminogen/plasmin system contributing to regulate sperm entry into the oocyte?  

PubMed

Plasminogen is present in the oviduct, on the zona pellucida (ZP) and on oolemma, and reduces the number of sperm penetrating the oocyte during in vitro fertilization in pig and cow. It is unknown how this reduction occurs. We tested whether plasminogen (1) changed the ZP resistance to enzymatic digestion thus making the passage of the spermatozoa across it difficult; (2) reduced the sperm functionality, assessed by sperm viability, motility, spontaneous acrosome reaction and membrane lipid disorder; or (3) affected the sperm-ZP binding before or after sperm-ZP interaction. The mechanism by which plasminogen/plasmin system contributes to regulate sperm entry into the oocyte is not inducing a ZP hardening or a decrease in sperm functionality but detaching more than 50% of sperm bound to the ZP. It is suggested that the fertilizing spermatozoon activates plasminogen into plasmin at the oocyte surface and that plasmin removes additional spermatozoa attached to the ZP. PMID:23420828

Grullón, Luis A; Gadea, Joaquín; Mondéjar, Irene; Matás, Carmen; Romar, Raquel; Coy, Pilar

2013-09-01

61

[Controversy in ART: should we cryopreserve oocytes or embryos? Do prefer oocytes].  

PubMed

Since the beginning of IVF, cryopreservation concern spermatozoa or embryos due to the poor efficiency of oocyte freezing. To date, oocyte vitrification allows changing our practice privileging female gamete vitrification instead of human embryo freezing. PMID:25153440

Boyer, P

2014-09-01

62

The oocyte-to-embryo transition : regulation of oocyte maturation and egg activation in Drosophila  

E-print Network

In oogenesis, meiosis must be highly regulated to ensure that growth of the oocyte and chromosomal segregation are coordinated properly. To do this, meiosis arrests at two points to permit oocyte differentiation and ...

Weingarten, Lisa Suzanne

2013-01-01

63

Comparison of in-vitro outcomes from cryopreserved oocytes and sibling fresh oocytes  

Microsoft Academic Search

In Italy, the restrictive IVF law generalizes the indication for oocyte freezing for surplus oocytes in 78.5% of in-vitro assisted reproductive cycles. With a view to understanding better what the prospects for intracytoplasmic sperm injection (ICSI) on frozen–thawed oocytes might be, the consequences of freeze–thaw procedures on fertilization, cleavage rates and embryo quality obtained from frozen–thawed oocytes were studied and

S Chamayou; C Alecci; C Ragolia; G Storaci; E Maglia; E Russo; A Guglielmino

2006-01-01

64

Effects of (-)-epigallocatechin gallate on the motility and penetrability of frozen-thawed boar spermatozoa incubated in the fertilization medium.  

PubMed

Epigallocatechin gallate (EGCG) is the major polyphenol in green tea (Camellia sinensis) and is known for its antioxidant effects. The objective of the present study was to examine the effects of EGCG during in vitro fertilization (IVF) on the sperm quality and penetrability into oocytes. In the first experiment, the effects of concentration and incubation period of EGCG on the motility and penetrability of spermatozoa were examined. When frozen-thawed spermatozoa were incubated in IVF medium supplemented with 0 (control), 1, 50 and 100 ?m EGCG for 1, 3 and 5 h, supplementation with 50 and 100 ?m EGCG improved motility of the spermatozoa (p < 0.05), but not viability, as compared with the control group. When frozen-thawed spermatozoa were co-incubated with in vitro-matured (IVM) oocytes in IVF medium supplemented with 50 and 100 ?m EGCG for 5 h, supplementation of EGCG had positive effects on sperm penetration rates. In the second experiment, the effects of supplementation of EGCG in IVF medium on penetrability of sperm from different boars and development of fertilized oocytes were evaluated. When frozen-thawed spermatozoa from six boars were co-incubated with IVM oocytes in IVF medium supplemented with 50 ?m EGCG, the effect of EGCG on sperm penetration and development of oocytes after fertilization was found to vary with individual boar. Our results indicate that motility and penetrability of boar spermatozoa are improved by co-incubation with 50 ?m EGCG, but the effects vary with individual boars. PMID:22299777

Kaedei, Y; Naito, M; Naoi, H; Sato, Y; Taniguchi, M; Tanihara, F; Kikuchi, K; Nagai, T; Otoi, T

2012-12-01

65

Fourier analysis of mitochondrial distribution in oocytes  

NASA Astrophysics Data System (ADS)

This paper describes a novel approach to quantifying mitochondrial patterns which are typically described using the qualitative terms "diffuse" "aggregated" and are potentially key indicators for an oocyte's health and survival potential post-implantation. An oocyte was isolated in a confocal image and a coarse grid was superimposed upon it. The spatial spectrum was calculated and an aggregation factor was generated. A classifier for healthy cells was developed and verified. The aggregation factor showed a clear distinction between the healthy and unhealthy oocytes. The ultimate goal is to screen oocytes for viability preimplantation, thus improving the outcome of in vitro fertilization (IVF) treatments.

Hollmann, Joseph L.; Brooks, Dana H.; Newmark, Judith A.; Warner, Carol M.; DiMarzio, Charles A.

2011-03-01

66

Stringent regulation of oocyte donation in China.  

PubMed

Currently in China, health regulations permit oocyte donation only from IVF/ICSI patients who have 20 or more mature oocytes retrieved from a single cycle, of which at least 15 must be kept for their own treatment. Oocyte donation from non-patients and commercial transaction of human gametes are strictly prohibited by law. Additionally, embryos derived from donated oocytes must be cryopreserved and cannot be transferred to prospective recipients, until donors have been screened to be free of communicable diseases after 6 months. Such overly stringent regulation has in turn led to a severe shortage of available donor oocytes in China. The situation is made worse by a cultural aversion to oocyte donation by the majority of patients, because biological kinship and blood relations are viewed as sacrosanct in traditional Chinese culture. The harsh social stigmatization of childlessness in Chinese society, increasing incidence of age-related female infertility in recent years and growing numbers of bereaved older women who have lost their only child to accidents, natural disasters and suicides would make it imperative to reconsider liberalizing the regulation of oocyte donation in China. In particular, the blanket ban on oocyte donation by non-patients should be lifted, as it is anticipated that there are many young healthy women in China who are generous and open-minded enough to consider altruistically donating their oocytes to childless couples. PMID:18854406

Heng, Boon Chin

2009-01-01

67

Directed Student Inquiry: Modeling in Roborovsky Hamsters  

ERIC Educational Resources Information Center

In this inquiry-based activity, Roborovsky hamsters are used to provide students with an opportunity to develop their skills of analysis, inquiry, and design. These hamsters are easy to maintain, yet offer students a means to use conventional techniques and those of their own design to make further observations through measuring, assessing, and…

Elwess, Nancy L.; Bouchard, Adam

2007-01-01

68

FATE OF INHALED FLY ASH IN HAMSTERS  

EPA Science Inventory

To determine pulmonary deposition, translocation, and clearance of inhaled fly ash, hamsters received a single 95-min nose-only exposure to neutron-activated fly ash. Over a period of 99 days postexposure, the hamsters were sacrificed in groups of six animals. Lungs, liver, kidne...

69

Mammalian oocyte growth and development in vitro  

Microsoft Academic Search

This paper is a review of the current status of technology for mammalian oocyte growth and development in vitro. It compares and contrasts the characteristics of the various culture systems that have been devised for the culture of either isolated preantral follicles or the oocyte-granulosa cell complexes form preantral follicles. The advantages and disadvantages of these various systems are discussed.

John J. Eppig; M OBrien; Karen Wigglesworth

1996-01-01

70

Apoptosis in Batch Cultures of Chinese Hamster Ovary Cells  

E-print Network

Apoptosis in Batch Cultures of Chinese Hamster Ovary Cells J. Goswami,1 A. J. Sinskey,2 H. Steller of the main problems in the culture of Chinese Hamster Ovary (CHO) cells continues to be the inability. Keywords: cell culture; Chinese Hamster Ovary; apopto- sis; caspase; bcl-2 INTRODUCTION Chinese Hamster

Sinskey, Anthony J.

71

Activation of oocytes after nuclear transfer.  

PubMed

After nuclear transfer, the recipient oocyte must be stimulated to initiate development. This stimulation is achieved by inducing changes in the oocyte cytoplasm that normally are triggered by the sperm during fertilization. In most cases, such changes include a transient increase in the intracellular-free calcium concentration induced by an electrical pulse or alternatively, by chemical agents. Many times, particularly in aged oocytes, this calcium signal is sufficient to stimulate the oocyte developmental program. Other activation protocols were designed to target pathways downstream of the initial calcium signal to affect the activity of regulatory proteins that play central roles in maintaining developmental arrest. This is achieved by the application of protein kinase or protein synthesis inhibitors; combined with a calcium stimulus such inhibitors are widely used for oocyte activation after nuclear transfer and are able to support embryonic development to term. PMID:16988371

Macháty, Zoltán

2006-01-01

72

Improvement in bovine embryo production in vitro by treatment with green tea polyphenols during in vitro maturation of oocytes.  

PubMed

The present study examined the effect of green tea polyphenols (GTP) during in vitro maturation (IVM) of bovine oocytes on in vitro fertilization (IVF) parameters, intracellular glutathione (GSH) concentration and subsequent embryo development. Cumulus-oocyte complexes were aspirated from the ovaries derived from slaughterhouse and cultured in modified synthetic oviduct fluid (m-SOF) supplemented with 0-25 microM GTP for 24h. After IVM, cumulus-free oocytes were coincubated with frozen-thawed spermatozoa for 15-18 h. Putative embryos were transferred to m-SOF and cultured for 8 days (Experiment 1). In comparison with the absence of GTP, treatment with GTP at a concentration of 15 microM showed a significant increase in the proportion of pronuclear (PN) formation after sperm penetration (65% versus 80%, P<0.05). No significant differences in the rates of sperm penetration and polyspermic fertilization were found among treatments. The cleavage rate at 48 h of in vitro insemination showed no difference in oocytes matured with or without GTP. However, compared to no addition (23.5%), the presence of 15 and 20 microM GTP during IVM significantly (P<0.05) increased the proportion of blastocysts (38.1% and 36.4%) on day 9 of in vitro insemination. A further increase from 20 to 25 microM GTP reduced (P<0.05) the proportion of blastocysts. In Experiment 2, after IVM, oocytes were fixed to analyze the GSH concentration. Compared to no addition, a higher (P<0.05) level of GSH was found in oocytes matured with 15 microM GTP and compared with 15 microM GTP, GSH was low (P<0.05) at 20 and 25 microM GTP. The results suggest that at certain concentrations of GTP (15 microM) in IVM medium has beneficial effects on subsequent embryo development, and is correlated with intracellular GSH level in bovine oocytes. PMID:16870363

Wang, Zheng-guang; Yu, Song-dong; Xu, Zi-rong

2007-07-01

73

Cryopreservation of human oocytes and ovarian tissue.  

PubMed

Oocyte cryopreservation has the potential to be an important adjunct to assisted reproductive technologies and bypasses some ethical, moral, and religious dilemmas posed by human embryo cryopreservation. The success of human oocyte cryopreservation depends on morphological and biophysical factors that could influence oocyte survival after thawing. Among the morphological factors, the maturity, quality, size of the oocyte, the presence or the absence of the cumulus oophorus seems to play an important role in oocyte survival after thawing. The main biophysical factor of cellular disruption during cryopreservation process in the intracellular ice formation that can be avoided by an adequate cell dehydration; thus reducing the intracellular water by increasing the dehydration process we can limit the damages of the cryopreservation procedure. The dehydration process can be affected by the presence and concentration of the cryoprotectants in the freezing solutions (equilibration and loading solutions), and by the freezing and thawing rate. Two additional properties of cryoprotectants help to protect cells during slow cooling, when the cells are very dehydrated and are surrounded by concentrated salts. The cryoprotectants appear to reduce damage caused by high levels of salt, a property known as salt buffering. Some events occurring to the oocyte during cryopreservation procedure has been found to be a premature exocitosis of cortical granules, leading to an intempestive zona hardening and consequently to a reduction of fertilization rate, and the cryoinjury to the zona pellucida leading to a polispermic fertilization. ICSI is an efficient method to by pass these two events and to achieve a satisfactory outcome in terms of normal fertilization of cryopreserved oocytes. The application of the ICSI to cryopreserved oocytes did not seem to increase the degeneration rate after insemination with respect to fresh oocytes. The increased oocyte survival rate and the use of ICSI have facilitated the recent increase in the number of pregnancies and live birth. PMID:16732414

Fabbri, Raffaella

2006-01-01

74

Oocyte cryopreservation: advances and drawbacks.  

PubMed

The field of oocyte cryopreservation (OC) had advanced dramatically since the first reported birth from cryopreserved oocytes in 1986, with a significant increase in pregnancy rates described over the past 5 years due to improvements in vitrification technology, a cryopreservation method which virtually means to achieve a "glass-like" state through avoidance of ice formation. The potential clinical benefits of achieving efficient OC protocols have long been recognized. Specifically, OC can be offered to women who face fertility-threatening situations such as therapy for cancer or rheumatologic disease, premature ovarian insufficiency, or need for ovarian surgery as a measure to preserve fertility. Moreover, many women who plan to delay childbearing are interested in pursuing OC in order to protect against age-related fertility decline. For infertility practices, efficient OC technology stands to dramatically streamline donor egg programs, and is a helpful adjuvant in situations where sperm is unexpectedly unavailable at the time of egg retrieval and for couples who do not wish to cryopreserve supernumerary embryos created from in vitro fertilization for moral / ethical reasons. This review will describe the history of OC technology over the past three decades, discuss clinical circumstances for its implementation, and address areas where more research is needed. Given the remarkable improvements in pregnancy rates witnessed over the past five years, OC is certain to play a much larger role in reproductive medicine over the coming decades. PMID:23232533

Dovey, S

2012-12-01

75

MULTIDRUG RESISTANT TRANSPORT ACTIVITY PROTECTS OOCYTES FROM CHEMOTHERAPEUTIC AGENTS AND CHANGES DURING OOCYTE MATURATION  

PubMed Central

Objective To determine the multidrug resistant (MDR) transporter activity in oocytes and their potential role in oocyte susceptibility to chemotherapy. Design Experimental laboratory study Setting University and Academic Center for reproductive medicine. Patients/Animals Women with eggs retrieved for ICSI cycles and adult female FVBN and B6C3F1 mouse strains. Intervention Inhibition of MDR activity in oocytes. Main Outcome measure(s) Efflux activity of MDRs using quantitative fluorescent dye efflux and oocyte cell death when exposed to chemotherapy. Results Oocytes effluxed fluorescent reporters and this activity was significantly reduced in the presence of the MDR inhibitor PSC 833. GV oocytes are more efficient at efflux compared to M2 oocytes. Human oocytes exposed to cyclophosphamide and PSC 833 showed cell death using two different viability assays compared to controls and those exposed to cyclophosphamide alone. Immunoblots detected MDR-1 in all oocytes with the greatest accumulation in the GV stage. Conclusions Oocytes have a vast repertoire of active MDRs. The implications of this study are that these protective mechanisms are important during oogenesis, and these activities change with maturation, increasing susceptibility to toxicants. Future directions may exploit the up regulation of these transporters during gonadotoxic therapy. PMID:23953328

Brayboy, Lynae M.; Oulhen, Nathalie; Witmyer, Jeannine; Robins, Jared; Carson, Sandra; Wessel, Gary M.

2013-01-01

76

Maturation arrest of human oocytes at germinal vesicle stage.  

PubMed

Maturation arrest of human oocytes may occur at various stages of the cell cycle. A total failure of human oocytes to complete meiosis is rarely observed during assisted conception cycles. We describe here a case of infertile couples for whom all oocytes repeatedly failed to mature at germinal vesicle (GV) stage during in vitro fertilization/Intra cytoplasmic sperm injection (IVF/ICSI). The patient underwent controlled ovarian stimulation followed by oocyte retrieval and IVF/ICSI. The oocytes were stripped off cumulus cells prior to the ICSI procedure and their maturity status was defined. The oocyte maturation was repeatedly arrested at the GV. Oocyte maturation arrest may be the cause of infertility in this couple. The recognition of oocyte maturation arrest as a specific medical condition may contribute to the characterization of the currently known as "oocyte factor." The cellular and genetic mechanisms causing oocyte maturation arrest should be the subject for further investigation. PMID:21234179

Chen, Zhi Qin; Ming, Teng Xiao; Nielsen, Hans Ingolf

2010-09-01

77

The DNA damage response in mammalian oocytes  

PubMed Central

DNA damage is one of the most common insults that challenge all cells. To cope, an elaborate molecular and cellular response has evolved to sense, respond to and correct the damage. This allows the maintenance of DNA fidelity essential for normal cell viability and the prevention of genomic instability that can lead to tumor formation. In the context of oocytes, the impact of DNA damage is not one of tumor formation but of the maintenance of fertility. Mammalian oocytes are particularly vulnerable to DNA damage because physiologically they may lie dormant in the ovary for many years (>40 in humans) until they receive the stimulus to grow and acquire the competence to become fertilized. The implication of this is that in some organisms, such as humans, oocytes face the danger of cumulative genetic damage for decades. Thus, the ability to detect and repair DNA damage is essential to maintain the supply of oocytes necessary for reproduction. Therefore, failure to confront DNA damage in oocytes could cause serious anomalies in the embryo that may be propagated in the form of mutations to the next generation allowing the appearance of hereditary disease. Despite the potential impact of DNA damage on reproductive capacity and genetic fidelity of embryos, the mechanisms available to the oocyte for monitoring and repairing such insults have remained largely unexplored until recently. Here, we review the different aspects of the response to DNA damage in mammalian oocytes. Specifically, we address the oocyte DNA damage response from embryonic life to adulthood and throughout oocyte development. PMID:23805152

Carroll, John; Marangos, Petros

2013-01-01

78

Halide transport in Xenopus oocytes.  

PubMed Central

1. Radioisotopes and intracellular microelectrodes were used to characterize the permeability of Xenopus oocytes to chloride and other halides. 2. Uptake of 36Cl had a half-time for equilibration of approximately 3 h, with an initial rate of Cl- entry corresponding to a permeability coefficient of 3.9 x 10(-7) cm/s, and an equilibrium uptake of 36Cl of 33 mM. 3. Replacement of bathing Na+ by K+ depolarized the oocytes from -46 to -7 mV and stimulated influx approximately 3-fold. 4. Influx was linearly dependent on bathing [Cl-] and was temperature dependent with an activation energy of 46 kJ/mol. Influx of 125I of 36Cl was not affected by the presence of equal concentrations of other halides or thiocyanate. These results are consistent with a channel-mediated entry mechanism. 5. Diphenylamine-2-carboxylate (DPAC) and 9-anthracene carboxylate (9-AC), blockers of Cl- channels in other cells, inhibited Cl- entry with dissociation constants (Kds) of approximately 5 x 10(-4) and approximately 10(-3) M, respectively. Inhibitors of Cl(-)-HCO3- exchange or Na(+)-K(+)-2Cl- co-transport did not affect Cl- influx. 6. Attempts to lower or raise intracellular Ca2+ with BAPTA or A23187, respectively, were also without effect on Cl- influx. 7. The halide selectivity sequence determined with isotopes was I- (3.2) greater than Br- (1.3) greater than Cl- (1.0). However, DPAC inhibited almost all of the 36Cl influx but only a small fraction of 125I influx. 8. Replacement of bathing Cl- by I- or Br-resulted in hyperpolarizations, from which the same selectivity sequence was determined. 9. Replacement of bathing Cl- by gluconate caused a marked depolarization, which was inhibited by DPAC and, less potently, by 9-AC. Images Fig. 9 PMID:1822540

Katayama, Y; Widdicombe, J H

1991-01-01

79

Spontaneous endomyometrial neoplasms in aging Chinese hamsters  

SciTech Connect

Twenty-one endomyometrial neoplasms among 93 nulliparous noninbred Chinese hamsters were evaluated. The median survival time of the 93 females was 1040 days. The median age of hamsters with endomyometrial neoplasms was 1200 days. Neoplasms were classified as carcinomas or malignant mixed muellerian tumors of the endometrium and benign or malignant myometrial neoplasms. There were 13 endometrial adenocarcinomas. Three tumors were mixed adenosquamous carcinomas, which occurred in significantly older Chinese hamsters than did adenocarcinomas. Three malignant mixed muellerian tumors consisted of 2 carcinosarcomas and 1 mixed mesodermal tumor. The 2 myometrial neoplasms were a lelomyoma and a lelomyosarcoma. The classification and relative frequency of these neoplasms were similar to endomyometrial neoplasms of women, which makes Chinese hamsters useful subjects for studies of spontaneous endomyometrial cancers.

Brownstein, D.G.; Brooks, A.L.

1980-05-01

80

Induction of lyme arthritis in LSH hamsters  

SciTech Connect

In studies of experimental Lyme disease, a major obstacle has been the unavailability of a suitable animal model. We found that irradiated LSH/Ss Lak hamsters developed arthritis after injection of Borrelia burgdorferi in the hind paws. When nonirradiated hamsters were injected in the hind paws with B. burgdorferi, acute transient synovitis was present. A diffuse neutrophilic infiltrate involved the synovia and periarticular structures. The inflammation was associated with edema, hyperemia, and granulation tissue. Numerous spirochetes were seen in the synovial and subsynovial tissues. The histopathologic changes were enhanced in irradiated hamsters. The onset and duration of the induced swelling were dependent on the dose of radiation and the inoculum of spirochetes. Inoculation of irradiated hamsters with Formalin-killed spirochetes or medium in which B. burgdorferi had grown for 7 days failed to induce swelling. This animal model should prove useful for studies of the immune response to B. burgdorferi and the pathogenesis of Lyme arthritis.

Schmitz, J.L.; Schell, R.F.; Hejka, A.; England, D.M.; Konick, L.

1988-09-01

81

Directed Student Inquiry: Modeling in Roborovsky Hamsters  

NSDL National Science Digital Library

In this inquiry-based activity, Roborovsky hamsters are used to provide students with an opportunity to develop their skills of analysis, inquiry, and design. These hamsters are easy to maintain, yet offer students a means to use conventional techniques and those of their own design to make further observations through measuring, assessing, and data collection. Based on the premise that this is a directed rather than dictated student inquiry, the activity will vary based on discussions and recommendations suggested by each class.

Adam Bouchard

2007-04-01

82

Mouse oocyte, a paradigm of cancer cell  

PubMed Central

Oocytes undergo extremely asymmetric divisions in terms of size. Coordinating spindle assembly and positioning in the absence of canonical centrosomes appears to be a challenge for oocytes, which divide with an elevated rate of errors in chromosome segregation. Here we highlight recent work on the characteristics of oocyte meiotic divisions, giving special emphasis on MTOCs clustering, generation of aneuploidy, and cortex softening, properties shared by cancer cells. While the loss of canonical centrosomes in oocytes might favor the asymmetry in size of meiotic divisions by reducing the distance between spindle poles and the cortex, we propose that this acentrosomal pathway might also render meiotic spindles less robust and, so, be responsible for the high error rate of female meiosis. PMID:24091531

Terret, Marie-Emilie; Chaigne, Agathe; Verlhac, Marie-Hélène

2013-01-01

83

Quantitative Microinjection of Mouse Oocytes and Eggs  

NASA Astrophysics Data System (ADS)

Quantitative microinjection is used to introduce known quantities of molecules or probes into single cells to examine cellular function. The relatively large mammalian oocyte or egg is easily manipulated and can be injected with impermeant reagents including a variety of signaling molecules and fluorescent probes. Techniques have been developed to inject picoliter quantities of solution into oocytes and eggs with precision and reliability. The methods described here outline the quantitative injection procedures as they are used to inject mouse oocytes and eggs in a culture dish on the stage on an inverted microscope. The techniques are applicable to the oocytes, eggs, and early embryos of most mammalian species. Included are some general instructions on fabrication of transfer pipettes, holding pipettes, beveled injection pipettes, and equipment for quantitative injection.

Kline, Douglas

84

Repetitive oocyte donation: a committee opinion.  

PubMed

This Committee Opinion concludes that donors be advised of the number of cycles/donations that a given oocyte donor may undergo. Although existing data cannot permit conclusive recommendations, a concern for the issues of the safety and well-being of oocyte donors warrants consideration. This document replaces the document of the same name, previously published in 2008 (Fertil Steril 2008; 90:S194-5). PMID:25064399

2014-10-01

85

Verification of buffalo ( Bubalus bubalisi) oocytes  

Microsoft Academic Search

The objective of the present study was to develop a method for the cryopreservation of buffalo oocytes by vitrification. Cumulus-oocyte complexes (COCs) were obtained from slaughterhouse ovaries. Prior to vitrification of COCs in the vitrification solution (VS) consisting of 4.5 M ethylene glycol, 3.4 M dimethyl sulfoxide, 5.56 mM glucose, 0.33 mM sodium pyruvate and 0.4% w\\/v bovine serum albumin

A. Dhali; R. S. Manik; S. K. Das; S. K. Singla; P. Palta

2000-01-01

86

Mitochondrial functions on oocytes and preimplantation embryos  

Microsoft Academic Search

Oocyte quality has long been considered as a main limiting factor for in vitro fertilization (IVF). In the past decade, extensive\\u000a observations demonstrated that the mitochondrion plays a vital role in the oocyte cytoplasm, for it can provide adenosine\\u000a triphosphate (ATP) for fertilization and preimplantation embryo development and also act as stores of intracellular calcium\\u000a and proapoptotic factors. During the

Li-ya Wang; Da-hui Wang; Xiang-yang Zou; Chen-ming Xu

2009-01-01

87

Use of sugars in cryopreserving human oocytes.  

PubMed

In the last 20 years, a worldwide effort to cryopreserve oocytes has resulted in 40 infants and approximately 50 ongoing pregnancies being reported. While the ability to freeze human embryos has become a standard of practice in assisted reproductive technologies, obtaining reliable techniques for oocyte cryopreservation has been more difficult. The unique properties of the mature oocyte, such as the meiotic stage with sensitive spindle structure as well as the large cell volume, are responsible for the limited success obtained to date. There have been two approaches to cryopreserving the oocyte: (i) slow freeze-rapid thaw, and (ii) vitrification protocols with rapid cooling-rapid warming. Both methods have incorporated sugars (sucrose) as a beneficial non-permeating extracellular cryoprotectant. Studies of organisms that survive extreme conditions of freezing/dehydration have demonstrated the ability to accumulate intracellular sugars to afford protection and survival. A novel technique using microinjection of sugars into the oocyte for cryopreservation has been developed as an alternative approach to external addition of sugars. Freezing the human oocyte has been a challenging goal; however, developing research and efforts will, in the near future, provide women with an important option for their reproductive health. PMID:15333248

Wright, Diane L; Eroglu, Ali; Toner, Mehmet; Toth, Thomas L

2004-08-01

88

Glycolytic Metabolites Are Critical Modulators of Oocyte Maturation and Viability  

PubMed Central

The maturation of an oocyte into an egg is a key step in preparation for fertilization. In Xenopus, oocyte maturation is independent of transcription, being regulated at the level of translation and post-translational modifications of proteins. To identify factors involved in the maturation process we used two-dimensional differential gel electrophoresis to compare the proteome of oocytes and eggs. Protein abundance changes were observed in multiple cellular pathways during oocyte maturation. Most prominent was a general reduction in abundance of enzymes in the glycolytic pathway. Injection into oocytes of the glycolytic intermediates glyceraldehyde-3-phosphate, phosphoenolpyruvate and glucose-6-phosphate prevented oocyte maturation. Instead, these metabolites stimulated ROS production and subsequent apoptosis of the oocyte. In contrast, all other metabolites tested had no effect on oocyte maturation and did not induce apoptosis. These data suggest that a subset of glycolytic metabolites have the capacity to regulate oocyte viability. PMID:24167578

Berger, Lloyd; Wilde, Andrew

2013-01-01

89

The insemination of goldfish ( Carassium auratus) oocyte matured in vitro  

NASA Astrophysics Data System (ADS)

Full maturation of goldfish oocyte was induced in vitro by 17 ?-hydroxy-20?-dihydroprogesterone. The oocyte maturation involves GV migration to the periphery of the oocyte and germinal vesicle breakdown (GVBD). In the experiment, incubation duration for GVBD varied in different broods of oocytes. Generally, if the duration for GVBD was shorter than 6 h, oocytes would have a better chance to survive after maturation and insemination. The maturation of nucleus (GV) and cytoplasm are not synchronous. Cytoplasm maturation occurs several hs after GVBD. Oocytes inseminated 8 9 h after GVBD have the highest fertilizing and hatching rate. Fertilized ova matured in vitro can develop to sexually mature adults capable of reproduction.

Wang, Renxue; Wu, Xianhan; Zhou, Jing; Zhang, Shicui; Ma, Yingjie; Wu, Shangqin; Shi, Yingxian

1991-03-01

90

A Miniature Probe for Ultrasonic Penetration of a Single Cell  

PubMed Central

Although ultrasound cavitation must be avoided for safe diagnostic applications, the ability of ultrasound to disrupt cell membranes has taken on increasing significance as a method to facilitate drug and gene delivery. A new ultrasonic resonance driving method is introduced to penetrate rigid wall plant cells or oocytes with springy cell membranes. When a reasonable design is created, ultrasound can gather energy and increase the amplitude factor. Ultrasonic penetration enables exogenous materials to enter cells without damaging them by utilizing instant acceleration. This paper seeks to develop a miniature ultrasonic probe experiment system for cell penetration. A miniature ultrasonic probe is designed and optimized using the Precise Four Terminal Network Method and Finite Element Method (FEM) and an ultrasonic generator to drive the probe is designed. The system was able to successfully puncture a single fish cell. PMID:22412314

Wu, Ting; Zhou, Zhaoying; Wang, Qun; Yang, Xing; Xiao, Mingfei

2009-01-01

91

Proteomes of Animal Oocytes: What Can We Learn for Human Oocytes in the In Vitro Fertilization Programme?  

PubMed Central

Oocytes are crucial cells for mammalian reproduction, yet the molecular principles underlying oocyte development are only partially understood. Therefore, contemporary proteomic approaches have been used increasingly to provide new insights into oocyte quality and maturation in various species such as mouse, pig, and cow. Especially, animal studies have helped in elucidating the molecular status of oocytes during in vitro maturation and other procedures of assisted reproduction. The aim of this review is to summarize the literature on mammalian oocyte proteome and secretome research in the light of natural and assisted reproduction and on lessons to be learned for human oocytes, which have so far remained inaccessible for proteome analysis. PMID:24804254

Virant-Klun, Irma; Krijgsveld, Jeroen

2014-01-01

92

Apoptosis maintains oocyte quality in aging Caenorhabditis elegans females.  

PubMed

In women, oocytes arrest development at the end of prophase of meiosis I and remain quiescent for years. Over time, the quality and quantity of these oocytes decreases, resulting in fewer pregnancies and an increased occurrence of birth defects. We used the nematode Caenorhabditis elegans to study how oocyte quality is regulated during aging. To assay quality, we determine the fraction of oocytes that produce viable eggs after fertilization. Our results show that oocyte quality declines in aging nematodes, as in humans. This decline affects oocytes arrested in late prophase, waiting for a signal to mature, and also oocytes that develop later in life. Furthermore, mutations that block all cell deaths result in a severe, early decline in oocyte quality, and this effect increases with age. However, mutations that block only somatic cell deaths or DNA-damage-induced deaths do not lower oocyte quality. Two lines of evidence imply that most developmentally programmed germ cell deaths promote the proper allocation of resources among oocytes, rather than eliminate oocytes with damaged chromosomes. First, oocyte quality is lowered by mutations that do not prevent germ cell deaths but do block the engulfment and recycling of cell corpses. Second, the decrease in quality caused by apoptosis mutants is mirrored by a decrease in the size of many mature oocytes. We conclude that competition for resources is a serious problem in aging germ lines, and that apoptosis helps alleviate this problem. PMID:19057674

Andux, Sara; Ellis, Ronald E

2008-12-01

93

Apoptosis Maintains Oocyte Quality in Aging Caenorhabditis elegans Females  

PubMed Central

In women, oocytes arrest development at the end of prophase of meiosis I and remain quiescent for years. Over time, the quality and quantity of these oocytes decreases, resulting in fewer pregnancies and an increased occurrence of birth defects. We used the nematode Caenorhabditis elegans to study how oocyte quality is regulated during aging. To assay quality, we determine the fraction of oocytes that produce viable eggs after fertilization. Our results show that oocyte quality declines in aging nematodes, as in humans. This decline affects oocytes arrested in late prophase, waiting for a signal to mature, and also oocytes that develop later in life. Furthermore, mutations that block all cell deaths result in a severe, early decline in oocyte quality, and this effect increases with age. However, mutations that block only somatic cell deaths or DNA-damage–induced deaths do not lower oocyte quality. Two lines of evidence imply that most developmentally programmed germ cell deaths promote the proper allocation of resources among oocytes, rather than eliminate oocytes with damaged chromosomes. First, oocyte quality is lowered by mutations that do not prevent germ cell deaths but do block the engulfment and recycling of cell corpses. Second, the decrease in quality caused by apoptosis mutants is mirrored by a decrease in the size of many mature oocytes. We conclude that competition for resources is a serious problem in aging germ lines, and that apoptosis helps alleviate this problem. PMID:19057674

Andux, Sara; Ellis, Ronald E.

2008-01-01

94

Oocyte cryopreservation for donor egg banking.  

PubMed

Oocyte donation is an efficient alternative to using own oocytes in IVF treatment for different indications. Unfortunately, 'traditional' (fresh) egg donations are challenged with inefficiency, difficulties of synchronization, very long waiting periods and lack of quarantine measures. Given the recent improvements in the efficiency of oocyte cryopreservation, it is reasonable to examine if egg donation through oocyte cryopreservation has merits. The objective of the current manuscript is to review existing literature on this topic and to report on the most recent outcomes from two established donor cryobank centres. Reports on egg donation using slow freezing are scarce and though results are encouraging, outcomes are not yet comparable to a fresh egg donation treatment. Vitrification on the other hand appears to provide high survival rates (90%) of donor oocytes and comparable fertilization, embryo development, implantation and pregnancy rates to traditional (fresh) egg donation. Besides the excellent outcomes, the ease of use for both donors and recipients, higher efficiency, lower cost and avoiding the problem of synchronization are all features associated with the benefit of a donor egg cryobank and makes it likely that this approach becomes the future standard of care. Oocyte donation is one of the last resorts in IVF treatment for couples challenged with infertility problems. However, traditional (fresh) egg donation, as it is performed today, is not very efficient, as typically all eggs from one donor are given to only one recipient, it is arduous as it requires an excellent synchronization between the donor and recipient and there are months or years of waiting time. Because of the development of an efficient oocyte cryopreservation technique, it is now possible to cryo-store donor (as well as non-donor) eggs, maintaining their viability and allowing their use whenever there is demand. Therefore, creating a donor oocyte cryobank would carry many advantages. In the present manuscript, the current experience with oocyte donation using cryopreservation technology is reviewed. The outcomes of two recently established donor egg cryobanks at Instituto Valenciano de Infertilidad in Spain and Reproductive Biology Associates in the USA (involving a large number of cases) demonstrate that egg cryo-survival is high and that fertilization, embryo development, implantation and pregnancy rates are similar to those reported after fresh egg donation. It also provides additional advantages of being more efficient, more economical, easier for both donors and recipients and potentially also safer, because eggs can now be quarantined for 6 months (or longer) to retest for infectious diseases in the donors. It is the opinion of the authors, based on several advantages associated with the use of donor egg cryobanking, that in the future there will be fewer traditional egg donations and increasingly more cryo-egg donations. PMID:21767989

Cobo, Ana; Remohí, José; Chang, Ching-Chien; Nagy, Zsolt Peter

2011-09-01

95

Nonagonistic familiarity decreases aggression in male Turkish hamsters, Mesocricetus brandti  

Microsoft Academic Search

In laboratory studies, hamsters (Mesocricetus spp.) show intense male-male aggression, thus making them an excellent model system for studies of the functional and mechanistic bases of aggression. In a field study of golden hamsters, M. auratus, in the wild, however, the few documented male-male interactions were not highly aggressive. Thus, we tested the hypothesis that familiarity modulates aggression in hamsters.

Javier delBarco-Trillo; M. Elsbeth McPhee; Robert E. Johnston

2009-01-01

96

Nonrandom Sex Composition of Gerbil, Mouse, and Hamster  

E-print Network

Nonrandom Sex Composition of Gerbil, Mouse, and Hamster Litters Before and After Birth MERTICE M litters of Mongolian gerbils, and 854 vaginally delivered litters of golden hamsters, we determined hamsters is present in vaginally delivered infant Mongolian gerbils. but not in their Caesarean

Galef Jr., Bennett G.

97

Optical coherence tomography of malignancy in hamster cheek pouches  

E-print Network

Optical coherence tomography of malignancy in hamster cheek pouches Erin S. Matheny Nevine M. Hanna-time spatially resolved blood flow in mi- crovasculature. Hamster cheek pouches with induced dysplasia Golden Syrian hamsters, 0.5% 9,10-dimethyl-1,2-benzanthracene induces carcinogenesis over 10 weeks

Chen, Zhongping

98

Spielerisches Erlernen der Programmierung mit dem Java-Hamster-Modell  

E-print Network

Spielerisches Erlernen der Programmierung mit dem Java- Hamster-Modell Dietrich Boles Department Oldenburg boles@informatik.uni-oldenburg.de Abstract: Das Java-Hamster-Modell ist ein spezielles Programmierkonzepte und den Programmentwurf kennen, indem sie so genannte Hamster-Programme entwickeln, mit denen sie

Appelrath, Hans-Jürgen

99

Establishment and persistence of photoperiodic memory in hamsters  

E-print Network

Establishment and persistence of photoperiodic memory in hamsters Brian J. Prendergast* , Michael R reproduction in Siberian hamsters. By contrast, intermedi- ate-duration day lengths (12.5­14 h long) either of these outcomes transpires depends on an animal's photoperiodic history, suggesting that hamsters must encode

Zucker, Irving

100

Multidrug-resistant transport activity protects oocytes from  

E-print Network

-resistant (MDR) transporter, P-glycoprotein (MDR-1), oocyte, chemotherapy Discuss: You can discuss this article, and this activity was significantly reduced in the presence of the MDR inhibitor PSC 833. Geminal vesicle oocytes

Wessel, Gary M.

101

CYTOPLASMIC MICROTUBULAR DYNAMIC AND CHROMATIN ORGANIZATION DURING MAMMALIAN OOCYTE MATURATION  

EPA Science Inventory

Coordinated alterations in oocyte chromosome and microtubule disposition occur during oogenesis and oocyte maturation in the mammal. imely transitions in meiotic spindle and cytoplasmic microtubules, due to modifications in both the assembly competence of the tubulin pool and nuc...

102

Motility contrast imaging of live porcine cumulus-oocyte complexes  

NASA Astrophysics Data System (ADS)

Freshly-harvested porcine oocytes are invested with cumulus granulosa cells in cumulus-oocyte complexes (COCs). The cumulus cell layer is usually too thick to image the living oocyte under a conventional microscope. Therefore, it is difficult to assess the oocyte viability. The low success rate of implantation is the main problem for in vitro fertilization. In this paper, we demonstrate our dynamic imaging technique called motility contrast imaging (MCI) that provides a non-invasive way to monitor the COCs before and after maturation. MCI shows a change of intracellular activity during oocyte maturation, and a measures dynamic contrast between the cumulus granulosa shell and the oocytes. MCI also shows difference in the spectral response between oocytes that were graded into quality classes. MCI is based on shortcoherence digital holography. It uses intracellular motility as the endogenous imaging contrast of living tissue. MCI presents a new approach for cumulus-oocyte complex assessment.

An, Ran; Turek, John; Machaty, Zoltan; Nolte, David

2013-02-01

103

Histopathology of Lyme arthritis in LSH hamsters  

SciTech Connect

The authors studied the histopathologic evolution of arthritis in nonirradiated and irradiated hamsters infected with Borrelia burgdorferi. Nonirradiated hamsters injected in the hind paws with B. burgdorferi developed an acute inflammatory reaction involving the synovium, periarticular soft tissues, and dermis. This acute inflammatory reaction was short-lived and was replaced by a mild chronic synovitis as the number of detectable spirochetes in the synovium, periarticular soft tissues, and perineurovascular areas diminished. Exposing hamsters to radiation before inoculation with B. burgdorferi exacerbated and prolonged the acute inflammatory phase. Spirochetes also persisted longer in the periarticular soft tissues. A major histopathologic finding was destructive and erosive bone changes of the hind paws, which resulted in deformation of the joints. These studies should be helpful in defining the immune mechanism participating in the onset, progression, and resolution of Lyme arthritis.

Hejka, A.; Schmitz, J.L.; England, D.M.; Callister, S.M.; Schell, R.F.

1989-05-01

104

Pregnancy rate in an oocyte donation program.  

PubMed

Oocyte donation programs offer an alternative treatment for infertile women with ovarian failure or abnormal ovarian function. Seventeen cycles of in vitro fertilization and embryo transfer with donated oocytes were performed in 13 women, with a mean age of 34.8 years. The hormonal replacement therapy consisted of a fixed dose of oral estradiol valerate, 6 mg daily, and intramuscular progesterone in oil, 100 mg daily. Estrogen and progesterone were continued for 10 more weeks after embryo transfer if pregnancy was established. After 13 embryo transfers, 8 pregnancies were obtained, for a pregnancy rate per transfer of 61.5%. Today seven pregnancies are progressing normally, including one set of twins. This results suggest that an oocyte donation program using a fixed and simple hormonal replacement therapy is an adequate treatment for these infertile couples. PMID:1472813

Zegers-Hochschild, F; Fernández, E; Fabres, C; Mackenna, A; Prado, J; Roblero, L; Lopez, T; Altieri, E; Guadarrama, A; Escudero, F

1992-08-01

105

LINE-1 of evidence for fetal oocyte attrition by retrotransposon.  

PubMed

Fetal oocytes in mammals undergo extensive apoptosis during development. In this issue of Developmental Cell, Malki et al. (2014) provide insight into how and why such massive oocyte loss occurs through the demonstration that the expression level of LINE-1 retrotransposon defines the survival threshold and thus viability of fetal oocytes. PMID:24914555

Chuma, Shinichiro

2014-06-01

106

Congenital transmission of experimental leishmaniasis in a hamster model.  

PubMed

Little information is available on transplacental transmission of Leishmania spp. We determined the frequency and impact of congenital infection caused by Leishmania panamensis or L. donovani in experimentally infected hamsters. A polymerase chain reaction showed that congenital transmission occurred in 25.8% (24 of 93) of offspring born to L. panamensis-infected hamsters and 14.6% (11 of 75) offspring born to L. donovani-infected hamsters. Mortality during lactation was higher in offspring born to L. panamensis-infected hamsters and offspring born to L. donovani-infected hamsters than controls, and lymphoproliferation to Leishmania was more frequent in offspring born to L. panamensis-infected hamsters (17.4%, 11 of 63) than in offspring born to L. donovani-infected hamsters (8.5%, 3 of 35). After weaning, only offspring born to L. donovani-infected hamsters had lower weight gain (P < 0.001) and hematocrit levels (P = 0.0045) than controls. Challenge of offspring born to L. panamensis-infected hamsters with L. panamensis showed no differences in lesion evolution, and offspring born to L. donovani-infected hamsters were more susceptible to L. donovani challenge than controls. Consequently, prenatal exposure of hamsters to L. donovani significantly increased the mortality risk and susceptibility to secondary homologous infection. PMID:22556079

Osorio, Yaneth; Rodriguez, Luz D; Bonilla, Diana L; Peniche, Alex G; Henao, Hector; Saldarriaga, Omar; Travi, Bruno L

2012-05-01

107

Role of focal adhesion kinase in oocyte-follicle communication.  

PubMed

Germ cells require communication with associated somatic cells for normal gametogenesis, as exemplified by an oocyte that interacts with granulosa cells via paracrine factors as well as gap junctions located at sites of contact between these two cell types. The objective of the present study was to define the mechanisms by which cell-cell contact with the oocyte is controlled and to determine the extent that the oocyte actively participates in this association. Proline-rich tyrosine kinase 2 (PTK2), a focal adhesion kinase, was found to be activated at sites of contact between the oocyte and trans-zonal cell processes from the surrounding granulosa cells. In order to determine the functional significance of oocyte-derived PTK2 signaling in oocyte-follicle communication, an oocyte-specific Ptk2 knockout was produced through a breeding strategy pairing a floxed Ptk2-CAT-eGFP mouse with the Zp3-Cre line. Since Ptk2-null mice never develop to birth, this represents the first opportunity to define the role of PTK2 in oocyte-follicle communication. Ablation of Ptk2 within the developing oocyte resulted in lower fertility with reduced numbers of pups, lower rates of blastocyst formation, and reduced cell numbers per blastocyst. Follicles containing Ptk2-null oocytes exhibited reduced oocyte diameter, reduced numbers of connexin 37 and 43 foci at the oocyte surface, and impaired dye coupling between oocyte and granulosa cells. These findings are consistent with a model in which PTK2 plays a critical role in establishing or maintaining oocyte-granulosa cell contacts that are essential for gap junction-mediated communication between granulosa cells and the oocyte. Mol. Reprod. Dev. 82: 90-102, 2015. © 2014 Wiley Periodicals, Inc. PMID:25536210

McGinnis, Lynda K; Kinsey, William H

2015-02-01

108

Sperm and Oocyte Communication Mechanisms Controlling C. elegans Fertility  

PubMed Central

During sexual reproduction in many species, sperm and oocyte secrete diffusible signaling molecules to help orchestrate the biological symphony of fertilization. In the Caenorhabditis elegans gonad, bidirectional signaling between sperm and oocyte is important for guiding sperm to the fertilization site and inducing oocyte maturation. The molecular mechanisms that regulate sperm guidance and oocyte maturation are being delineated. Unexpectedly, these mechanisms are providing insight into human diseases, such as amyotrophic lateral sclerosis, spinal muscular atrophy, and cancer. Here we review sperm and oocyte communication in C. elegans and discuss relationships to human disorders. PMID:20034089

Han, Sung Min; Cottee, Pauline A.; Miller, Michael A.

2010-01-01

109

Characterization of diabetes in the Chinese hamster  

Microsoft Academic Search

Summary  Diabetes in Chinese hamster was initially detected by qualitative tests for urine glucose. The disease was characterized by quantitating urine glucose, glucose tolerance tests and measurement of fasting and nonfasting blood sugar, blood ketones, plasma free fatty acids (FFA), plasma insulin, pancreatic insulin and fasting levels of liver glycogen. In addition, basal levels of glucose utilization by diaphragms and epididymal

G. C. Gerritsen; W. E. Dulin

1967-01-01

110

IN VITRO CULTURE OF POSTIMPLANTATION HAMSTER EMBRYOS  

EPA Science Inventory

In vitro culture of intact rat and mouse embryos has been described extensively, but information on the culture of other species is sparse. The present study examined some culture requirements of early somite stage hamster embryos and assessed the embryotoxic effects of sodium sa...

111

Session: Hard Rock Penetration  

SciTech Connect

This session at the Geothermal Energy Program Review X: Geothermal Energy and the Utility Market consisted of five presentations: ''Hard Rock Penetration - Summary'' by George P. Tennyson, Jr.; ''Overview - Hard Rock Penetration'' by James C. Dunn; ''An Overview of Acoustic Telemetry'' by Douglas S. Drumheller; ''Lost Circulation Technology Development Status'' by David A. Glowka; ''Downhole Memory-Logging Tools'' by Peter Lysne.

Tennyson, George P. Jr.; Dunn, James C.; Drumheller, Douglas S.; Glowka, David A.; Lysne, Peter

1992-01-01

112

General Manual Cell Penetration  

E-print Network

General Manual Cell Penetration Supporting material for product series CPP-C, CPP-P, CPP-A, CPP-3641-6285 100 http://www.jenabioscience.com Last update: Jun 02, 2008 1 Introduction to Cell Penetration [1-12] Plasma membranes protect cells from environment. They are commonly permeable only for lipids and small

Lebendiker, Mario

113

Oocyte cryopreservation and ovarian tissue banking  

Microsoft Academic Search

Oocyte cryopreservation, despite its impact on conservation of genetic resources, is not yet an established technology. Several problems need to be solved before this technology can be applied regularly. Chilling membrane susceptibility and formation of ice due to the large volume of the cell are the major problems observed. However, during the last years, several studies were done to obtain

S. Ledda; G. Leoni; L. Bogliolo; S. Naitana

2001-01-01

114

Sensitivity of canine oocytes to low temperature.  

PubMed

This study compared the viability of canine oocytes after storage for 5 h at 4 or 38 degrees C. The ovaries were collected after ovariohysterectomy of bitches and transported to the laboratory within 5 h at 4 or 38 degrees C. The collected oocytes were matured in DMEM supplemented with 10% FBS, 0.6 mM/mL cysteine, 0.2 mM pyruvic acid, 20 ng/mL E2 and 1 microg/mL rbST, and incubated for 0, 24 and 48 h, at 38 degrees C and in 95% air with 5% CO2. The viability of the oocytes after 0 h did not differ significantly between 4 and 38 degrees C group (79.6% versus 83.9%), but after 24 and 48 h, significant differences were apparent (13.2% versus 77.8% after 24 h and 0.0% versus 72.9% after 48 h; P < 0.05). Therefore, canine oocytes were remarkably sensitive to low temperatures. PMID:16499959

Lee, H S; Yin, X J; Kong, I K

2006-10-01

115

A Cdc42-regulated actin cytoskeleton mediates Drosophila oocyte polarization.  

PubMed

Polarity of the Drosophila oocyte is essential for correct development of the egg and future embryo. The Par proteins Par-6, aPKC and Bazooka are needed to maintain oocyte polarity and localize to specific domains early in oocyte development. To date, no upstream regulator or mechanism for localization of the Par proteins in the oocyte has been identified. We have analyzed the role of the small GTPase Cdc42 in oocyte polarity. We show that Cdc42 is required to maintain oocyte fate, which it achieves by mediating localization of Par proteins at distinct sites within this cell. We establish that Cdc42 localization itself is polarized to the anterolateral cortex of the oocyte and that Cdc42 is needed for maintenance of oocyte polarity throughout oogenesis. Our data show that Cdc42 ensures the integrity of the oocyte actin network and that disruption of this network with Latrunculin A phenocopies loss of Cdc42 or Par protein function in early stages of oogenesis. Finally, we show that Cdc42 and Par proteins, as well as Cdc42/Par and Arp3, interact in the context of oocyte polarity, and that loss of Par proteins reciprocally affects Cdc42 localization and the actin network. These results reveal a mutual dependence between Par proteins and Cdc42 for their localization, regulation of the actin cytoskeleton and, consequently, for the establishment of oocyte polarity. This most likely allows for the robustness in symmetry breaking in the cell. PMID:23250210

Leibfried, Andrea; Müller, Sandra; Ephrussi, Anne

2013-01-15

116

Selection of Ovine Oocytes by Brilliant Cresyl Blue Staining  

PubMed Central

Sheep oocytes derived from the ovaries collected from the slaughterhouse are often used for research on in vitro embryo production, animal cloning, transgenesis, embryonic stem cells, and other embryo biotechnology aspects. Improving the in vitro culture efficiency of oocytes can provide more materials for similar studies. Generally, determination of oocyte quality is mostly based on the layers of cumulus cells and cytoplasm or cytoplasm uniformity and colors. This requires considerable experience to better identify oocyte quality because of the intense subjectivity involved (Gordon (2003), Madison et al. (1992) and De Loos et al. (1992)). BCB staining is a function of glucose-6-phosphate dehydrogenase (G6PD) activity, an enzyme synthesized in developing oocytes, which decreases in activity with maturation. Therefore, unstained oocytes (BCB?) are high in G6PD activity, while the less mature oocytes stains are deep blue (BCB+) due to insuffcient G6PD activity to decolorize the BCB dye. PMID:22675245

Wang, Liqin; Lin, Jiapeng; Huang, Juncheng; Wang, Jing; Zhao, Yuncheng; Chen, Tong

2012-01-01

117

Selection of ovine oocytes by brilliant cresyl blue staining.  

PubMed

Sheep oocytes derived from the ovaries collected from the slaughterhouse are often used for research on in vitro embryo production, animal cloning, transgenesis, embryonic stem cells, and other embryo biotechnology aspects. Improving the in vitro culture efficiency of oocytes can provide more materials for similar studies. Generally, determination of oocyte quality is mostly based on the layers of cumulus cells and cytoplasm or cytoplasm uniformity and colors. This requires considerable experience to better identify oocyte quality because of the intense subjectivity involved (Gordon (2003), Madison et al. (1992) and De Loos et al. (1992)). BCB staining is a function of glucose-6-phosphate dehydrogenase (G6PD) activity, an enzyme synthesized in developing oocytes, which decreases in activity with maturation. Therefore, unstained oocytes (BCB-) are high in G6PD activity, while the less mature oocytes stains are deep blue (BCB+) due to insuffcient G6PD activity to decolorize the BCB dye. PMID:22675245

Wang, Liqin; Lin, Jiapeng; Huang, Juncheng; Wang, Jing; Zhao, Yuncheng; Chen, Tong

2012-01-01

118

Oocyte apoptosis: like sand through an hourglass.  

PubMed

Although the study of germ cell death is arguably still in its infancy as a field, several recent breakthroughs have provided the fodder for a story, replete with episodes of apparent mass cellular suicide if not murder, that will undoubtedly serve as a research base for many laboratories over the next several years. Death is known to strike the male and female germlines with roughly equal intensity, but the innate feature of male germ cells being self-renewing while those of the female are not places the death of oocytes in a completely different light. Indeed, the functional life span of the female gonads is defined in most species, including humans, by the size and rate of depletion of the precious endowment of oocytes enclosed within follicles in the ovaries at birth. This continuous loss of oocytes throughout life, referred to by many as the female biological clock, appears to be driven by a genetic program of cell death that is composed of players and pathways conserved from worms to humans. It is on this genetic pathway, and the role of its constituent molecules in regulating female germ cell fate, that this review will focus. Emphasis will be placed on those studies using genetic-null or transgenic models to explore the functional requirement of proteins, such as Bcl-2 family members, Apaf-1, and caspases in vertebrates to CED-9, CED-4, and CED-3 in Caenorhabditis elegans, in oocyte survival and death. Furthermore, hypotheses regarding the potential impact of translating what is now known of the oocyte death pathway into new approaches for the clinical diagnosis and management of female infertility and the menopause will be offered as a means to stimulate further research in this new and exciting field. PMID:10452843

Morita, Y; Tilly, J L

1999-09-01

119

Role of focal adhesion kinase in oocyte-follicle communication  

PubMed Central

Germ cells require communication with associated somatic cells for normal gametogenesis as exemplified by the oocyte which interacts with granulosa cells via paracrine factors as well as gap junctions located at sites of contact between these two cell types. The objective of the present study was to define the mechanisms by which cell-cell contact with the oocyte is controlled and determine the extent to which the oocyte actively participates in this association. Focal adhesion kinase (PTK2) was found to be activated at sites of contact between the oocyte and trans-zonal cell processes from the surrounding granulosa cells. In order to determine the functional significance of oocyte-derived PTK2 signaling in oocyte-follicle communication, an oocyte-specific ptk2 knockout was produced through a breeding strategy pairing a floxed ptk2-CAT-eGFP mouse with the Zp3-cre line. Since ptk2-null mice never develop to birth, this represents the first opportunity to define the role of ptk2 in oocytefollicle communication. Ablation of ptk2 within the developing oocyte resulted in lower fertility with reduced numbers of pups, lower rates of blastocyst formation, and reduced cell numbers per blastocyst. Follicles containing ptk2-null oocytes exhibited reduced oocyte diameter, reduced numbers of connexin 37 and 43 foci at the oocyte surface, and impaired dye coupling between oocyte and granulosa cells. These findings are consistent with a model in which PTK2 plays a critical role in establishing or maintaining oocyte-granulosa cell contacts that are essential for gap junction - mediated communication between granulosa cells and the oocyte. PMID:25536210

McGinnis, Lynda K.; Kinsey, William H.

2015-01-01

120

Mitochondrial DNA rearrangements in human oocytes and embryos.  

PubMed

Human mitochondrial DNA (mtDNA) rearrangements, including more than 150 deletions and insertions, accumulate with age and are responsible for certain neuromuscular diseases. Human oocytes, arrested for up to 50 years, may express certain mtDNA rearrangements possibly affecting function. Investigations have previously shown a single mtDNA rearrangement (dmtDNA(4977)) in human oocytes. Sequencing of other rearrangements and their correlation with maternal age have not been performed in human oocytes or embryos. Here we use a nested PCR strategy of long followed by short polymerase chain reaction (PCR) that amplifies two-thirds of the mitochondrial genome. mtDNA rearrangements were detected in 50.5% of the oocytes (n = 295) and 32.5% of the embryos (n = 197). This represents a significant difference in the percentage of mtDNA rearrangements between oocytes and embryos (P < 0.0001). Twenty-three novel mtDNA rearrangements with deletions, insertions and duplications were found. There was no significant age-related increase in the percentage of human oocytes or embryos that contained mtDNA rearrangements. Significant reductions in the number of oocytes containing mtDNA rearrangements occurred as oocyte development progressed from germinal vesicle to the mature metaphase II oocyte (P < 0.05). These findings are discussed as they relate to mitochondria, mtDNA, and ATP production in human oocytes and embryos. PMID:10508220

Barritt, J A; Brenner, C A; Cohen, J; Matt, D W

1999-10-01

121

Accurate dispensing system for single oocytes using air ejection  

PubMed Central

In this study, we propose a new approach to increase the success rate of single-oocyte dispensing and investigate the subsequent viability of the dispensed oocytes. We used a pair of capacitance sensors placed in a microfluidic chip to detect the oocyte, and custom-designed a special buffer zone in the microchannel to decelerate the flow velocity and reduce the hydraulic pressure acting on the oocyte. In the buffer zone, a semicircular bay, formed by equally spaced micro-pillars, is used to stop the oocyte at the dispensing nozzle hole. Finally, the oocyte is ejected by airflow to the culture array. The novel feature of the developed microfluidic system is that the extraordinary improvement in success rate is accompanied by a lack of change in oocyte survival rate (as assessed by a comparison of survival rates before and after the dispensing procedure). By using this device, we achieved a highly accurate single-oocyte dispensing process with a success rate of 100%. The oocyte survival rate is approximately 70%, regardless of whether or not the oocyte is dispensed. The newly proposed system has the advantages of high operation speed and potential usage for two-dimensional micropatterning. PMID:24404076

Feng, Lin; Sun, Yiling; Ohsumi, Chisato; Arai, Fumihito

2013-01-01

122

Cryosurvival of ex situ and in situ feline oocytes.  

PubMed

Cryosurvival of feline oocytes preserved as isolated cells (ex situ) or enclosed in ovarian follicles (in situ) has been demonstrated, and significant advances have recently been achieved. However, an ideal protocol for oocyte cryopreservation has not been established to date because of extreme sensitivity of the structural complex to chilling injury. Several factors, such as stage of maturation, membrane permeability and plasticity of the cytoskeleton, affect cryosurvival of the oocyte. Also, intercellular communications between cumulus cells and oocyte are compromised after freezing or vitrification of ex situ or in situ cumulus-oocyte complexes, which has a detrimental effect on oocyte maturational competence. Despite these issues, embryo development, pregnancies and live kittens have been obtained after in vitro fertilization (by ICSI) and transfer of embryos derived from cryopreserved oocytes. It is a general belief that the efficiency of cryopreservation would increase through a better understanding of oocyte responses to cryoprotectants, cooling rates and all the physical events occurring during the exposure of feline oocytes to low temperatures. Cryobanking of feline oocytes would significantly contribute to the preservation of rare genotypes and to the maintenance of a valuable source of genetic material for research applications. PMID:23279515

Luvoni, G C

2012-12-01

123

The human cumulus--oocyte complex gene-expression profile  

PubMed Central

BACKGROUND The understanding of the mechanisms regulating human oocyte maturation is still rudimentary. We have identified transcripts differentially expressed between immature and mature oocytes, and cumulus cells. METHODS Using oligonucleotides microarrays, genome wide gene expression was studied in pooled immature and mature oocytes or cumulus cells from patients who underwent IVF. RESULTS In addition to known genes such as DAZL, BMP15 or GDF9, oocytes upregulated 1514 genes. We show that PTTG3 and AURKC are respectively the securin and the Aurora kinase preferentially expressed during oocyte meiosis. Strikingly, oocytes overexpressed previously unreported growth factors such as TNFSF13/APRIL, FGF9, FGF14, and IL4, and transcription factors including OTX2, SOX15 and SOX30. Conversely, cumulus cells, in addition to known genes such as LHCGR or BMPR2, overexpressed cell-tocell signaling genes including TNFSF11/RANKL, numerous complement components, semaphorins (SEMA3A, SEMA6A, SEMA6D) and CD genes such as CD200. We also identified 52 genes progressively increasing during oocyte maturation, comprising CDC25A and SOCS7. CONCLUSION The identification of genes up and down regulated during oocyte maturation greatly improves our understanding of oocyte biology and will provide new markers that signal viable and competent oocytes. Furthermore, genes found expressed in cumulus cells are potential markers of granulosa cell tumors. PMID:16571642

Assou, Said; Anahory, Tal; Pantesco, Véronique; Le Carrour, Tanguy; Pellestor, Franck; Klein, Bernard; Reyftmann, Lionel; Dechaud, Hervé; De Vos, John; Hamamah, Samir

2006-01-01

124

Sex aneuploidy of unfertilized human oocytes after intracytoplasmic sperm injection  

SciTech Connect

Intracytoplasmic sperm injection (ICSI) has recently achieved successful fertilization and pregnancy in human in vitro fertilization, particularly in cases of severe male factor infertility. One criticism of this novel clinical technique is that it bypasses the natural selection process of fertilization. We use fluorescence in situ hybridization (FISH) to analyze oocytes which fail to fertilize after ICSI in the Yale IVF Program. The purpose of this study is to determine whether failed fertilization after ICSI can be attributed to sex chromosome aneuploidy in the oocyte. Fertilization of oocytes is determined by the presence of two pronuclei on light microscopic examination (400X). Multi-probe FISH with DAPI (4,6,-diamino-2-phenyl-indole) counterstain is then performed to determine oocyte ploidy and the presence of decondensed sperm. Centromeric probes for X, Y and 17 are used simultaneously in each oocyte for in situ hybridization to oocyte chromatin. In all oocytes examined after ICSI to date, unfertilized oocytes have decondensed sperm DNA present confirming appropriate intracytoplasmic placement of the sperm. Preliminary results obtained from 31 oocytes have not identified any sex chromosome aneuploidies. The FISH technique used in post-ICSI oocytes is a model system for delineating genetic causes of failed fertilization in the human.

Lee, G.; Ward, D.C.; Jones, E.E. [Yale Univ., New Haven, CT (United States)

1994-09-01

125

Developmental potential of in vitro or in vivo matured oocytes.  

PubMed

This study compared the embryological features of mature and immature oocytes (different stages) collected from stimulated cycles of in vitro fertilization (IVF). Immature oocytes were identified, classified as PI (prophase I - germinal vesicle, GV) or MI (metaphase I), were matured in vitro and fertilized using the intra-cytoplasmic sperm injection (ICSI) technique. Fertilization potential, cleavage, and subsequent transfer/cryopreservation of the embryos derived from these in vitro matured oocytes were compared with those of in vivo matured oocytes (collected at the MII stage). The characteristics of embryos derived from gametes recovered in the MI and MII stages were similar. The fertilization rate of immature oocytes recovered in PI was significantly lower than that of MII oocytes (P = 0.031), and the cleavage rate of the PI group was also lower than that of the MI (P = 0.004) and MII (P < 0.001) groups. In vitro maturation of MI oocytes is a suitable alternative when immature oocytes are recovered, as their characteristics and development are similar to those of in vivo matured oocytes. Optimization of outcomes for PI oocytes will require development of techniques that can distinguish which of these gametes will mature and fertilize. PMID:23735140

Alcoba, Diego D; Pimentel, Anita M; Brum, Ilma S; Corleta, Helena E

2015-02-01

126

9 CFR 3.36 - Primary enclosures used to transport live guinea pigs and hamsters.  

Code of Federal Regulations, 2010 CFR

...enclosures used to transport live guinea pigs and hamsters. 3.36 Section 3...Treatment, and Transportation of Guinea Pigs and Hamsters Transportation Standards...enclosures used to transport live guinea pigs and hamsters. No person...

2010-01-01

127

Motility induction in hamster spermatozoa from caput epididymidis : effects of forward motility protein  

E-print Network

Motility induction in hamster spermatozoa from caput epididymidis : effects of forward motility observed is very often anarchic. In the hamster, we have previously provided evidence that forward motility characteristics and behaviour of hamster caput spermatozoa were investigated. Spermatozoa were diluted

Paris-Sud XI, Université de

128

Pantothenic Acid Deficiency in Cholesterol-fed Hamsters1  

Microsoft Academic Search

A 5-week study of hamsters fed a pantothenic acid-deficient diet, with and without cholesterol, is reported. The adverse effect of pantothenate deficiency on growth was greater in the cholesterol-fed female hamster than in the male. Pantothenic acid deficiency did not prevent the accumulation of lipid in the liver of the hamster as it did in the rat. However, cholesterol feeding

NINA L. COHEN; LOTTE ARNRICH

129

Calcium waves in the the maturing oocyte  

NASA Astrophysics Data System (ADS)

Calcium waves in oocytes are sustained by release of Ca2+ from the endoplasmic reticulum (ER) through clustered release channels. As the oocytes matures, a) the calcium waves slow down by about a factor of two, b) the overall duration of Ca2+ elevation grows substantially, and c) the cell is more susceptible to wave initiation. At the same time, the kinetics of release of Ca2+ from a single cluster is changed only insignificantly. Based on a computational model that accurately reproduces elemental Ca2+ release kinetics from channel clusters, we propose that the changing spatial organization of signaling effectors is a common underlying cause for all the above described observations as the Ca2+ signaling machinery matures.

Ullah, Aman; Ullah, Ghanim; Jung, Peter; Machaca, Khaled

2009-03-01

130

Nuclear transfer in the mouse oocyte.  

PubMed

The nuclear transfer (NT) technique in the mouse has enabled us to generate cloned mice and to establish NT embryonic stem (ntES) cells. Direct nuclear injection into mouse oocytes with a piezo impact drive unit can aid in the bypass of several steps of the original cell fusion procedure. It is important to note that only the NT approach can reveal dynamic and global modifications in the epigenome without using genetic modification as well as generating live animals from single cells. Thus, these techniques could also be applied to the preservation of genetic material from any mouse strain instead of preserving embryos or gametes. Moreover, with this technique, we can use not only living cells but also the nuclei of dead cells from frozen mouse carcasses for NT. This chapter describes our most recent protocols of NT into the mouse oocyte for cloning mice and for the establishment of ntES cells from cloned embryos. PMID:23138960

Mizutani, Eiji; Ogura, Atsuo; Wakayama, Teruhiko

2013-01-01

131

Ultrarapid vitrification of mouse oocytes and embryos.  

PubMed

Cryopreservation facilitates long-term storage of gametes and embryos for numerous purposes. For example, cryobanking of unique mouse strains, particularly transgenic mice, offers important protection of valuable genetics. It also provides a practical solution for facilities trying to house large numbers of research animals or those looking to relocate without the risk of introducing an animal-derived pathogen. Furthermore, cryopreservation is currently being used for fertility preservation both in humans and as a safeguard for endangered animals. Ultrarapid vitrification offers an elegant, quick, and very reliable method for cryopreservation of mouse oocytes and embryos. Furthermore, research into the effects on mouse oocyte and embryo physiology has indicated that ultrarapid vitrification is superior to conventional slow freezing. High survival rates, embryo development, and viability are routinely achieved with the ultrarapid vitrification method described in this chapter. PMID:24318819

Larman, Mark G; Gardner, David K

2014-01-01

132

Vitrification of oocytes, embryos and blastocysts.  

PubMed

In assisted reproductive technology, cryopreservation of human oocytes and embryos has been significantly improved by refined slow-cooling and the new vitrification method. The slow-cooling method requires a programmed cryo-machine, and usually takes several hours. It is, however, difficult to eliminate injuries resulting from ice formation completely. Vitrification has become a reliable strategy because it is simple, can lead to high survival rates and viability, and has better clinical outcome. Vitrification transforms cells into an amorphous glassy state inside and outside the vitrified cell with ultra-rapid cooling and warming steps by plunging the oocytes and embryos into liquid nitrogen, instead of ice-crystal formation. Over the past decade, several advances in vitrification technologies have improved clinical efficiency and outcome. In this chapter, we focus on vitrification technologies for cryopreservation in human assisted reproductive technology. PMID:22940094

Mukaida, Tetsunori; Oka, Chikahiro

2012-12-01

133

SV40 induces mesotheliomas in hamsters.  

PubMed Central

In the course of studies to elucidate the relative contribution of simian virus 40 (SV40) large T and small t proteins during oncogenesis, we observed the appearance of pericardial and pleural tumors in 100% of Syrian hamsters injected in the pleural space with wild type SV40. When SV40 was injected via the intracardiac or intraperitoneal routes, more than 50% of hamsters developed mesothelial tumors. Macroscopic, microscopic, ultramicroscopic, and histochemical characteristics identify these neoplasms and derived cell lines as mesotheliomas and mesothelioma-derived cell lines. The SV40 genome was integrated and expressed in the mesotheliomas and derived cell lines. The absence of mesotheliomas in hamsters injected with SV40 small t deletion mutants indicates that the small t protein plays an important role in the development of SV40-induced mesotheliomas. To the best of our knowledge, this is the first definitive report of virus-induced mesotheliomas in mammals. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:8388174

Cicala, C.; Pompetti, F.; Carbone, M.

1993-01-01

134

In-vitro Maturation of Human Oocytes  

Microsoft Academic Search

\\u000a Since the birth of the first in-vitro fertilization (IVF) baby in 1978 [1], assisted reproductive technologies (ART) have\\u000a helped thousands of couples to have children through the use of ovarian stimulation [2]. IVF was first performed in a natural\\u000a cycle without any hormonal stimulation, and the oocyte retrieval was performed laparoscopically. However, natural cycle IVF\\u000a was soon replaced by ovarian

Ezgi Demirtas; Hananel Holzer; Weon-Young SON; Ri-Cheng Chain; Seang Lin Tan

135

Ultrasonic backscatter coefficient quantitative estimates from Chinese hamster ovary cell pellet biophantoms  

E-print Network

Ultrasonic backscatter coefficient quantitative estimates from Chinese hamster ovary cell pellet to the ultrasonic backscatter coefficient BSC estimate using Chinese hamster ovary CHO cells. Also introduced

Illinois at Urbana-Champaign, University of

136

Social defeat differentially affects immune responses in Siberian hamsters (Phodopus sungorus)  

E-print Network

Social defeat differentially affects immune responses in Siberian hamsters (Phodopus sungorus concentration as well as innate and acquired immune responses in adult male Siberian hamsters (Phodopus sungorus

Demas, Greg

137

Cryopreservation of human embryos and oocytes.  

PubMed

The success rate of human embryo cryopreservation depends on technical and embryonic parameters. First of all, the cryoprotectant can affect embryo survival as we found by comparing two freeze-thaw procedures using propanediol (PROH) (1.5 mol) alone or with sucrose (0.1 mol). Embryo survival was significantly enhanced with sucrose (62 versus 32%). Embryo quality is another major parameter involved in the success of freezing; the rates of positive survival were found to be 67% for morphologically normal embryos versus 49% for embryos with fragments (P less than 0.001). The efficiency of embryo cryopreservation in an IVF programme could be estimated in 1986: a woman with extra embryos, stored after transfer of 3-4 fresh embryos (16% of all cycles), can expect a 22% pregnancy rate per transfer of fresh embryos and a 32% pregnancy rate per collection after transfer of the stored eggs. A comparative study of the cryopreservability of immature or mature oocytes was performed in humans. Human oocytes have a low survival rate (36%) whatever the cryopreservation protocol or the initial maturation stage. Immature human oocytes could survive freezing and thawing, mature and be fertilized in vitro, but with a very low efficiency. PMID:3350932

Mandelbaum, J; Junca, A M; Plachot, M; Alnot, M O; Salat-Baroux, J; Alvarez, S; Tibi, C; Cohen, J; Debache, C; Tesquier, L

1988-01-01

138

A rational approach to oocyte cryopreservation.  

PubMed

Reports of clinical pregnancies from cryopreserved human oocytes have been steadily increasing in recent years. However, success in terms of births per thawed oocyte remains poor. A wide variety of freezing techniques has been used lately, but modifications to protocols are made on an empirical basis. Methods of cryopreservation are often poorly described or protocols are not strictly adhered to, resulting in variability of outcome. The first stage of a freezing protocol is exposure to cryoprotectant. If performed inappropriately, such exposure can result in damage due to chemical toxicity and/or osmotic stress. Measurement of cell volume change during exposure to cryoprotectants demonstrates the extent of osmotic stress experienced by that cell. Such measurements have been performed during perfusion of murine and human oocytes with cryoprotectant concentrations commonly used for cryopreservation of these cells. It has been demonstrated that changes in the cryoprotectant type, concentration and temperature of exposure can dramatically affect the extent of cell volume change. Even small changes in duration of exposure to cryoprotectant prior to cooling can result in drastic changes in cellular hydration. Such factors will potentially influence the ability of the cell to survive the stresses experienced during the subsequent stages of the cryopreservation protocol. PMID:15949213

Paynter, S J

2005-05-01

139

Molecular and immunological characterization of the first allergenic lipocalin in hamster: the major allergen from Siberian hamster (Phodopus sungorus).  

PubMed

The most frequent pet allergy is to cat and dog, but in recent years, it has become increasingly popular to have other pets, and the risk of exposure to new allergens is more prevalent. The list of new pets includes hamsters, and one of the most popular hamsters is the Siberian hamster (Phodopus sungorus). The aim of this study was the characterization and cloning of the major allergen from this hamster. The study of its allergenicity and cross-reactivity could improve the specific diagnosis and treatment for hamster-allergic patients. Thirteen Siberian hamster-allergic patients were recruited at the outpatient clinic. Protein extracts were prepared from the hair, urine, and salivary glands of four hamster species (European, golden, Siberian, and Roborovski). IgE-binding proteins were detected by immunoblotting and identified by mass spectrometry. The recombinant protein was produced in Escherichia coli and then purified by metal chelate affinity chromatography. The allergenic properties of the recombinant protein were tested by ELISA and immunoblotting, and biological activity was tested according to capacity for basophil activation. Three IgE-binding proteins were identified in extracts obtained from Siberian hamster hair, urine, and salivary glands. All proteins corresponded to the same protein, which was identified as a lipocalin. This lipocalin had no cross-reactivity with common and golden hamsters. The recombinant allergen was cloned and purified, showing similar IgE reactivity in vitro to Siberian hamster protein extracts. Also, the recombinant allergen was capable of producing biological activation in vivo. The major Siberian hamster allergen was cloned, and allergenic properties were characterized, providing a new tool for specific diagnosis of allergy to Siberian hamster. PMID:24993820

Torres, José Alberto; de Las Heras, Manuel; Maroto, Aroa Sanz; Vivanco, Fernando; Sastre, Joaquín; Pastor-Vargas, Carlos

2014-08-22

140

Oocyte quality in patients with severe ovarian hyperstimulation syndrome  

Microsoft Academic Search

Objective: To study the oocyte quality in patients with ovarian hyperstimulation syndrome (OHSS).Design: Retrospective study.Setting: The Egyptian IVF-ET Center.Patient(s): Forty-two patients who developed severe OHSS (group A) were studied for the mean number of oocytes retrieved, percentage of high-quality oocytes, embryo quality, and fertilization, implantation, and pregnancy rates; these patients were compared with an agematched control group who did not

Mohamed A Aboulghar; Ragaa T Mansour; Gamal I Serour; Abdel Maguid Ramzy; Yahia M Amin

1997-01-01

141

Aging-related Changes in In Vitro-matured Bovine Oocytes: Oxidative Stress, Mitochondrial Activity and ATP Content After Nuclear Maturation  

PubMed Central

The objective of this research was to clarify the aging-related changes in in vitro-matured bovine oocytes. Firstly, we examined the fertilization and embryonic development of bovine oocytes after 22 and 30–34 h of in vitro maturation (IVM). The oocytes after 30–34 h of IVM (penetrated by sperm at around 40 h after starting IVM) showed a lower developmental rate to blastocysts (P<0.01), although normal fertilization rates were similar regardless of IVM duration. In the next experiment, reactive oxygen species (ROS), mitochondrial activity and ATP content in oocytes after 20, 30 and 40 h of IVM were examined. The lowest level of ROS was found in the group subjected to 30 h of IVM. The mitochondrial activity and ATP content in the group subjected to 40 h of IVM were higher than in the group subjected to 20 h of IVM (P<0.01), and those in the group subjected to 30 h of IVM showed intermediate values. Thereafter, the mitochondrial activities at 3 days after in vitro fertilization in embryos derived from the oocytes subjected to 22 and 34 h of IVM were evaluated. In the group subjected to 34 h of IVM, high-polarized mitochondria were frequently observed at the periphery of blastomeres. The present results suggest that high mitochondrial activity observed in oocytes after prolonged IVM culture and localization of high-polarized mitochondria at the periphery of blastomeres during early embryonic development may be associated with the low developmental competence in aged bovine oocytes. PMID:24492658

KOYAMA, Keisuke; KANG, Sung-Sik; HUANG, Weiping; YANAGAWA, Yojiro; TAKAHASHI, Yoshiyuki; NAGANO, Masashi

2014-01-01

142

Why is Chromosome Segregation Error in Oocytes Increased With Maternal Aging?- Figure 2  

NSDL National Science Digital Library

This cartoon illustrates the comparison of the cohesion-related molecular basis and chromosome behaviors in oocytes from young (Â?young oocyteÂ?) and old (Â?old oocyteÂ?) mammals, based on mouse model

Zhen-Bo Wang (Chinese Academy of Sciences State Key Laboratory of Reproductive Biology, Institute of Zoology)

2011-10-01

143

Deployable Wireless Camera Penetrators  

NASA Technical Reports Server (NTRS)

A lightweight, low-power camera dart has been designed and tested for context imaging of sampling sites and ground surveys from an aerobot or an orbiting spacecraft in a microgravity environment. The camera penetrators also can be used to image any line-of-sight surface, such as cliff walls, that is difficult to access. Tethered cameras to inspect the surfaces of planetary bodies use both power and signal transmission lines to operate. A tether adds the possibility of inadvertently anchoring the aerobot, and requires some form of station-keeping capability of the aerobot if extended examination time is required. The new camera penetrators are deployed without a tether, weigh less than 30 grams, and are disposable. They are designed to drop from any altitude with the boost in transmitting power currently demonstrated at approximately 100-m line-of-sight. The penetrators also can be deployed to monitor lander or rover operations from a distance, and can be used for surface surveys or for context information gathering from a touch-and-go sampling site. Thanks to wireless operation, the complexity of the sampling or survey mechanisms may be reduced. The penetrators may be battery powered for short-duration missions, or have solar panels for longer or intermittent duration missions. The imaging device is embedded in the penetrator, which is dropped or projected at the surface of a study site at 90 to the surface. Mirrors can be used in the design to image the ground or the horizon. Some of the camera features were tested using commercial "nanny" or "spy" camera components with the charge-coupled device (CCD) looking at a direction parallel to the ground. Figure 1 shows components of one camera that weighs less than 8 g and occupies a volume of 11 cm3. This camera could transmit a standard television signal, including sound, up to 100 m. Figure 2 shows the CAD models of a version of the penetrator. A low-volume array of such penetrator cameras could be deployed from an aerobot or a spacecraft onto a comet or asteroid. A system of 20 of these penetrators could be designed and built in a 1- to 2-kg mass envelope. Possible future modifications of the camera penetrators, such as the addition of a chemical spray device, would allow the study of simple chemical reactions of reagents sprayed at the landing site and looking at the color changes. Zoom lenses also could be added for future use.

Badescu, Mircea; Jones, Jack; Sherrit, Stewart; Wu, Jiunn Jeng

2008-01-01

144

Penetration of yawed projectiles  

SciTech Connect

We used computer simulations and experiment to study the penetration of tungsten-alloy projectiles into a thick, armored steel target. These projectiles, with length-to-diameter ratios of 4, strike the target with severe yaws, up to 90{degree}(side-on-impact), such as might be induced in an originally longer projectile by a multiple-spaced plate array. In this study, we focus on the terminal ballistics of these projectiles and ignore how the yaw was induced. We found that the minimum penetration depth occurs at 90{degree}yaw. This case is well approximated by the two-dimensional plane-strain penetration of a side-on cylinder. The ratio of penetration depth to diameter, P:D, for this case is larger than that for a sphere because the plane-strain geometry lacks hoop stress, which is activated in axisymmetric geometry. A more surprising result of work is that the penetration at 60{degree} yaw is only slightly deeper than that of the side-on impact. 8 refs., 15 figs., 3 tabs.

Reaugh, J.E.

1990-10-08

145

Ultrastructure of immature and mature human oocytes after cryotop vitrification  

PubMed Central

In vitro maturation of vitrified immature germinal vesicle (GV) oocytes is a promising fertility preservation option. We analyzed the ultrastructure of human GV oocytes after Cryotop vitrification (GVv) and compared it with fresh GV (GVc), fresh mature metaphase II (MIIc) and Cryotop-vitrified mature (MIIv) oocytes. By phase contrast microscopy and light microscopy, the oolemmal and cytoplasmic organization of fresh and vitrified oocytes did not show significant changes. GVv oocytes showed significant ultrastructural alterations of the microvilli in 40% of the samples; small vacuoles and occasional large/isolated vacuoles were abnormally present in the ooplasm periphery of 50% of samples. The ultrastructure of nuclei and mitochondria-vesicle (MV) complexes, as well as the distribution and characteristics of cortical granules (CGs), were comparable with those of GVc oocytes. MIIv oocytes showed an abnormal ultrastructure of microvilli in 30% of the samples and isolated large vacuoles in 70% of the samples. MV complexes were normal, but mitochondria-smooth endoplasmic reticulum aggregates appeared to be of reduced size. CGs were normally located under the oolemma but presented abnormalities in distribution and matrix electron density. In conclusion, Cryotop vitrification preserved main oocyte characteristics in the GV and MII stages, even if peculiar ultrastructural alterations appeared in both stages. This study also showed that the GV stage appears more suitable for vitrification than the MII stage, as indicated by the good ultrastructural preservation of important structures that are present only in immature oocytes, like the nucleus and migrating CGs.

PALMERINI, Maria Grazia; ANTINORI, Monica; MAIONE, Marta; CERUSICO, Fabrizio; VERSACI, Caterina; NOTTOLA, Stefania Annarita; MACCHIARELLI, Guido; KHALILI, Mohammad Ali; ANTINORI, Severino

2014-01-01

146

STEROID INHIBITION OF PROTEIN INCORPORATION BY ISOLATED AMPHIBIAN OOCYTES  

PubMed Central

The relationship between blood protein (vitellogenin) incorporation and nuclear maturation was studied in individual amphibian oocytes after in vitro exposure to desoxycorticosterone acetate (DOCA). Isolated Rana pipiens oocytes were incubated in vitro with radioactively labeled oocyte yolk precursor ([3H]vitellogenin) obtained from estrogenized Xenopus laevis. Incorporation of labeled vitellogenin into the oocytes continued over a 24-h period. Oocytes simultaneously exposed to DOCA and to labeled vitellogenin exhibited both inhibition of vitellogenin incorporation and stimulation of nuclear maturation and cortical changes. Inhibition of vitellogenin incorporation was observed after approximately 9 h of incubation and was correlated with the time of nuclear breakdown. Preincubation of oocytes in steroid for 9 h essentially terminated vitellogenin incorporation. Incorporation of vitellogenin occurred after removal of follicle cells from the oocyte by a short treatment with EDTA. These results demonstrate the macromolecular vitellogenin transport system remains operative in oocytes which can undergo nuclear maturation and that the steroid DOCA can affect its function. Evidence suggests that the mechanism of steroid inhibition is in part the result of inhibition of the micropinocytotic process in the oocyte cortex. PMID:4544841

Schuetz, Allen W.; Wallace, Robin A.; Dumont, James N.

1974-01-01

147

Oocyte triplet pairing for electrophysiological investigation of gap junctional coupling  

PubMed Central

Gap junctions formed by expressing connexin subunits in Xenopus oocytes provide a valuable tool for revealing the gating properties of intercellular gap junctions in electrically coupled cells. We describe a new method that consists of simultaneous triple recordings from 3 apposed oocytes expressing exogenous connexins. The advantages of this method is that in one single experiment, one oocyte serves as control while a pair of oocytes, which have been manipulated differently, may be tested for different gap junctional properties. Moreover, we can study simultaneously the gap junctional coupling of 3 different pairs of oocytes in the same preparation. If the experiment consists of testing the effect of a single drug, this approach will reduce the time required, as background coupling in control pairs of oocytes does not need to be measured separately as with the conventional 2 oocyte pairing. The triplet approach also increases confidence that any changes seen in junctional communication are due to the experimental treatment and not variation in the preparation of oocytes or execution of the experiment. In this study, we show the example of testing the gap junctional properties among three oocytes, two of which are expressing rat connexin36. PMID:20230857

Hayar, Abdallah; Charlesworth, Amanda; Garcia-Rill, Edgar

2010-01-01

148

In vitro oocyte maturation and preantral follicle culture from the luteal-phase baboon ovary produce mature oocytes.  

PubMed

Female cancer patients who seek fertility preservation but cannot undergo ovarian stimulation and embryo preservation may consider 1) retrieval of immature oocytes followed by in vitro maturation (IVM) or 2) ovarian tissue cryopreservation followed by transplantation or in vitro follicle culture. Conventional IVM is carried out during the follicular phase of menstrual cycle. There is limited evidence demonstrating that immature oocyte retrieved during the luteal phase can mature in vitro and be fertilized to produce viable embryos. While in vitro follicle culture is successful in rodents, its application in nonhuman primates has made limited progress. The objective of this study was to investigate the competence of immature luteal-phase oocytes from baboon and to determine the effect of follicle-stimulating hormone (FSH) on baboon preantral follicle culture and oocyte maturation in vitro. Oocytes from small antral follicle cumulus-oocyte complexes (COCs) with multiple cumulus layers (42%) were more likely to resume meiosis and progress to metaphase II (MII) than oocytes with a single layer of cumulus cells or less (23% vs. 3%, respectively). Twenty-four percent of mature oocytes were successfully fertilized by intracytoplasmic sperm injection, and 25% of these developed to morula-stage embryos. Preantral follicles were encapsulated in fibrin-alginate-matrigel matrices and cultured to small antral stage in an FSH-independent manner. FSH negatively impacted follicle health by disrupting the integrity of oocyte and cumulus cells contact. Follicles grown in the absence of FSH produced MII oocytes with normal spindle structure. In conclusion, baboon luteal-phase COCs and oocytes from cultured preantral follicles can be matured in vitro. Oocyte meiotic competence correlated positively with the number of cumulus cell layers. This study clarifies the parameters of the follicle culture system in nonhuman primates and provides foundational data for future clinical development as a fertility preservation option for women with cancer. PMID:21123815

Xu, Min; Fazleabas, Asgerally T; Shikanov, Ariella; Jackson, Erin; Barrett, Susan L; Hirshfeld-Cytron, Jenny; Kiesewetter, Sarah E; Shea, Lonnie D; Woodruff, Teresa K

2011-04-01

149

An oocyte-specific ELAVL2 isoform is a translational repressor ablated from meiotically competent antral oocytes  

PubMed Central

At the end of the growth phase, mouse antral follicle oocytes acquire full developmental competence. In the mouse, this event is marked by the transition from the so-called non-surrounded nucleolus (NSN) chromatin configuration into the transcriptionally quiescent surrounded nucleolus (SN) configuration, which is named after a prominent perinucleolar condensed chromatin ring. However, the SN chromatin configuration alone is not sufficient for determining the developmental competence of the SN oocyte. There are additional nuclear and cytoplamic factors involved, while a little is known about the changes occurring in the cytoplasm during the NSN/SN transition. Here, we report functional analysis of maternal ELAVL2 an AU-rich element binding protein. Elavl2 gene encodes an oocyte-specific protein isoform (denoted ELAVL2°), which acts as a translational repressor. ELAVL2° is abundant in fully grown NSN oocytes, is ablated during the NSN/SN transition and remains low during the oocyte-to-embryo transition (OET). ELAVL2° overexpression during meiotic maturation causes errors in chromosome segregation, indicating the significance of naturally reduced ELAVL2° levels in SN oocytes. On the other hand, during oocyte growth, prematurely reduced Elavl2 expression results in lower yields of fully grown and meiotically matured oocytes, suggesting that Elavl2 is necessary for proper oocyte maturation. Moreover, Elavl2 knockdown showed stimulating effects on translation in fully grown oocytes. We propose that ELAVL2 has an ambivalent role in oocytes: it functions as a pleiotropic translational repressor in efficient production of fully grown oocytes, while its disposal during the NSN/SN transition contributes to the acquisition of full developmental competence. PMID:24553115

Chalupnikova, Katerina; Solc, Petr; Sulimenko, Vadym; Sedlacek, Radislav; Svoboda, Petr

2014-01-01

150

Hydrogen Sulfide Donor Protects Porcine Oocytes against Aging and Improves the Developmental Potential of Aged Porcine Oocytes  

PubMed Central

Porcine oocytes that have matured in in vitro conditions undergo the process of aging during prolonged cultivation, which is manifested by spontaneous parthenogenetic activation, lysis or fragmentation of aged oocytes. This study focused on the role of hydrogen sulfide (H2S) in the process of porcine oocyte aging. H2S is a gaseous signaling molecule and is produced endogenously by the enzymes cystathionine-?-synthase (CBS), cystathionine-?-lyase (CSE) and 3-mercaptopyruvate sulfurtransferase (MPST). We demonstrated that H2S-producing enzymes are active in porcine oocytes and that a statistically significant decline in endogenous H2S production occurs during the first day of aging. Inhibition of these enzymes accelerates signs of aging in oocytes and significantly increases the ratio of fragmented oocytes. The presence of exogenous H2S from a donor (Na2S.9H2O) significantly suppressed the manifestations of aging, reversed the effects of inhibitors and resulted in the complete suppression of oocyte fragmentation. Cultivation of aging oocytes in the presence of H2S donor positively affected their subsequent embryonic development following parthenogenetic activation. Although no unambiguous effects of exogenous H2S on MPF and MAPK activities were detected and the intracellular mechanism underlying H2S activity remains unclear, our study clearly demonstrates the role of H2S in the regulation of porcine oocyte aging. PMID:25615598

Krejcova, Tereza; Smelcova, Miroslava; Petr, Jaroslav; Bodart, Jean-Francois; Sedmikova, Marketa; Nevoral, Jan; Dvorakova, Marketa; Vyskocilova, Alena; Weingartova, Ivona; Kucerova-Chrpova, Veronika; Chmelikova, Eva; Tumova, Lenka; Jilek, Frantisek

2015-01-01

151

Preliminary characterization of calcium binding to melano-somes isolated from amphibian oocytes  

E-print Network

Preliminary characterization of calcium binding to melano- somes isolated from amphibian oocytes. The steroid hormone, progesterone, induces meiotic maturatilon of the full- grown amphibian oocyte (Smith

Paris-Sud XI, Université de

152

Interactions between oocytes and cumulus cells during in vitro maturation of porcine cumulus-oocyte complexes in a chemically defined medium: effect of denuded oocytes on cumulus expansion and oocyte maturation.  

PubMed

The aim of the present study was to clarify interactions between oocytes and cumulus cells (CCs) on the level of cumulus expansion and oocyte maturation during IVM of cumulus-oocyte complexes (COCs) in a chemically defined medium using a system that allows individual tracking of oocytes. Especially, the influence of oocyte-secreted factors was investigated by the aid of addition of denuded oocytes (DOs) as a possible approach to improve the IVM system. The basic maturation medium was porcine oocyte medium with addition of gonadotropins only during the first 20 hours of IVM. During IVM, COCs were kept fixed to the bottom of culture dish by adhesive Cell-Tak coating, which enabled individual tracking of COCs during IVM. Size changes in COCs during IVM were measured by digital image analysis. Cumulus expansion in a porcine oocyte medium of intact COCs increased in a typical manner until 20 hours and decreased in size subsequently until 48 hours of IVM (P < 0.05). Removal of oocytes from COCs by oocytectomy allowed the expansion of CCs to some extent, although their expansion ability was lower than that of COCs (P < 0.05). Addition of DOs (COCs to DOs ratio of 9:16) did not improve cumulus expansion and oocyte maturation rates of intact COCs (P > 0.05) but did enhance cumulus expansion of oocytectomized complexes (P < 0.05). Furthermore, removal of CCs before IVM increased oocyte maturation rates compared with COCs (52.3% and 32.9%, respectively) (P < 0.05) and a similar effect was observed in COCs when the gap junction inhibitor carbenoxolone was added to the IVM medium: carbenoxolone repressed the expansion of COCs at 20 hours of IVM. In conclusion, the porcine oocyte enhances cumulus expansion both by gap junctional communications and presumably by oocyte-secreted factor production. Nevertheless, the presence of oocytes is not a prerequisite for this process. In return, CCs maintain meiotic arrest in cumulus-enclosed oocytes during the initial culture through gap junctions. On the basis of these findings, future research could investigate if coculture with DOs during IVM is beneficial for fertilization and embryo development. PMID:25467769

Appeltant, R; Somfai, T; Nakai, M; Bodó, S; Maes, D; Kikuchi, K; Van Soom, A

2015-03-01

153

Cryopreservation of zebrafish (Danio rerio) oocytes by vitrification.  

PubMed

Cryopreservation of fish oocytes is challenging because these oocytes have low membrane permeability to water and cryoprotectant and are highly chilling sensitive. Vitrification is considered to be a promising approach for their cryopreservation as it involves rapid freezing and thawing of the oocytes and therefore minimising the chilling injury. In the present study, vitrification properties and the toxicity of a range of vitrification solutions containing different concentrations of Me2SO, methanol, propylene glycol and ethylene glycol were investigated. Two different base media and vitrification methods were compared. The effect of different post-thaw dilution solutions together with incubation periods on oocyte viability were also investigated. Stage III zebrafish oocytes were equilibrated in increasing concentrations of cryoprotectants for 30 min in 3 steps. Oocytes were thawed rapidly in a water bath and cryoprotectants were removed in 4 steps. Oocyte viability was assessed using trypan blue staining. The results showed that vitrification solutions V3 and V4 in KCl buffer had low toxicity and vitrified well. The survivals of oocytes after stepwise dilution using solutions containing permeable cryoprotectants were significant higher than those diluted in 0.5M glucose, and the use of CVA65 vitrification system improved oocyte survival when compared with plastic straws after 30 min at 22 degrees C post-thawing. Cryopreservation of zebrafish oocytes by vitrification is reported here for the first time, although oocyte survivals after cryopreservation assessed by trypan blue staining were relatively high shortly after thawing, they became swollen and translucent after incubation in KCl buffer. Further studies are needed to optimise the post-thaw culturing conditions. PMID:20919452

Guan, M; Rawson, D M; Zhang, T

2010-01-01

154

The molecular clock ticks regularly in muroid rodents and hamsters  

Microsoft Academic Search

Summary Extensive DNA sequence data are used to compare the rates of nucleotide substitution in the mouse, rat, and hamster lineages. A relative rate test using hamster sequences as references shows that the rates of synonymous and nonsynonymous substitution in the mouse and rat lineages are nearly equal and a test using human sequences as references shows that the rates

Colin O'hUigin; Wen-Hsiung Li

1992-01-01

155

Serum Melatonin Response to Melatonin Administration in the Syrian Hamster  

Microsoft Academic Search

Chronic daily administration of melatonin (MT) can have potent effects on reproduction in the hamster. Various theories have been elaborated to explain these effects but little information has been available on circulating levels of MT following MT administration. We have examined the serum MT response in the male hamster to a single dose of 25 ?g MT administered in the

Gregory M. Brown; J. Seggie; Lee J. Grota

1985-01-01

156

Lipid Deposition in Oocytes of Teleost Fish During Secondary Oocyte Growth  

Microsoft Academic Search

Research pertaining to the deposition of lipids into the maturing oocytes of teleost fish has progressed significantly since the review by Wiegand (1996). Studies on broodstock diets have elucidated the importance of incorporating n-3 and n-6 highly unsaturated fatty acids (HUFAs) into diets at the proper levels and ratios for optimal egg viability and survival of progeny. The vitellogenin receptor,

Ronald B. Johnson

2009-01-01

157

A penetration mechanics database  

NASA Astrophysics Data System (ADS)

A penetration mechanics database has been compiled that contains experimental results from a variety of researchers of different nationalities (German, French, English, Canadian, and American), during different decades (the 1960's to the 1990's), and with different purposes or objectives (space debris impact, armor design and evaluation, penetrator design and evaluation, and theoretical verification). The data fall naturally into three subgroupings: (1) penetration into semi-infinite targets; (2) perforation of finite-thickness targets; and (3) penetration from multiple impact (segmented rods). Data collection was primarily limited to experiments conducted against generic type targets, and emphasis was placed on data that would be more relevant to heavy armor issues. This diverse collection of data for metallic targets has been tabulated; data tables include the initial impact conditions, the projectile and target geometries and materials, and the measured response. Information on the materials, e.g., hardness of yield strength, along with impact information such as yaw has been tabulated if available. Graphical displays of the data have been used to summarize the data, and cross-plotting that combines data from different sources has also been provided.

Anderson, Charles E., Jr.; Morris, Bruce L.; Littlefield, David L.

1992-01-01

158

Radiation penetration test  

Microsoft Academic Search

Radiation penetration tests are one of the nondestructive testing ; methods which are employed as a method of detecting internal defects with ; ultrasonic flaw detection. Of these, radiography is the most widely used method ; for defect detection, which produces two dimensional images of defects on x-ray ; films. Experimental investigations have been carried out for the defects to

Kannoo

1973-01-01

159

Single wall penetration equations  

NASA Technical Reports Server (NTRS)

Five single plate penetration equations are compared for accuracy and effectiveness. These five equations are two well-known equations (Fish-Summers and Schmidt-Holsapple), two equations developed by the Apollo project (Rockwell and Johnson Space Center (JSC), and one recently revised from JSC (Cour-Palais). They were derived from test results, with velocities ranging up to 8 km/s. Microsoft Excel software was used to construct a spreadsheet to calculate the diameters and masses of projectiles for various velocities, varying the material properties of both projectile and target for the five single plate penetration equations. The results were plotted on diameter versus velocity graphs for ballistic and spallation limits using Cricket Graph software, for velocities ranging from 2 to 15 km/s defined for the orbital debris. First, these equations were compared to each other, then each equation was compared with various aluminum projectile densities. Finally, these equations were compared with test results performed at JSC for the Marshall Space Flight Center. These equations predict a wide variety of projectile diameters at a given velocity. Thus, it is very difficult to choose the 'right' prediction equation. The thickness of a single plate could have a large variation by choosing a different penetration equation. Even though all five equations are empirically developed with various materials, especially for aluminum alloys, one cannot be confident in the shield design with the predictions obtained by the penetration equations without verifying by tests.

Hayashida, K. B.; Robinson, J. H.

1991-01-01

160

cAMP modulation during sheep in vitro oocyte maturation delays progression of meiosis without affecting oocyte parthenogenetic developmental competence.  

PubMed

Removal of oocytes from their natural inhibitory follicular environment results in spontaneous resumption of meiosis independent of normal signaling events that occur in vivo. Controlling the onset of meiotic resumption via maintenance of elevated oocyte cAMP levels with adenylyl cyclase (AC) activation and phosphodiesterase (PDE) inhibition, and subsequent hormone stimulation with follicle FSH has been shown to dramatically improve developmental competence of bovine and murine IVM oocytes. This study evaluated the effect of cAMP modulation during IVM of sheep oocytes on meiotic progression and development to blastocyst after parthenogenetic activation. Changes in oocyte cAMP levels were quantified during the first 2h of in vitro maturation in control or cAMP-modulating medium. No significant changes in intra-oocyte cAMP were observed under control conditions, though a slight and transient drop was noticed at 15 min of maturation. Addition of the AC stimulator Forskolin and the PDE inhibitors IBMX altered the cAMP profile, resulting in 10-fold elevation of cAMP by 15 min and sustained >3-fold elevated levels from 30 to 120 min. The effect of cAMP elevation on meiotic resumption was measured by completion of germinal vesicle breakdown. Modulated oocytes were significantly delayed when compared to control media oocytes. Also, progression to MII was significantly delayed in modulated versus control oocytes at 20 and 24h, though no differences persisted to 28 h. Lastly, when control and modulated oocytes were parthenogenetically activated, no differences in blastocyst formation were observed. Thus, while cAMP modulation delayed meiotic progression, it did not improve developmental competence of sheep IVM oocytes. PMID:25595334

Buell, Margaret; Chitwood, James L; Ross, Pablo J

2015-03-01

161

Original article The germinal vesicle of the mouse oocyte contains  

E-print Network

Original article The germinal vesicle of the mouse oocyte contains elements of the phosphoinositide in the germinal vesicle (GV). Using confocal laser scanning microscopy, we analysed the effects of the nuclear/Elsevier, Paris oocyte / nucleus / calcium / IP3 receptors / PLC Résumé ― La vésicule germinative de l

Boyer, Edmond

162

Selective protein incorporation by vitellogenic Salmo gairdneri oocytes in vitro  

E-print Network

the selectivity of protein incorporation. Materials and methods. Donor fish and preparation of oocytes. HatcherySelective protein incorporation by vitellogenic Salmo gairdneri oocytes in vitro C. M. CAMPBELL B Josas, France. Summary. An in vitro method is presented for investigation of protein incorporation

Boyer, Edmond

163

The type and extent of injuries in vitrified mouse oocytes.  

PubMed

To improve the vitrification of mouse oocytes using straws, we attempted to estimate the type and extent of injuries during vitrification with a vitrification solution EAFS10/10. Injuries in oocytes were assessed based on cellular viability, the integrity of the plasma membrane, the status of the meiotic spindle/chromosomes, and morphological appearance. For morphologically normal oocytes, the ability to be fertilized and to develop into blastocysts was examined. Morphological assessment revealed 15% of oocytes to be injured by intracellular ice formed during vitrification, and 10% by osmotic swelling during removal of the cryoprotectant. When assessed by the status of spindles/chromosomes, the most sensitive criterion, damage was found in 16% of oocytes without any treatment. This value was similar to the proportion of fresh oocytes that did not cleave after insemination (13%). On exposure to EAFS10/10, the spindles/chromosomes were affected in 33% of oocytes. The exposure reduced the rate of cleavage by 18% points and the rate of development into blastocysts by 19 points. Vitrification reduced these rates by 15% and 36% points, respectively. Although the mechanism responsible for this moderate toxic effect on developmental ability is not known, information obtained in the present study will be useful to develop a practical method for the vitrification of mouse oocytes using straws. PMID:22202671

Liang, Yang; Ning, Fang-Yong; Du, Wen-Jing; Wang, Chun-Sheng; Piao, Shan-Hua; An, Tie-Zhu

2012-04-01

164

Original article Cytogenetic analysis of horse oocytes matured  

E-print Network

by in vitro fertilization (IVF) has become a source of embryos for embryo transfer (ET) and geneticOriginal article Cytogenetic analysis of horse oocytes matured in vitro for different periods by aspiration and maturation in vitro for 24, 30, 36 or 42 h. A total of 522 oocytes were recovered from 2211

Paris-Sud XI, Université de

165

CYTOPLASMIC AND NUCLEAR MATURATION OF RABBIT OOCYTES IN VITRO  

E-print Network

of oocytes having thus finished nuclear matu- ration in vitro when analyzed shows that fertilization developsCYTOPLASMIC AND NUCLEAR MATURATION OF RABBIT OOCYTES IN VITRO C. THIBAULT Micheline GÃ?RARD Station by normal fertilization and growth of the male pro- nucleus. Thus, two steps may be considered in complete

Boyer, Edmond

166

On-chip enucleation of an oocyte by untethered microrobots  

NASA Astrophysics Data System (ADS)

We propose a novel on-chip enucleation of an oocyte with zona pellucida by using a combination of untethered microrobots. To achieve enucleation within the closed space of a microfluidic chip, two microrobots, a microknife and a microgripper were integrated into the microfluidic chip. These microrobots were actuated by an external magnetic force produced by permanent magnets placed on the robotic stage. The tip of the microknife was designed by considering the biological geometric feature of an oocyte, i.e. the oocyte has a polar body in maturation stage II. Moreover, the microknife was fabricated by using grayscale lithography, which allows fabrication of three-dimensional microstructures. The microgripper has a gripping function that is independent of the driving mechanism. On-chip enucleation was demonstrated, and the enucleated oocytes are spherical, indicating that the cell membrane of the oocytes remained intact. To confirm successful enucleation using this method, we investigated the viability of oocytes after enucleation. The results show that the production rate, i.e. the ratio between the number of oocytes that reach the blastocyst stage and the number of bovine oocytes after nucleus transfer, is 100%. The technique will contribute to complex cell manipulation such as cell surgery in lab-on-a-chip devices.

Ichikawa, Akihiko; Sakuma, Shinya; Sugita, Masakuni; Shoda, Tatsuro; Tamakoshi, Takahiro; Akagi, Satoshi; Arai, Fumihito

2014-09-01

167

In Vitro Growth and Maturation of Vitrified-Warmed Bovine Oocytes Collected from Early Antral Follicles  

PubMed Central

Abstract. Cryopreservation of growing oocytes enriches the choice of timing and location of artificial embryo production. However, completion of oocyte growth after warming is crucial when using such cryopreserved oocytes. Our research objective was to develop a sequential system that incorporates cryopreservation of growing bovine oocytes and their subsequent in vitro growth. Oocyte-granulosa cell complexes with a mean oocyte diameter of approximately 100 µm were vitrified-warmed and then cultured for 14 days. The percentage of surviving oocytes following cryopreservation and 14-day culture was approximately 80%. More than half of the surviving oocytes were capable of maturing to metaphase II after in vitro maturation; the rate was comparable to that of control oocytes grown in vitro without cryopreservation. Taken together, the combined protocols for vitrification-warming of growing oocytes and subsequent in vitro growth can produce oocytes capable of undergoing meiotic maturation. PMID:24126072

HIRAO, Yuji; SOMFAI, Tamás; NARUSE, Kenji

2013-01-01

168

Mode of oocyte maturation affects EGF-like peptide function and oocyte competence.  

PubMed

The function and impact of epidermal growth factor (EGF)-like peptide signalling during ovulation and in vivo oocyte maturation (IVV) has been recently characterized, however, little is currently known about the effect of oocyte in vitro maturation (IVM) on this pathway. The aim of this study was to examine expression and functional aspects of three EGF-like peptides (amphiregulin, epiregulin and betacellulin) and their common receptor (EGFR) in cumulus cells during mouse oocyte IVM compared with IVV. Cumulus-oocyte complexes (COCs) were collected from prepubertal mice either 46 h post-eCG (IVM) or 46 h post-eCG plus 0.5-12 h post-hCG (IVV). Time course experiments showed mRNA expression of all three EGF-like peptides and amphiregulin protein in IVM media were significantly lower for the majority of FSH-supplemented IVM compared with IVV. The supplementation of EGF during IVM yielded EGF-like peptide expression levels comparable with IVV and amphiregulin/epiregulin supplemented IVM. However, despite this, EGF activation of the COC EGFR remained significantly lower at 3 and 6 h of IVM than in vivo, and levels were similar to those observed during FSH-supplemented IVM. The addition of exogenous epiregulin during IVM significantly increased blastocyst rates, and epiregulin and amphiregulin improved blastocyst quality, compared with FSH or EGF. In conclusion, findings from this study suggest that the widely used IVM additives, FSH and EGF, are inadequate propagators of the essential EGF-like peptide signalling cascade. In contrast, the use of epiregulin and/or amphiregulin during IVM leads to improved oocyte developmental competence and therefore may be preferable IVM additives than FSH or EGF. PMID:23594928

Richani, D; Ritter, L J; Thompson, J G; Gilchrist, R B

2013-08-01

169

Kinetics of Leptospira interrogans Infection in Hamsters after Intradermal and Subcutaneous Challenge  

PubMed Central

Background Leptospirosis is a zoonosis caused by highly motile, helically shaped bacteria that penetrate the skin and mucous membranes through lesions or abrasions, and rapidly disseminate throughout the body. Although the intraperitoneal route of infection is widely used to experimentally inoculate hamsters, this challenge route does not represent a natural route of infection. Methodology/Principal Findings Here we describe the kinetics of disease and infection in hamster model of leptospirosis after subcutaneous and intradermal inoculation of Leptospira interrogans serovar Copenhageni, strain Fiocruz L1-130. Histopathologic changes in and around the kidney, including glomerular and tubular damage and interstitial inflammatory changes, began on day 5, and preceded deterioration in renal function as measured by serum creatinine. Weight loss, hemoconcentration, increased absolute neutrophil counts (ANC) in the blood and hepatic dysfunction were first noted on day 6. Vascular endothelial growth factor, a serum marker of sepsis severity, became elevated during the later stages of infection. The burden of infection, as measured by quantitative PCR, was highest in the kidney and peaked on day 5 after intradermal challenge and on day 6 after subcutaneous challenge. Compared to subcutaneous challenge, intradermal challenge resulted in a lower burden of infection in both the kidney and liver on day 6, lower ANC and less weight loss on day 7. Conclusions/Significance The intradermal and subcutaneous challenge routes result in significant differences in the kinetics of dissemination and disease after challenge with L. interrogans serovar Copenhageni strain Fiocruz L1-130 at an experimental dose of 2×106 leptospires. These results provide new information regarding infection kinetics in the hamster model of leptospirosis. PMID:25411782

Coutinho, Mariana L.; Matsunaga, James; Wang, Long-Chieh; de la Peña Moctezuma, Alejandro; Lewis, Michael S.; Babbitt, Jane T.; Aleixo, Jose Antonio G.; Haake, David A.

2014-01-01

170

Male Syrian hamsters demonstrate a conditioned place preference for sexual behavior and female chemosensory stimuli  

E-print Network

Male Syrian hamsters demonstrate a conditioned place preference for sexual behavior and female induce a CPP in male Syrian hamsters. As male Syrian hamsters are an animal model commonly used. Experiment 1 tested the prediction that male hamsters show a CPP for sexual behavior. Female chemosensory

Sisk, Cheryl

171

Sympathoadrenal System Differentially Affects Photoperiodic Changes in Humoral Immunity of Siberian Hamsters  

E-print Network

Hamsters (Phodopus sungorus) G. E. Demas*, D. L. Drazen², A. M. Jasnow³, T. J. Bartness³§ and R. J. Nelson, antibodies, catecholamines, norepinephrine. Abstract Siberian hamsters (Phodopus sungorus) rely catecholamines, to photoperiodic changes in immune function in male Siberian hamsters. In Experiment 1, hamsters

Demas, Greg

172

Social interactions differentially affect reproductive and immune responses of Siberian hamsters  

E-print Network

Social interactions differentially affect reproductive and immune responses of Siberian hamsters in Siberian hamsters (Phodopus sungorus). Male and female hamsters were housed alone, in same-sex pairsM, and mitogen-stimulated splenocyte proliferation. Male hamsters housed with a female had increased testosterone

Demas, Greg

173

Zona-free hamster ovum penetration by human spermatozoa : influence of various sperm factors  

E-print Network

. Higher in vitro fertilization rates were obtained after sperm selection by « swim-up» migration, 4-h-ovum interaction seem to be analogous in heterospecific in vitro fertilization. In 1976, Yanagimachi et al. Mammalian fertilization needs intraspecific gamete recognition. However, some stages of sperm

Boyer, Edmond

174

Oocyte-specific expression of Gpr3 is required for the maintenance of meiotic arrest in mouse oocytes  

E-print Network

Oocyte-specific expression of Gpr3 is required for the maintenance of meiotic arrest in mouse, at least in large part, by the Gs-linked G-protein-coupled receptor, GPR3. Gpr3 is localized in the mouse oocyte but is also present throughout the follicle. To investigate whether Gpr3 in the follicle cells

Terasaki, Mark

175

Selection of follicles, preculture oocyte evaluation, and duration of culture for in vitro maturation of equine oocytes  

Microsoft Academic Search

Equine oocytes (n = 537) were collected from slaughterhouse ovaries (n = 118 mares) by scraping the internal follicular wall. Preculture record was made of the appearance of oocyte investments (no cumulus, corona radiata only, compact cumulus, expanded cumulus), appearance of cytoplasm (homogeneous, condensed heterogeneous\\/fragmented), and nuclear maturation stages (germinal vesicle, germinal-vesicle breakdown, metaphase I, metaphase II, degenerated). There was

M. R. Del Campo; X. Donoso; J. J. Parrish; O. J. Ginther

1995-01-01

176

In Vitro Matured Oocytes Are More Susceptible than In Vivo Matured Oocytes to Mock ICSI Induced Functional and Genetic Changes  

PubMed Central

Background Concerns regarding the safety of ICSI have been intensified recently due to increased risk of birth defects in ICSI born children. Although fertilization rate is significantly higher in ICSI cycles, studies have failed to demonstrate the benefits of ICSI in improving the pregnancy rate. Poor technical skill, and suboptimal in vitro conditions may account for the ICSI results however, there is no report on the effects of oocyte manipulations on the ICSI outcome. Objective The present study elucidates the influence of mock ICSI on the functional and genetic integrity of the mouse oocytes. Methods Reactive Oxygen Species (ROS) level, mitochondrial status, and phosphorylation of H2AX were assessed in the in vivo matured and IVM oocytes subjected to mock ICSI. Results A significant increase in ROS level was observed in both in vivo matured and IVM oocytes subjected to mock ICSI (P<0.05-0.001) whereas unique mitochondrial distribution pattern was found only in IVM oocytes (P<0.01-0.001). Importantly, differential H2AX phosphorylation was observed in both in vivo matured and IVM oocytes subjected to mock ICSI (P <0.001). Conclusion The data from this study suggests that mock ICSI can alter genetic and functional integrity in oocytes and IVM oocytes are more vulnerable to mock ICSI induced changes. PMID:25786120

Salian, Sujit Raj; Singh, Vikram Jeet; Kalthur, Guruprasad; Adiga, Satish Kumar

2015-01-01

177

Clostridium difficile infection in adult hamsters.  

PubMed

Diarrhea was encountered in a group of adult female golden Syrian hamsters (Mesocricetus auratus) used for titrating the scrapie agent. Ninety percent of the cases occurred in animals over 210 days old even though animals of all age groups lived in the colony concurrently. The cause of diarrhea was investigated in both uninoculated animals and those receiving greater than a limiting dilution of scrapie infectivity, i.e., animals that were not expected to contract the experimental scrapie disease. Three forms of diarrhea were observed. The most commonly encountered was profuse and watery. A chronic form presented with semiformed, thin fecal material smearing the retroperitoneal region. Hemorrhagic diarrhea was observed rarely. Mortality was high among animals with acute watery or hemorrhagic diarrhea. Animals with semiformed soft stools were dehydrated, had a roughened hair-coat, and hunched back. Cardinal lesions were necrosis, inflammation, and mucosal hyperplasia of the cecum and colon and cholangiohepatitis with amyloid deposition. Diffuse renal amyloidosis was present in chronic cases. Toxigenic, cytotoxin B-positive Clostridium difficile was isolated from a majority of affected animals. Cytotoxin B was also present in cecal homogenates of diarrheic animals with C. difficile. The pathological and microbiologic findings indicated a typhlitis and colitis in adult hamsters that was associated with C. difficile infection. PMID:1667195

Chang, J; Rohwer, R G

1991-12-01

178

Wee1B depletion promotes nuclear maturation of canine oocytes.  

PubMed

Most mammalian oocytes are arrested at the germinal vesicle stage by activation of Wee1B. Meiotic resumption is regulated by inactivation of Wee1B and activation of cell division cycle 25B. The aim of this study was to determine whether treatment with Wee1B-targeting small interfering RNA (Wee1B-siRNA) promotes nuclear maturation of canine oocytes from germinal vesicle stage to metaphase II (MII) stage. In experiment 1, the percentage of canine oocytes that matured to MII stage was higher (P < 0.05) among oocytes cultured in vitro for 72 hours than among those cultured for 24 and 48 hours (5.4 ± 2.5% vs. 0.0 ± 0.0% and 1.4 ± 1.0%, respectively). Furthermore, the percentage of oocytes that matured to metaphase I (MI) stage was higher (P < 0.05) among oocytes cultured for 48 and 72 hours than among those cultured for 24 hours (14.9 ± 10.0% and 22.4 ± 8.1%, respectively, vs. 5.7 ± 6.0%). In experiment 2, canine oocytes were intracytoplasmically microinjected with Wee1B-siRNA (50 ?M) at various culture time points (0, 24, 48, or 72 hours). The nuclear configuration of the exception of oocytes in the 72-hour group was examined after 84 hours of culture. The percentage of oocytes that matured to the MII stage was higher (P < 0.05) among those treated with Wee1B-siRNA at 0 hours than among control oocytes and those injected at 72 hours (18.0 ± 1.7% vs. 2.1 ± 2.8% and 0.0 ± 0.0%, respectively). Moreover, the percentage of oocytes that matured to the MI stage was higher (P < 0.05) among those injected at 0 hours than among control oocytes and those injected at 24 and 72 hours (45.9 ± 6.8% vs. 22.1 ± 3.5%, 22.8 ± 10.0%, and 10.0 ± 4.4%, respectively). In experiment 3, oocytes were intracytoplasmically microinjected with Wee1B-siRNA at 0 hours of IVM and cultured for 0, 24, 48, or 72 hours. Thereafter, maturation-related gene expression was analyzed by quantitative real-time polymerase chain reaction. Messenger RNA expression of cAMP and cell division cycle 25B was lower (P < 0.05) in oocytes injected at 48 hours than in the other groups. Messenger RNA expression of cAMP was lower (P < 0.05) in oocytes injected at 0 hours than in control oocytes and those injected at 72 hours. Messenger RNA expression of mitogen-activated protein kinase 1 and mitogen-activated protein kinase 3 was higher (P < 0.05) in oocytes injected at 72 hours than in the other groups. In conclusion, we confirmed that Wee1B-siRNA microinjection enhances the percentages of canine oocytes that reach the MI and MII stages. These data suggest that Wee1B-siRNA microinjection could be a useful strategy to obtain mature canine oocytes for research and assisted canine reproduction. PMID:25457679

Kim, Yu-Gon; Kim, Dong-Hoon; Song, Seok-Hwan; Lee, Kyeong-Lim; Yang, Byoung-Chul; Oh, Jeong Su; Lee, Sang-Ryeul; Kong, Il-Keun

2015-03-01

179

Mars penetrator: Subsurface science mission  

NASA Technical Reports Server (NTRS)

A penetrator system to emplace subsurface science on the planet Mars is described. The need for subsurface science is discussed, and the technologies for achieving successful atmospheric entry, Mars penetration, and data retrieval are presented.

Lumpkin, C. K.

1974-01-01

180

Levels of salivary lysozyme, lactoperoxidase, and lactoferrin in diabetic hamsters.  

PubMed Central

In an attempt to clarify the mechanism(s) of increased susceptibility to oral infection in diabetics, we examined the levels of salivary antibacterial factors, including lysozyme, lactoperoxidase, and lactoferrin, in diabetic hamsters whose condition was induced with streptozotocin. Saliva was collected from these hamsters periodically for 19 weeks after the administration of streptozotocin. Diabetes persisted with significant hyperglycemia throughout the experiment after a single injection of streptozotocin. There was no significant difference between groups in the amount of saliva secreted. In diabetic hamsters, lysozyme activity decreased by 56% and lactoperoxidase activity decreased by 53% compared with the control hamsters 19 weeks after the administration of streptozotocin. There was no significant difference between groups in the amount of salivary lactoferrin. However, the ratio of lactoferrin to total protein increased to approximately double the amount of that of the control hamsters. Insulin treatment had a significant effect on lysozyme and lactoperoxidase activity, recovering 73 and 74% those of the controls, respectively, and the ratio of lactoferrin to total salivary protein reverted to normal values. Growth inhibition of Lactobacillus plantarum ATCC 8014 with whole saliva and amylase activity significantly decreased in diabetic hamsters. The position of each protein band of whole saliva on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was almost the same for control and diabetic hamsters; however, there was some variability in band intensity. Images PMID:2580790

Muratsu, K; Morioka, T

1985-01-01

181

Control of oocyte growth and maturation by follicular cells and molecules present in follicular fluid.  

E-print Network

Review Control of oocyte growth and maturation by follicular cells and molecules present the interactions between the oocyte and its sur- rounding granulosa cells which are involved in the control of oocyte growth or apoptosis as well as those playing a key role in the ability of the oocyte to undergo

Paris-Sud XI, Université de

182

The nuclear envelope in the developing oocytes of the Tunicate, Boltenia villosa  

Microsoft Academic Search

1.In the developing oocyte of Boltenia villosa, five distinct morphological manifestations have been observed, all involving the nuclear envelope. None of them is continuous throughout the growth of an oocyte but each occupies a definite period of oocyte development. These different manifestations reflecting different activities at the nucleo-cytoplasmic boundary in B. villosa oocytes are listed as follows, starting from very

W. Siang Hsu

1962-01-01

183

E. Rodrguez-Gonzlez et al.BCB-selected-goat oocytes matured withcysteamine Original article  

E-print Network

E. Rodríguez-González et al.BCB-selected-goat oocytes matured withcysteamine Original article Developmental competence of prepubertal goat oocytes selected with brilliant cresyl blue and matured maturation (IVM) medium to improve the in vitro embryo development of prepubertal goat oocytes. The oocytes

Paris-Sud XI, Université de

184

[Source of asymmetry in ontogeny: early polarization of the germline cyst and oocyte in Drosophila].  

PubMed

One or more body axes are already formed in the egg in many vertebrates and invertebrates. In Drosophila, the anteroposterior and dorsoventral axes are determined during oogenesis owing to the asymmetric localization of the bicoid, oskar, and gurken mRNAs in the oocyte (prospective egg). The localization of these transcripts depends on the polarized organization of the oocyte cytoskeleton and, consequently, the oocyte polarity. Initial asymmetry, leading to the oocyte polarity, is established in early ontogeny, during oocyte determination. The review considers the steps of early polarization and oocyte differentiation in Drosophila, the genetic control of these processes, and the findings that suggest an early oocyte polarity for vertebrates. PMID:18846812

Simonova, O B; Vorontsova, Iu E

2008-09-01

185

Murine follicular development requires oocyte DICER, but not DROSHA.  

PubMed

Both DICER and DROSHA are RNase III enzymes involved in the biogenesis of small noncoding RNAs. DROSHA cleaves the stem-loop portion of the primary miRNAs and produces precursor miRNAs in the nucleus, whereas DICER processes double-stranded RNA precursors into mature miRNAs and endogenous small interference RNAs in the cytoplasm. Selective inactivation of Dicer in growing oocytes of primary follicles leads to female infertility due to oocyte spindle defects. However, it remains unknown if oocyte Dicer expression in the fetal ovary is required for proper follicular development in the postnatal ovary. Moreover, the role of Drosha in folliculogenesis has never been investigated. Here, we report that conditional knockout of Dicer in prophase I oocytes of the fetal ovary led to compromised folliculogenesis, premature ovarian failure, and female infertility in the adult ovary, whereas selective inactivation of Drosha in oocytes of either the fetal or the developing ovary had no effects on normal folliculogenesis and female fertility in adulthood. Our data indicate that oocyte DICER expression in the fetal ovary is required, and oocyte DROSHA is dispensable, for postnatal follicular development and female fertility in adulthood. PMID:24990804

Yuan, Shuiqiao; Ortogero, Nicole; Wu, Qiuxia; Zheng, Huili; Yan, Wei

2014-08-01

186

Mitochondrial DNA deletion in human oocytes and embryos.  

PubMed

Mitochondrial DNA (mtDNA) deletions are present in both human oocytes and embryos. It has been found that these tissues contain a mtDNA mutation which is present in high amounts in patients with Kearns-Sayre syndrome (KSS) and progressive external ophthalmoplegia. In the present study, the frequency of this KSS deletion was investigated in human oocytes and embryos. Using a nested primer polymerase chain chian reaction (PCR) strategy, the frequency of the KSS deletion in 74 human oocytes and 137 embryos was found to be 32.8 and 8.0% respectively. Using a 'long PCR-short PCR' nested primer strategy, the frequency of the KSS deletion in 181 human oocytes and 104 embryos was found to be 47.0 and 20.2% respectively. There was no statistical correlation between the age of the patients at the time of oocyte retrieval and the presence of the deleted molecules. There was a statistical difference between the presence of the deleted molecules in oocytes versus embryos using either technique (P < 0.0001). The relevance of these findings to the accumulation of low levels of deleted mtDNA in both oocytes and embryos is discussed in this study. PMID:9783850

Brenner, C A; Wolny, Y M; Barritt, J A; Matt, D W; Munné, S; Cohen, J

1998-09-01

187

Polarity and asymmetry during mouse oogenesis and oocyte maturation.  

PubMed

Cell polarity and asymmetry play a fundamental role in embryo development. The unequal segregation of determinants, cues, and activities is the major event in the differentiation of cell fate and function in all multicellular organisms. In oocytes, polarity and asymmetry in the distribution of different molecules are prerequisites for the progression and proper outcome of embryonic development. The mouse oocyte, like the oocytes of other mammals, seems to apply a less stringent strategy of polarization than other vertebrates. The mouse embryo undergoes a regulative type of development, which permits the full rectification of development even if the embryo loses up to half of its cells or its size is experimentally doubled during the early stages of embryogenesis. Such pliability is strongly related to the proper oocyte polarization before fertilization. Thus, the molecular mechanisms leading to the development and maintenance of oocyte polarity must be included in any fundamental understanding of the principles of embryo development. In this chapter, we provide an overview of current knowledge regarding the development and maintenance of polarity and asymmetry in the distribution of organelles and molecules in the mouse oocyte. Curiously, the mouse oocyte becomes polarized at least twice during ontogenesis; the question of how this phenomenon is achieved and what role it might play is addressed in this chapter. PMID:22918799

Kloc, Malgorzata; Ghobrial, Rafik M; Borsuk, Ewa; Kubiak, Jacek Z

2012-01-01

188

Cryopreservation of immature bovine oocytes by vitrification in straws.  

PubMed

The aim of this study was to cryopreserve by vitrification by ethylene glycol (EG) and dimethyl sulfoxide (DMSO) immature bovine oocytes in straws and to investigate the effects of vitrification on post-thaw oocyte maturation. A total of 575 cumulus oocyte complexes were obtained by follicle aspiration from 238 ovaries of cows slaughtered at a local abattoir. Following selection, oocytes with compacted cumulus cells and evenly granulated ooplasm were vitrified using one of the three different solutions with a non-vitrified group served as control. The first step vitrification solution contained 20% EG while the second step solution contained 40% EG+1M sucrose in a basic media used in group EG. Oocytes were matured in N-2-hidroxyethyl piperazine-N-2-ethanosulfonic acid (HEPES) buffered tissue culture medium (TCM) 199 for 24h at 39 degrees C in a humidified atmosphere of 5% CO2 in air. Oocytes were fixed following evaluation for polar body formation, stained with Giemsa solution and nuclear maturation was examined. The numbers of oocytes which were observed at Metaphase II (MII) stage were 41 (34.1%), 17 (14.9%), 29 (20.7%) and 78 (79.6%) in groups EG, DMSO, Mix and Control, respectively. Maturation rate distribution in group Mix was not statistically different when compared to maturation rate distributions in groups EG and DMSO (p>0.05). Differences between other groups were significant (p<0.001). However, better results were obtained in EG group compared to DMSO and mix groups. Maturation rates were lower in all treatment groups than the control group. The lowest maturation result was obtained in DMSO group. Maturation rate in group Mix was between maturation rates of EG and DMSO groups. Immature bovine oocytes can be vitrified in straws, but maturation success differs with the cryoprotectant and it seems that to obtain better maturation rates, new cryopreservation techniques specific for immature bovine oocytes are needed. PMID:16019167

Cetin, Yunus; Bastan, Ayhan

2006-03-01

189

Protein patterns of pig oocytes during in vitro maturation.  

PubMed

In vitro maturation (IVM) of fully grown mammalian oocytes is characterized by initial germinal vesicle (GV) breakdown and rearrangement of microtubule network during the first meiosis (MI), followed by extrusion of the first polar body and block of the oocytes in metaphase of the second meiosis (MII). Only fully matured oocytes are capable of undergoing fertilization and the initiation of zygotic development. These observations are mostly based on morphological evaluation; however, the molecular events responsible for these processes are not known. In this study, we have launched the analysis of pig oocytes during in vitro maturation using a proteomics approach. First, oocyte proteins have been separated by two-dimensional gel electrophoresis and identified by mass spectrometry. Remarkably, several proteins, including peroxiredoxins, ubiquitin carboxyl-terminal hydrolase isozyme L1, and spermine synthase, are even more abundant than actin, usually the most abundant protein in somatic cells. Furthermore, we have initiated comparative analysis of the oocytes at different stages of maturation to characterize candidate proteins, which are differentially expressed during in vitro maturation. To date, we have identified antiquitin (D7A1), the member of aldehyde dehydrogenase family7 that has been significantly increased in MI and MII stages compared with GV oocytes. To our knowledge, this is the first pig oocyte proteome available so far that may be used as a reference map. The proteins that are differentially regulated during IVM may present potential biomarkers of oocyte maturation and quality. It is a useful inventory toward a deeper understanding of the mechanisms underlying reproduction and development. PMID:15229143

Ellederova, Zdenka; Halada, Petr; Man, Petr; Kubelka, Michal; Motlik, Jan; Kovarova, Hana

2004-11-01

190

THE KINETICS OF PENETRATION  

PubMed Central

When the only solute present is a weak acid, HA, which penetrates as molecules only into a living cell according to a curve of the first order and eventually reaches a true equilibrium we may regard the rate of increase of molecules inside as See PDF for Equation where PM is the permeability of the protoplasm to molecules, Mo, denotes the external and Mi the internal concentration of molecules, Ai denotes the internal concentration of the anion A- and See PDF for Equation (It is assumed that the activity coefficients equal 1.) Putting PMFM = VM, the apparent velocity constant of the process, we have See PDF for Equation where e denotes the concentration at equilibrium. Then See PDF for Equation where t is time. The corresponding equation when ions alone enter is See PDF for Equation. where K is the dissociation constant of HA, PA is the permeability of the protoplasm to the ion pair H+ + A-, and Aie denotes the internal concentration of Ai at equilibrium. Putting PAKFM = VA, the apparent velocity constant of the process, we have See PDF for Equation and See PDF for Equation When both ions and molecules of HA enter together we have See PDF for Equation where Si = Mi + Ai and Sie is the value of Si at equilibrium. Then See PDF for Equation VM, VA, and VMA depend on FM and hence on the internal pH value but are independent of the external pH value except as it affects the internal pH value. When the ion pair Na+ + A- penetrates and Nai = BAi, we have See PDF for Equation and See PDF for Equation where PNaA is the permeability of the protoplasm to the ion pair Na+ + A-, Nao and Nai are the external and internal concentrations of Na+, See PDF for Equation, and VNa is the apparent velocity constant of the process. Equations are also given for the penetration of: (1) molecules of HA and the ion pair Na+ + A-, (2) the ion pairs H+ + A- and Na+ + A-, (3) molecules of HA and the ion pairs Na+ + A- and H+ + A-. (4) The penetration of molecules of HA together with those of a weak base ZOH. (5) Exchange of ions of the same sign. When a weak electrolyte HA is the only solute present we cannot decide whether molecules alone or molecules and ions enter by comparing the velocity constants at different pH values, since in both cases they will behave alike, remaining constant if FM is constant and falling off with increase of external pH value if FM falls off. But if a salt (e.g., NaA) is the only substance penetrating the velocity constant will increase with increase of external pH value: if molecules of HA and the ions of a salt NaA. penetrate together the velocity constant may increase or decrease while the internal pH value rises. The initial rate See PDF for Equation (i.e., the rate when Mi = 0 and Ai = 0) falls off with increase of external pH value if HA alone is present and penetrates as molecules or as ions (or in both forms). But if a salt (e.g., NaA) penetrates the initial rate may in some cases decrease and then increase as the external pH value increases. At equilibrium the value of Mi equals that of Mo (no matter whether molecules alone penetrate, or ions alone, or both together). If the total external concentration (So = Mo + Ao) be kept constant a decrease in the external pH value will increase the value of Mo and make a corresponding increase in the rate of entrance and in the value at equilibrium no matter whether molecules alone penetrate, or ions alone, or both together. What is here said of weak acids holds with suitable modifications for weak bases and for amphoteric electrolytes and may also be applied to strong electrolytes. PMID:19872523

Osterhout, W. J. V.

1929-01-01

191

Donor motivations, associated risks and ethical considerations of oocyte donation.  

PubMed

Three decades after the first reported successful cases, oocyte donation continues to grow in popularity and regard as an established method to aid women in achieving their reproductive goals. As a result of the increased demand for donated oocytes, many young women in the U.S. volunteer to undergo complex medical procedures to donate their oocytes in return for financial compensation. To best care for these women before, during and after donation, it is important to explore donor characteristics and motivations, discuss the safety of the donation procedure and examine the ethical issues related to this process. PMID:24750650

Boutelle, Amy L

2014-01-01

192

A role for glucose in hypothermic hamsters  

NASA Technical Reports Server (NTRS)

Hypothermic hamsters at a rectal temperature of 7 C showed a fivefold increase in survival times from 20 to 100.5 hr when infused with glucose which maintained a blood level at about 45 mg/100 ml. A potential role for osmotic effects of the infusion was tested and eliminated. There was no improvement in survival of 3-O-methylglucose or dextran 40-infused animals. The fact that death eventually occurs even in the glucose-infused animal after about 4 days and that oxygen consumption undergoes a slow decrement in that period suggests that hypothermic survival is not wholly substrate limited. Radioactive tracer showed that localization of the C-14 was greatest in brain tissue and diaphragm, intermediate in heart and kidney, and lowest in skeletal muscle and liver. The significance of the label at sites important to respiration and circulation was presented.

Resch, G. E.; Musacchia, X. J.

1976-01-01

193

Deep penetration calculations. [LMFBR  

SciTech Connect

Several Monte Carlo techniques are compared in the transport of neutrons of different source energies through two different deep-penetration problems each with two parts. The first problem involves transmission through a 200-cm concrete slab. The second problem is a 90/sup 0/ bent pipe jacketed by concrete. In one case the pipe is void, and in the other it is filled with liquid sodium. Calculations are made with two different Los Alamos Monte Carlo codes: the continuous-energy code MCNP and the multigroup code MCMG.

Thompson, W.L.; Deutsch, O.L.; Booth, T.E.

1980-04-01

194

NEONATAL CHIORDECONE EXPOSURE ALTERS BEHAVIORAL SEX DIFFERENTIATION IN FEMALE HAMSTERS  

EPA Science Inventory

The present study was designed in order to determine if exposure to the weakly estrogenic pesticide Chlordecone during a critical period of behavioral sex differentiation of the brain could masculinize and defeminize the behavior of female hamsters....

195

Micro-bioreactor design for Chinese hamster ovary cells  

E-print Network

The research objective is to design a micro-bioreactor for the culture of Chinese Hamster Ovary (CHO) cells. There is an increasing demand for upstream development in high-throughput micro-bioreactors specifically for the ...

Goh, Shireen

2013-01-01

196

Adenovirus type 12 DNA sequences in primary hamster tumors.  

PubMed Central

In five out of six primary hamster tumors induced by adenovirus type 12, less than 55% of the adenovirus type 12 genome is present. Various fragments of the integrated viral DNA were present in non-equimolar amounts. PMID:904032

Lee, K C; Mak, S

1977-01-01

197

Dancing Hamsters and Marble Statues: Characterizing Student Visualizations of Algorithms  

E-print Network

Dancing Hamsters and Marble Statues: Characterizing Student Visualizations of Algorithms Teresa-created visualizations. However, constructionist and situated theories of learning suggest that students should develop studies in which students engaged in these activities are discussed. The resulting learning benefits

Narayanan, N. Hari

198

Evidence for a metabolic limitation of survival in hypothermic hamsters.  

NASA Technical Reports Server (NTRS)

The underlying factors limiting survival in the hypothermic state are studied. Hamsters of both sexes, clipped and unclipped, were inducted into profound hypothermia by the helium cold method until they reached a temperature between 7 and 10 C. It appears that the primary cause of death is failure of respiration due to the depletion of carbohydrate energy supplies and may explain why survival time in hypothermia is shorter than the normal hibernation time of the hamster.

Prewitt, R. L.; Anderson, G. L.; Musacchia, X. J.

1972-01-01

199

Monolithic ballasted penetrator  

DOEpatents

The present invention is a monolithic ballasted penetrator capable of delivering a working payload to a hardened target, such as reinforced concrete. The invention includes a ballast made from a dense heavy material insert and a monolithic case extending along an axis and consisting of a high-strength steel alloy. The case includes a nose end containing a hollow portion in which the ballast is nearly completely surrounded so that no movement of the ballast relative to the case is possible during impact with a hard target. The case is cast around the ballast, joining the two parts together. The ballast may contain concentric grooves or protrusions that improve joint strength between the case and ballast. The case further includes a second hollow portion; between the ballast and base, which has a payload fastened within this portion. The penetrator can be used to carry instrumentation to measure the geologic character of the earth, or properties of arctic ice, as they pass through it.

Hickerson, Jr., James P. (Cedar Crest, NM); Zanner, Frank J. (Sandia Park, NM); Baldwin, Michael D. (Albuquerque, NM); Maguire, Michael C. (Worcester, MA)

2001-01-01

200

Regulation of hamster splenocyte reactivity to concanavalin A during pregnancy  

SciTech Connect

The survival to term of mammalian fetuses regardless of their expression of paternal or embryonic developmental antigens suggests that some alteration in the immune capabilities of a female occur during pregnancy. The immunocompetence of female Syrian golden hamsters during pregnancy was investigated with respect to the blastogenic response of spleen cells to the T-cell mitogen concanavalin A (Con A). The blastogenic response of spleen cells from pregnant hamsters during mid- or late gestation is 10% of that observed for spleen cells from age-matched, virgin female animals. The spleen cells from pregnant hamsters are not capable of suppressing the proliferative response of spleen cells from virgin females to Con A. However, the serum from pregnant hamsters, in comparison with serum from virgin female animals, is capable of reducing this mitogenic response. Extensive washing of the splenocytes from pregnant hamsters does reduce the degree of suppression. These results suggest that the hamster is an excellent animal model for the investigation of the mechanism(s) of immune regulation that operate during pregnancy.

Weppner, W.A.; Coggin, J.H. Jr.

1980-08-15

201

Regulation of tonic gonadotropin release in prepubertal female hamsters  

SciTech Connect

Basal serum gonadotropin levels were monitored weekly in female hamsters from birth to 10 weeks of age. Hamsters raised on three different photoperiods presented uniform pre- and postpubertal patterns of serum LH and FSH, suggesting that gonadotropin release in the young hamster occurs independently of ambient photoperiod. In all groups, serum LH levels increased gradually in animals up to 4 weeks of age, after which levels plateaued at 50--100 ng/ml. Serum FSH was markedly elevated in 2- and 3-week-old hamsters (800--1200 ng/ml), but remained at 200--400 ng/ml in all other groups. We next examined the change in the responsiveness of the pituitary to exogenous gonadotropin-releasing hormone (GnRH) challenge. Female hamsters 2 days of age failed to respond to any dose (0.025--1000 ng) of GnRH, while 10-day old females responded in typical dose-dependent fashion. GnRH-stimulated LH release first occurred in 6-day-old hamsters and was maximal by day 9, whereas FSH release first occurred on day 8 and was maximal by day 9. The prepubertal pattern of gonadotropin release can, in part, be explained on the basis of the development of pituitary GnRH sensitivity, which occurs independently of photoperiod.

Smith, S.G.; Matt, K.S.; Prestowitz, W.F.; Stetson, M.H.

1982-04-01

202

Pressure Measurement during Penetration Experiments  

NASA Astrophysics Data System (ADS)

Penetration experiments are common tools for the investigation of physical surface properties. Additionally penetration experiments will find several applications in exploration missions in the near future. A penetration test stand has been flown for the investigation of penetration force reduction under reduced gravity in the 2nd Joint European Partial-G Parabolic Flight Campaign (JEPPF-2) of ESA, CNES and DLR [1]. The main contribution to the bearing resistance of a soil is combined of shaft and base resistance. During the penetration the grains of the granular material will be squeezed into the surrounding material. The penetration will cause a change in the pressure distribution inside the surrounding soil [2],[3]. An experimental setup has been designed and built for understanding and measurement of this induced pressure distribution. In the last year the parabolic flight test stand has been further developed for the measurement of pressure during the penetration process. The main part of the experiments stayed the same with a steel rod penetration into a sample cell measuring the penetration force and recording it in relation to the depth. The sample cell is equipped with a supporting sieving mechanism for sample preparation. The pressure sensors are mounted at the sample cell. During the last test campaigns the principle of measurement has been investigated and first measurements have been performed. In the presentation the measurement principle will be shown and its implementation into the parabolic flight setup. Pressure measurement results on ground tests of different penetrator and tip configurations will be presented.

Krause, C.; Demming, J.; Flecht, T.; Heller, S.

2014-04-01

203

Recent advances in oocyte and ovarian tissue cryopreservation and transplantation  

PubMed Central

Options for preserving fertility in women include well-established methods such as fertility-sparing surgery, shielding to reduce radiation damage to reproductive organs, and emergency in-vitro fertilisation after controlled ovarian stimulation, with the aim of freezing embryos. The practice of transfering frozen or thawed embryos has been in place for over 25 years, and today is a routine clinical treatment in fertility clinics. Oocytes may also be frozen unfertilised for later thawing and fertilisation by intracytoplasmic sperm injection in vitro. In recent years, oocyte cryopreservation methods have further developed, reaching promising standards. More than 1000 children are born worldwide after fertilisation of frozen and thawed oocytes. Nevertheless, this technique is still considered experimental. In this chapter, we focus on options for fertility preservation still in development that can be offered to women. These include freezing of oocytes and ovarian cortex and the transplantation of ovarian tissue. PMID:22301053

Rodriguez-Wallberg, Kenny A.; Oktay, Kutluk

2012-01-01

204

Active diffusion positions the nucleus in mouse oocytes.  

PubMed

In somatic cells, the position of the cell centroid is dictated by the centrosome. The centrosome is instrumental in nucleus positioning, the two structures being physically connected. Mouse oocytes have no centrosomes, yet harbour centrally located nuclei. We demonstrate how oocytes define their geometric centre in the absence of centrosomes. Using live imaging of oocytes, knockout for the formin 2 actin nucleator, with off-centred nuclei, together with optical trapping and modelling, we discover an unprecedented mode of nucleus positioning. We document how active diffusion of actin-coated vesicles, driven by myosin Vb, generates a pressure gradient and a propulsion force sufficient to move the oocyte nucleus. It promotes fluidization of the cytoplasm, contributing to nucleus directional movement towards the centre. Our results highlight the potential of active diffusion, a prominent source of intracellular transport, able to move large organelles such as nuclei, providing in vivo evidence of its biological function. PMID:25774831

Almonacid, Maria; Ahmed, Wylie W; Bussonnier, Matthias; Mailly, Philippe; Betz, Timo; Voituriez, Raphaël; Gov, Nir S; Verlhac, Marie-Hélène

2015-04-01

205

Oocyte donation program using a simplified hormonal regimen.  

PubMed

It has been well recognized that both the synchronization of luteinizing hormone (LH) surge between the donor and the recipient for normally cycling women and the complex steroid replacement regimen given on a sequential and incremental basis for women with primary or secondary ovarian failure are two important aspects in oocyte donation. In oocyte donation program at SNUH, a simplified hormonal regimen applicable both to normally cycling women and to those with ovarian failure which consisted of administering 2 mg estradiol (E2) valerate orally 3 times a day augmented with 100 mg progesterone (P) in oil intramuscularly daily starting on the day preceding the oocyte retrieval from the donor was utilized. From July 1988 to December 1989 at SNUH, 11 cycles of oocyte donation program in 10 infertile patients were undertaken and 5 patients succeeded in pregnancy. PMID:2128445

Chang, Y S; Kim, S H; Choi, Y M; Moon, S Y; Lee, J Y

1990-09-01

206

Functional expression of murine multidrug resistance in Xenopus laevis oocytes  

SciTech Connect

The development of multidrug resistance (MDR) is associated with the overproduction of a plasma membrane glycoprotein, P glycoprotein. Here the authors report the functional expression of a member of the murine MDR family of proteins and show that Xenopus oocytes injected with RNA encoding the mouse mdr1b P glycoprotein develop a MDR-like phenotype. Immunological analysis indicated that oocytes injected with the mdr1b RNA synthesized a protein with the size and immunological characteristics of the mouse mdr1b P glycoprotein. These oocytes exhibited a decreased accumulation of ({sup 3}H)vinblastine and showed an increased capacity to extrude the drug compared to control oocytes not expressing the P glycoprotein. In addition, competition experiments indicated that verapamil, vincristine, daunomycin, and quinidine, but not colchicine, can overcome the rapid drug efflux conferred by the expression of the mouse P glycoprotein.

Castillo, G.; Vera, J.C.; Rosen, O.M. (Memorial Sloan-Kettering Cancer Research Center, New York, NY (USA)); Yang, Chiaping Huang; Horwitz, S.B. (Albert Einstein College of Medicine, Bronx, NY (USA))

1990-06-01

207

Hypotonicity activates a native chloride current in Xenopus oocytes  

PubMed Central

Xenopus oocytes are frequently utilized for in vivo expression of cellular proteins, especially ion channel proteins. A thorough understanding of the endogenous conductances and their regulation is paramount for proper characterization of expressed channel proteins. Here we detail a novel chloride current (ICl.swell) responsive to hypotonicity in Xenopus oocytes using the two-electrode voltage clamp technique. Reducing the extracellular osmolarity by 50% elicited a calcium-independent chloride current having an anion conductivity sequence identical with swelling-induced chloride currents observed in epithelial cells. The hypotonicity-activated current was blocked by chloride channel blockers, trivalent lanthanides, and nucleotides. G- protein, cAMP-PKA, and arachidonic acid signaling cascades were not involved in ICl.swell activation. ICl.swell is distinct from both stretch-activated nonselective cation channels and the calcium- activated chloride current in oocytes and may play a critical role in volume regulation in Xenopus oocytes. PMID:8189203

1994-01-01

208

Syrian Hamster model of postmenopausal hypercholesterolemia atherosclerosis and the development of plaques as imaged by high field MRI  

E-print Network

Syrian Hamster model of postmenopausal hypercholesterolemia atherosclerosis and the development of flaxseed in a hamster model of postmenopausal atherosclerosis. The focus is to examine in vivo identification of plaques in cerebral arteries and aortas of overiectomized (ovx) hamsters with endogenous

McQuade, D. Tyler

209

Suppression of hamster lymphocyte reactivity to simian virus 40 tumor surface antigens by spleen cells from pregnant hamsters  

SciTech Connect

SV40-transformed tumor cells in hamsters have been found to have cell surface antigens cross-reactive with antigens temporally expressed on fetal tissues. Using a lymphocyte transformation assay, spleen cells from pregnant hamsters were found to be incapable of responding to preparations of either hamster fetal tissue or SV40-transformed cells. However, a suppressor component can be demonstrated in spleen cell populations of both primi-and multiparous hamsters during pregnancy that is capable of reducing the response of lymphocytes sensitized against SV40 tumor-associated antigens. The degree of suppression is proportional to the ratio of responder cells to spleen cells from pregnant animals. These results suggest there is a subpopulation of spleen cells involved in immunoregulation during pregnancy that has the ability to suppress the reactivity of lymphocytes sensitized against SV40-associated oncofetal antigens.

Weppner, W.A.; Adkinson, L.R.; Coggin, J.H.Jr

1980-09-01

210

OOCYTE DIFFERENTIATION AND VITELLOGENESIS IN THE ROACH PERIPLANETA AMERICANA  

PubMed Central

The ovary of the roach Periplaneta americana has been studied by techniques of light and electron microscopy. Each ovariole (panoistic type) contains a linear array of oocytes in varying stages of development. Newly formed oocytes become encased by a layer of follicle cells and begin pinocytosis. All subsequent growth stages of the oocytes are dependent, in part, on this phenomenon. All of the pinocytotic caveolae show an unique surface modification; i.e., on their internal surface they have an amorphous or filamentous substance and their external surface is studded with many fine radially oriented spike-like projections. The pinosomes of early oocytes do not contain a demonstrable internal structure; they are thought to contain nutritive substances for the developing oocytes rather than yolk precursors. When the oocyte enters its last stage of growth, characterized by yolk deposition, the caveolae become filled with a dense material which is thought to be the precursors of yolk. Hence the conclusion is drawn that yolk formation is independent of any cytoplasmic organelle system of the oocyte and that the precursors of this deutoplasmic substance are manufactured outside the ovary and are internalized by the process of pinocytosis. Under the phase-contrast microscope the nucleoli of early oocytes are large irregular masses and show the phenomenon of nucleolar emission (fragmentation). These "emissions" become randomly dispersed in the nucleoplasm and some of them come to be intimately associated with the fenestrated nuclear envelope. After this process ceases, the main nucleolar mass becomes vacuolated. Electron micrographs suggest that the constituent particles of the nucleolar emissions migrate from the nucleus through patent pores of the nuclear envelope. PMID:14105205

Anderson, Everett

1964-01-01

211

Electrophysiological responses of crayfish oocytes to biogenic amines  

Microsoft Academic Search

Intracellular recordings were made from immature, growing oocytes of the crayfish Pacifastacus leniusciulus. Oocytes had a relatively negative resting potential of ?74.7±2.2 mV (n=26; range ?53 to ?90) and a mean input resistance of 0.86±0.19 M? (n=22; range 0.17–3.3). Octopamine induced a long-lasting response involving biphasic changes in input resistance, together with bi- or multiphasic changes in membrane potential. The

Peter Skorupski; Richard Melarange

2000-01-01

212

Factors affecting in vitro maturation of alpaca (Lama paco) oocytes.  

PubMed

The present study utilized a 2×2×2 factorial design examining age (old vs. young), follicle size (?2mm vs. <2mm) and media supplementation (with or without fetal bovine serum [FBS]) to determine factors that might affect in vitro maturation of alpaca oocytes. We hypothesized that oocytes collected from follicles ?2mm from young alpacas and incubated in maturation media supplemented with FBS would have greater maturation rates than those incubated in any other factorial combination. Oocytes were collected from the ovaries of 11 young alpacas (<10 years old) and 14 old alpacas (>11 years old). Oocytes were classified as morphologically normal oocytes (MNO) and deemed suitable for incubation if ?3 compact layers of cumulus cells and a homogeneous, evenly granulated cytoplasm were observed. Oocytes from each group of follicle sizes were incubated separately and halves of each group were randomly divided and incubated 24h in chemically defined maturation media with or without 10% FBS. Maturation was defined as the visualization of a polar body at the end of the incubation period. Overall, a greater proportion of MNO were collected from follicles ?2mm than that obtained from smaller follicles, 55% (136/247) vs. 29.6% (162/547), respectively (P<0.05). A greater proportion of oocytes reached maturation when collected from ?2mm follicles 36% (49/136) than from <2mm follicles 8% (13/162) (P<0.05). For oocytes obtained from ?2mm follicles of old alpacas, a greater proportion reached maturation when incubated in media supplemented with FBS than when incubated without FBS; 57.6% (19/33) vs. 18.2% (6/33), respectively (P<0.05). PMID:25261077

Leisinger, Ca; Coffman, Ea; Coutinho da Silva, Ma; Forshey, Bs; Pinto, Crf

2014-11-10

213

Expression of epithelial Na channels in Xenopus oocytes  

Microsoft Academic Search

Epithelial Na channel activity was expressed in oocytes from Xenopus laev\\/s after injection of mRNA from A6 cells, derived from Xenopus kidney. Poly A(+) RNA was extracted from confluent cell monolayers grown on either plastic or permeable supports. 1-50 ng RNA was injected into stage 5-6 oocytes. Na channel activity was assayed as amiloride-sensitive current (Isa) under voltage-clamp condi- tions

LAWRENCE G. PALMER; IRENE CORTHESY-THEULAZ; HANS-PETER GAEGGELER; JEAN-PIERRE KRAEHENBUHL; BERNARD ROSSIER

1990-01-01

214

The use of transmission electron microscopy and oocyte transfer to evaluate in vitro maturation of equine oocytes in different culture conditions  

Microsoft Academic Search

The current study evaluates the ability of equine oocytes matured in different conditions to undergo nuclear and cytoplasmic maturation. After oocyte transfer, embryonic development was diagnosed at 15 and 90 days of gestation. For each group, immature oocytes obtained from slaughterhouse ovaries were matured in vitro (5 replicates). In experiment I, three different media were tested, HTF:BME, SOFaa, and TCM

C. B. Fernandes; K. R. Peres; M. A. Alvarenga; F. C. Landim-Alvarenga

2006-01-01

215

Effect of leptin during in vitro maturation of prepubertal calf oocytes: Embryonic development and relative mRNA abundances of genes involved in apoptosis and oocyte competence  

Microsoft Academic Search

During the in vitro maturation of adult bovine oocytes, leptin has beneficial effects on blastocyst development, apoptosis and transcription levels of developmentally important genes. The present study analyzes the differential effects of leptin on prepubertal bovine oocytes and cumulus cells. Effects were determined of leptin treatment during oocyte maturation on their developmental capacity after fertilization (Exp. 1), incidence of apoptosis

Bladimir Córdova; Roser Morató; Celia de Frutos; Pablo Bermejo-Álvarez; Teresa Paramio; Alfonso Gutiérrez-Adán; Teresa Mogas

2011-01-01

216

Control of Oocyte Growth and Meiotic Maturation in C. elegans  

PubMed Central

In sexually reproducing animals, oocytes arrest at diplotene or diakinesis and resume meiosis (meiotic maturation) in response to hormones. Chromosome segregation errors in female meiosis I are the leading cause of human birth defects, and age-related changes in the hormonal environment of the ovary are a suggested cause. C. elegans is emerging as a genetic paradigm for studying hormonal control of meiotic maturation. The meiotic maturation processes in C. elegans and mammals share a number of biological and molecular similarities. Major sperm protein (MSP) and luteinizing hormone (LH), though unrelated in sequence, both trigger meiotic resumption using somatic G?s-adenylate cyclase pathways and soma-germline gap-junctional communication. At a molecular level, the oocyte responses apparently involve the control of conserved protein kinase pathways and post-transcriptional gene regulation in the oocyte. At a cellular level, the responses include cortical cytoskeletal rearrangement, nuclear envelope breakdown, assembly of the acentriolar meiotic spindle, chromosome segregation, and likely changes important for fertilization and the oocyte-to-embryo transition. This chapter focuses on signaling mechanisms required for oocyte growth and meiotic maturation in C. elegans and discusses how these mechanisms coordinate the completion of meiosis and the oocyte-to-embryo transition. PMID:22872481

Kim, Seongseop; Spike, Caroline; Greenstein, David

2013-01-01

217

Greatwall kinase is required for meiotic maturation in porcine oocytes.  

PubMed

Meiotic maturation in many species is initiated by the activation of maturation-promoting factor (MPF) with concomitant inactivation of counteracting phosphatases, most notably protein phosphatase 2A (PP2A). Recently, Greatwall (GWL) has been identified as a cell cycle regulator that inhibits PP2A activity. In this study, we demonstrate that GWL is required for meiotic maturation in porcine oocytes. GWL expression increases from germinal vesicle (GV) to metaphase II (MII) stages of porcine oocytes and dramatically decreases with progression of the meiotic cell cycle. GWL is initially localized in the nucleus of GV oocytes and is associated with spindle fibers following GV breakdown. Depletion of GWL inhibited or delayed meiotic maturation secondary to defects in chromosome congression and spindle formation. Conversely, overexpression of GWL overcame meiotic arrest and initiated progression to MII stage. However, these oocytes had severe spindle defects. Furthermore, MII oocytes depleted of GWL progressed to pronuclear formation. Taken together, our data demonstrate that GWL is required not only for meiotic maturation but also for maintenance of MII arrest in porcine oocytes. PMID:23843240

Li, Ying-Hua; Kang, Hyoeun; Xu, Yong-Nan; Heo, Young-Tae; Cui, Xiang-Shun; Kim, Nam-Hyung; Oh, Jeong Su

2013-09-01

218

Regulation of Greatwall kinase during Xenopus oocyte maturation.  

PubMed

Greatwall kinase has been identified as a key element in M phase initiation and maintenance in Drosophila, Xenopus oocytes/eggs, and mammalian cells. In M phase, Greatwall phosphorylates endosulfine and related proteins that bind to and inhibit protein phosphatase 2A/B55, the principal phosphatase for Cdk-phosphorylated substrates. We show that Greatwall binds active PP2A/B55 in G2 phase oocytes but dissociates from it when progesterone-treated oocytes reach M phase. This dissociation does not require Greatwall kinase activity or phosphorylation at T748 in the presumptive T loop of the kinase. A mutant K71M Greatwall, also known as Scant in Drosophila, induces M phase in the absence of progesterone when expressed in oocytes, despite its reduced stability and elevated degradation by the proteasome. M phase induction by Scant Greatwall requires protein synthesis but is not associated with altered binding or release of PP2A/B55 as compared to wild-type Greatwall. However, in vitro studies with Greatwall proteins purified from interphase cells indicate that Scant, but not wild-type Greatwall, has low but detectable activity against endosulfine. These results demonstrate progesterone-dependent regulation of the PP2A/B55-Greatwall interaction during oocyte maturation and suggest that the cognate Scant Greatwall mutation has sufficient constitutive kinase activity to promote M phase in Xenopus oocytes. PMID:21551066

Yamamoto, Tomomi M; Blake-Hodek, Kristina; Williams, Byron C; Lewellyn, Andrea L; Goldberg, Michael L; Maller, James L

2011-07-01

219

Resveratrol Protects Mouse Oocytes from Methylglyoxal-Induced Oxidative Damage  

PubMed Central

Methylglyoxal, a reactive dicarbonyl compound, is mainly formed from glycolysis. Methylglyoxal can lead to the dysfunction of mitochondria, the depletion of cellular anti-oxidation enzymes and the formation of advanced glycation ends. Previous studies showed that the accumulation of methylglyoxal and advanced glycation ends can impair the oocyte maturation and reduce the oocyte quality in aged and diabetic females. In this study, we showed that resveratrol, a kind of phytoalexin found in the skin of grapes, red wine and other botanical extracts, can alleviate the adverse effects caused by methylglyoxal, such as inhibition of oocyte maturation and disruption of spindle assembly. Besides, methylglyoxal-treated oocytes displayed more DNA double strands breaks and this can also be decreased by treatment of resveratrol. Further investigation of these processes revealed that methylglyoxal may affect the oocyte quality by resulting in excessive reactive oxygen species production, aberrant mitochondrial distribution and high level lipid peroxidation, and resveratrol can block these cytotoxic changes. Collectively, our results showed that resveratrol can protect the oocytes from methylglyoxal-induced cytotoxicity and this was mainly through the correction of the abnormity of cellular reactive oxygen species metabolism. PMID:24194906

Liu, Yu; He, Xiao-Qin; Huang, Xin; Ding, Lu; Xu, Lin; Shen, Yu-Ting; Zhang, Fei; Zhu, Mao-Bi; Xu, Bai-Hui; Qi, Zhong-Quan; Wang, Hai-Long

2013-01-01

220

Advanced ground penetrating radar  

SciTech Connect

An advanced Ground Penetrating Radar (GPR) system has the potential for efficiently and reliably providing high resolution images for inspecting concrete civil structures for defects and damage assessment. To achieve the required performance, improvements in radar hardware, and development and adaptation of advanced 2- and 3-dimensional synthetic aperture imaging techniques are needed. Recent and continuing advancement in computer and computer-related technology areas have made it possible to consider more complex and capable systems for a variety of imaging applications not previously conceived. The authors developed conceptual designs, analyzed system requirements, and performed experiments, modeling, and image reconstructions to study the feasibility of improving GPR technology for non-destructive evaluation of bridge decks and other high-value concrete structures. An overview and summary of practical system concepts and requirements, are presented.

Warhus, J.P.; Mast, J.E.; Johansson, E.M.; Nelson, S.D. [Lawrence Livermore National Lab., CA (United States). Electronics Engineering Dept.

1994-07-26

221

Projectile penetration into representative targets  

NASA Astrophysics Data System (ADS)

The differential equation representing the penetration of a 'hard' projectile into semi-infinite, homogeneous target materials is solved for several generic combinations of the target material/projectile characteristics. A 'hard' projectile is defined as one that does not change size or shape and does not lose mass during the penetration process. The target materials evaluated range from the structurally 'soft' materials (liquids) to structurally 'hard' materials (armor plate) with viscous and fluid dynamic drag considered. The solutions to the differential equation(s) are expanded in series form to demonstrate the underlying parameters governing projectile penetration and the way they interact to limit penetration in a given target material. It is shown that the fundamental parameter governing projectile penetration into structurally 'firm' materials is the initial kinetic energy of the projectile divided by the frontal area of the projectile and the inherent structural characteristic of the target. Experimental data on the penetration of steel spheres into ballistic gelatin and for armor piercing bullets into armor plate materials are used to verify the characteristics of the solutions to the equation of motion for the projectile and to demonstrate how penetration can vary with projectile size and target characteristics. The penetration equation for a single 'hard' target material is used to develop a solution for the penetration of multilayered 'hard' target materials.

Stone, George W.

1994-10-01

222

Developmental stage of the oocyte during antral follicle growth and cumulus investment determines in vitro embryo development of sow oocytes  

Microsoft Academic Search

The aim of the study was to determine the contribution of cumulus cells on the developmental competence of porcine oocytes during follicle growth. Oocytes from large (5–8mm) and small (2–3mm) follicles were cultured with or without follicle stimulating hormone (FSH), subsequently examined for nuclear stage and spindle morphology, or fertilized and cultured for embryo development, or analyzed for glutathione content.

E. J. Schoevers; B. Colenbrander; B. A. J. Roelen

2007-01-01

223

Universal penetration test apparatus with fluid penetration sensor  

DOEpatents

A universal penetration test apparatus for measuring resistance of a material to a challenge fluid. The apparatus includes a pad saturated with the challenge fluid. The apparatus includes a compression assembly for compressing the material between the pad and a compression member. The apparatus also includes a sensor mechanism for automatically detecting when the challenge fluid penetrates the material.

Johnson, Phillip W. (Rochester, MN); Stampfer, Joseph F. (Santa Fe, NM); Bradley, Orvil D. (Santa Fe, NM)

1999-01-01

224

Universal penetration test apparatus with fluid penetration sensor  

DOEpatents

A universal penetration test apparatus is described for measuring resistance of a material to a challenge fluid. The apparatus includes a pad saturated with the challenge fluid. The apparatus includes a compression assembly for compressing the material between the pad and a compression member. The apparatus also includes a sensor mechanism for automatically detecting when the challenge fluid penetrates the material. 23 figs.

Johnson, P.W.; Stampfer, J.F.; Bradley, O.D.

1999-02-02

225

Quantitative proteomics reveals the dynamics of protein changes during Drosophila oocyte maturation and the oocyte-to-embryo transition  

PubMed Central

The onset of development is marked by two major, posttranscriptionally controlled, events: oocyte maturation (release of the prophase I primary arrest) and egg activation (release from the secondary meiotic arrest). Using quantitative mass spectrometry, we previously described proteome remodeling during Drosophila egg activation. Here, we describe our quantitative mass spectrometry-based analysis of the changes in protein levels during Drosophila oocyte maturation. This study presents the first quantitative survey, to our knowledge, of proteome changes accompanying oocyte maturation in any organism and provides a powerful resource for identifying both key regulators and biological processes driving this critical developmental window. We show that Muskelin, found to be up-regulated during oocyte maturation, is required for timely nurse cell nuclei clearing from mature egg chambers. Other proteins up-regulated at maturation are factors needed not only for late oogenesis but also completion of meiosis and early embryogenesis. Interestingly, the down-regulated proteins are predominantly involved in RNA processing, translation, and RNAi. Integrating datasets on the proteome changes at oocyte maturation and egg activation uncovers dynamics in proteome remodeling during the change from oocyte to embryo. Notably, 66 proteins likely act uniquely during late oogenesis, because they are up-regulated at maturation and down-regulated at activation. We find down-regulation of this class of proteins to be mediated partially by APC/CCORT, a meiosis-specific form of the E3 ligase anaphase promoting complex/cyclosome (APC/C). PMID:25349405

Kronja, Iva; Whitfield, Zachary J.; Yuan, Bingbing; Dzeyk, Kristina; Kirkpatrick, Joanna; Krijgsveld, Jeroen; Orr-Weaver, Terry L.

2014-01-01

226

Quantitative proteomics reveals the dynamics of protein changes during Drosophila oocyte maturation and the oocyte-to-embryo transition.  

PubMed

The onset of development is marked by two major, posttranscriptionally controlled, events: oocyte maturation (release of the prophase I primary arrest) and egg activation (release from the secondary meiotic arrest). Using quantitative mass spectrometry, we previously described proteome remodeling during Drosophila egg activation. Here, we describe our quantitative mass spectrometry-based analysis of the changes in protein levels during Drosophila oocyte maturation. This study presents the first quantitative survey, to our knowledge, of proteome changes accompanying oocyte maturation in any organism and provides a powerful resource for identifying both key regulators and biological processes driving this critical developmental window. We show that Muskelin, found to be up-regulated during oocyte maturation, is required for timely nurse cell nuclei clearing from mature egg chambers. Other proteins up-regulated at maturation are factors needed not only for late oogenesis but also completion of meiosis and early embryogenesis. Interestingly, the down-regulated proteins are predominantly involved in RNA processing, translation, and RNAi. Integrating datasets on the proteome changes at oocyte maturation and egg activation uncovers dynamics in proteome remodeling during the change from oocyte to embryo. Notably, 66 proteins likely act uniquely during late oogenesis, because they are up-regulated at maturation and down-regulated at activation. We find down-regulation of this class of proteins to be mediated partially by APC/C(CORT), a meiosis-specific form of the E3 ligase anaphase promoting complex/cyclosome (APC/C). PMID:25349405

Kronja, Iva; Whitfield, Zachary J; Yuan, Bingbing; Dzeyk, Kristina; Kirkpatrick, Joanna; Krijgsveld, Jeroen; Orr-Weaver, Terry L

2014-11-11

227

Successful pregnancy and delivery after ICSI with artificial oocyte activation by calcium ionophore in in-vitro matured oocytes: a case report.  

PubMed

The achievement of a successful pregnancy and delivery after oocyte activation with calcium ionophore is reported in a couple having low fertilization rates after intracytoplasmic sperm injection (ICSI) of in-vitro matured oocytes. A couple, in which the wife had polycystic ovary syndrome and the husband had moderate oligoteratozoospermia, showed a low fertilization rate in a previous in-vitro maturation cycle (2/11 [18.2%]). The most likely cause of complete fertilization failure or low fertilization rates is failure of oocyte activation. Therefore, artificial oocyte activation by calcium ionophore was combined with ICSI to achieve viable fertilized oocytes. Oocytes were stimulated with calcium ionophore for 30?min after ICSI. The fertilization rate of oocytes activated with calcium ionophore (13/15 [86.7%] and 7/9 [77.8%]) was higher than that of the non-activated oocytes. In the latest cycle, three embryos derived from the activated oocytes were transferred into the uterus on day 3. Subsequently, two gestational sacs were identified on ultrasound. The patient delivered dizygotic twins (girl 2260?g and boy 2760?g) at 35 weeks and 6 days gestation by caesarean section. This result suggests that calcium ionophore could be useful for oocyte fertilization in couples with low fertilization rates after ICSI of in-vitro matured oocytes. PMID:25592974

Kim, Jun-Woo; Yang, Seong-Ho; Yoon, San-Hyun; Kim, Sang-Don; Jung, Jae-Hoon; Lim, Jin-Ho

2015-04-01

228

Trichloroethylene Metabolism in the Rat Ovary Reduces Oocyte Fertilizability  

PubMed Central

Exposure to trichloroethylene (TCE, an environmental toxicant) reduced oocyte fertilizability in the rat. In vivo, TCE may be metabolized by cytochrome P450 dependent oxidation or glutathione conjugation in the liver or kidneys, respectively. Cytochrome P450 dependent oxidation is the higher affinity pathway. The primary isoform of cytochrome P450 to metabolize TCE in the liver, cytochrome P450 2E1, is present in the rodent ovary. Ovarian metabolism of TCE by the oxidative pathway and the production of reactive oxygen species may occur given the presence of the metabolizing enzyme. The objectives of this study were to define the sensitive interval of oocyte growth to TCE exposure, and to determine if TCE exposure resulted in the formation of ovarian protein carbonyls, an indicator of oxidative damage. Rats were exposed to TCE in drinking water (0.45% TCE (v/v) in 3% Tween) or 3% Tween (vehicle-control) during three 4–5 day intervals of oocyte development preceding ovulation. Oocytes from TCE-exposed females were less fertilizable compared with vehicle-control oocytes. Immunohistochemical labeling of ovaries and Western blotting of ovarian proteins demonstrated TCE treatment induced a greater incidence of protein carbonyls compared with vehicle controls. Protein carbonyl formation in the ovary is consistent with TCE metabolism by the cytochrome P450 pathway. Oxidative damage following ovarian TCE metabolism or the presence of TCE metabolites may contribute to reduced oocyte fertilizability. In summary, these results indicate maturing oocytes are susceptible to very short in vivo exposures to TCE. PMID:17673192

Wu, Katherine Lily; Berger, Trish

2007-01-01

229

Lipoprotein mediated lipid uptake in oocytes of polychaetes (Annelida).  

PubMed

The uptake of the 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI)-labeled sex-unspecific Nereis lipoprotein was investigated in oocytes of the nereidid polychaetes Nereis virens and Platynereis dumerilii. The fluorescence label was first observed in endocytic vesicles (<1 microm diameter), which later fused to larger vesicles (2-3 microm); these were finally incorporated into existing unlabeled yolk granules (5-6 microm). In Platynereis oocytes, the fusion of endocytic vesicles was delayed in oocytes at their final stage of development compared with those at an early stage of development. Lipoprotein double-labeled with fluorescein isothiocyanate (FITC) and DiI revealed that both the protein and the lipid moiety remained co-localized during incorporation into the yolk granules of the oocyte. No labeling of the cytoplasmic lipid droplets was observed. In N. virens, unlabeled Nereis lipoprotein was effective as a competitive inhibitor of DiI-labeled Nereis lipoprotein. Ligand blot experiments demonstrated the presence of a lipoprotein receptor with an apparent molecular mass of 120 kDa, which is different from that of the known yolk protein receptor. This indicates the presence, in the polychaete oocyte, of two distinct receptors mediating yolk protein and lipoprotein uptake, respectively. Thus, the sex-unspecific lipoprotein contributes to the lipid supply of the growing oocyte in addition to the known uptake of the yolk-protein-associated lipids. The absence of label in the cytoplasmic lipid droplets, even after prolonged incubation with labeled lipoprotein, suggests that these lipids arise either by the breakdown and resynthesis of lipoprotein-derived lipids and/or by de novo synthesis within the oocyte. PMID:19533173

Schenk, Sven; Hoeger, Ulrich

2009-08-01

230

Projectile penetration into representative targets  

Microsoft Academic Search

The differential equation representing the penetration of a 'hard' projectile into semi-infinite, homogeneous target materials is solved for several generic combinations of the target material\\/projectile characteristics. A 'hard' projectile is defined as one that does not change size or shape and does not lose mass during the penetration process. The target materials evaluated range from the structurally 'soft' materials (liquids)

George W. Stone

1994-01-01

231

Penetration of bacteria into meat.  

PubMed Central

Bacteria are confined to the surface of meat during the logarithmic phase of growth. When proteolytic bacteria approach their maximum cell density, extracellular proteases secreted by the bacteria apparently break down the connective tissue between muscle fibers, allowing the bacteria to penetrate the meat. Non-proteolytic bacteria do not penetrate meat, even when grown in association with proteolytic species. Images PMID:406846

Gill, C O; Penney, N

1977-01-01

232

Ground-penetrating radar methods  

Technology Transfer Automated Retrieval System (TEKTRAN)

Ground-penetrating radar geophysical methods are finding greater and greater use in agriculture. With the ground-penetrating radar (GPR) method, an electromagnetic radio energy (radar) pulse is directed into the subsurface, followed by measurement of the elapsed time taken by the radar signal as it ...

233

Sidewall penetrator for oil wells  

NASA Technical Reports Server (NTRS)

Penetrator bores horizontal holes in well casing to increase trapped oil drainage. Several penetrators operated by common drive are inserted into well at once. Shaft, made from spiraling cable, rotates and thrusts simultaneously through rigid curvilinear guide tube forcing bit through casing into strata. Device pierces more deeply than armor-piercing bullets and shaped explosive charges.

Collins, E. R., Jr.

1981-01-01

234

Proteomic Analysis of Chinese Hamster Ovary Cells  

PubMed Central

In order to complement the recent genomic sequencing of Chinese hamster ovary (CHO) cells, proteomic analysis was performed on CHO including the cellular proteome, secretome, and glycoproteome using tandem mass spectrometry (MS/MS) of multiple fractions obtained from gel electrophoresis, multi-dimensional liquid chromatography, and solid phase extraction of glycopeptides (SPEG). From the 120 different mass spectrometry analyses generating 682,097 MS/MS spectra, 93,548 unique peptide sequences were identified with at most a 0.02 false discovery rate (FDR). A total of 6164 grouped proteins were identified from both glycoproteome and proteome analysis, representing an 8-fold increase in the number of proteins currently identified in the CHO proteome. Furthermore, this is the first proteomic study done using CHO genome exclusively which provides for more accurate identification of proteins. From this analysis, the CHO codon frequency was determined and found to be distinct from humans, which will facilitate expression of human proteins in CHO cells. Analysis of the combined proteomic and mRNA data sets indicated the enrichment of a number of pathways including protein processing and apoptosis but depletion of proteins involved in steroid hormone and glycosphingolipid metabolism. 504 of the detected proteins included N-acetylation modifications and 1292 different proteins were observed to be N-glycosylated. This first large-scale proteomic analysis will enhance the knowledge base about CHO capabilities for recombinant expression and provide information useful in cell engineering efforts aimed at modifying CHO cellular functions. PMID:22971049

Baycin-Hizal, Deniz; Tabb, David L.; Chaerkady, Raghothama; Chen, Lily; Lewis, Nathan E.; Nagarajan, Harish; Sarkaria, Vishaldeep; Kumar, Amit; Wolozny, Daniel; Colao, Joe; Jacobson, Elena; Tian, Yuan; O'Meally, Robert N.; Krag, Sharon S.; Cole, Robert N.; Palsson, Bernhard O.; Zhang, Hui; Betenbaugh, Michael

2013-01-01

235

Cryopreservation of mammalian oocytes by using sugars: Intra- and extracellular raffinose with small amounts of dimethylsulfoxide yields high cryosurvival, fertilization, and development rates.  

PubMed

Accumulation of intra- and extracellular sugars such as trehalose, glucose, and raffinose is central to survival strategies of a variety of organisms coping with extreme conditions including freezing and almost complete drying. The objective of the present study was to investigate the potential application of intra- and extracellular raffinose in combination with low concentrations of dimethylsulfoxide (Me(2)SO) to mammalian oocyte cryopreservation. To this end, the fertilization and embryonic development of cryopreserved metaphase II (M II) mouse oocytes were studied in comparison to unfrozen controls. For cryopreservation, M II oocytes were microinjected with 0.1M raffinose, and then cooled to -196 degrees C in the presence of either 0.3M raffinose and 0.5M Me(2)SO (cryopreservation group 1) or 0.3M raffinose and 1.0M Me(2)SO (cryopreservation group 2). The control groups included untreated oocytes (untreated control) and oocytes microinjected with raffinose, but not frozen (injection control). The post-thaw survival rates were 83.9% and 80.6% for the cryopreservation group 1 and 2, respectively. The fertilization and blastocyst rates in the cryopreservation group 1 (90.0% and 77.8%, respectively) and 2 (94.6% and 72.5%, respectively) were also high and similar to the ones of the injection controls (97.8% and 78.5%, respectively) and untreated controls (98.8% and 83.6%, respectively). These results are consistent with the findings of our earlier studies and support the use of sugars as intra- and extracellular cryoprotectants. Furthermore, the results of the present study indicate that the presence of intra- and extracellular sugars alleviates high concentrations of conventional penetrating cryoprotectants, and thus minimizes their toxicity. PMID:19596315

Eroglu, Ali

2010-07-01

236

Cryopreservation of Mammalian Oocytes by Using Sugars: Intra- and extracellular raffinose with small amounts of dimethylsulfoxide yields high cryosurvival, fertilization, and development rates  

PubMed Central

Accumulation of intra- and extracellular sugars such as trehalose, glucose, and raffinose is central to survival strategies of a variety of organisms coping with extreme conditions including freezing and almost complete drying. The objective of the present study was to investigate the potential application of intra- and extracellular raffinose in combination with low concentrations of dimethylsulfoxide (Me2SO) to mammalian oocyte cryopreservation. To this end, the fertilization and embryonic development of cryopreserved metaphase II (M II) mouse oocytes were studied in comparison to unfrozen controls. For cryopreservation, M II oocytes were microinjected with 0.1 M raffinose, and then cooled to -196°C in the presence of either 0.3 M raffinose and 0.5 M Me2SO (cryopreservation group 1) or 0.3 M raffinose and 1.0 M Me2SO (cryopreservation group 2). The control groups included untreated oocytes (untreated control) and oocytes microinjected with raffinose, but not frozen (injection control). The post-thaw survival rates were 83.9% and 80.6% for the cryopreservation group 1 and 2, respectively. The fertilization and blastocyst rates in the cryopreservation group 1 (90.0% and 77.8%, respectively) and 2 (94.6% and 72.5%, respectively) were also high and similar to the ones of the injection controls (97.8% and 78.5%, respectively) and untreated controls (98.8% and 83.6%, respectively). These results are consistent with the findings of our earlier studies and support the use of sugars as intra- and extracellular cryoprotectants. Furthermore, the results of the present study indicate that the presence of intra- and extracellular sugars alleviates high concentrations of conventional penetrating cryoprotectants, and thus minimizes their toxicity. PMID:19596315

Eroglu, Ali

2009-01-01

237

Time-lapse dynamics of the mouse oocyte chromatin organisation during meiotic resumption.  

PubMed

In the mammalian oocyte, distinct patterns of centromeres and pericentromeric heterochromatin localisation correlate with the gamete's developmental competence. Mouse antral oocytes display two main types of chromatin organisation: SN oocytes, with a ring of Hoechst-positive chromatin surrounding the nucleolus, and NSN oocytes lacking this ring. When matured to MII and fertilised, only SN oocytes develop beyond the 2-cell, and reach full term. To give detailed information on the dynamics of the SN or NSN chromatin during meiosis resumption, we performed a 9 hr time-lapse observation. The main significant differences recorded are: (1) reduction of the nuclear area only in SN oocytes; (2) ~17 min delay of GVBD in NSN oocytes; (3) chromatin condensation, after GVBD, in SN oocytes; (4) formation of 4-5 CHCs in SN oocytes; (5) increase of the perivitelline space, ~57 min later in NSN oocytes; (6) formation of a rosette-like disposition of CHCs, ~84 min later in SN oocytes; (7) appearance of the MI plate ~40 min later in NSN oocytes. Overall, we described a pathway of transition from the GV to the MII stage that is punctuated of discrete recordable events showing their specificity and occurring with different time kinetics in the two types of oocytes. PMID:24864231

Belli, Martina; Vigone, Giulia; Merico, Valeria; Redi, Carlo Alberto; Garagna, Silvia; Zuccotti, Maurizio

2014-01-01

238

Time-Lapse Dynamics of the Mouse Oocyte Chromatin Organisation during Meiotic Resumption  

PubMed Central

In the mammalian oocyte, distinct patterns of centromeres and pericentromeric heterochromatin localisation correlate with the gamete's developmental competence. Mouse antral oocytes display two main types of chromatin organisation: SN oocytes, with a ring of Hoechst-positive chromatin surrounding the nucleolus, and NSN oocytes lacking this ring. When matured to MII and fertilised, only SN oocytes develop beyond the 2-cell, and reach full term. To give detailed information on the dynamics of the SN or NSN chromatin during meiosis resumption, we performed a 9?hr time-lapse observation. The main significant differences recorded are: (1) reduction of the nuclear area only in SN oocytes; (2) ~17?min delay of GVBD in NSN oocytes; (3) chromatin condensation, after GVBD, in SN oocytes; (4) formation of 4-5 CHCs in SN oocytes; (5) increase of the perivitelline space, ~57?min later in NSN oocytes; (6) formation of a rosette-like disposition of CHCs, ~84?min later in SN oocytes; (7) appearance of the MI plate ~40?min later in NSN oocytes. Overall, we described a pathway of transition from the GV to the MII stage that is punctuated of discrete recordable events showing their specificity and occurring with different time kinetics in the two types of oocytes. PMID:24864231

Redi, Carlo Alberto; Zuccotti, Maurizio

2014-01-01

239

Pineal melatonin synthesis in Syrian hamsters: A summary  

NASA Astrophysics Data System (ADS)

During the past decade there has been ample documentation of the proposition that the pineal gland mediates photoperiodic influences upon reproductive behavior of hamsters. It is commonly hypothesized that the pineal gland expresses its activity by transformation of photoperiodic information into an hormonal output, that hormone being melatonin. If this hypothesis is correct, there must be some essential diffrence in melatonin's output when hamsters are exposed to different photoperiodic environments. The experiments summarized in this communication analyze pineal melatonin contents in Syrian hamsters maintained in a variety of photoperiodic conditions during different physiological states. The results demonstrate that adult hamsters have a daily surge in pineal melatonin content throughout their lifetime when exposed to simulated annual photoperiodic cycles. There is some fluctuation in the amount of pineal melatonin produced during different physiological states and photoperiodic environments, but these fluctuations seem small when compared to those normally found for other regulatory hormones. When hamsters are exposed to different photoperiodic regimens, the daily melatonin surge maintains a relatively constant phase relationship with respect to the onset of daily activity. There is a concomitant change in its phase relationship with respect to light-dark transitions.

Rollag, M. D.

1982-12-01

240

Hamster and murine models of severe destructive Lyme arthritis.  

PubMed

Arthritis is a frequent complication of infection in humans with Borrelia burgdorferi. Weeks to months following the onset of Lyme borreliosis, a histopathological reaction characteristic of synovitis including bone, joint, muscle, or tendon pain may occur. A subpopulation of patients may progress to a chronic, debilitating arthritis months to years after infection which has been classified as severe destructive Lyme arthritis. This arthritis involves focal bone erosion and destruction of articular cartilage. Hamsters and mice are animal models that have been utilized to study articular manifestations of Lyme borreliosis. Infection of immunocompetent LSH hamsters or C3H mice results in a transient synovitis. However, severe destructive Lyme arthritis can be induced by infecting irradiated hamsters or mice and immunocompetent Borrelia-vaccinated hamsters, mice, and interferon-gamma- (IFN-?-) deficient mice with viable B. burgdorferi. The hamster model of severe destructive Lyme arthritis facilitates easy assessment of Lyme borreliosis vaccine preparations for deleterious effects while murine models of severe destructive Lyme arthritis allow for investigation of mechanisms of immunopathology. PMID:22461836

Munson, Erik; Nardelli, Dean T; Du Chateau, Brian K; Callister, Steven M; Schell, Ronald F

2012-01-01

241

The hamster flank organ model: Is it relevant to man  

SciTech Connect

The critical role that androgens play in the etiology of acne has led to a search for topically active antiandrogens and the frequent use of the flank organ of the golden Syrian hamster as an animal model. 17-alpha-propyltestosterone (17-PT) has been identified as having potent antiandrogenic activity in the hamster model, and this report describes its clinical evaluation. Two double-blind placebo controlled studies comparing 4% 17-PT in 80% alcohol versus vehicle alone were conducted. One study examined 17-PT sebosuppressive activity in 20 subjects. The second study examined its efficacy in 44 subjects having mild to moderate acne. A third study measured in vitro percutaneous absorption of 17-PT through hamster flank and monkey skin, and human face skin in-vivo, using radioactive drug. 17-PT was found to be ineffective in reducing either the sebum excretion rate or the number of inflammatory acne lesions. Failure of 17-PT to show clinical activity was not a result of poor percutaneous absorption. Total absorption in man was 7.7% of the dose and only 1.0% in the hamster. The sebaceous gland of hamster flank organ is apparently more sensitive to antiandrogens than the human sebaceous gland.

Franz, T.J.; Lehman, P.A.; Pochi, P.; Odland, G.F.; Olerud, J. (Univ. of Washington, Seattle (USA))

1989-10-01

242

Gastric metabolism of ethanol in Syrian golden hamster.  

PubMed

First-pass metabolism (FPM) of orally ingested alcohol has been attributed to gastric alcohol dehydrogenase (ADH) activity in both humans and rats. To determine whether gastric alcohol dehydrogenase is essential for alcohol FPM, we sought a species lacking this enzyme. We found that Syrian golden hamsters have negligible gastric ADH yet alcohol FPM (265 +/- 25 mg ethanol/kg) was comparable to that of rats (251 +/- 31 mg/kg). To determine whether hamster gastric mucosal cells metabolize sufficient alcohol to account for this FPM, primary cultures were established, and these cells metabolized 1.99 +/- 0.84 mumol ethanol/10(6) cells/hr, an amount sufficient to account for the bulk of alcohol FPM. In contrast to alcohol dehydrogenase, catalase activity in hamster gastric mucosa (870 +/- 93 units/g tissue) was eightfold higher than in rat gastric mucosa (111 +/- 9 units/g tissue; P < 0.0001). FPM in hamsters treated with 3-aminotriazole was reduced from 242 +/- 24 to 130 +/- 22 mg/kg (P < 0.05) but was not reduced in rats. The results imply that catalase participates in gastric alcohol metabolism of hamsters. PMID:8536535

Batra, S C; Haber, P S; Mirmiran-Yazdy, F S; Korsten, M A; Gentry, R T; Lieber, C S

1995-12-01

243

Isolation, antimicrobial activities, and primary structures of hamster neutrophil defensins.  

PubMed Central

Hamster (Mesocricetus auratus) neutrophil granules contain at least four microbicidal peptides belonging to the defensin family. These compounds were purified from granule acid extracts by reverse-phase chromatography and termed HaNP-1 to -4 (hamster neutrophil peptide). HaNP-1 and HaNP-3 revealed the most bactericidal activity, with a 50% inhibitory concentration of 0.3 to 0.8 microg/ml for Staphylococcus aureus and Streptococcus pyogenes strains. The HaNP-4 was always isolated in concentrations exceeding about 10 times the concentrations of other hamster peptides, but its antibacterial activity as well as that of HaNP-2 was relatively lower, probably as a result of conserved Arg residue substitutions. Other microorganisms were also tested, and generally, hamster defensins exhibited less potency against gram-negative bacteria. The amino acid sequence of hamster defensins showed a high percentage of identity to the sequence of mouse enteric defensins, reaching about 60% identical residues in the case of HaNP-3 and cryptdin 3. PMID:8890190

Mak, P; Wójcik, K; Thogersen, I B; Dubin, A

1996-01-01

244

Penetration resistance and penetrability in pyramidal (nano)indentations.  

PubMed

Pyramidal nanoindentation loading curves were linearly plotted, normal force versus (penetration depth)(3/2) . The slope is penetration resistance k, its inverse penetrability. Linear correlations verify. All contributions to the indentation are included in the penetrability. Dependencies and uses of the extrapolation tools are exemplified, identified, and discussed. In the case of phase transition including twinning within the loading range a sharp kink occurs, again with verifying correlation in both branches of the linear plot. The exponent 3/2 applies to all types of materials upon conical or pyramidal indentations onto normal flat surfaces, independent of the various mechanistic responses. While common curve fitting procedures of loading curves and finite element (FE) calculations miss phase transitions, gradients, surface effects, elbows, (nano)pores, and change from tip rounding to cone (at very low penetrations), these are recognized by the penetration resistance analysis. Also prominent undisturbed pyramidal or conical micro- and macroindentations provide linear plots with exponent 3/2. Numerous FE simulations create experimentally unsupported "loading curves." This is discussed with typical published examples. An explanation for the deviation from Sneddon's and Love's theory is given by correction for the shear-force part that does not participate in the penetration depth. The validation of the exponent 3/2 instead of previously assumed 2 requires adjustment of mechanical parameters that were defined by using the nonsupported exponent. PMID:22886889

Kaupp, Gerd; Naimi-Jamal, Mohammad Reza

2013-01-01

245

[Fertilizing capacity of the ejaculate of nutria (Myocastor coypus) after the removal of the seminal vesicles as evaluated by the penetration test and natural mating].  

PubMed

The fertility of male coypu sperm following seminal vesicle extirpation was investigated using the penetration test into the egg of Syrian golden hamster (Mesocricetus auratus). Ejaculates were obtained from five males by means of electro-ejaculation under halothane narcosis. The results of the zona-free hamster eggs (ZFHE) penetration test showed that the ejaculates of all the surgically treated coypu males were fertile and that ZFHE value fluctuated from 54 to 76.6%. The results obtained in experiments with natural mating revealed that the extirpation of male coypu seminal vesicles did not affect their fertility. In total 47 foetuses were found post mortem in ten coypu females covered by surgically treated males, which on average represented 4.7 foetuses per female. PMID:2678717

Jakubicka, I; Barta, M; Babusík, P

1989-07-01

246

Adrenal hormones mediate melatonin-induced increases in aggression in male Siberian hamsters (Phodopus sungorus)  

E-print Network

Adrenal hormones mediate melatonin-induced increases in aggression in male Siberian hamsters of melatonin, as well as adrenal hormones, in the regulation of seasonal aggression in male Siberian hamsters of melatonin (15 Ag/day) or saline 2 h before lights out for 10 consecutive days. In Experiment 2, hamsters

Demas, Greg

247

The hamster as a model system for the study of influenza vaccines  

Microsoft Academic Search

A series of experiments was carried out in hamsters to determine their value as an experimental animal for the study of influenza virus infection and immunization. Hamsters could be infected intranasally with approximately 100 EID50 of unadapted influenza A\\/Port Chalmers\\/73 virus; infection produced serum HI antibody and virus was recovered from both nasal washings and from lungs. Inoculation of hamsters

C. W. Potter; R. Jennings

1976-01-01

248

Replacement of hamsters with physiochemical analytical methods for Leptospira vaccine batch  

E-print Network

1 Replacement of hamsters with physiochemical analytical methods for Leptospira vaccine batch and Veterinary Laboratories Agency (AHVLA). Histological processing and analysis of hamster tissue was performed serovar Canicola vaccines requires the use of a large number of hamsters and has severe effects; whilst

Chittka, Lars

249

Acute and Chronic Social Defeat Suppresses Humoral Immunity of Male Syrian Hamsters  

E-print Network

Acute and Chronic Social Defeat Suppresses Humoral Immunity of Male Syrian Hamsters (Mesocricetus examined the role of a stressor, social defeat, on humoral immune function of Syrian hamsters (Me- vated in both acutely and chronically defeated hamsters compared with control animals. In contrast

Demas, Greg

250

Long-term persistence of male copulatory behavior in castrated and photo-inhibited Siberian hamsters  

E-print Network

hamsters Jin Ho Park,a,* Nana Takasu,a Maria I. Alvarez,a Kathryn Clark,a Rahim Aimaq,a and Irving Zuckera the persistence of the ejaculatory reflex 19 weeks after orchidectomy in 40% of male Siberian hamsters maintained hamsters transferred from a long to a short photoperiod underwent gonadal regression: 50% of these animals

Zucker, Irving

251

Photoperiod Affects Neuronal Nitric Oxide Synthase and Aggressive Behaviour in Male Siberian Hamsters (Phodopus  

E-print Network

Hamsters (Phodopus sungorus) J. C. Wen,*a A. K. Hotchkiss,* G. E. Demas and R. J. Nelson* *Departments University, Bloomington, IN, USA. Key words: photoperiod, Siberian hamster, seasonal, aggression, nitric including photoperiod (day length). Male Siberian hamsters (Phodopus sungorus) housed in short photoperiod

Demas, Greg

252

Photoperiod-dependent effects of neuronal nitric oxide synthase inhibition on aggression in Siberian hamsters  

E-print Network

in Siberian hamsters Tracy A. Bedrosian a, , Laura K. Fonken a , Gregory E. Demas b , Randy J. Nelson and aggression. Seasonally-breeding Siberian hamsters, however, are paradoxically more aggressive in short, is associated with increased aggression in male short-day hamsters. In the present study, we hypothesized

Demas, Greg

253

Behavioural and Neuroendocrine Adaptations to Repeated Stress during Puberty in Male Golden Hamsters  

E-print Network

Hamsters J. C. Wommack,* A. Salinas,* R. H. Melloni Jr, and Y. Delville* *Psychology Department, the consequences of stress are often severe and long lasting. Repeated subjugation in adult male golden hamsters-pubertal changes in stress hormones may explain why juvenile hamsters are more resilient to social stress than

Delville, Yvon

254

9 CFR 3.36 - Primary enclosures used to transport live guinea pigs and hamsters.  

Code of Federal Regulations, 2014 CFR

...used to transport live guinea pigs and hamsters. 3.36 Section 3.36 Animals...and Transportation of Guinea Pigs and Hamsters Transportation Standards § 3.36...used to transport live guinea pigs and hamsters. No person subject to the...

2014-01-01

255

Vasopressin/Serotonin Interactions in the Anterior Hypothalamus Control Aggressive Behavior in Golden Hamsters  

E-print Network

in Golden Hamsters Craig F. Ferris,1 Richard H. Melloni Jr,1 Gary Koppel,2 Kenneth W. Perry,2 Ray W. Fuller) in the control of offensive aggres- sion in Syrian golden hamsters. First, specific V1A and 5-HT1B binding sites/intruder paradigm, resident hamsters treated with fluoxetine, a selective 5-HT reuptake inhibitor, have

Delville, Yvon

256

Reproduction (2003) 125, 397407 Seasonal control of penile development of Siberian hamsters  

E-print Network

hamsters (Phodopus sungorus) by daylength and testicular hormones J. H. Park1 , E. M. Spencer2 , N. J and androgens on penile develop- ment in the Siberian hamster (Phodopus sungorus). Adult penile masses were achieved at 18 and 8 weeks of age in hamsters maintained from birth under short (10 h light: 14 h dark

Breedlove, Marc

257

A NEURAL NETWORK UNDERLYING INDIVIDUAL DIFFERENCES IN EMOTION AND AGGRESSION IN MALE GOLDEN HAMSTERS  

E-print Network

HAMSTERS J. T. DAVID, M. C. CERVANTES, K. A. TROSKY, J. A. SALINAS AND Y. DELVILLE* Psychology Department a common neural network. Male golden hamsters were first screened for offensive aggression. Then. Similar protocols have been used to test behaviors associated with frustration. At first, all hamsters

Delville, Yvon

258

POUVOIR PATHOGNE EXPRIMENTAL DES MYCOBACTRIES DITES ATYPIQUES CHEZ LA SOURIS, LE HAMSTER ET LE MRION  

E-print Network

POUVOIR PATHOGÃ?NE EXPÃ?RIMENTAL DES MYCOBACTÃ?RIES DITES «ATYPIQUES» CHEZ LA SOURIS, LE HAMSTER ET LE, HAMSTER AND MERION. ― Three animal species were used for the experimental pathogenicity of fourteen of healthy pigs in the surrounding area of Montevideo. White mice, hamsters and merions were inoculated

Paris-Sud XI, Université de

259

Neuropeptide Y Activates Protein Kinase C in Hamster Suprachiasmatic Nuclei Brain Slices  

E-print Network

Neuropeptide Y Activates Protein Kinase C in Hamster Suprachiasmatic Nuclei Brain Slices Kathryn M in the SCN of a variety of animal species including the hamster (Biello et al., 1997; Van der Zee & Bult to test the hypothesis that neuropeptide Y should increase PKC activity in the hamster SCN. Material

Harrington, Mary

260

Analysis of superovulation in the hamster : 1962-1978 G. S. GREENWALD  

E-print Network

Analysis of superovulation in the hamster : 1962-1978 G. S. GREENWALD Department of Physiology in the cyclic hamster by single injections of PMSG. Four principal approaches have been used : 1) ovulation rate the hamster for studies of follicular growth and atresia. Introduction. For the past 16 yearsI have been

Paris-Sud XI, Université de

261

Photoperiodic regulation of gene expression in brown and white adipose tissue of Siberian hamsters  

E-print Network

Photoperiodic regulation of gene expression in brown and white adipose tissue of Siberian hamsters expression in brown and white adipose tissue of Siberian hamsters (Phodopus sungorus). Am J Physiol Regulatory Integrative Comp Physiol 282: R114­R121, 2002.--Siberian hamsters exhibit seasonal fluctuations

Demas, Greg

262

CELL-MEDIATED CYTOTOXICITY AGAINST HAMSTER CELLS TRANSFORMED BY AVIAN SARCOMA VIRUSES  

E-print Network

CELL-MEDIATED CYTOTOXICITY AGAINST HAMSTER CELLS TRANSFORMED BY AVIAN SARCOMA VIRUSES 2 CYTOTOXICITE A MEDIATION CELLULAIRE ENVERS DES CELLULES DE HAMSTER TRANS- FORMEES PAR DES VIRUS DE SARCOMES apparus à la surface de différents clones de cellules de hamster transformées par des virus de sarcomes

Paris-Sud XI, Université de

263

In vivo fertilization after initiation of sperm motility in the hamster epididymis  

E-print Network

In vivo fertilization after initiation of sperm motility in the hamster epididymis Marie P. et M. Curie, 4, place Jussieu, 75230 Paris Cedex 05, France. Summary. Hamster spermatozoa ; hamster : Kann and Serres, 1980). Moreover, cinematographic studies by these same authors revealed

Paris-Sud XI, Université de

264

Differential Cytokine Gene Expression According to Outcome in a Hamster Model of Leptospirosis  

E-print Network

Differential Cytokine Gene Expression According to Outcome in a Hamster Model of Leptospirosis Fre studied in a hamster model. Methodology/Principal Findings: Using an LD50 model of leptospirosis in hamsters, we first determined that 3 days post- infection was a time-point that allowed studying

Paris-Sud XI, Université de

265

Behavioral and Neurobiological Consequences of Social Subjugation during Puberty in Golden Hamsters  

E-print Network

Behavioral and Neurobiological Consequences of Social Subjugation during Puberty in Golden Hamsters Department, University of Massachusetts Medical Center, Worcester, Massachusetts 01655 In golden hamsters subjugation) during puberty. Male golden hamsters were weaned at postna- tal day 25 (P25), exposed daily

Delville, Yvon

266

Response to exogenous kisspeptin varies according to sex and reproductive condition in Siberian hamsters (Phodopus sungorus)  

E-print Network

hamsters (Phodopus sungorus) Timothy J. Greives, Kimberly L. Long, Christine M. Bergeon Burns, Gregory E Siberian hamsters held on reproductively inhibitory or stimulatory photoperiods. In Experiment 2, we asked in this response, with males showing greater LH responses to kisspeptin than females. Hamsters responded

Demas, Greg

267

TUDE DE LA CYTOTOXICIT MDIATION CELLULAIRE DANS LE SARCOME DE ROUS CHEZ LE HAMSTER  

E-print Network

Ã?TUDE DE LA CYTOTOXICITÃ? Ã? MÃ?DIATION CELLULAIRE DANS LE SARCOME DE ROUS CHEZ LE HAMSTER Ginette, France Summary CELL-MEDIATED CYTOTOXICITY AGAINST ROUS SARCOMA IN THE HAMSTER. - The cytotoxic response of lymphoid cells from hamsters immunized against RS 2/3 cells, transformed by Rous sarcoma virus

Paris-Sud XI, Université de

268

Pubertal growth of the medial amygdala delayed by short photoperiods in the Siberian hamster, Phodopus sungorus  

E-print Network

Pubertal growth of the medial amygdala delayed by short photoperiods in the Siberian hamster nucleus of the amygdala (MeA) by comparing Siberian hamsters (Phodopus sungorus) that had been raised from birth in either long day (LD; 16:8 h light:dark) or short day (SD; 8:16) photoperiods. Hamsters were

Breedlove, Marc

269

CELL-MEDIATED CYTOTOXICITY AGAINST HAMSTER CELLS TRANSFORMED BY AVIAN SARCOMA VIRUSES.  

E-print Network

CELL-MEDIATED CYTOTOXICITY AGAINST HAMSTER CELLS TRANSFORMED BY AVIAN SARCOMA VIRUSES. 1 A MEDIATION CELLULAIRE ENVERS DES CELLULES DE HAMSTER TRANS- FORMEES PAR DES VIRUS DE SARCOMES AVIAIRES. 1. Description de la réaction. - La présence de nouveaux antigènes apparus à la surface de cellules de hamster

Boyer, Edmond

270

Male-induced estrus synchronization in the female Siberian hamster (Phodopus sungorus sungorus)  

E-print Network

Male-induced estrus synchronization in the female Siberian hamster (Phodopus sungorus sungorus, affect the reproductive physiology of the female Siberian hamster (Phodopus sungorus sungorus). This lack estrous cyclicity in the female Siberian hamster and, if so, whether the production of these priming

Carr, Leslie

271

Development and initiation of sperm motility in the hamster Marie-Louise KANN, Catherine SERRES  

E-print Network

Development and initiation of sperm motility in the hamster epididymis Marie-Louise KANN, Catherine Bicêtre, 78 rue du Général Leclerc 94270 Kremlin-Bicêtre, France. Summary. Hamster spermatozoa were of the head movements. It is shown that during epididymal transit of hamster spermatozoa, the induction

Paris-Sud XI, Université de

272

Synchrotron FT-IR Analysis of Collagen Localization in Normal, Cardiomyopathic and Losartan-treated Hamsters  

E-print Network

-treated Hamsters K.M. Gough, I. Dixon, P. Bromberg, D. Zielinski Department of Chemistry, University of Manitoba scale). Figure 1. Photomicrograph of stained cardiomyopathic heart tissue from Syrian hamsters. Blue and localization of collagen in model animals: the UM-X7.1 strain of Syrian hamsters. In cases where a collagen

273

9 CFR 3.36 - Primary enclosures used to transport live guinea pigs and hamsters.  

Code of Federal Regulations, 2012 CFR

...used to transport live guinea pigs and hamsters. 3.36 Section 3.36 Animals...and Transportation of Guinea Pigs and Hamsters Transportation Standards § 3.36...used to transport live guinea pigs and hamsters. No person subject to the...

2012-01-01

274

Leptin Effects on Immune Function and Energy Balance Are Photoperiod Dependent in Siberian Hamsters  

E-print Network

Leptin Effects on Immune Function and Energy Balance Are Photoperiod Dependent in Siberian Hamsters- sters (Phodopus sungorus) housed in long or short days for a total of 12 weeks. Short-day hamsters days. In Exp 2, when the leptin-induced increase in food intake in short-day hamsters was prevented

Demas, Greg

275

Gender differences in the development of hyperlipemia and atherosclerosis in hybrid hamsters  

Microsoft Academic Search

In response to a diet enriched in saturated fat and cholesterol (CH), male Syrian hamsters develop hyperlipemia and changes of early atherosclerosis. However, it has not been determined if female hamsters are equally susceptible to an atherogenic diet. Male and female hamsters of the F1B hybrid strain (Bio Breeders, Fitchburg, MA) were fed either a chow diet or this diet

Sander J. Robins; Joan M. Fasulo; George M. Patton; Ernst J. Schaefer; Donald E. Smith; Jose M. Ordovas

1995-01-01

276

Circadian Clock Resetting by Sleep Deprivation without Exercise in Syrian Hamsters: Dark Pulses Revisited  

Microsoft Academic Search

Circadian rhythms in Syrian hamsters can be phase shifted by procedures that stimulate wheel running (“exercise”) in the mid-subjective day (the hamster's usual sleep period). The authors recently demonstrated that keeping hamsters awake by gentle handling, without continuous running, is sufficient to mimic this effect. Here, the authors assessed whether wakefulness, independent of wheel running, also mediates phase shifts to

Ralph E. Mistlberger; Jodi Belcourt; Michael C. Antle

2002-01-01

277

Altered somatosensory receptive fields in hamster colliculus after infraorbital nerve section and xylocaine injection.  

PubMed Central

The effects of acute infraorbital (i.o.) nerve section upon the responses of somatosensory cells in the rostral part of the deep layers of the hamster's superior colliculus were studied using standard extracellular single-unit recording and receptive field mapping techniques. In nine animals a given cell's receptive field was determined both before and after i.o. nerve section and, in all cases, new areas of sensitivity were unmasked within 15 min after the nerve was cut. In a given electrode penetration where the i.o. nerve was sectioned (n = 13), somatosensory cells recorded after the nerve was cut, as the electrode was being withdrawn from the colliculus, exhibited receptive fields considerably different from those of somatosensory cells isolated during the descent of the recording electrode. Seventeen deep-layer somatosensory cells (in eight hamsters) were tested before and after subcutaneous injections of xylocaine into their receptive fields. This manipulation unmasked new areas of cutaneous sensitivity for sixteen units. Of these, the new receptive fields of nine cells disappeared as sensitivity in the original receptive field returned; five ultimately retained both the new and old receptive fields; in two instances, sensitivity in the original receptive field never returned over the 3 h of testing. Control experiments (n = 7) demonstrated that the changes observed did not result from spontaneous alterations in receptive field borders, changes induced by variations in the level of general anaesthesia, or non-specific trauma associated with the xylocaine injections or the surgery required to expose the i.o. nerve. Images Fig. 7 Fig. 8 Plate 1 PMID:6716292

Jacquin, M F; Mooney, R D; Rhoades, R W

1984-01-01

278

PTK2b function during fertilization of the mouse oocyte.  

PubMed

Fertilization triggers rapid changes in intracellular free calcium that serve to activate multiple signaling events critical to the initiation of successful development. Among the pathways downstream of the fertilization-induced calcium transient is the calcium-calmodulin dependent protein tyrosine kinase PTK2b or PYK2 kinase. PTK2b plays an important role in fertilization of the zebrafish oocyte and the objective of the present study was to establish whether PTK2b also functions in mammalian fertilization. PTK2b was activated during the first few hours after fertilization of the mouse oocyte during the period when anaphase resumption was underway and prior to the pronuclear stage. Suppression of PTK2b kinase activity in oocytes blocked sperm incorporation and egg activation although sperm-oocyte binding was not affected. Oocytes that failed to incorporate sperm after inhibitor treatment showed no evidence of a calcium transient and no evidence of anaphase resumption suggesting that egg activation did not occur. The results indicate that PTK2b functions during the sperm-egg fusion process or during the physical incorporation of sperm into the egg cytoplasm and is therefore critical for successful development. PMID:24667605

Luo, Jinping; McGinnis, Lynda K; Carlton, Carol; Beggs, Hilary E; Kinsey, William H

2014-08-01

279

Drosha protein levels are translationally regulated during Xenopus oocyte maturation.  

PubMed

MicroRNAs (miRNAs) are ?21-nucleotide-long, single-stranded noncoding RNAs that regulate gene expression. Biogenesis of miRNAs is mediated by the two RNase III-like enzymes, Drosha and Dicer. Here we study miRNA biogenesis during maturation of Xenopus oocytes to eggs using microinjection of pri-miRNAs. We show that processing of exogenous and endogenous primary miRNAs (pri-miRNAs) is strongly enhanced upon maturation of oocytes to eggs. Overexpression of cloned Xenopus Drosha in oocytes, however, boosts pri-miRNA processing dramatically, indicating that Drosha is a rate-limiting factor in Xenopus oocytes. This developmental regulation of Drosha is controlled by poly(A) length addition to the Drosha mRNA, which boosts translation upon transition from oocytes to eggs. Processing of pri-miRNAs by Drosha and Dicer has been shown to be affected by adenosine-to-inosine deamination-type RNA editing. Using activated Xenopus eggs for microinjection experiments, we demonstrate that RNA editing can reduce pri-miRNA processing in vivo. This processing block is determined by the structural but not sequence changes introduced by RNA editing. PMID:24829383

Muggenhumer, Dominik; Vesely, Cornelia; Nimpf, Simon; Tian, Nan; Yongfeng, Jin; Jantsch, Michael F

2014-07-01

280

Drosha protein levels are translationally regulated during Xenopus oocyte maturation  

PubMed Central

MicroRNAs (miRNAs) are ?21-nucleotide-long, single-stranded noncoding RNAs that regulate gene expression. Biogenesis of miRNAs is mediated by the two RNase III-like enzymes, Drosha and Dicer. Here we study miRNA biogenesis during maturation of Xenopus oocytes to eggs using microinjection of pri-miRNAs. We show that processing of exogenous and endogenous primary miRNAs (pri-miRNAs) is strongly enhanced upon maturation of oocytes to eggs. Overexpression of cloned Xenopus Drosha in oocytes, however, boosts pri-miRNA processing dramatically, indicating that Drosha is a rate-limiting factor in Xenopus oocytes. This developmental regulation of Drosha is controlled by poly(A) length addition to the Drosha mRNA, which boosts translation upon transition from oocytes to eggs. Processing of pri-miRNAs by Drosha and Dicer has been shown to be affected by adenosine-to-inosine deamination–type RNA editing. Using activated Xenopus eggs for microinjection experiments, we demonstrate that RNA editing can reduce pri-miRNA processing in vivo. This processing block is determined by the structural but not sequence changes introduced by RNA editing. PMID:24829383

Muggenhumer, Dominik; Vesely, Cornelia; Nimpf, Simon; Tian, Nan; Yongfeng, Jin; Jantsch, Michael F.

2014-01-01

281

Meiotic maturation of vitrified immature chousingha (Tetracerus quadricornis) oocytes recovered postmortem.  

PubMed

The ability to recover and cryopreserve oocytes from postmortem ovaries of endangered or wildlife species holds tremendous potential for conservation using assisted reproductive technologies. The objective of this study was to assess the in vitro meiotic maturation of chousingha (four-horned antelope) oocytes following vitrification using open pulled straw (OPS) method. The average number of oocytes recovered per ovary was 65.6. The proportion of oocytes that matured was significantly lower in vitrified oocytes (29.4%) when compared with fresh oocytes (69.3%). The study provides evidence that it is possible to cryopreserve immature oocytes by vitrification collected from the ovaries of chousingha at postmortem and also demonstrates that these cryopreserved oocytes retain their potential to undergo in vitro meiotic maturation. PMID:21168399

Rao, Brahmasani Sambasiva; Mahesh, Yelisetti Uma; Suman, Komjeti; Charan, Katari Venu; Lakshmikantan, Uthandaraman; Gibence, Henderson Rose Winnie; Shivaji, Sisinthy

2011-02-01

282

Helminth fauna of the golden hamster Mesocricetus auratus in Brazil.  

PubMed

Helminth fauna of conventionally maintained hamsters from institutional animal houses that supply the research community with laboratory animals and from an openly kept control group, randomly purchased in a pet shop in the State of Rio de Janeiro, were evaluated and compared. Necropsied animals from institutional suppliers were infected with the oxyurid nematodes Syphacia criceti and S. mesocriceti and with the cestode Rodentolepis nana; those from the pet shop were infected with S. mesocriceti and R. nana. These are the first morphometric data that are based on Brazilian samples of these species parasitizing hamsters. Mesocricetus auratus is a newly recorded host for S. criceti, previously recovered from Oryzomys subflavus and Calomys callosus in Brazil. The potential of pet and laboratory hamsters in the spreading of helminth infections to humans is also considered. PMID:11300683

Pinto, R M; Gonçalves, L; Gomes, D C; Noronha, D

2001-03-01

283

Characteristics of cadmium-induced nephrotoxicity in Syrian hamsters.  

PubMed

Male Syrian hamsters were used to evaluate cadmium (Cd)-induced nephrotoxicity. Furthermore, they were treated with polyaspartic acid (PAA) in an attempt to prevent renal damage due to Cd. To induce renal proximal tubular damage, the hamsters were administered a single subcutaneous injection of cadmium chloride (CdCl2) at the dose of 3 mg/kg body weight. Within 24 hours, they exhibited significant proteinuria and an increased urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG). Renal histological changes consisted of degenerative changes in renal tubule cells, irregularly shaped nuclei with marginated chromatin and rounded mitochondria. The administration of PAA did not improve creatinine clearance or urinary protein excretion. The concentration of Cd in the renal tissue showed a gradual increase on day 3 following cadmium exposure. Cadmium nephrotoxicity appears to be more severe in Syrian hamsters than in rats or mice. Thus, this animal model appears to be excellent for studying Cd-induced nephrotoxicity. PMID:8255000

Shibasaki, T; Ohno, I; Ishimoto, F; Sakai, O

1993-08-01

284

Pathogenesis of Modoc virus (Flaviviridae; Flavivirus) in persistently infected hamsters.  

PubMed

The long-term persistence of Modoc virus (MODV) infection was investigated in a hamster model. Golden hamsters (Mesocricetus auratus) were infected by subcutaneous inoculation with MODV, in which fatal encephalitis developed in 12.5% (2 of 16). Surviving hamsters shed infectious MODV in their urine during the first five months after infection, and infectious MODV was recovered by co-cultivation of kidney tissue up to eight months after infection. There were no histopathologic changes observed in the kidneys despite detection of viral antigen for 250 days after infection. Mild inflammation and neuronal degeneration in the central nervous system were the primary lesions observed during early infection. These findings confirm previous reports of persistent flavivirus infection in animals and suggest a mechanism for the maintenance of MODV in nature. PMID:23358636

Adams, A Paige; Travassos da Rosa, Amelia P A; Nunes, Marcio R; Xiao, Shu-Yuan; Tesh, Robert B

2013-03-01

285

Pathogenesis of Modoc Virus (Flaviviridae; Flavivirus) in Persistently Infected Hamsters  

PubMed Central

The long-term persistence of Modoc virus (MODV) infection was investigated in a hamster model. Golden hamsters (Mesocricetus auratus) were infected by subcutaneous inoculation with MODV, in which fatal encephalitis developed in 12.5% (2 of 16). Surviving hamsters shed infectious MODV in their urine during the first five months after infection, and infectious MODV was recovered by co-cultivation of kidney tissue up to eight months after infection. There were no histopathologic changes observed in the kidneys despite detection of viral antigen for 250 days after infection. Mild inflammation and neuronal degeneration in the central nervous system were the primary lesions observed during early infection. These findings confirm previous reports of persistent flavivirus infection in animals and suggest a mechanism for the maintenance of MODV in nature. PMID:23358636

Adams, A. Paige; Travassos da Rosa, Amelia P. A.; Nunes, Marcio R.; Xiao, Shu-Yuan; Tesh, Robert B.

2013-01-01

286

Experimental investigation of wavelength dependence of penetration depth and imaging contrast for ultrahigh-resolution optical coherence tomography  

NASA Astrophysics Data System (ADS)

Optical coherence tomography (OCT) is a non invasive optical imaging technology for micron-scale cross-sectional imaging of biological tissue and materials. Although OCT has many advantages in medical equipments, low penetration depth is a serious limitation for other applications. To realize the ultrahigh resolution and the high penetration depth at the same time, it is effective to choose the proper wavelength to maximize the light penetration and enhance the image contrast at deeper depths. Recently, we have demonstrated ultrahigh resolution and high penetration depth OCT by use of all-fiber based Gaussian shaped supercontinuum source at 1.7 ?m center wavelength. Gaussian-like supercontinuum with 360 nm bandwidth at center wavelength of 1.7 ?m was generated by ultrashort pulse Er doped fiber laser based system. In this paper, using 0.8 ?m and 1.3 ?m SC sources in addition to the 1.7 ?m SC source, we have investigated the wavelength dependence of ultrahigh resolution OCT in terms of penetration depth. Longitudinal resolutions at each wavelength region are almost 4.6 ?m in air. The obtained sensitivity was 95 dB for all wavelength regions. We confirmed the difference of imaging contrast and penetration depth with hamster's cheek pouch and so on. As the wavelength was increased, the magnitude of penetration depth was increased for these samples.

Ishida, S.; Nishizawa, N.; Itoh, K.

2011-03-01

287

Projectile penetration into ballistic gelatin.  

PubMed

Ballistic gelatin is frequently used as a model for soft biological tissues that experience projectile impact. In this paper we investigate the response of a number of gelatin materials to the penetration of spherical steel projectiles (7 to 11mm diameter) with a range of lower impacting velocities (<120m/s). The results of sphere penetration depth versus projectile velocity are found to be linear for all systems above a certain threshold velocity required for initiating penetration. The data for a specific material impacted with different diameter spheres were able to be condensed to a single curve when the penetration depth was normalised by the projectile diameter. When the results are compared with a number of predictive relationships available in the literature, it is found that over the range of projectiles and compositions used, the results fit a simple relationship that takes into account the projectile diameter, the threshold velocity for penetration into the gelatin and a value of the shear modulus of the gelatin estimated from the threshold velocity for penetration. The normalised depth is found to fit the elastic Froude number when this is modified to allow for a threshold impact velocity. The normalised penetration data are found to best fit this modified elastic Froude number with a slope of 1/2 instead of 1/3 as suggested by Akers and Belmonte (2006). Possible explanations for this difference are discussed. PMID:24184862

Swain, M V; Kieser, D C; Shah, S; Kieser, J A

2014-01-01

288

Autonomic Nervous Dysfunction in Hamsters Infected with West Nile Virus  

PubMed Central

Clinical studies and case reports clearly document that West Nile virus (WNV) can cause respiratory and gastrointestinal (GI) complications. Other functions controlled by the autonomic nervous system may also be directly affected by WNV, such as bladder and cardiac functions. To investigate how WNV can cause autonomic dysfunctions, we focused on the cardiac and GI dysfunctions of rodents infected with WNV. Infected hamsters had distension of the stomach and intestines at day 9 after viral challenge. GI motility was detected by a dye retention assay; phenol red dye was retained more in the stomachs of infected hamsters as compared to sham-infected hamsters. The amplitudes of electromygraphs (EMGs) of intestinal muscles were significantly reduced. Myenteric neurons that innervate the intestines, in addition to neurons in the brain stem, were identified to be infected with WNV. These data suggest that infected neurons controlling autonomic function were the cause of GI dysfunction in WNV-infected hamsters. Using radiotelemetry to record electrocardiograms and to measure heart rate variability (HRV), a well-accepted readout for autonomic function, we determined that HRV and autonomic function were suppressed in WNV-infected hamsters. Cardiac histopathology was observed at day 9 only in the right atrium, which was coincident with WNV staining. A subset of WNV infected cells was identified among cells with hyperplarization-activated cyclic nucleotide-gated potassium channel 4 (HCN4) as a marker for cells in the sinoatrial (SA) and atrioventricular (AV) nodes. The unique contribution of this study is the discovery that WNV infection of hamsters can lead to autonomic dysfunction as determined by reduced HRV and reduced EMG amplitudes of the GI tract. These data may model autonomic dysfunction of the human West Nile neurological disease. PMID:21573009

Wang, Hong; Siddharthan, Venkatraman; Hall, Jeffery O.; Morrey, John D.

2011-01-01

289

Maternal Photoperiodic History Affects Offspring Development in Syrian Hamsters  

PubMed Central

During the first 7 weeks of postnatal life, short day lengths inhibit the onset of puberty in many photoperiodic rodents, but not in Syrian hamsters. In this species, timing of puberty and fecundity are independent of the early postnatal photoperiod. Gestational day length affects postnatal reproductive development in several rodents; its role in Syrian hamsters has not been assessed. We tested the hypothesis that cumulative effects of pre- and postnatal short day lengths would restrain gonadal development in male Syrian hamsters. Males with prenatal short day exposure were generated by dams transferred to short day lengths 6 weeks, 3 weeks, and 0 weeks prior to mating. Additional groups were gestated in long day lengths and transferred to short days at birth, at 4 weeks of age, or not transferred (control hamsters). In pups of dams exposed to short day treatment throughout gestation, decreased testis growth was apparent by 3 weeks and persisted through 9 weeks of age, at which time maximum testis size was attained. A subset of males (14%), whose dams had been in short days for 3 to 6 weeks prior to mating displayed pronounced delays in testicular development, similar to those of other photoperiodic rodents. This treatment also increased the percentage of male offspring that underwent little or no gonadal regression postnatally (39%). By 19 weeks of age, males housed in short days completed spontaneous gonadal development. After prolonged long day treatment to break refractoriness, hamsters that initially were classified as nonregressors underwent testicular regression in response to a 2nd sequence of short day lengths. The combined action of prenatal and early postnatal short day lengths diminishes testicular growth of prepubertal Syrian hamsters no later than the 3rd week of postnatal life, albeit to a lesser extent than in other photoperiodic rodents. PMID:18838610

Beery, Annaliese K.; Paul, Matthew J.; Routman, David M.; Zucker, Irving

2009-01-01

290

Birth of a healthy female after intracytoplasmic sperm injection of cryopreserved human oocytes  

Microsoft Academic Search

Objective: To describe the first birth achieved after intracytoplasmic sperm injection (ICSI) of cryopreserved human oocytes.Design: Case report.Setting: University of Bologna Hospital, Department of Obstetrics and Gynecology, Reproductive Endocrinology Unit, IVF and Infertility Center.Patient(s): One patient undergoing IVF.Intervention(s): Transvaginal ultrasound-guided oocyte retrieval followed by oocyte freezing. Artificial preparation of the endometrium with E2 and P, oocyte thawing, and ICSI.Result(s): Four

Eleonora Porcu; Raffaella Fabbri; Renato Seracchioli; Patrizia M. Ciotti; Otello Magrini; Carlo Flamigni

1997-01-01

291

Discovery and Characterization of a New Cell-Penetrating Protein  

PubMed Central

We describe a new cell-penetrating protein, B1, capable of delivering conjugated proteins and nucleic acids into mammalian cells. B1 is a 244-amino-acid product of a single-base frameshift in the gene encoding enhanced green fluorescent protein (eGFP). The molecule has a net positive charge of 43 and a very high charge-to-mass ratio of 1.5. eGFP-fused B1 potently penetrates both adherent and suspension cells with >80% of cells taking up the protein when exposed to concentrations as low as 1 ?M. The protein was found to cluster in the paranuclear region of TZM-bl cells. Most importantly, we show that B1 not only facilitates cellular uptake, but allows biomolecular cargo to reach sites of biological relevance. For example, baby hamster kidney cells underwent DNA recombination when exposed to B1-tagged Cre recombinase at protein concentrations as low as 2.5 ?M, indicating potent nuclear delivery of functional protein cargos. Additionally, B1 delivers non-covalently conjugated RNA and DNA across the cell membrane to cytosolic and nuclear sites accessible to the cellular translation and transcription machinery, as gauged by detection of encoded reporter functions, with efficiency comparable to commercially available cationic lipid reagents. B1 appears to utilize cell-surface glycans and multiple competing endocytic pathways to enter and traffic through cells. These studies provide both a new tool for intracellular delivery of biomolecules and insights that could aid in the design of more effective cell penetrating proteins. PMID:24047285

Simeon, Rudo L.; Chamoun, Ana Maria; McMillin, Thomas; Chen, Zhilei

2013-01-01

292

Retrospective comparison of two media for invitro maturation of oocytes.  

PubMed

In-vitro maturation of oocytes (IVM) is a new IVF technology developed in order to avoid iatrogenic complications of standard IVF treatments. This technique is particularly useful in patients suffering from polycystic ovary syndrome (PCOS) who are concerned with the risk of ovarian hyperstimulation syndrome. This technique is nowadays routinely practised in many international centres. However, the efficiency of this technique needs to be improved for a better support of maturation conditions to maximize oocyte developmental competence. In order to improve IVM results, the efficiency of two IVM media was retrospectively compared. Ninety-three PCOS candidates undergoing their first IVM cycle were included in this study, and IVM was conducted with TCM-199 or IVM-Medicult medium. This is the first study comparing two maturation media. Both media resulted in the same results concerning total oocyte maturation, fertilization, early embryo development and pregnancy rates. PMID:18284882

Filali, M; Hesters, L; Fanchin, R; Tachdjian, G; Frydman, R; Frydman, N

2008-02-01

293

Production of Sry knockout mouse using TALEN via oocyte injection  

PubMed Central

Recently developed transcription activator-like effector nuclease (TALEN) technology has enabled the creation of knockout mice, even for genes on the Y chromosome. In this study, we generated a knockout mouse for Sry, a sex-determining gene on the Y chromosome, using microinjection of TALEN RNA into pronuclear stage oocytes. As expected, the knockout mouse had female external and internal genitalia, a female level of blood testosterone and a female sexually dimorphic nucleus in the brain. The knockout mouse exhibited an estrous cycle and performed copulatory behavior as females, although it was infertile or had reduced fertility. A histological analysis showed that the ovary of the knockout mouse displayed a reduced number of oocytes and luteinized unruptured follicles, implying that a reduced number of ovulated oocytes is a possible reason for infertility and/or reduced fertility in the KO mouse. PMID:24190364

Kato, Tomoko; Miyata, Kohei; Sonobe, Miku; Yamashita, Satoshi; Tamano, Moe; Miura, Kento; Kanai, Yoshiakira; Miyamoto, Shingo; Sakuma, Tetsushi; Yamamoto, Takashi; Inui, Masafumi; Kikusui, Takefumi; Asahara, Hiroshi; Takada, Shuji

2013-01-01

294

Cumulus and granulosa cell markers of oocyte and embryo quality  

PubMed Central

Lack of an objective, accurate, and noninvasive embryo assessment strategy remains one of the major challenges encountered in in vitro fertilization. Cumulus and mural granulosa cells reflect the characteristics of the oocyte, providing a noninvasive means to assess oocyte quality. Specifically, transcriptomic profiling of follicular cells may help identify biomarkers of oocyte and embryo competence. Current transcriptomics technologies include quantitative reverse transcriptase–polymerase chain reaction (qRT-PCR) for analysis of individual genes and microarrays and high-throughput deep sequencing for whole genome expression profiling. Recently, using qRT-PCR and microarray technologies, a multitude of studies correlated changes in cumulus or granulosa cell gene expression with clinically relevant outcome parameters, including in vitro embryo development and pregnancy. While the initial findings are promising, a clinical benefit from the use of identified biomarker genes remains to be demonstrated in randomized controlled trials. PMID:23498999

Uyar, Asli; Torrealday, Saioa; Seli, Emre

2013-01-01

295

Isolation and identification of normal killer cells from Syrian hamsters  

SciTech Connect

This paper gives data on isolation of normal killer cells from the blood and various tissues of Syrian hamsters in a Percoll density gradient and their identification on the basis of morphologic criteria and cytotoxic activity (CTA). CTA of the isolated cells was studied in the cytotoxic test with target cells of a human MOLT-4 thymoma cell labeled with /sup 51/Cr. Isolation of large granular lymphocytes from blood, spleen, and bone marrow of Syrian hamsters in Percoll density gradient is shown in the results of five experiments used for cells of each type.

Matveeva, V.A.; Klyuchareva, T.E.

1986-09-01

296

Expression of TGF? superfamily components and other markers of oocyte quality in oocytes selected by brilliant cresyl blue staining: Relevance to early embryonic development  

PubMed Central

Brilliant cresyl blue (BCB) is a super vital stain that has been used to select competent oocytes in different species. The objectives of present studies were to determine mRNA abundance for select TGF? superfamily components, SMAD2/3 and SMAD1/5 phosphorylation levels and transcript abundance for other oocyte (JY1) and cumulus cell (CTSB, CTSK, CTSS and CTSZ) markers of oocyte quality in bovine oocytes and or adjacent cumulus cells classified based on developmental potential using BCB staining. The ability of exogenous FST, JY1, or cathepsin inhibitor treatment to enhance development of embryos derived from poor quality oocytes selected based on BCB staining was also determined. Cumulus oocyte complexes (COCs) from abattoir derived ovaries were subjected to BCB staining and GV stage oocytes and cumulus cells harvested from control, BCB+ and BCB- (poor oocyte quality) groups for real time PCR or Western blot analysis. Remaining COCs underwent in vitro maturation, in vitro fertilization and embryo culture in presence or absence of above described treatments. Levels of FST, JY1, BMP15 and SMAD1, 2, 3 and 5 transcripts were higher in BCB+ oocytes whereas abundance of CTSB, CTSK, CTSS and CTSZ mRNAs was higher in cumulus cells surrounding poor quality BCB- oocytes. Western blot analysis revealed SMAD1/5 and SMAD2/3 phosphorylation were higher in BCB+ than BCB? oocytes. Embryo culture studies demonstrated that follistatin and cathepsin inhibitor treatment but not JY-1 treatment can promote developmental competence of BCB- oocytes. Results provide further understanding of molecular indices of oocyte competence. PMID:25704641

Ashry, Mohamed; Lee, KyungBon; Mondal, Mohan; Datta, Tirtha K.; Folger, Joseph K.; Rajput, Sandeep K.; Zhang, Kun; Hemeida, Nabil A.; Smith, George W.

2015-01-01

297

AMS 102: QUIZ 2 1. The population of long-tailed hamster in eastern Texas has been de-  

E-print Network

AMS 102: QUIZ 2 SOLUTIONS 1. The population of long-tailed hamster in eastern Texas has been de of the hamster's natural habitat. Two statistical studies were conducted that, on the year-by-year basis (1) compared hamster population with the area of human settlement, (2) compared hamster population with annual

Retakh, Alexander

298

Cholesterol Depletion Disorganizes Oocyte Membrane Rafts Altering Mouse Fertilization  

PubMed Central

Drastic membrane reorganization occurs when mammalian sperm binds to and fuses with the oocyte membrane. Two oocyte protein families are essential for fertilization, tetraspanins and glycosylphosphatidylinositol-anchored proteins. The firsts are associated to tetraspanin-enriched microdomains and the seconds to lipid rafts. Here we report membrane raft involvement in mouse fertilization assessed by cholesterol modulation using methyl-?-cyclodextrin. Cholesterol removal induced: (1) a decrease of the fertilization rate and index; and (2) a delay in the extrusion of the second polar body. Cholesterol repletion recovered the fertilization ability of cholesterol-depleted oocytes, indicating reversibility of these effects. In vivo time-lapse analyses using fluorescent cholesterol permitted to identify the time-point at which the probe is mainly located at the plasma membrane enabling the estimation of the extent of the cholesterol depletion. We confirmed that the mouse oocyte is rich in rafts according to the presence of the raft marker lipid, ganglioside GM1 on the membrane of living oocytes and we identified the coexistence of two types of microdomains, planar rafts and caveolae-like structures, by terms of two differential rafts markers, flotillin-2 and caveolin-1, respectively. Moreover, this is the first report that shows characteristic caveolae-like invaginations in the mouse oocyte identified by electron microscopy. Raft disruption by cholesterol depletion disturbed the subcellular localization of the signal molecule c-Src and the inhibition of Src kinase proteins prevented second polar body extrusion, consistent with a role of Src-related kinases in fertilization via signaling complexes. Our data highlight the functional importance of intact membrane rafts for mouse fertilization and its dependence on cholesterol. PMID:23638166

Buschiazzo, Jorgelina; Ialy-Radio, Come; Auer, Jana; Wolf, Jean-Philippe; Serres, Catherine

2013-01-01

299

Optimization of cryoprotectant loading into murine and human oocytes.  

PubMed

Loading of cryoprotectants into oocytes is an important step of the cryopreservation process, in which the cells are exposed to potentially damaging osmotic stresses and chemical toxicity. Thus, we investigated the use of physics-based mathematical optimization to guide design of cryoprotectant loading methods for mouse and human oocytes. We first examined loading of 1.5 M dimethyl sulfoxide (Me(2)SO) into mouse oocytes at 23°C. Conventional one-step loading resulted in rates of fertilization (34%) and embryonic development (60%) that were significantly lower than those of untreated controls (95% and 94%, respectively). In contrast, the mathematically optimized two-step method yielded much higher rates of fertilization (85%) and development (87%). To examine the causes for oocyte damage, we performed experiments to separate the effects of cell shrinkage and Me(2)SO exposure time, revealing that neither shrinkage nor Me(2)SO exposure single-handedly impairs the fertilization and development rates. Thus, damage during one-step Me(2)SO addition appears to result from interactions between the effects of Me(2)SO toxicity and osmotic stress. We also investigated Me(2)SO loading into mouse oocytes at 30°C. At this temperature, fertilization rates were again lower after one-step loading (8%) in comparison to mathematically optimized two-step loading (86%) and untreated controls (96%). Furthermore, our computer algorithm generated an effective strategy for reducing Me(2)SO exposure time, using hypotonic diluents for cryoprotectant solutions. With this technique, 1.5 M Me(2)SO was successfully loaded in only 2.5 min, with 92% fertilizability. Based on these promising results, we propose new methods to load cryoprotectants into human oocytes, designed using our mathematical optimization approach. PMID:24246951

Karlsson, Jens O M; Szurek, Edyta A; Higgins, Adam Z; Lee, Sang R; Eroglu, Ali

2014-02-01

300

Microinjection of Xenopus laevis oocytes as a system for studying nuclear transport of viruses  

Microsoft Academic Search

Microinjection of Xenopus laevis oocytes is an excellent system for studying nuclear transport because of the large size of the oocyte and its high nuclear pore complex (NPC) density. In addition, the fact that Xenopus oocytes are not permissive for most mammalian viruses makes this system especially useful for studying nuclear transport of viruses in the absence of the confounding

Shelly Au; Sarah Cohen; Nelly Panté

2010-01-01

301

Maintenance of meiotic prophase arrest in vertebrate oocytes by a G s protein-mediated pathway  

Microsoft Academic Search

Maintenance of meiotic prophase arrest in fully grown vertebrate oocytes depends on an elevated level of cAMP in the oocyte. To investigate how the cAMP level is regulated, we examined whether the activity of an oocyte G protein of the family that stimulates adenylyl cyclase, Gs, is required to maintain meiotic arrest. Microinjection of a dominant negative form of Gs

Rebecca R. Kalinowski; Catherine H. Berlot; Teresa L. Z. Jones; Lavinia F. Ross; Laurinda A. Jaffe; Lisa M. Mehlmann

2004-01-01

302

Precocious induction of oocyte maturation and ovulation in rainbow trout (Salmo gairdneri) : problems when using  

E-print Network

Precocious induction of oocyte maturation and ovulation in rainbow trout (Salmo gairdneri of 3 mg/kg at a 2-day interval) induced oocyte maturation in 94 p. 100 of the fish, but only 25 p. 100 fish, 59 p. 100 of which ovulated. In both cases, fish in which ovulation did not follow oocyte

Paris-Sud XI, Université de

303

The influence of strain, maternal age, and method of maturation on mouse oocyte aneuploidy  

Microsoft Academic Search

The chromosome complement of metaphase II oocytes matured by different methods was determined. These oocytes were collected from young and old Swiss-Webster random bred and CBA inbred mice. The results from the two strains were remarkably similar. Superovulation caused no increase in aneuploidy frequencies; however, in vitro maturation resulted in increased hyperploidy rates for oocytes from younger females of both

M. S. Golbus

1981-01-01

304

Vitrification of oocytes from endangered Mexican gray wolves ( Canis lupus baileyi)  

Microsoft Academic Search

Careful genetic management, including cryopreservation of genetic material, is central to conservation of the endangered Mexican gray wolf. We tested a technique, previously used to vitrify human and domestic animal oocytes, on oocytes from domestic dogs as a model and from the endangered Mexican wolf. This method provided a way to conserve oocytes from genetically valuable older female Mexican wolves

S. Boutelle; K. Lenahan; R. Krisher; K. L. Bauman; C. S. Asa; S. Silber

2011-01-01

305

A Method for Viewing the Germinal Vesicle in Oocytes of Commercial Catfishes  

Microsoft Academic Search

Position of the germinal vesicle is a key indicator of stage of oocyte maturation, and thus it is a valuable assessment tool in studies of the reproductive biology of fishes. The germinal vesicle of untreated oocytes of commercially reared catfish species cannot be seen because oocytes of these species are opaque. However, submerging them in a bath of Serra's fixative

Joseph N. Stoeckel

2000-01-01

306

Volume changes of mature human oocytes on exposure to cryoprotectant solutions used in slow cooling procedures  

Microsoft Academic Search

BACKGROUND: Despite the recent increase in pregnancies from cryopreserved human oocytes, success in terms of births per thawed oocyte is still poor. Modifications to cryopreservation protocols have not been based on measurement of the osmotic response of oocytes, and methodologies are often poorly described or protocols not strictly adhered to, inevitably resulting in variability. METHODS: Volume change of mature human

S. J. Paynter; A. Borini; V. Bianchi; L. De Santis; C. Flamigni; G. Coticchio

2005-01-01

307

Survival of human oocytes cryopreserved with or without the cumulus in 1,2-propanediol  

Microsoft Academic Search

Background Although cryopreservation of human preembryos has been carried out with success, the cryostorage of oocytes, which pose fewer controversial moral, ethical, and legal problems has been much less successful. Various attempts to cryopreserve human oocytes have been mostly unsuccessful and the search for an optimal protocol for oocyte cryopreservation remains elusive. We therefore undertook this study to determine the

Daniel G. Imoedemhe; Alejandro B. Sigue

1992-01-01

308

Fertilizable oocytes reconstructed from patient's somatic cell nuclei and donor ooplasts  

Microsoft Academic Search

The only assisted reproduction treatment now available for women with ovarian failure or irreparable oocyte defects is oocyte donation. However, some women experience psychological barriers to the recourse to donor oocytes, related to the lack of contribution of their proper genes to the progeny. A pilot study in humans suggests that this problem may be overcome by the development of

J Tesarik; ZP Nagy; M Sousa; C Mendoza; R Abdelmassih

2001-01-01

309

INTRODUCTION The end-point of mammalian oocyte development is in the pro-  

E-print Network

fertile oocyte. The mouse oocyte grows from an initial diameter of 20 µm to 70 µm in size, during which be stimulated in vitro by releasing the oocyte from the follicle into a suitable culture medium (Pincus metaphase II where it arrests, awaiting fertilization. At fertilization the sperm triggers a series

Newcastle upon Tyne, University of

310

Human oocyte and ovarian tissue cryopreservation and its application  

Microsoft Academic Search

Purpose  To review the recent progress in human oocyte and ovarian tissue cryopreservation, and in the application of these two technologies\\u000a for preserving female fertility of patients who are undergoing cancer treatment.\\u000a \\u000a \\u000a \\u000a Design  The literature on human oocyte and ovarian tissue freezing was searched with PubMed. The scientific background, current developments\\u000a and potential future applications of these two methods were reviewed.\\u000a \\u000a \\u000a \\u000a Results  Chemotherapy

Tao Tao; Alfonso Del Valle

2008-01-01

311

Investigations into Monochloramine Biofilm Penetration  

EPA Science Inventory

Biofilm in drinking water systems is undesirable. Free chlorine and monochloramine are commonly used as secondary drinking water disinfectants, but monochloramine is perceived to penetrate biofilm better than free chlorine. However, this hypothesis remains unconfirmed by direct b...

312

Ground Penetrating Radar, Barrow, Alaska  

DOE Data Explorer

This is 500 MHz Ground Penetrating Radar collected along the AB Line in Intensive Site 1 beginning in October 2012 and collected along L2 in Intensive Site 0 beginning in September 2011. Both continue to the present.

John Peterson

313

Inspecting the reactor vessel penetrations  

SciTech Connect

The susceptibility of Alloy 600 to Primary Water Stress Corrosion Cracking (PWSCC) continues to plague nuclear power plants. Recently, the problem of PWSCC cracking has manifested itself in Control Rod Drive Mechanism (CRDM) head penetrations in nuclear plants in Europe. Framatome has been extensively involved in the performance of both inspections and repairs of CRDM head penetrations at Electricite de France (EdF) plants. B and W Nuclear Technologies (BWNT), building on Framatome technology, has developed a fully integrated service package and robotic manipulator to inspect and repair CRDM head penetrations for US utilities. Reactor vessel bottom penetration are also made of Alloy 600 and to tackle this potential PWSCC problem at EdF plants, Framatome has been performing specific inspections in order to detect the appearance of the phenomenon. This paper describes the overall range of inspection techniques and toolings developed to address these issues.

Bodson, F. [Framatome, Chalon-Sur-Saone (France); Fleming, K.W. [BWNT, Lynchburg, VA (United States)

1995-08-01

314

Particle Penetration Through Building Cracks  

Microsoft Academic Search

Particle penetration into buildings influences human exposure to particles of ambient origin. In this study, we present the results of laboratory experiments measuring particle penetration through surrogates of cracks in building envelopes. Rectangular slots were prepared, with crack heights of 0.25 and 1 mm and flow-path lengths of 4-10 cm, using common building materials: aluminum, brick, concrete, plywood, redwood lumber,

De-Ling Liu; William W. Nazaroff

2003-01-01

315

Efficient Gene Targeting in Golden Syrian Hamsters by the CRISPR/Cas9 System  

PubMed Central

The golden Syrian hamster is the model of choice or the only rodent model for studying many human diseases. However, the lack of gene targeting tools in hamsters severely limits their use in biomedical research. Here, we report the first successful application of the CRISPR/Cas9 system to efficiently conduct gene targeting in hamsters. We designed five synthetic single-guide RNAs (sgRNAs)—three for targeting the coding sequences for different functional domains of the hamster STAT2 protein, one for KCNQ1, and one for PPP1R12C—and demonstrated that the CRISPR/Cas9 system is highly efficient in introducing site-specific mutations in hamster somatic cells. We then developed unique pronuclear (PN) and cytoplasmic injection protocols in hamsters and produced STAT2 knockout (KO) hamsters by injecting the sgRNA/Cas9, either in the form of plasmid or mRNA, targeting exon 4 of hamster STAT2. Among the produced hamsters, 14.3% and 88.9% harbored germline-transmitted STAT2 mutations from plasmid and mRNA injection, respectively. Notably, 10.4% of the animals produced from mRNA injection were biallelically targeted. This is the first success in conducting site-specific gene targeting in hamsters and can serve as the foundation for developing other genetically engineered hamster models for human disease. PMID:25299451

Meng, Qinggang; Shi, Bi; Bunch, Thomas D.; White, Kenneth L.; Kong, Il-Keun; Wang, Zhongde

2014-01-01

316

Demonstration of Survivable Space Penetrator  

NASA Astrophysics Data System (ADS)

This work was performed in support of MoonLITE which is a proposed UK space mission to the moon. The basic premise is to deploy 4 instrumented penetrators, one each on the near-side, far-side and at the poles of the moon, with an impact velocity of approximately 300m/s. The primary science aims are to set up a passive seismometer network, investigate the presence of water and volatiles and determine thermal gradients in the lunar soil (i.e. regolith). A key requirement is that the penetrator shell survives the impact together with the instrument payload and supporting subsystems. The material chosen for the penetrator shell was 7075 aluminum alloy, which is a good compromise between high compressive strength and low mass. The baseline penetrator design was evaluated and refined using the DYNA3D hydrocode to determine the survivability of the penetrator in sand at an impact velocity of 300m/s and an attack angle of 8 degrees. The simulations predicted that the penetrator design would survive this severe impact condition which was confirmed by experiments on the Pendine rocket test track.

Church, P.; Huntington-Thresher, W.; Penny, N.; Bruce, A.; Smith, A.; Gowan, R.

2009-06-01

317

Molecular cloning of hamster brain and atrial natriuretic peptide cDNAs. Cardiomyopathic hamsters are useful models for brain and atrial natriuretic peptides.  

PubMed Central

Brain and atrial natriuretic peptides (BNP and ANP) are cardiac hormones with diuretic, natriuretic, and vasodilatory activities. Cardiomyopathic hamsters are widely used animal models of heart failure. Due to the structural divergence of BNP among species, examination on pathophysiological roles of BNP using cardiomyopathic hamsters is so far impossible. We therefore isolated hamster BNP and ANP cDNAs, and investigated synthesis and secretion of these peptides in normal and cardiomyopathic hamsters. The COOH-terminal 32-residue peptide of cloned hamster preproBNP with 122 amino acids, preceded by a single arginine residue, supposedly represents hamster BNP showing < 50% homology to rat BNP. Alpha-hamster ANP, 28-residue peptide, is identical to alpha-rat ANP. In hamsters, BNP and ANP occur mainly in the ventricle and the atrium, respectively. The 32-wk-old hypertrophic cardiomyopathic BIO14.6 strain exhibited ventricular hypertrophy. The 32-wk-old dilated cardiomyopathic BIO53.58 strain remained at the stage without apparent heart failure. In BIO14.6 and BIO53.58 strains at this age, ventricular BNP and ANP gene expressions are augmented, and the plasma BNP concentration is elevated to 136 and 108 fmol/ml, respectively, three times greater than the elevated plasma ANP concentration, which well mimics changes of the plasma BNP and ANP concentrations in human heart failure. Cardiomyopathic hamsters, therefore, are useful models to investigate the implication of BNP in human cardiovascular diseases. Images PMID:8083346

Tamura, N; Ogawa, Y; Itoh, H; Arai, H; Suga, S; Nakagawa, O; Komatsu, Y; Kishimoto, I; Takaya, K; Yoshimasa, T

1994-01-01

318

Vitrification of oocytes from endangered Mexican gray wolves (Canis lupus baileyi).  

PubMed

Careful genetic management, including cryopreservation of genetic material, is central to conservation of the endangered Mexican gray wolf. We tested a technique, previously used to vitrify human and domestic animal oocytes, on oocytes from domestic dogs as a model and from the endangered Mexican wolf. This method provided a way to conserve oocytes from genetically valuable older female Mexican wolves as an alternative to embryos for preserving female genes. Oocytes were aspirated from ovaries of 36 female dogs in December and March (0 to 65 oocytes per female) and from six female wolves (4 to 73 per female) during their physiologic breeding season, or following stimulation with the GnRH agonist deslorelin. Oocytes from dogs were pooled; half were immediately tested for viability and the remainder vitrified, then warmed and tested for viability. All oocytes were vitrified by being moved through media of increasing cryoprotectant concentration, placed on Cryotops, and plunged into liquid nitrogen. There was no difference in viability (propidium iodide staining) between fresh and vitrified, warmed dog oocytes (65.7 and 61.0%, respectively, P = 0.27). Oocyte viability after warming was similarly assessed in a subset of wolves (4 to 15 oocytes from each of three females; total 29 oocytes). Of these, 57.1% of the post-thaw intact oocytes were viable, which was 41.4% of all oocytes warmed. These were the first oocytes from a canid or an endangered species demonstrated to have maintained viability after vitrification and warming. Furthermore, our results demonstrated that vitrification of oocytes with the Cryotop technique was an option for preserving female gametes from Mexican wolves for future use in captive breeding programs, although in vitro embryo production techniques must first be developed in canids for this technique to be used. PMID:21111469

Boutelle, S; Lenahan, K; Krisher, R; Bauman, K L; Asa, C S; Silber, S

2011-03-01

319

Nuclear Inclusions in the Liver Cells of the Golden Hamster  

PubMed Central

Intranuclear inclusion bodies were observed in the liver cells of 21 of 104 hamsters. The cause of these inclusions remains unknown but they probably represent infoldings of the nuclear membrane in which the cytoplasm has become segregated. An account is given of the age incidence, morphology and histo-chemical features of these inclusions. ImagesFig. 1.Fig. 2. PMID:4234790

Lussier, G.; Pavilanis, V.

1968-01-01

320

RELATIONSHIP BETWEEN AUTONOMIC AND BEHAVIORAL THERMOREGULATION IN THE GOLDEN HAMSTER  

EPA Science Inventory

Preferred ambient temperature (Ta) of male golden hamsters (Mesocricitus auratus) was measured repeatedly by placing the animals in a temperature gradient for 80 min. A total of 180 observations were made during the last 20 min of treatment in the gradient. The mean preferred Ta ...

321

Stimulation of Hamster Sebaceous Glands by Epidermal Growth Factor  

Microsoft Academic Search

Subcutaneous injection of epidermal growth factor (EGF) into the pinna of adult female and castrated male Syrian hamsters resulted in an increase in the number of cells per sebaceous gland unit. The effect of EGF on the sebaceous cell number was localized to the treated ear and accompanied by epidermal hyperplasia. The injection of testosterone into the ear also produced

Jonathan R. Matias; Norman Orentreich

1983-01-01

322

Melatonin Production Accompanies Arousal from Daily Torpor in Siberian Hamsters  

E-print Network

577 Melatonin Production Accompanies Arousal from Daily Torpor in Siberian Hamsters Jennie E is accompanied by a transient rise of melatonin (Mel) in circulation; there are no comparable analyses of Mel of Chicago. All rights reserved. 1522-2152/2003/7604-2092$15.00 Introduction Production of melatonin (Mel

Zucker, Irving

323

Cigarette smoke inhalation affects the reproductive system of female hamsters  

Microsoft Academic Search

The purpose of this study was to determine if inhalation of mainstream (MS) or sidestream (SS) smoke affects the reproductive organs of female hamsters. Females inhaled smoke from one or two cigarettes twice per day for 30 d prior to mating using a smoking machine equipped for nose only breathing. Serum cotinine levels were within the ranges found in active

T. Magers; P. Talbot; G. DiCarlantonio; M. Knoll; D. Demers; I. Tsai; T. Hoodbhoy

1995-01-01

324

CARCINOGENIC POTENTIAL OF ROTENONE. PHASE I: DIETARY ADMINISTRATION TO HAMSTERS  

EPA Science Inventory

Studies were performed to evaluate the potential carcinogenicity rotenone in the Syrian Golden hamster. Several ancillary range-finding studies were carried out including 14-day feeding trials and a reproduction experiment. The latter experiment indicated that rotenone at a level...

325

Potent Circadian Effects of Dim Illumination at Night in Hamsters  

Microsoft Academic Search

Conventional wisdom holds that the circadian pacemaker of rodents and humans is minimally responsive to light of the intensity provided by dim moonlight and starlight. However, dim illumination (,0.005 lux) provided during the daily dark periods markedly alters entrainment in hamsters. Under dimly lit scotophases, compared to completely dark ones phases, the upper range of entrainment is increased by 4h

Michael R. Gorman; Jennifer A. Evans; Jeffrey A. Elliott

2006-01-01

326

Testicular Hypoplasia and Epididymal Cysts in the Syrian Hamster  

Microsoft Academic Search

WE have observed certain definite changes in the testes of some inbred hamsters. These changes are characterized by a striking testicular hypoplasia, mostly bilateral. Such testes, represented sometimes by structures which are almost invisible to the naked eye, appeared very pale, with an apparent lack of normal vascularization. Their individual weight was only 2-306 mgm., as compared with a testicular

Humberto Granados; Henrik Dam

1948-01-01

327

A Mutation of the Circadian System in Golden Hamsters  

Microsoft Academic Search

A mutation has been found that dramatically shortens the period of the circadian locomotor rhythm of golden hamsters. The pattern of inheritance of this mutation suggests that it occurred at a single, autosomal locus (tau). Wild-type animals have rhythms with free-running periods averaging about 24 hours; animals heterozygous for the mutation have periods of about 22 hours, whereas homozygous animals

Martin R. Ralph; Michael Menaker

1988-01-01

328

Adenylylcyclase Supersensitization in -Opioid Receptor-transfected Chinese Hamster Ovary  

E-print Network

Adenylylcyclase Supersensitization in -Opioid Receptor-transfected Chinese Hamster Ovary Cells Following Chronic Opioid Treatment* (Received for publication, September 13, 1995) Tomer Avidor with rat -opioid receptor cDNA, we show that the -agonists morphine and [D-Ala2 , N-methyl-Phe4 ,Gly-ol5

Vogel, Zvi

329

DOSE RESPONSE OF ELASTASE-INDUCED EMPHYSEMA IN HAMSTERS  

EPA Science Inventory

Elastase-induced emhysema in hamsters was studied using pulmonary function tests in an effort to develop techniques for determining the effects of air pollutants on the progression of this disease. It appears that as little as 6 units of elastase produces mild emphysema in hamste...

330

PULMONARY CELL POPULATIONS IN HAMSTERS MAINTAINED UNDER EGYPTIAN LABORATORY CONDITIONS  

EPA Science Inventory

The study was conducted to obtain baseline values for pulmonary cells in golden hamsters (Mesocricetus auratus) bred and maintained under the laboratory conditions of Al-Azhar University in Egypt. An improvised technique is presented for measuring pulmonary cells obtained by lung...

331

Effects of oocyte collection techniques and maturation media on in vitro maturation and subsequent embryo development in Boer goat  

Microsoft Academic Search

The oocytes (experiment 1) were harvested by one of the four collection techniques (slicing, punc - ture, aspiration I and aspiration II) and the total number and the number of each grade of oocytes were counted, respectively. The good-quality oocytes (good and fair grade) were cultured for maturation. In experiment 2, the oocytes were matured in TCM-199 supplemented with 10

Z. G. Wang; Z. R. Xu; S. D. Yu

2007-01-01

332

Gene-specific timing and epigenetic memory in oocyte imprinting.  

PubMed

Imprinted genes are differentially marked during germ cell development to allow for their eventual parent-of-origin specific expression. A subset of imprinted genes becomes methylated during oocyte growth in both mouse and human. However the timing and mechanisms of methylation acquisition are unknown. Here, we examined the methylation of the Snrpn, Igf2r, Peg1 and Peg3 differentially methylated regions in postnatal growing mouse oocytes. Our findings indicate that methylation was acquired asynchronously at these different genes. Further analysis of Snrpn DMR1 revealed that parental alleles retain an epigenetic memory of their origin as the two alleles were recognized in a parental-specific manner in the absence of DNA methylation. In addition, we show that methylation acquisition was probably related to oocyte diameter and coincided with the accumulation of Dnmt3a, Dnmt3b and Dnmt3L transcripts. Methylation of the repetitive retroviral-like intracisternal A particle also occurred during this same window of oocyte growth. These findings contribute to our understanding of the epigenetic mechanisms underlying imprint acquisition during female germ cell development and have implications for the practice of assisted reproductive technologies. PMID:14998934

Lucifero, Diana; Mann, Mellissa R W; Bartolomei, Marisa S; Trasler, Jacquetta M

2004-04-15

333

FT-IR Microspectroscopy on molecular building of Zebrafish oocytes  

NASA Astrophysics Data System (ADS)

Zebrafish oocytes growth causes relevant modifications in protein components; in fact, the proteolytic cleavage of vitellogenin, a large sex-specific phospholipoglycoprotein synthesized in the female liver, leads to three main yolk protein components (phosphovitin and lipovitellins 1, 2), present as a complex in the oocyte. FT-IR Microspectroscopy could have the potentiality of monitoring these biochemical changes during the maturation process. Representative spectra for I-II, IIIA, IIIB and IV classes oocytes (Hierarchical Clustering Analysis and Principal Component Analysis) were investigated to find specific vibrational patterns corresponding to different maturation degrees. On going from I-II to IV class oocytes, relevant spectral differences were found with III class exhibiting an intermediate spectroscopic behaviour. In particular, the increase of the convoluted band at 2925 cm -1 (CH 2 and CH 3 stretching modes), as well as of intensity band ratios at 2926/2954 cm -1 ( ?asym CH 2/CH 3), 2854/2873 cm -1 ( ?sym CH 2/CH 3) and 1452/1392 cm -1 ( ?CH/?COO), suggest longer lipidic chains; broadening of Amide I and II bands and absorptions at 1737 ( ?C dbnd O, phospholipids) and 1157 cm -1 ( ?C sbnd O and ?C sbnd OH carbohydrates) in lipovitellin and vitellogenin spectra, are present in III and IV classes.

Carnevali, Oliana; Conti, Carla; Ferraris, Paolo; Garavaglia, Maria Grazia; Gioacchini, Giorgia; Giorgini, Elisabetta; Rubini, Corrado; Sabbatini, Simona; Tosi, Giorgio

2009-12-01

334

[Progress in proteomics of mammalian oocyte and early embryo].  

PubMed

The development of female germ cell is the cornerstone for animal reproduction. Mammalian oocyte and early embryo have many distinct phenomena and mechanisms during their growth and development, involving series dynamic changes of protein synthesis/degradation and phosphorylation. Research on the regulatory mechanism of oocyte division, maturation, and developmental principle of pre-implantation embryo is an important topic in the field of animal developmental biology. Proteomics using all of proteins expressed by a cell or tissue as research object, systematically identify, quantify and study the function of all these proteins. With the rapid development of protein separation and identification technology, proteomics provide some new methods and the research contents on fields of oogenesis, differentiation, maturation and quality control, such as protein quantification, modification, location and interaction important information which other omics technology can not provide. These information will contribute to uncover the molecular mechanisms of mammalian oocyte maturation and embryonic development. And it is great significant for improving the culture system of oocyte in vitro maturation, the efficiency of embryo production in vitro, somatic cell clone and transgenic animal production. PMID:25345004

Chen, Lingsheng; Xu, Ping; Shi, Deshun; Li, Xiangping

2014-07-01

335

In vitro follicular growth affects oocyte imprinting establishment in mice  

Microsoft Academic Search

In vitro folliculogenesis of cryopreserved ovarian tissue could be an effective method for insuring fertility for patients who receive gonadotoxic treatment. Although several culture systems have been described for growing female gametes in vitro, the production of competent oocytes for further development remains a considerable challenge. The purpose of our study was to determine whether maternal primary imprinting progresses normally

Antoine Kerjean; Philippe Couvert; Thomas Heams; Céline Chalas; Karine Poirier; Jamel Chelly; Pierre Jouannet; Andras Paldi; Catherine Poirot

2003-01-01

336

Electrophysiological responses of crayfish oocytes to biogenic amines.  

PubMed

Intracellular recordings were made from immature, growing oocytes of the crayfish Pacifastacus leniusciulus. Oocytes had a relatively negative resting potential of -74.7+/-2.2 mV (n=26; range -53 to -90) and a mean input resistance of 0.86+/-0.19 MOmega (n=22; range 0.17-3.3). Octopamine induced a long-lasting response involving biphasic changes in input resistance, together with bi- or multiphasic changes in membrane potential. The resistance-decreasing phase involved (in different oocytes) membrane hyperpolarization, depolarization or both. The resistance-increasing phase was usually a depolarization. The hyperpolarizing form of the resistance-decreasing response, and the depolarizing resistance-increasing response reversed in polarity at membrane potentials of (respectively) -90 and -92 mV, suggesting increases and decreases in K(+) conductance underly the biphasic changes in input resistance. The threshold concentration for the response was remarkably low (>10(-12) M) and showed little or no dose-dependence over the concentration range 10(-12)-10(-6) M. Similar responses were evoked by dopamine and serotonin (at 10(-9) M), although a higher proportion of oocytes responded to octopamine and/or dopamine than to serotonin. PMID:10908853

Skorupski, P; Melarange, R

2000-05-01

337

Gonadotrophin regimens and oocyte quality in women with polycystic ovaries  

Microsoft Academic Search

The systemic endocrine environment during the later stages of follicle development has a crucial role in co-ordinating follicular and oocyte maturation before ovulation. Polycystic ovary syndrome (PCOS) is associated with abnormal circulating hormones, abnormal peri-follicular vascularity and significant abnormalities of granulosa cell function. After induction of ovulation, fertilization rates in vivo in women with PCOS are normal, but there is

Stephen Franks; Ruth Roberts; Kate Hardy

2003-01-01

338

Protein tyrosine phosphorylation during meiotic divisions of starfish oocytes  

SciTech Connect

We have used an antibody specific for phosphotyrosine to investigate protein phosphorylation on tyrosine during hormone-induced maturation of starfish oocytes. Analysis of immunoprecipitates from cortices of in vivo labeled Marthasterias glacialis oocytes revealed the presence of labeled phosphotyrosine-containing proteins only after hormone addition. Six major phosphoproteins of 195, 155, 100, 85, 45, and 35 kDa were detected. Total activity in immunoprecipitates increased until first polar body emission and was greatly reduced upon completion of meiosis but some proteins exhibited different kinetics. The labeling of the 155-kDa protein reached a maximum at germinal vesicle breakdown, while the 35-kDa appeared later and disappeared after polar body emission. Similar results were obtained with Asterias rubens oocytes. In vitro phosphorylation of cortices showed that tyrosine kinase activity is a major protein kinase activity in this fraction, the main endogenous substrate being a 68-kDa protein. The proteins phosphorylated on tyrosine in vitro were almost similar in extracts from oocytes treated or not with the hormone.

Peaucellier, G.; Andersen, A.C.; Kinsey, W.H. (Univ. of Miami School of Medicine, FL (USA))

1990-04-01

339

INTRANUCLEAR AND CYTOPLASMIC ANNULATE LAMELLAE IN TUNICATE OOCYTES  

Microsoft Academic Search

Electron microscope studies were made on various tunicate oocytes at different stages of growth and development. Both the inner and outer lamellae of the perforated nuclear en- velope demonstrate considerable blebbing activity. The blebs of the inner lamella detach into the nucleoplasrn where they undergo a special type of fusion process resulting in the formation of numerous, usually single, differentiated

R. G. Kessel

1965-01-01

340

Intranuclear annulate lamellae in oocytes of the tunicate, Styela partita  

Microsoft Academic Search

Intranuclear annulate lamellae have been observed with the electron microscope in oocytes of the tunicate, Styela partita. Morphological evidence suggests that the annulate lamellae may arise by a specialized fusion process of individual vesicles. Intranuclear vesicles appear to be formed, in time, before differentiated annulate lamellae. It is also suggested that the position and structure of an annulus is in

R. G. Kessel

1964-01-01

341

A Simplified Approach for Oocyte Enucleation in Mammalian Cloning  

PubMed Central

Abstract Despite its success in almost all farm and laboratory animals, somatic cell nuclear transfer (SCNT) is still a low-efficiency technique. In this investigation, we determined the impact of each enucleation step on oocyte viability (assessed by parthenogenetic activation): Hoechst (HO) staining, cytochalasin B, ultraviolet (UV) exposure, and demecolcine. Our data showed that of all the factors analyzed, UV exposure impaired oocyte development (cleavage, 59% for untreated oocytes vs. 8% UV exposed; blastocyst stage, 32% untreated vs. 0% UV exposed). A minor toxicity was detected following demecolcine treatment (cleavage, 62%; blastocyst stage, 13%). Next, we compared HO/UV (canonical) and demecolcine-assisted enucleation (DAE), with a straight removal of metaphase chromosomes without any chemical or physical aid (straight enucleation). DAE improved the preimplantation development of sheep cloned embryos compared to HO/UV enucleation (cleavage, 38% vs. 19%; blastocysts, 17% vs. 4%), yet straight enucleation resulted in the highest cleavage and blastocysts rates (61% and 30%, respectively). We concluded that: (1) UV exposure harms sheep oocyte and embryo development; (2) DAE may represent an alternative approach, especially for unskilled operators; and (3) straight enucleation remains, in our estimation, the most reliable and least harmful protocol for SCNT. PMID:24219576

Iuso, Domenico; Czernik, Marta; Zacchini, Federica

2013-01-01

342

Imaging 103 Imaging Ca2+ Signals in Xenopus Oocytes  

E-print Network

Parker Summary Xenopus oocytes have become a favored preparation in which to study the spatiotempo- ral © Humana Press Inc., Totowa, NJ #12;104 Dargan, Demuro, and Parker 2. Materials 2.1. Preparation-222 (Sigma, St. Louis, MO). Store at -20°C in a desiccator. This is a possible car- cinogen; wear

Parker, Ian

343

Original article Bovine cumulus expansion and corona-oocyte  

E-print Network

Original article Bovine cumulus expansion and corona-oocyte disconnection during culture in vitro J into 2 groups: complexes in which a dark rim of corona cells were visible around the zona pellucida (group 1and those in which the corona displayed the same density as the rest of the cumulus cell mass

Boyer, Edmond

344

Granulosa cell-oocyte interactions: the phosphorylation of specific proteins in mouse oocytes at the germinal vesicle stage is dependent upon the differentiative state of companion somatic cells  

SciTech Connect

The role of granulosa cells in the regulation of mouse ovarian oocyte metabolism was investigated. Fully grown antral oocytes, isolated from surrounding cumulus cells, were cultured on monolayers of preantral granulosa cells in the presence of dbcAMP to prevent the resumption of meiosis. Under these conditions metabolic cooperativity was established between the two cell types as early as 1 hr after seeding. Moreover, cocultured oocytes phosphorylated two polypeptides of 74 and 21 kDa which are normally phosphorylated in follicle-enclosed growing oocytes but not in cumulus cell-enclosed fully grown oocytes at the germinal vesicle stage. When cocultured oocytes were allowed to resume meiosis, the 74 and 21 kDa proteins were synthesized but no longer phosphorylated even though intercellular coupling between the two cell types was maintained during radiolabeling. It appears therefore: (a) that the different protein kinase activity of growing and fully grown germinal vesicle-stage mouse oocytes is related to the differentiative state of granulosa cells, and (b) that the regulation of oocyte protein phosphorylation activity by granulosa cells is dependent on the meiotic stage of the oocyte.

Cecconi, S.; Tatone, C.; Buccione, R.; Mangia, F.; Colonna, R. (Universita dell'Aquila, (Italy))

1991-05-01

345

Advances in oocyte cryopreservation technology will eventually blur the ethical and moral boundaries between compensated egg sharing and commercialized oocyte donation.  

PubMed

Compensated egg sharing was originally conceived as a patient self-help co-operative scheme to avoid overt 'commodification' of donor oocytes and prevent doctors and medical institutions from acting as 'brokers' of donated human material. As such, egg sharing is an ethically justifiable and much-preferred alternative to commercialized oocyte donation. However, recent advances in oocyte cryopreservation technology are likely to blur the ethical and moral boundaries between compensated egg sharing and commercialized oocyte donation. The banking of cryopreserved oocytes would negate the requirement for donors and recipients in egg sharing to have co-ordinated and synchronized treatment cycles. Instead, fertility doctors and medical institutions can now offer subsidized fertility treatment upfront to any patient willing to donate a portion of her retrieved cohort of oocytes for banking and subsequent donation. Thus, more opportunity is now open for them to act as 'middle-man' to broker the transaction of oocytes between donor and recipient, which would inevitably result in overt 'commodification' of donated human material. Administrative and processing fees will de finitely be billed to prospective recipients, for banking and storage of the cryopreserved oocytes, which would mean that a direct profit can now be made from the transaction of oocytes between donor and recipient. PMID:16569309

Heng, Boon Chin

2006-03-01

346

CD81 and CD9 work independently as extracellular components upon fusion of sperm and oocyte  

PubMed Central

Summary When a sperm and oocyte unite into one cell upon fertilization, membranous fusion between the sperm and oocyte occurs. In mice, Izumo1 and a tetraspanin molecule CD9 are required for sperm-oocyte fusion as one of the oocyte factors, and another tetraspanin molecule CD81 is also thought to involve in this process. Since these two tetraspanins often form a complex upon cell-cell interaction, it is probable that such a complex is also formed in sperm-oocyte interaction; however, this possibility is still under debate among researchers. Here we assessed this problem using mouse oocytes. Immunocytochemical analysis demonstrated that both CD9 and CD81 were widely distributed outside the oocyte cell membrane, but these molecules were separate, forming bilayers, confirmed by immunobiochemical analysis. Electron-microscopic analysis revealed the presence of CD9- or CD81-incorporated extracellular structures in those bilayers. Finally, microinjection of in vitro-synthesized RNA showed that CD9 reversed a fusion defect in CD81-deficient oocytes in addition to CD9-deficient oocytes, but CD81 failed in both oocytes. These results suggest that both CD9 and CD81 independently work upon sperm-oocyte fusion as extracellular components. PMID:23213457

Ohnami, Naoko; Nakamura, Akihiro; Miyado, Mami; Sato, Masahiro; Kawano, Natsuko; Yoshida, Keiichi; Harada, Yuichirou; Takezawa, Youki; Kanai, Seiya; Ono, Chihiro; Takahashi, Yuji; Kimura, Ken; Shida, Toshio; Miyado, Kenji; Umezawa, Akihiro

2012-01-01

347

Effects of Postmortem Interval on Mouse Ovary Oocyte Survival and Maturation  

PubMed Central

To study the time- and temperature-dependent survival of ovarian oocytes collected from postmortem carcass, ICR mice were killed and placed for different periods (0, 1, 2, 4, 6, 8 and 10 h) at different temperatures (25°C, 4°C and 37°C). After preservation, oocyte morphology, germinal vesicle (GV) oocyte number, oocyte meiotic maturation percentage, mitochondrial distribution and intracellular glutathione (GSH) level were evaluated. The results showed no surviving oocytes could be collected by 2h, 6h, and 12 h after carcass preservation at 37°C, 25°C and 4°C, respectively. The number of collected GV oocytes in the ovary deceased as the preservation time lasted at the same temperature. Meanwhile at the same point in time, the ratio of germinal vesicle breakdown (GVBD) and the first polar body emission (PBE) gradually reduced as preservation temperature increased. In addition, the percentage of abnormal mitochondrial distribution in the preserved oocytes was obviously higher than that in the control oocytes, while GSH level was not altered in collected oocytes. Unexpectedly, neither chromosome arrangement nor spindle organization was affected as long as the oocytes from preserved carcasses could complete maturation. These data are helpful for proper use of ovary oocytes from postmortem carcass of valuable individuals. PMID:24874949

Zhang, Guang-Li; Ma, Jun-Yu; Sun, Quan; Hu, Meng-Wen; Yang, Xiu-yan; Gao, Si-Hua; Jiang, Guang-Jian

2014-01-01

348

Raman Micro-Spectroscopy Can Be Used to Investigate the Developmental Stage of the Mouse Oocyte  

PubMed Central

In recent years, the uptake of assisted reproductive techniques such as in vitro fertilisation has risen exponentially. However, there is much that is still not fully understood about the biochemical modifications that take place during the development and maturation of the oocyte. As such, it is essential to further the understanding of how oocyte manipulation during these procedures ultimately affects its developmental potential; yet, there are few methods currently available which are capable of providing a quantitative measure of oocyte quality. Raman spectroscopy enables investigation of the global biochemical profile of intact cells without the need for labelling. Here, Raman spectra were acquired from the ooplasm of mouse oocytes at various stages of development, from late pre-antral follicles, collected after in vitro maturation within their ovarian follicles and from unstimulated and stimulated ovulatory cycles. Using a combination of univariate and multivariate statistical methods, it was found that ooplasm lipid content could be used to discriminate between different stages of oocyte development. Furthermore, the spectral profiles of mature oocytes revealed that oocytes which have developed in vitro are protein-deficient when compared to in vivo grown oocytes. Finally, the ratio of two Raman peak intensities, namely 1605?1447 cm?1, used as a proxy for the protein-to-lipid ratio of the ooplasm, was shown to be indicative of the oocyte’s quality. Together, results indicate that Raman spectroscopy may present an alternative analytical tool for investigating the biochemistry of oocyte developmental stage and quality. PMID:23840882

Davidson, Bryony; Murray, Alison A.

2013-01-01

349

Thioredoxin-Interacting Protein Regulates Glucose Metabolism and Affects Cytoplasmic Streaming in Mouse Oocytes  

PubMed Central

Thioredoxin-interacting protein (Txnip) regulates intracellular redox state and prompts oxidative stress by binding to and inhibiting Thioredoxin (Trx). In addition, via a Trx-independent mechanism, Txnip regulates glucose metabolism and thus maintains intracellular glucose levels. Previously, we found Txnip mRNA highly expressed in immature germinal vesicle (GV) oocytes, but currently there is no report describing the role of Txnip in oocytes. Therefore, we conducted the present study to determine the function of Txnip in mouse oocytes' maturation and meiosis by using RNA interference (RNAi) method. Upon specific depletion of Txnip, 79.5% of oocytes were arrested at metaphase I (MI) stage. Time-lapse video microscopy analysis revealed that the formation of granules in the oocyte cytoplasm increased concurrent with retarded cytoplasmic streaming after Txnip RNAi treatment. Txnip RNAi-treated oocytes had upregulated glucose uptake and lactate production. To confirm the supposition that mechanism responsible for these observed phenomena involves increased lactate in oocytes, we cultured oocytes in high lactate medium and observed the same increased granule formation and retarded cytoplasmic streaming as found by Txnip RNAi. The MI-arrested oocytes exhibited scattered microtubules and aggregated chromosomes indicating that actin networking was disturbed by Txnip RNAi. Therefore, we conclude that Txnip is a critical regulator of glucose metabolism in oocytes and is involved in maintaining cytoplasmic streaming in mouse oocytes. PMID:23976953

Lee, Su-Yeon; Lee, Hyun-Seo; Kim, Eun-Young; Ko, Jung-Jae; Yoon, Tae Ki; Lee, Woo-Sik; Lee, Kyung-Ah

2013-01-01

350

Developmental competence of IVM pig oocytes after SCNT in relation to the shrinkage pattern induced by hyperosmotic treatment.  

PubMed

The objective of this study was to examine the developmental competence of IVM pig oocytes in relation to the pattern of morphologic changes after exposure to hyperosmotic medium to select oocytes of a higher quality. IVM oocytes were treated with a hyperosmotic (593 mOsm) medium containing NaCl, sorbitol, or sucrose. Oocytes that shrunk spherically (SSP oocytes) or in irregular shapes (SIR oocytes) were collected separately, and washed for 15 minutes in an isotonic (297 mOsm) medium for recovery. Irrespective of the chemicals used, hyperosmotic treatment of oocytes for 1 hour or 15 minutes did not alter embryonic development after parthenogenesis (PA) and SCNT. A significantly higher proportion of SSP oocytes developed to the blastocyst stage (34.0%) compared with SIR oocytes (15.8%) after PA. The intracellular glutathione content was significantly higher in SSP oocytes than in SIR oocytes. Conversely, the reactive oxygen species level was significantly higher in SIR oocytes than in SSP oocytes. The maturation promoting factor level as measured by p34(cdc2) kinase activity was not influenced by hyperosmotic treatment itself but was 1.3-fold higher (P < 0.05) in SSP oocytes than in SIR oocytes. When IVM oocytes were divided into two groups according to their diameters (large and small), and treated separately in hyperosmotic medium, significantly more SSP oocytes (71.4%) were found in the large oocytes than in the small oocytes (51.4%). Moreover, the proportion of metaphase II oocytes was significantly higher in SSP oocytes than in SIR oocytes in both groups (98.5% vs. 73.1% in large oocytes, and 92.2% vs. 48.0% in small oocytes). After SCNT, a significantly higher proportion of SSP oocytes displayed blastocyst formation (36.4%) than untreated (29.0%) and SIR oocytes (22.1%). Our results demonstrated that SSP oocytes were of a higher quality than SIR oocytes, which was shown by higher intracellular glutathione and maturation promoting factor levels, lower reactive oxygen species levels, and improved embryonic development to the blastocyst stage after PA and SCNT. PMID:24576710

Lee, Joohyeong; Lee, Yongjin; Park, Bola; Elahi, Fazle; Jeon, Yubyeol; Hyun, Sang-Hwan; Lee, Eunsong

2014-04-15

351

Role of animal pole protuberance and microtubules during meiosis in sea cucumber Apostichopus japonicus oocytes  

NASA Astrophysics Data System (ADS)

Fully grown oocytes of Apostichopus japonicus have a cytoplasmic protuberance where the oocyte attaches to the follicle. The protuberance and the oolamina located on the opposite side of the oocyte indicate the animal-vegetal axis. Two pre-meiotic centrosomes are anchored to the protuberance by microtubules between centrosomes and protuberance. After meiosis reinitiation induced by DTT solution, the germinal vesicle (GV) migrates towards the protuberance. The GV breaks down after it migrates to the oocyte membrane on the protuberance side. The protuberance then contracts back into the oocyte and the first polar body extrudes from the site of the former protuberance. The second polar body forms beneath the first. Thus the oocyte protuberance indicates the presumptive animal pole well before maturation of the oocyte.

Pang, Zhenguo; Chang, Yaqing; Sun, Huiling; Yu, Jiaping

2010-05-01

352

Double-Plate Penetration Equations  

NASA Technical Reports Server (NTRS)

This report compares seven double-plate penetration predictor equations for accuracy and effectiveness of a shield design. Three of the seven are the Johnson Space Center original, modified, and new Cour-Palais equations. The other four are the Nysmith, Lundeberg-Stern-Bristow, Burch, and Wilkinson equations. These equations, except the Wilkinson equation, were derived from test results, with the velocities ranging up to 8 km/sec. Spreadsheet software calculated the projectile diameters for various velocities for the different equations. The results were plotted on projectile diameter versus velocity graphs for the expected orbital debris impact velocities ranging from 2 to 15 km/sec. The new Cour-Palais double-plate penetration equation was compared to the modified Cour-Palais single-plate penetration equation. Then the predictions from each of the seven double-plate penetration equations were compared to each other for a chosen shield design. Finally, these results from the equations were compared with test results performed at the NASA Marshall Space Flight Center. Because the different equations predict a wide range of projectile diameters at any given velocity, it is very difficult to choose the "right" prediction equation for shield configurations other than those exactly used in the equations' development. Although developed for various materials, the penetration equations alone cannot be relied upon to accurately predict the effectiveness of a shield without using hypervelocity impact tests to verify the design.

Hayashida, K. B.; Robinson, J. H.

2000-01-01

353

Penetration Experiments under Reduced Gravity  

NASA Astrophysics Data System (ADS)

Penetration experiments will find several applications in exploration missions in the near future. Penetrators are common tools for the investigation of physical surface properties. The techniques and theories are widely applied under 1g condition on Earth and the results are used by engineers and scientists. The main contribution to the bearing resistance of a soil is combined of shaft and base resistance [1]. The theories show, that the resistance scales with gravity. Penetration experiments during a parabolic flight campaign have been performed for evaluating this gravity scaling of the bearing resistance in different materials during a parabolic flight campaign in December 2012. The main part of the experiment is composed of a steel rod penetrating into a sample cell. Depth and penetration force are recorded during this process. A sieving mechanism provided the ability of sample preparation during flight. Different compaction regimes of the sample material could be created with a ruttler mounted underneath the sample cell. The parabolic flight campaign consisted of 4 flight days. On each day 13 parabolas with Martian gravity, 12 parabolas with lunar gravity and 6 microgravity parabolas could be performed. Three different sample materials have been examined within the 4 flight days: glass spheres, glass corn and Mojawe sand. The glass spheres and glass corn samples were made of the same material, but with different shape. The Mojawe sand is a natural soil from the Mojawe desert in California (US). The experimental description and the first results will be presented.

Krause, C.; Gehlen, M.; Jaquemet, A.; Heller, S.; Sperl, M.; Willnecker, R.

2013-09-01

354

PENETRATION OF 5FLUOROURACIL IN EXCISED SKIN  

Microsoft Academic Search

Total penetration of 5-fluorouracil (FU) through human and hairless mouse skin was measured in vitro and the results were compared using radioactively labelled drug and a gas chromatographic method specific for the free FU molecule. Both methods were used to determine whether metabolism, either in the skin during penetration or in the penetration wells, could have affected the percent penetration

Jordan L. Cohen; Richard B. Stoughton

1974-01-01

355

In Vitro Grown Sheep Preantral Follicles Yield Oocytes with Normal Nuclear-Epigenetic Maturation  

PubMed Central

Background Assisted reproductive technologies allow to utilize a limited number of fully grown oocytes despite the presence in the ovary of a large pool of meiotically incompetent gametes potentially able to produce live births. In vitro folliculogenesis could be useful to recruit these oocytes by promoting their growth and differentiation. Methodology/Principal Findings In vitro folliculogenesis was performed starting from sheep preantral (PA) follicles to evaluate oocyte nuclear/epigenetic maturation. Chromatin configuration, quantification of global DNA methylation, and epigenetic remodelling enzymes were evaluated with immunocytochemistry, telomere elongation was assessed with the Q-FISH technique, while the DNA methylation status at the DMRs of maternally IGF2R and BEGAIN, and paternally H19 methylated imprinted genes was determined by bisulfite sequencing and COBRA. Specifically, 70% of PA underwent early antrum (EA) differentiation and supported in culture oocyte global DNA methylation, telomere elongation, TERT and Dnmt3a redistribution thus mimicking the physiological events that involve the oocyte during the transition from secondary to tertiary follicle. Dnmt1 anticipated cytoplasmic translocation in in vitro grown oocytes did not impair global and single gene DNA methylation. Indeed, the in vitro grown oocytes acquired a methylation profile of IGF2R and BEGAIN compatible with the follicle/oocyte stage reached, and maintained an unmethylated status of H19. In addition, the percentage of oocytes displaying a condensed chromatin configuration resulted lower in in vitro grown oocytes, however, their ability to undergo meiosis and early embryo development after IVF and parthenogenetic activation was similar to that recorded in EA follicle in vivo grown oocytes. Conclusions/Significance In conclusion, the in vitro folliculogenesis was able to support the intracellular/nuclear mechanisms leading the oocytes to acquire a meiotic and developmental competence. Thus, the in vitro culture may increase the availability of fertilizable oocytes in sheep, and become an in vitro translational model to investigate the mechanisms governing nuclear/epigenetic oocyte maturation. PMID:22132111

Barboni, Barbara; Russo, Valentina; Cecconi, Sandra; Curini, Valentina; Colosimo, Alessia; Garofalo, Maria Luigia A.; Capacchietti, Giulia; Di Giacinto, Oriana; Mattioli, Mauro

2011-01-01

356

The involvement of neurofilament heavy chain phosphorylation in the maturation and degeneration of rat oocytes.  

PubMed

Neurofilaments (NF) are intermediate filament proteins that were originally found to be expressed in neurons and are involved in the maintenance of axonal structure. However, there has not been much research on the expression and physiological significance of NF in other organs. In the present study, we examined the expression of NF in rat ovaries and found that NF heavy chain (NF-H) was expressed in oocytes of follicles from the primary to mature stages, ovulated oocytes, fertilized zygotes, and degenerative oocytes of atretic follicles. Cytoplasmic NF-H disappeared at the two-cell stage of embryonic development, whereas degenerative oocytes of atretic follicles continued to express NF-H even after fragmentation. An antibody that specifically recognizes phosphorylated NF-H (pNF-H) was used to examine the pattern of NF-H phosphorylation in oocytes. pNF-H was detected in the cytoplasm and nuclei of oocytes of mature and atretic follicles, ovulated oocytes, and one-cell zygotes. Treatment with 3,3'-iminodipropionitrile, which induces aberrant phosphorylation of NF in the perikarya of neurons and causes neuropathy, induced oocyte degeneration with follicular atresia, phosphorylation of NF-H in oocytes, and ovarian gene expression of cyclin-dependent kinase 5, a candidate kinase of NF-H. However, an indicator of neuron degeneration, Fluoro-Jade C, failed to stain the pNF-H-immunopositive oocytes. Our results indicate that NF-H expressed in oocytes may be involved in the maintenance of oocyte structure during follicular growth and that the phosphorylation of NF-H in ephemeral oocytes may contribute to the degeneration process of oocytes. PMID:22315443

Takahashi, Noriyuki; Ishizuka, Bunpei

2012-04-01

357

Effect of Hyaluronan on Developmental Competence and Quality of Oocytes and Obtained Blastocysts from In Vitro Maturation of Bovine Oocytes  

PubMed Central

The objective of the present study was to evaluate the effect of hyaluronan (HA) during IVM on meiotic maturation, embryonic development, and the quality of oocytes, granulosa cells (GC), and obtained blastocysts. COCs were matured in vitro in control medium and medium with additional 0.035% or 0.07% of exogenous HA. The meiotic maturity did not differ between the analysed groups. The best rate and the highest quality of obtained blastocysts were observed when 0.07% HA was used. A highly significant difference (P < 0.001) was noted in the mean number of apoptotic nuclei per blastocyst and in the DCI between the 0.07% HA and the control blastocysts (P < 0.01). Our results suggest that addition of 0.035% HA and 0.07% HA to oocyte maturation media does not affect oocyte nuclear maturation and DNA fragmentation. However, the addition of 0.07% HA during IVM decreases the level of blastocysts DNA fragmentation. Finally, our results suggest that it may be risky to increase the HA concentration during IVM above 0.07% as we found significantly higher Bax mRNA expression levels in GC cultured with 0.07% HA. The final concentration of HA being supplemented to oocyte maturation media is critical for the success of the IVP procedure. PMID:24689043

Opiela, Jolanta; Romanek, Joanna; Lipi?ski, Daniel; Smor?g, Zdzis?aw

2014-01-01

358

Oocyte insemination techniques are related to alterations of embryo developmental timing in an oocyte donation model.  

PubMed

Because of the different intrinsic characteristics of the classic IVF and intracytoplasmic sperm injection (ICSI) techniques, the timing of zygote development can be influenced by the method of fertilization. However, there is no information about the relevance of the insemination procedure on embryo-quality parameters as measured through their developmental dynamics. The aim of this work was to determine if the insemination technique, IVF or ICSI, influences embryo developmental kinetics by examining 1203 embryos from 178 couples undergoing oocyte donation with IVF or ICSI. Using time-lapse information, this work calculated several developmental kinetic variables, from pronuclear fading (PNF) to expanded blastocyst, and also the proportion of optimal embryos in a best time range with a predicted higher implantation potential. Embryo development after ICSI was slightly faster than after IVF; however, when PNF, rather than time of insemination, was established as t0, the differences between the two procedures disappeared. The percentage of optimal embryos showed a trend towards higher values in IVF-derived embryos; however, the difference was not statistically significant. With these results and through the time-lapse monitoring system, it is concluded that it is the fertilization method which determines embryo developmental kinetics if insemination time is used as the starting point. A key step in assisted reproduction is the assessment of oocyte and embryo viability to determine the embryo(s) most likely to implant. Current embryo assessment strategies in clinical settings largely rely on embryo morphology and cleavage rates, and although these systems have been successful in improving pregnancy rates, their precision is far from ideal as they are based on visual information. In contrast, automated image analysis may add objectivity to the process of embryo selection and consequently, lead to an improvement in the implantation rates. Timing of zygote development can be influenced by the method of fertilization or by in-vitro culture conditions, so it has been suggested that the fertilization procedure influences the length of time elapsed between fertilization and the first cleavage. In this report, we show the results from using a time-lapse monitoring system to determine the timing of key events during embryo development both in IVF and ICSI. Due to the different intrinsic characteristics of the classic IVF and ICSI techniques, the selection of a critical time point is essential so as to maximize the differences between the two methods. For that reason, we searched for evidence in the data obtained from the image analysis for a link connecting embryo cleavage and the fertilization technique and also to find whether the kinetic of development derived from classic IVF or ICSI is also related to a predicted higher implantation. PMID:23953584

Cruz, María; Garrido, Nicolás; Gadea, Blanca; Muñoz, Manuel; Pérez-Cano, Inmaculada; Meseguer, Marcos

2013-10-01

359

Lunar regolith penetrators and cutters  

NASA Technical Reports Server (NTRS)

An apparatus was designed and built for conducting simulation experiments on cutting tool penetration in the centrifuge. This equipment is mounted on the laminar container which is used for the regolith densification study, so that the end product of the latter, i.e., a regolith bed with the proper density profile, can be used directly for the penetration tests. In this apparatus, an etching tool is suspended through a pulley system by the action of a double acting air cylinder. By adjusting the air pressure acting on each side of the cylinder, the net downward force acting on the tool can be controlled. The penetration of the tool is measured by an LVDT. This apparatus was proof-tested in the centrifuge and is ready for use in conjunction with the regolith densification experiments.

Barnes, Frank; Sture, Stein

1991-01-01

360

Mars surface penetrator: System description  

NASA Technical Reports Server (NTRS)

A point design of a penetrator system for a Mars mission is described. A strawman payload which is to conduct measurements of geophysical and meteorological parameters is included in the design. The subsystems used in the point design are delineated in terms of power, mass, volume, data, and functional modes. The prospects for survival of the rigors of emplacement are described. Data handling and communications plans are presented to allow consideration of the requirements placed by the penetrator on the orbiter and ground operations. The point design is technically feasible and the payload selection scientifically desirable.

Manning, L. A. (editor)

1977-01-01

361

[Penetrating transorbital intracranial foreign body].  

PubMed

We report a seven year-old boy who suffered left orbital penetration of an industrial sewing machine needle. The needle passing through the left orbit and sphenoid bone at the posterior was extending into the layers of the dura of the left temporal lobe. In this patient, we preferred surgical approach and there was no complication after surgery. Penetrating intraorbital foreign materials with intracranial extension may lead to complications such as intracerebral hematoma, brain abscess, CSF fistula, proptosis of the eye, diplopia, orbital cellulitis and periorbital abscess. They have to be removed by surgical approach to prevent potential complications. PMID:16850365

Civelek, Erdinç; Bilgiç, Salih; Kabata?, Serdar; Hepgül, Kemal Tanju

2006-07-01

362

?-hexosaminidase from Xenopus laevis eggs and oocytes: from gene to immunochemical characterization.  

PubMed

Glycosidases are present both in sperm and eggs in vertebrates and have been associated with different fertilization steps as gamete binding, egg coat penetration, and polyspermy prevention. In this manuscript, we have analyzed the activity of different glycosidases of Xenopus laevis eggs. The main activity corresponded to N-acetyl-?-D-glucosaminidase (Hex), which was reported to participate both in gamete binding and polyspermy prevention among phylogenetically distant animals. We have raised homologous antibodies against a recombinant N-terminal fragment of a X. laevis Hex, and characterized egg's Hex both by Western blot and immunohistochemical assays. Noteworthy, Hex was mainly localized to the cortex of animal hemisphere of full-grown oocytes and oviposited eggs, and remained unaltered after fertilization. Hex is constituted by different pair arrangements of two subunits (? and ?), giving rise to three possible Hex isoforms: A (??), B (??), and S (??). However, no information was available regarding molecular identity of Hex in amphibians. We present for the first time the primary sequences of two isoforms of X. laevis Hex. Interestingly, our results suggest that ?- and ?-like subunits that constitute Hex isoforms could be synthesized from a same gene in Xenopus, by alternative exon use. This finding denotes an evolutionary divergence with mammals, where ? and ? Hex subunits are synthesized from different genes on different chromosomes. PMID:22753314

Morales, Enrique S; Krapf, Darío; Botta, Pablo E; Cabada, Marcelo O; Arranz, Silvia E

2012-12-01

363

Follicular fluid content and oocyte quality: from single biochemical markers to metabolomics  

PubMed Central

The assessment of oocyte quality in human in vitro fertilization (IVF) is getting increasing attention from embryologists. Oocyte selection and the identification of the best oocytes, in fact, would help to limit embryo overproduction and to improve the results of oocyte cryostorage programs. Follicular fluid (FF) is easily available during oocyte pick-up and theorically represents an optimal source on non-invasive biochemical predictors of oocyte quality. Unfortunately, however, the studies aiming to find a good molecular predictor of oocyte quality in FF were not able to identify substances that could be used as reliable markers of oocyte competence to fertilization, embryo development and pregnancy. In the last years, a well definite trend toward passing from the research of single molecular markers to more complex techniques that study all metabolites of FF has been observed. The metabolomic approach is a powerful tool to study biochemical predictors of oocyte quality in FF, but its application in this area is still at the beginning. This review provides an overview of the current knowledge about the biochemical predictors of oocyte quality in FF, describing both the results coming from studies on single biochemical markers and those deriving from the most recent studies of metabolomics PMID:19413899

Revelli, Alberto; Piane, Luisa Delle; Casano, Simona; Molinari, Emanuela; Massobrio, Marco; Rinaudo, Paolo

2009-01-01

364

The study of mammalian oocyte competence by transcriptome analysis: progress and challenges.  

PubMed

Various morphological and cytological traits of oocytes and their surrounding cumulus cells may be used to select oocytes for assisted reproduction. However, even with careful selection, successful IVF and subsequent embryo development remain uncertain. The factors that ensure oocyte competence are unclear and other approaches to assessing developmental potential must be explored. With the constant development of the molecular toolbox, genomic/transcriptomic analysis is becoming a more and more interesting approach to understand oocyte quality on the basis of RNA composition. Using bovine and mouse models as well as human oocytes of known developmental potential, various efforts are underway to characterize the mRNA profile of the competent oocyte using microarray technology. The proliferation of gene expression data sets raises new opportunities to identify the mechanisms involved in this complex phenotype, which should lead to improved techniques of assisted reproduction. Although several molecular markers of oocyte quality are known, translating these into cellular functions remains challenging, largely due to the poor correlation between mRNA level and protein synthesis. Unlike most somatic cells, the oocyte can store mRNA for days, with transcriptional activity remaining at a halt during the 4-5 days beginning before ovulation and ending with embryonic genome activation. This review provides an overview of the transcriptomic data obtained from oocytes of different quality as well as interesting avenues to explore in order to improve our understanding of oocyte competence. PMID:24233546

Labrecque, Rémi; Sirard, Marc-André

2014-02-01

365

Actin synthesis is not regulated by granulosa cells in mouse growing and preovulatory oocytes.  

PubMed

The synthesis and intracellular distribution of actin were studied in isolated dictyate and metaphase II mouse oocytes by 1) sodium dodecyl sulfate-polyacrylamide gel electrophoresis of newly synthetized oocyte protein and 2) cytochemical F-actin labeling by fluorescent phalloidin. Unpermeabilized, fully grown oocytes bound phalloidin intensely at the level of the zona pellucida (ZP), such ZP-associated actin representing a significant portion of total actin found in these cells. In contrast, phalloidin binding to ZP was very low in growing oocytes and was undetectable in ovulated, metaphase II eggs. When ZP-associated actin of fully grown oocytes was removed by prolongedly exposing oocytes to alpha-chymotrypsin, the amount of newly synthesized actin displayed by cumulus-enclosed oocytes was reduced to a level comparable to that shown by oocytes isolated from granulosa cells. We demonstrate that ZP-associated actin belongs to granulosa cell processes that remain within the ZP as a consequence of oocyte isolation procedures. We conclude that actin synthesis of mouse oocytes is not regulated by granulosa cells. PMID:3235031

Canipari, R; Bevilacqua, A; Colonna, R; De Felici, M; Mangia, F

1988-06-01

366

Metabolic control of oocyte development: linking maternal nutrition and reproductive outcomes  

PubMed Central

Obesity, diabetes, and related metabolic disorders are major health issues worldwide. As the epidemic of metabolic disorders continues, the associated medical comorbidities, including the detrimental impact on reproduction, increase as well. Emerging evidence suggests that the effects of maternal nutrition on reproductive outcomes are likely to be mediated, at least in part, by oocyte metabolism. Well-balanced and timed energy metabolism is critical for optimal development of oocytes. To date, much of our understanding of oocyte metabolism comes from the effects of extrinsic nutrients on oocyte maturation. In contrast, intrinsic regulation of oocyte development by metabolic enzymes, intracellular mediators, and transport systems is less characterized. Specifically, decreased acid transport proteins levels, increased glucose/lipid content and elevated reactive oxygen species in oocytes have been implicated in meiotic defects, organelle dysfunction and epigenetic alteration. Therefore, metabolic disturbances in oocytes may contribute to the diminished reproductive potential experienced by women with metabolic disorders. In-depth research is needed to further explore the underlying mechanisms. This review also discusses several approaches for metabolic analysis. Metabolomic profiling of oocytes, the surrounding granulosa cells, and follicular fluid will uncover the metabolic networks regulating oocyte development, potentially leading to the identification of oocyte quality markers and prevention of reproductive disease and poor outcomes in offspring. PMID:25280482

Liu, Honglin; Gu, Xi; Boots, Christina; Moley, Kelle H.

2015-01-01

367

Absence of cumulus cells during in vitro maturation affects lipid metabolism in bovine oocytes.  

PubMed

Cumulus cells (CC) surround the oocyte and are coupled metabolically through regulation of nutrient intake. CC removal before in vitro maturation (IVM) decreases bovine oocyte developmental competence without affecting nuclear meiotic maturation. The objective was to investigate the influence of CC on oocyte cytoplasmic maturation in relation to energy metabolism. IVM with either cumulus-enclosed (CEO) or -denuded (DO) oocytes was performed in serum-free metabolically optimized medium. Transmission electron microscopy revealed different distribution of membrane-bound vesicles and lipid droplets between metaphase II DO and CEO. By Nile Red staining, a significant reduction in total lipid level was evidenced in DO. Global transcriptomic analysis revealed differential expression of genes regulating energy metabolism, transcription, and translation between CEO and DO. By Western blot, fatty acid synthase (FAS) and hormone-sensitive phospholipase (HSL) proteins were detected in oocytes and in CC, indicating a local lipogenesis and lypolysis. FAS protein was significantly less abundant in DO that in CEO and more highly expressed in CC than in the oocytes. On the contrary, HSL protein was more abundant in oocytes than in CC. In addition, active Ser??³-phosphorylated HSL was detected in the oocytes only after IVM, and its level was similar in CEO and DO. In conclusion, absence of CC during IVM affected lipid metabolism in the oocyte and led to suboptimal cytoplasmic maturation. Thus, CC may influence the oocyte by orienting the consumption of nutritive storage via regulation of local fatty acid synthesis and lipolysis to provide energy for maturation. PMID:23321473

Auclair, Sylvain; Uzbekov, Rustem; Elis, Sébastien; Sanchez, Laura; Kireev, Igor; Lardic, Lionel; Dalbies-Tran, Rozenn; Uzbekova, Svetlana

2013-03-15

368

Prolonged exposure to hyaluronidase decreases the fertilization and development rates of fresh and cryopreserved mouse oocytes  

PubMed Central

Hyaluronidase is generally used to remove cumulus cells from mouse oocytes before oocyte cryopreservation, intracytoplasmic sperm injection or DNA injection. In general, use of cumulus-free mouse oocytes decreases in vitro fertilizing ability compared with cumulus-surrounded oocytes. The effect of hyaluronidase exposure on the quality of mouse oocytes is not fully understood. Here, we investigated the effect of hyaluronidase exposure time on the fertilization rate of fresh and vitrified mouse oocytes and their subsequent developmental ability in vitro. We found that the fertilization rate decreased with hyaluronidase treatments. This reduction in the fertilization rate following treatment with hyaluronidase was fully reversed by removal of the zona pellucida. In addition, oocytes treated with hyaluronidase for 5 min or longer had a reduced capacity to develop to the morula and blastocyst stage. The survival, fertilization, and developmental rates of vitrified-warmed oocytes were also reduced by longer exposure to hyaluronidase. In conclusion, these results suggest that prolonged exposure to hyaluronidase decreases the quality of mouse oocytes and shorter hyaluronidase treatment times may help achieve a stable and high fertilization rate in fresh and cryopreserved oocytes. PMID:25225080

ISHIZUKA, Yuta; TAKEO, Toru; NAKAO, Satohiro; YOSHIMOTO, Hidetaka; HIROSE, Yumiko; SAKAI, Yuki; HORIKOSHI, Yuka; TAKEUJI, Shiori; TSUCHIYAMA, Shuuji; NAKAGATA, Naomi

2014-01-01

369

Protein deubiquitination during oocyte maturation influences sperm function during fertilisation, antipolyspermy defense and embryo development.  

PubMed

Ubiquitination is a covalent post-translational modification of proteins by the chaperone protein ubiquitin. Upon docking to the 26S proteasome, ubiquitin is released from the substrate protein by deubiquitinating enzymes (DUBs). We hypothesised that specific inhibitors of two closely related oocyte DUBs, namely inhibitors of the ubiquitin C-terminal hydrolases (UCH) UCHL1 (L1 inhibitor) and UCHL3 (L3 inhibitor), would alter porcine oocyte maturation and influence sperm function and embryo development. Aberrant cortical granule (CG) migration and meiotic spindle defects were observed in oocytes matured with the L1 or L3 inhibitor. Embryo development was delayed or blocked in oocytes matured with the general DUB inhibitor PR-619. Aggresomes, the cellular stress-inducible aggregates of ubiquitinated proteins, formed in oocytes matured with L1 inhibitor or PR-619, a likely consequence of impaired protein turnover. Proteomic analysis identified the major vault protein (MVP) as the most prominent protein accumulated in oocytes matured with PR-619, suggesting that the inhibition of deubiquitination altered the turnover of MVP. The mitophagy/autophagy of sperm-contributed mitochondria inside the fertilised oocytes was hindered by DUB inhibitors. It is concluded that DUB inhibitors alter porcine oocyte maturation, fertilisation and preimplantation embryo development. By regulating the turnover of oocyte proteins and mono-ubiquitin regeneration, the DUBs may promote the acquisition of developmental competence during oocyte maturation. PMID:24848520

Yi, Young-Joo; Sutovsky, Miriam; Song, Won-Hee; Sutovsky, Peter

2014-05-22

370

Metabolic control of oocyte development: linking maternal nutrition and reproductive outcomes.  

PubMed

Obesity, diabetes, and related metabolic disorders are major health issues worldwide. As the epidemic of metabolic disorders continues, the associated medical co-morbidities, including the detrimental impact on reproduction, increase as well. Emerging evidence suggests that the effects of maternal nutrition on reproductive outcomes are likely to be mediated, at least in part, by oocyte metabolism. Well-balanced and timed energy metabolism is critical for optimal development of oocytes. To date, much of our understanding of oocyte metabolism comes from the effects of extrinsic nutrients on oocyte maturation. In contrast, intrinsic regulation of oocyte development by metabolic enzymes, intracellular mediators, and transport systems is less characterized. Specifically, decreased acid transport proteins levels, increased glucose/lipid content and elevated reactive oxygen species in oocytes have been implicated in meiotic defects, organelle dysfunction and epigenetic alteration. Therefore, metabolic disturbances in oocytes may contribute to the diminished reproductive potential experienced by women with metabolic disorders. In-depth research is needed to further explore the underlying mechanisms. This review also discusses several approaches for metabolic analysis. Metabolomic profiling of oocytes, the surrounding granulosa cells, and follicular fluid will uncover the metabolic networks regulating oocyte development, potentially leading to the identification of oocyte quality markers and prevention of reproductive disease and poor outcomes in offspring. PMID:25280482

Gu, Ling; Liu, Honglin; Gu, Xi; Boots, Christina; Moley, Kelle H; Wang, Qiang

2015-01-01

371

Cumulus cells accelerate oocyte aging by releasing soluble Fas Ligand in mice  

PubMed Central

Although previous studies have suggested that cumulus cells (CCs) accelerate oocyte aging by secreting soluble and heat-sensitive paracrine factors, the factors involved are not well characterized. Because Fas-mediated apoptosis represents a major pathway in induction of apoptosis in various cells, we proposed that CCs facilitate oocyte aging by releasing soluble Fas ligand (sFasL). In this study, we reported that when the aging of freshly ovulated mouse oocytes were studied in vitro, both the apoptotic rates of CCs and the amount of CCs produced sFasL increased significantly with the culture time. We found that oocytes expressed stable levels of Fas receptors up to 24?h of in vitro aging. Moreover, culture of cumulus-denuded oocytes in CCs-conditioned CZB medium (CM), in CZB supplemented with recombinant sFasL, or in CM containing sFasL neutralizing antibodies all showed that sFasL impaired the developmental potential of the oocytes whereas facilitating activation and fragmentation of aging oocytes. Furthermore, CCs from the FasL-defective gld mice did not accelerate oocyte aging due to the lack of functional FasL. In conclusion, we propose that CCs surrounding aging oocytes released sFasL in an apoptosis-related manner, and the released sFasL accelerated oocyte aging by binding to Fas receptors. PMID:25731893

Zhu, Jiang; Zhang, Jie; Li, Hong; Wang, Tian-Yang; Zhang, Chuan-Xin; Luo, Ming-Jiu; Tan, Jing-He

2015-01-01

372

Are zona pellucida laser drilling and polar body biopsy safe for in vitro matured oocytes?  

PubMed Central

Introduction Preconception diagnosis requires first polar body biopsy. When the hole in the zona pellucida is made with a laser beam, heat propagation could, like the biopsy itself, be deleterious. Our aim was to evaluate the effect of this technique on human in vitro matured oocyte and embryo development. Methods One hunded fifty five retrieved immature oocytes from 75 women, matured in vitro, were distributed in 3 groups: 50 oocytes in a control group, without laser drilling and first polar body biopsy, 52 oocytes in a group with only laser drilling, and 53 oocytes in a group with both laser drilling and first polar body biopsy. Safety was evaluated using four criteria: [1] oocyte lysis rate, [2] oocyte activation rate, [3] oocyte development after calcium ionophore treatment, [4] and embryo chromosome breakage incidence after Tarkowski preparation. Results No difference in the four criteria was observed between the 3 oocyte groups. Conclusions We did not find evidence of deleterious effect of laser drilling and first polar body biopsy on in vitro matured oocytes, according to our criteria. PMID:20495883

Hammoud, Ibrahim; Molina-Gomes, Denise; Albert, Martine; Bergere, Marianne; Bailly, Marc; Wainer, Robert; Selva, Jacqueline

2010-01-01

373

Optimising vitrification of human oocytes using multiple cryoprotectants and morphological and functional assessment.  

PubMed

Oocyte vitrification is a clinical practice that allows preservation of fertility potential in women. Vitrification involves quick cooling using high concentrations of cryoprotectants to minimise freezing injuries. However, high concentrations of cryoprotectants have detrimental effects on oocyte quality and eventually the offspring. In addition, current assessment of oocyte quality after vitrification is commonly based only on the morphological appearance of the oocyte, raising concerns regarding its efficiency. Using both morphological and functional assessments, the present study investigated whether combinations of cryoprotectants at lower individual concentrations result in better cryosurvival rates than single cryoprotectants at higher concentrations. Surplus oocytes from IVF patients were vitrified within 24h after retrieval using the Cryotop method with several cryoprotectants, either individually or in combination. The morphological and functional quality of the vitrified oocytes was investigated using light microscopy and computer-based quantification of mitochondrial integrity, respectively. Oocyte quality was significantly higher using a combination of cryoprotectants than vitrification with individual cryoprotectants. In addition, the quality of vitrified oocyte varied depending on the cryoprotectants and type of combination used. The results of the present study indicate that observations based purely on the morphological appearance of the oocyte to assess the cryosurvival rate are insufficient and sometimes misleading. The outcome will have a significant implication in the area of human oocyte cryopreservation as an important approach for fertility preservation. PMID:22967503

Seet, V Y K; Al-Samerria, S; Wong, J; Stanger, J; Yovich, J L; Almahbobi, G

2013-01-01

374

Oocyte development and fecundity type of the skipjack, Katsuwonus pelamis, in the Western Indian Ocean  

NASA Astrophysics Data System (ADS)

The study aims to define the oogenesis pattern of the skipjack (Katsuwonus pelamis) of the Western Indian Ocean basin in terms of oocyte growth and recruitment style. The main objective is to define the type of fecundity regulation (i.e. determinate or indeterminate) based on four lines of evidence: (a) oocyte size-frequency distribution; (b) seasonal variation of the relative number and percentage of oocyte stages, (c) diameter of the advanced vitellogenic oocytes in females in the spawning capable phase; and (d) incidence of atresia throughout the spawning season. The samples were collected from 2009 to 2010 in the Western Indian Ocean, and 673 ovaries were classified in the different reproductive phases using histological staging. Moreover, the oocyte size distribution of 93 mature individuals was described by the newly implemented image analysis method. This species showed a broad oocyte size frequency distribution with no gap formation between the primary and secondary oocyte growth stages. There was no seasonal variation in the percentage of oocyte stages in ovaries in the spawning capable phase, and the diameter of those oocytes at the most advanced vitellogenic stage was approximately constant during the sampling period. These facts provide evidence of continuous oocyte recruitment into the standing stock of developing oocytes. Moreover, when reaching the end of the active reproductive period (i.e. February and March) the prevalence of atresia increased. This is a mechanism adopted by fishes of the indeterminate fecundity type to reabsorb the surplus oocyte production. Based on the findings, we state that the skipjack in the Western Indian Ocean shows asynchronous oocyte growth and an indeterminate fecundity type.

Grande, Maitane; Murua, Hilario; Zudaire, Iker; Korta, Maria

2012-10-01

375

G-protein coupled estrogen receptor (GPER) inhibits final oocyte maturation in common carp, Cyprinus carpio.  

PubMed

GPR-30, now named as GPER (G protein-coupled estrogen receptor) was first identified as an orphan receptor and subsequently shown to be required for estrogen-mediated signaling in certain cancer cells. Later studies demonstrated that GPER has the characteristics of a high affinity estrogen membrane receptor on Atlantic croaker and zebra fish oocytes and mediates estrogen inhibition of oocyte maturation in these two distantly related teleost. To determine the broad application of these findings to other teleost, expression of GPER mRNA and its involvement in 17?-estradiol mediated inhibition of oocyte maturation in other cyprinid, Cyprinus carpio was investigated. Carp oocytes at pre-vitellogenic, late-vitellogenic and post-vitellogenic stages of development contained GPER mRNA and its transcribed protein with a maximum at late-vitellogenic oocytes. Ovarian follicular cells did not express GPER mRNA. Carp oocytes GPER mRNA was essentially identical to that found in other perciformes and cyprinid fish oocytes. Both spontaneous and 17,20?-dihydroxy-4-pregnen-3-one (17,20?-P)-induced oocyte maturation in carp was significantly decreased when they were incubated with either E2, or GPER agonist G-1. On the other hand spontaneous oocyte maturation was significantly increased when carp ovarian follicles were incubated with an aromatase inhibitor, fadrozole, GPER antagonist, G-15 and enzymatic removal of the ovarian follicle cell layers. This increase in oocyte maturation was partially reversed by co-treatment with E2. Consistent with previous findings with human and fish GPR30, E2 treatment in carp oocytes caused increase in cAMP production and simultaneously decrease in oocyte maturation, which was inhibited by the addition of 17,20?-P. The results suggest that E2 and GPER play a critical role in regulating re-entry in to meiotic cell cycle in carp oocytes. PMID:25485460

Majumder, Suravi; Das, Sumana; Moulik, Sujata Roy; Mallick, Buddhadev; Pal, Puja; Mukherjee, Dilip

2015-01-15

376

CRL4DCAF1 is required in activated oocytes for follicle maintenance and ovulation.  

PubMed

In mammals, oocytes within the primordial follicles require a number of essential factors to maintain their survival. However, the survival factors for activated oocytes have been poorly characterized. Recently we reported that damaged DNA binding protein-1 (DDB1), the linker subunit of the cullin ring-finger ubiquitin E3 ligase-4 (CRL4) complex, and its substrate adaptor, DDB1-CUL4 associated factor-1 (DCAF1), were essential for primordial follicle maintenance. In this study we specifically deleted these in the oocytes of growing follicles, to investigate if DDB1 and DCAF1 were also survival factors for activated oocytes. In the ovaries of Ddb1(fl/fl);Zp3-Cre mice, the primordial follicle pool was intact, but awakened oocytes and growing follicles beyond the primary stage were rapidly depleted. In the ovaries of Dcaf1(fl/fl);Pten(fl/fl);Gdf9-Cre and Ddb1(fl/fl);Pten(fl/fl);Gdf9-Cre mice, global primordial follicle activation was stimulated by enhanced PI3K signaling, but the awakened oocytes were rapidly lost due to no CRL4(DCAF1) activity. These mouse models provided original evidence that CRL4(DCAF1) was essential for maintaining oocyte survival, not only those in dormancy at the primordial follicle stage, but also naturally awakened oocytes and those awakened by hyper-activation of PI3K signaling. Interestingly, the oocyte-specific Ddb1 or Dcaf1 knockout mice had ovulation defects even before oocyte exhaustion. CRL4(DCAF1) within oocytes was required for cumulus expansion and ovulation-related somatic gene expression in a cell non-autonomous manner. Granulosa cells that surrounded these Ddb1 or Dcaf1-deleted oocytes exhibited increased rates of apoptosis and showed poor responses to ovulation signals. These results suggested that CRL4 in oocytes also regulated granulosa cell functions in a cell non-autonomous manner. PMID:25371539

Yu, Chao; Xu, Yi-Wen; Sha, Qian-Qian; Fan, Heng-Yu

2015-02-01

377

The Effect of Vitrification on Mouse Oocyte Apoptosis by Cryotop Method  

PubMed Central

Background : Oocyte cryopreservation is one of the most important topics in the field of assisted reproductive technology to preserve women fertility, but relationship between cryopreservation and apoptosis is still a matter of debate. The present study was aimed to investigate the effects of vitrification on apoptosis in mouse oocytes by Cryotop method. Method: A total of 200 germinal vesicle (GV) and 200 metaphase II (MII) oocytes were obtained from ovaries and fallopian tubes of NMRI mice, respectively and divided into control and experimental groups. Oocytes in experimental group were vitrified by Cryotop using vitrification medium and were kept in liquid nitrogen for one month. The survival rate of oocytes was evaluated after 2 hour incubation time. Then, the oocyte apoptosis was evaluated by TUNEL technique and compared with those in control group. The data was compared statistically using SPSS software and chi-square test. Results: The survival rates of vitrified GV (93%) and MII oocytes (88%) showed a significant decrease compared with the control group (P<0.05), but there was no significant difference in survival rate of both vitrified oocyte groups. The incidence of apoptosis in vitrified and control GV oocytes showed no significant difference (13% vs. 7%), but the rate of apoptosis in vitrified MII oocytes increased significantly not only in comparison with MII control group (25% vs. 5%) but also with vitrified GV oocytes (P<0.05). Conclusion: The results indicate that vitrification increases apoptosis in mouse MII oocytes and apoptosis may play a role in MII oocyte injury after vitrification. PMID:23999716

Rajaei, Farzad; Abedpour, Neda; Salehnia, Mojdeh; Jahanihashemi, Hassan

2013-01-01

378

Birth of a domestic cat kitten produced by vitrification of lipid polarized in vitro matured oocytes.  

PubMed

The ability to cryopreserve oocytes is an effective method to retain valuable genetic material of mammals, including that of endangered animals. Embryos of domestic cats are amenable to cryopreservation, whereas their oocytes are much less cryo-tolerant. The capability of oocytes to survive cryopreservation is affected by several factors, one of which has been hypothesized to be the high concentration of intracellular lipids. To test this hypothesis, in this study we polarized lipids of cat oocytes and tested their cooling and freezing sensitivity. We found that the sensitivity of oocytes to cooling and cryopreservation does appear to be related to their high intracellular lipid content, as indicated by higher cryosurvival and development into blastocysts when intracellular lipids of in vitro matured oocytes were polarized before vitrification. However, polarization of all intracellular lipids was detrimental to development of embryos. Cell numbers in blastocysts derived from fully polarized/vitrified oocytes were significantly lower than those of partially polarized/vitrified or non-vitrified/fresh oocytes. Although embryos derived from fully polarized/vitrified oocytes developed to the blastocyst stage at higher rates than those of partially polarized/vitrified or non-centrifuged/vitrified oocytes, their in vivo developmental competence was compromised. When embryos derived from fully polarized/vitrified oocytes were transferred, although two recipients became pregnant, all implanted embryos were reabsorbed. In contrast, when embryos derived from oocytes that were only partially lipid polarized before vitrification and then were transferred, one recipient did become pregnant and produced a live healthy kitten. The present results suggest that other approaches to altering intra-cellular lipid levels in cat oocytes should be evaluated to improve their functional survival after cryopreservation. PMID:24631204

Galiguis, Jason; Gómez, Martha C; Leibo, S P; Pope, C Earle

2014-06-01

379

Translocation and Endocytosis for Cell-penetrating Peptide Internalization  

PubMed Central

Cell-penetrating peptides (CPPs) share the property of cellular internalization. The question of how these peptides reach the cytoplasm of cells is still widely debated. Herein, we have used a mass spectrometry-based method that enables quantification of internalized and membrane-bound peptides. Internalization of the most used CPP was studied at 37 °C (endocytosis and translocation) and 4 °C (translocation) in wild type and proteoglycan-deficient Chinese hamster ovary cells. Both translocation and endocytosis are internalization pathways used by CPP. The choice of one pathway versus the other depends on the peptide sequence (not the number of positive changes), the extracellular peptide concentration, and the membrane components. There is no relationship between the high affinity of these peptides for the cell membrane and their internalization efficacy. Translocation occurs at low extracellular peptide concentration, whereas endocytosis, a saturable and cooperative phenomenon, is activated at higher concentrations. Translocation operates in a narrow time window, which implies a specific lipid/peptide co-import in cells. PMID:19833724

Jiao, Chen-Yu; Delaroche, Diane; Burlina, Fabienne; Alves, Isabel D.; Chassaing, Gérard; Sagan, Sandrine

2009-01-01

380

Heat and cold acclimation in helium-cold hypothermia in the hamster.  

NASA Technical Reports Server (NTRS)

A study was made of the effects of acclimation of hamsters to high (34-35 C) and low (4-5 C) temperatures for periods up to 6 weeks on the induction of hypothermia in hamsters. Hypothermia was achieved by exposing hamsters to a helox mixture of 80% helium and 20% oxygen at 0 C. Hypothermic induction was most rapid (2-3 hr) in heat-acclimated hamsters and slowest (6-12 hr) in cold-acclimated hamsters. The induction period was intermediate (5-8 hr) in room temperature nonacclimated animals (controls). Survival time in hypothermia was relatable to previous temperature acclimations. The hypothesis that thermogenesis in cold-acclimated hamsters would accentuate resistance to induction of hypothermia was substantiated.

Musacchia, X. J.

1972-01-01

381

Restoration of immune responses of aging hamsters by treatment with isoprinosine.  

PubMed Central

Immune competence declines with advanced age in hamsters, as in other laboratory mammals and in humans. We found significant alterations in the functional parameters of different populations of immunocytes (natural killer cells, T cells, monocytes, and suppressor cells) in aging hamsters, beginning at approximately 14 mo of age. Natural killer cytotoxicity, phytohemagglutinin-induced lymphocyte stimulation, and monocyte chemotaxis were decreased in aging Lak:LvG(Syr) outbred hamsters. When old hamsters were given a single injection (5 mg/kg body wt) of isoprinosine, a chemical immune potentiator, these three immune parameters increased almost to the levels found in young adult hamsters but returned to pretreatment levels after 7 d. Suppressor cell activity for the lymphocyte response to phytohemagglutinin, which increased with age, was decreased after treatment. In old hamsters treated with weekly injections of isoprinosine, these four immunological parameters remained at or near the levels found in young adults. PMID:6190840

Tsang, K Y; Fudenberg, H H; Gnagy, M J

1983-01-01

382

GROUND PENETRATING RADAR ON MARS  

Microsoft Academic Search

In the past decade and in the next decade, several ground penetrating radar systems have been or will be launched toward Mars. GPR systems have been built or proposed for study of Mars and the Martian satellite, Phobos, from orbit, balloons, and rovers. The scientific problems include those of finding evidence of past or present life on Mars, unraveling the

Gary R. Olhoeft

1998-01-01

383

FAA Fluorescent Penetrant Laboratory Inspections  

SciTech Connect

The Federal Aviation Administration Airworthiness Assurance NDI Validation Center currently assesses the capability of various non-destructive inspection (NDI) methods used for analyzing aircraft components. The focus of one such exercise is to evaluate the sensitivity of fluorescent liquid penetrant inspection. A baseline procedure using the water-washable fluorescent penetrant method defines a foundation for comparing the brightness of low cycle fatigue cracks in titanium test panels. The analysis of deviations in the baseline procedure will determine an acceptable range of operation for the steps in the inspection process. The data also gives insight into the depth of each crack and which step(s) of the inspection process most affect penetrant sensitivities. A set of six low cycle fatigue cracks produced in 6.35-mm thick Ti-6Al-4V specimens was used to conduct the experiments to produce sensitivity data. The results will document the consistency of the crack readings and compare previous experiments to find the best parameters for water-washable penetrant.

WINDES,CONNOR L.; MOORE,DAVID G.

2000-08-02

384

Milder Etchant For Penetrant Inspection  

NASA Technical Reports Server (NTRS)

New etching solution for chemical penetrant inspection of Inconel(R) 718 castings and weldments. Etchant does not introduce artifacts mistaken for defects. Applied by swabbing or by immersion. Used to detect unwanted residues of Nioro(R) (or equivalent) gold brazing alloy on type 347 stainless steel.

O'Tousa, Joseph E.; Thomas, Clark S.

1990-01-01

385

Drug penetration in solid tumours  

Microsoft Academic Search

To be most effective anticancer drugs must penetrate tissue efficiently, reaching all the cancer cells that comprise the target population in a concentration sufficient to exert a therapeutic effect. Most research into the resistance of cancers to chemotherapy has concentrated on molecular mechanisms of resistance, whereas the role of limited drug distribution within tumours has been neglected. We summarize the

Andrew I. Minchinton; Ian F. Tannock

2006-01-01

386

Nonhost Root Penetration by Soybean Cyst Nematode  

PubMed Central

A total of 66 plants in 50 species were inoculated with eggs and juveniles of soybean cyst nematode, Heterodera glycines. Roots were stained and observed for penetration and development of the nematode. Twenty-six plants were not penetrated; twenty-three were penetrated, but there was no development of the nematode; eight were penetrated with some nematode development; two were penetrated and had considerable nematode development, but few nematodes, if any, matured; and seven were penetrated with many nematodes maturing. The penetration of nonhosts may imply some susceptibility and that populations eventually would build up on the penetrated plants. Plants not penetrated may be useful as rotation plants because no reproduction would occur. PMID:19290137

Riggs, R. D.

1987-01-01

387

Penetration below a convective zone  

NASA Technical Reports Server (NTRS)

Two-dimensional numerical simulations are used to investigate how fully compressible nonlinear convection penetrates into a stably stratified zone beneath a stellar convection zone. Estimates are obtained of the extent of penetration as the relative stability S of the stable to the unstable zone is varied over a broad range. The model deals with a perfect gas possessing a constant dynamic viscosity. The dynamics is dominated by downward-directed plumes which can extend far into the stable material and which can lead to the excitation of a broad spectrum of internal gravity waves in the lower stable zone. The convection is highly time dependent, with the close coupling between the lateral swaying of the plumes and the internal gravity waves they generate serving to modulate the strength of the convection. The depth of penetration delta, determined by the position where the time-averaged kinetic flux has its first zero in the stable layer, is controlled by a balance between the kinetic energy carried into the stable layer by the plumes and the buoyancy braking they experience there. A passive scalar is introduced into the unstable layer to evaluate the transport of chemical species downward. Such a tracer is effectively mixed within a few convective overturning times down to a depth of delta within the stable layer. Analytical estimates based on simple scaling laws are used to interpret the variation of delta with S, showing that it first involves an interval of adiabatic penetration if the local Peclet number of the convection exceeds unity, followed by a further thermal adjustment layer, the depths of each interval scaling in turn as S(exp -1) and S(exp -1/4). These estimates are in accord with the penetration results from the simulations.

Hurlburt, Neal E.; Toomre, Juri; Massaguer, Josep M.; Zahn, Jean-Paul

1994-01-01

388

Hibernation, stress, intestinal functions, and catecholoamine turnover rate in hamsters and gerbils  

NASA Technical Reports Server (NTRS)

Bioenergetic studies on hamsters during depressed metabolic states are reported. External support of blood glucose extended the survival times of hibernating animals. Radioresistance increased in hibernating as well as in hypothermic hamsters. Marked changes in hamster catecholamine turnover rates were observed during acclimatization to high temperature stress. High radioresistance levels of the gerbil gastrointestinal system were attributed in part to the ability of the gut to maintain functional integrity.

Musacchia, X. J.

1973-01-01

389

Young hamsters are more resistant than adults to endotracheally instilled porcine pancreatic elastase.  

PubMed

We measured the physiologic and stereologic response to 0.1, 0.2, and 0.4 microgram of porcine pancreatic elastase instilled in a volume of 0.25 ml 0.9% NaCl/100 g body weight into the trachea of groups of young and adult hamsters. The young hamsters averaged 50 g and the adult hamsters 116 g in initial body weight. Twenty-one days after administration of elastase, lung volumes, static lung compliance, maximum expiratory flow, the whole section mean linear intercepts (MLI) were measured. The degree of emphysema increased in all animals as a function of dose. Examination of the lung volume and compliance dose-response characteristics indicated that young hamsters developed less physiologic change with increasing elastase dose than did adult hamsters. Maximum expiratory flow and whole section MLI dose-response were similar in the young and adult elastase-treated groups. However, the MLI in young hamsters treated with the 0.4 microgram elastase dose was decreased in the outer third of the lung compared to adult emphysematous hamsters. Also, mean airspace density relative to saline control values in young hamsters was double that found in adult hamsters treated with the 0.4 microgram elastase dose. Although serum alpha 1-globulin levels were equivalent in both young and adult normal hamsters, values normalized for lung elastin content were significantly increased in young animals. We conclude that young hamsters show less change in lung function as a function of elastase dose twenty-one days after elastase instillation. Possible reasons for this include an increased ratio of lung alpha 1-globulin/lung elastin in young hamsters, their continued ability to grow new alveoli, and age related differences in airway size favoring a central distribution of enzyme. PMID:3640708

Karlinsky, J B; Goldstein, R H; Catanese, A; Snider, G L

1986-01-01

390

Serotonin modulates offensive attack in adolescent anabolic steroid-treated hamsters  

Microsoft Academic Search

Chronic anabolic–androgenic steroid (AAS) treatment during adolescence facilitates offensive aggression in male Syrian hamsters (Mesocricetus auratus). The current study assessed whether adolescent AAS-facilitated offensive attack was modulated by serotonin (5-HT) and if AAS exposure during this developmental period influenced 5-HT innervation to areas of hamster brain implicated in aggressive behavior. In a first experiment, hamsters were administered high-dose AAS throughout

Jill M. Grimes; Richard H. Melloni Jr

2002-01-01

391

Glutamic acid decarboxylase (GAD 65) immunoreactivity in brains of aggressive, adolescent anabolic steroid-treated hamsters  

Microsoft Academic Search

Chronic anabolic–androgenic steroid (AAS) treatment during adolescence facilitates offensive aggression in male Syrian hamsters (Mesocricetus auratus). The current study assessed whether adolescent AAS exposure influenced the immunohistochemical localization of glutamic acid decarboxylase (GAD65), the rate-limiting enzyme in the synthesis of ?-aminobutyric acid (GABA), in areas of hamster brain implicated in aggressive behavior. Hamsters were administered high dose AAS throughout adolescence,

Jill M. Grimes; Lesley A. Ricci; Richard H. Melloni Jr

2003-01-01

392

Microsurgical Injection of Spermatozoa into Hamster Eggs with Subsequent Transformation of Sperm Nuclei into Male Pronuclei  

Microsoft Academic Search

Isolated nuclei of hamster spermatozoa develop into male pronuclei when injected into hamster eggs. The nuclei of fresh, frozen-thawed and freeze-dried human spermatozoa are equally capable of developing into male pronuclei when injected into hamster eggs. These results indicate that sperm nuclei are stable organdIes and the egg cytoplasmic factors controlling the transformation of sperm nuclei into male pronuclei are

T. UEHARA; R. YANAGIMACHI

1976-01-01

393

Conjugated linoleic acid isomers reduce blood cholesterol levels but not aortic cholesterol accumulation in hypercholesterolemic hamsters  

Microsoft Academic Search

The aim of the present study was to characterize plasma lipids and lipoprotein cholesterol and glucose concentrations in hamsters\\u000a fed either cis-9, trans-11 CLA (9c, 11t CLA); trans-10, cis-12 CLA (10t, 12c CLA); or linoleic acid (LA) on the accumulation of aortic cholesterol in hypercholesterolemic hamsters. One hundred male\\u000a F1B strain Syrian Golden Hamsters (Mesocricetus auratus) (BioBreeders Inc., Watertown, MA)

Thomas A. Wilson; Robert J. Nicolosi; Andrew Saati; Timothy Kotyla; David Kritchevsky

2006-01-01

394

[Investigation on the prevalence of gastrointestinal parasites in pet hamsters].  

PubMed

One hundred and fifty-three fecal samples of pet hamsters (Mesocricetus auratus, Phodopus sungorus, P. campbelli and P. roborovskii) were collected from a pet-market in Zhengzhou, and examined by Sheather's sugar flotation, modified acid-fast staining and Lugol's iodine-solution staining. The prevalence of parasites was 70.7% (41/58), 96.7% (59/61), 83.9% (26/31), and 100% (3/3) respectively, with an overall prevalence of 84.3%. Eggs, cysts or oocysts of Cryptosporidium sp. (15.0%), Giardia sp. (22.2%), coccidian (2.0%), Hymenolepis nana (31.4%), Hymenolepis diminuta (25.5%), Syphacia spp. (41.8%), Aspiculuris tetraptera (7.2%) and undetermined Trichurata nematode (18.3%) were found from the samples. The results suggest that pet hamsters may be infected and transmit several zoonotic parasites. PMID:19852376

Lv, Chao-Chao; Feng, Chao; Qi, Meng; Yang, Hong-Yu; Jian, Fu-Chun; Ning, Chang-Shen; Zhang, Long-Xian

2009-06-01

395

Sarcolemmal phospholipid N-methylation in genetically determined hamster cardiomyopathy  

SciTech Connect

The heart sarcolemmal phosphatidylethanolamine N-methylation in UM-X7.1 strain of cardiomyopathic hamsters was examined by using 0.055, 10 and 150 microM S-adenosyl-L-(methyl-/sup 3/H) methionine as methyl donor for sites I, II and III, respectively. In comparison with control values, methylation activities at site I was increased in 40, 120 and 250 days old cardiomyopathic hamsters. On the other hand, methylation activities at sites II and III in 120 and 250 days old cardiomyopathic animals were depressed without any change in the 40 days old group. The alterations in N-methylation activities were associated with kinetic changes in apparent Vmax values without any changes in the apparent Km. These results indicate a defect in the phospholipid N-methylation process in heart sarcolemma during the development of genetically determined cardiomyopathy.

Okumura, K.; Panagia, V.; Jasmin, G.; Dhalla, N.S.

1987-02-27

396

Autoradiography in fetal golden hamsters treated with tritiated diethylnitrosamine  

SciTech Connect

Tritiated diethylnitrosamine was administered to female Syrian golden hamsters on each of the last 4 days (days 12-15) of pregnancy. The distribution of bound radioactivity was monitored by light microscopic autoradiography of fetal tracheas and livers, the placentas, and the maternal livers. In the trachea, the fetal target organ, bound radioactivity was restricted to the respiratory epithelium, where diethylnitrosamine-induced tracheal tumors arise. Mucous cells and nonciliated stem cells were identified as the principal sites of binding; other cell types within the tracheal epithelium contained only small amounts of bound radioactivity. The level of binding observed in the fetal trachea increased steadily from day 12 to day 15, which correlated well with the levels of differentiation of this tissue during this period. This observation also agrees with the previously reported observation that tumor incidence increases from 40 to 95% in Syrian golden hamsters between days 12 and 15.

Reznik-Schueller, H.M.; Hague, B.F. Jr.

1981-04-01

397

Identification and characterization of the hamster polyomavirus middle T antigen.  

PubMed Central

Hamster polyomavirus (HaPV) is associated with lymphoid and hair follicle tumors in Syrian hamsters. The early region of HaPV has the potential to encode three polypeptides (which are related to the mouse polyomavirus early proteins) and can transform fibroblasts in vitro. We identified the HaPV middle T antigen (HamT) as a 45-kDa protein. Like its murine counterpart, HamT was associated with serine/threonine phosphatase, phosphatidylinositol-3 kinase, and protein tyrosine kinase activities. However, whereas mouse middle T antigen associates predominantly with pp60c-src and pp62c-yes, HamT was associated with a different tyrosine kinase, p59fyn. The ability of HaPV to cause lymphoid tumors may therefore reside in its ability to associate with p59fyn, a potentially important tyrosine kinase in lymphocytes. Images PMID:1709702

Courtneidge, S A; Goutebroze, L; Cartwright, A; Heber, A; Scherneck, S; Feunteun, J

1991-01-01

398

The pineal affects life span in hamsters with heart disease.  

PubMed

Cardiomyopathic hamsters (CMH) develop heart disease early in life which leads to congestive heart failure and death as these hamsters age. We have previously shown that living in constant light or other non-24-h light-dark (LD) cycles can increase longevity in these hamsters, and the current experiment examined potential mechanisms for this effect. Thus, CMH were orchidectomized, pinealectomized, or given melatonin treatment and then placed on either 1:23 or 1:23.6 LD cycles. Orchidectomy had no effect on longevity in either LD cycle, but in 1:23.6 it did lead to death with a greater degree of heart failure. On the other hand, pinealectomy of 1:23 CMH led to changes in life span similar to those produced by placing the hamsters in 1:23.6. Moreover, melatonin implant treatment of CMH in 1:23.6 led to changes in life span that were similar to those caused by life in 1:23, at least over the first half of the survival curves. Thus, it appears that the pineal gland and melatonin may be involved in mediating the effects of non-24-h LD cycles, whether these effects are beneficial or detrimental. In addition, the testes and testosterone appear to have no role in mediating these effects. These data suggest that inhibition, rather than stimulation, of pineal function might be beneficial for those with congestive heart failure, but further experiments are necessary to clarify when during the disease process potential treatments might be helpful. PMID:9333200

Natelson, B H; Ottenweller, J E; Tapp, W N; Heung, S; Beldowicz, D

1997-11-01

399

Self-administration of estrogen and dihydrotestosterone in male hamsters  

Microsoft Academic Search

Anabolic–androgenic steroids (AAS) are drugs of abuse. Previous studies have shown that male and female hamsters self-administer testosterone (T) and other AAS, suggesting that androgens are reinforcing in a context where athletic performance is irrelevant. AAS are synthetic derivatives of T, which may be aromatizable to estrogen and\\/or reducible to dihydrotestosterone (DHT). However, we do not know which metabolites of

Anita N. DiMeo; Ruth I. Wood

2006-01-01

400

Chromosome and chromatid exchanges in chinese hamster cells  

Microsoft Academic Search

Chromosomes were studied by autoradiography in a mixed culture of diploid and tetraploid cells, after having induced fusion with Sendai virus between two Chinese hamster cell populations, one labelled with 3H-, the other with 14C-thymidine; sister chromatid exchanges were studied in the 3H diploid cells and exchanges between chromosomes in the 3H-14C tetraploid synkaryons. In both cases, the frequency of

J. Rommelaere; M. Susskind; M. Errera

1973-01-01

401

Prognostic value of various spermatological attributes as predictors of zona binding and zona penetration of buffalo (Bubalus bubalis) semen.  

PubMed

Twenty-four ejaculates from six (four ejaculates each) Surti buffalo bulls aged 4-8 years were used to assess various attributes of spermatozoa influencing the zona-binding and zona-penetration tests. Ejaculates from each bulls were subjected to in vitro sperm--zona binding and sperm--zona penetration tests (four replicates per bull) using immature buffalo oocytes. The average number of spermatozoa bound per oocyte was 27.79 +/- 5.90. The average number of spermatozoa penetrated per oocyte was 3.35 +/- 0.64. The average number of zona-bound and -penetrated spermatozoa differed significantly between animals. Significant difference (p < 0.05) was observed between the plasmalemma integrity as assessed by eosin--nigrosin stain and hypo-osmotic swelling (HOS) test. Furthermore, the percentage of cells positive for the HOS test, i.e. functional membrane integrity (51.25 +/- 2.32) was significantly (p < 0.05) higher than hypo-osmotic swelling-Giemsa (HOS-G) test, i.e. the subpopulation of spermatozoa positive for functional membrane and acrosomal integrities (42.87 +/- 4.56). The HOS test had significant correlations with plasmalemma integrity as measured by the vital stain, eosin--nigrosin (r = 0.85, p < 0.05). The HOS-G test also had significant correlation with plasmalemma integrity measured by vital stains such as eosin--nigrosin (r = 0.90, p < 0.05) and fluorogenic stains [carboxyfluorescein diacetate (CFDA) and propidium iodide (PI); r = 0.92, p < 0.01] and HOS test (r = 0.93), acrosomal integrity (r = 0.86, p < 0.05) and mitochondrial membrane potential (r = 0.99, p < 0.01). The plasmalemma integrity (fluorogenic stain), functional membrane integrity (HOS test), subpopulation of spermatozoa positive for functional membrane and acrosomal integrities (HOS-G test) and mitochondrial membrane potential had significant (p < 0.05) correlation with sperm zona binding and penetration. The present study indicates that these parameters could represent important determinants of sperm quality influencing zona binding and penetration. PMID:18673329

Selvaraju, S; Ghosh, J; Ravindra, J P

2009-02-01

402

Electrical currents through full-grown and maturing Xenopus oocytes.  

PubMed Central

An extracellular vibrating electrode was used to map the current pattern around Xenopus laevis oocytes. Current was found to enter the animal hemisphere and leave the vegetal hemisphere; in fully grown oocytes from which the follicle cells had been removed, the maximal current density was about 1 microamperemeter/cm2. This current decreased to nearly zero in response to progesterone and several other maturation-producing agents. In the case of progesterone, the decline began within a few minutes of the addition of the hormone and proceeded with a half-time of about 20 min. An analysis of the effects on the current of the removal or addition of various ions and drugs led to the inference that the major current-carrying ion was chloride and that the chloride permeability was controlled by calcium. PMID:284407

Robinson, K R

1979-01-01

403

The extracellular calcium-sensing receptor is expressed in the cumulus-oocyte complex in mammals and modulates oocyte meiotic maturation.  

PubMed

The extracellular calcium-sensing receptor (CASR) plays an important role in cells involved in calcium (Ca2+) homeostasis by directly sensing changes in the extracellular Ca2+ ion concentration. We previously reported the localization and quantitative expression of CASR protein in human oocytes. In this study, we examined the expression and the functional role of CASR during oocyte meiotic maturation in a large mammal animal model, the horse. As in humans, CASR protein was found to be expressed in equine oocytes and cumulus cells. Western-blot analysis revealed a single 130 kDa band in denuded oocytes and a doublet of 130-120 kDa in cumulus cells. CASR labeling was observed by confocal microscopy in cumulus cells and in oocytes on the plasma membrane and within the cytoplasm at all examined stages of meiosis. Functionally, the CASR allosteric effector NPS R-467, in the presence of 2.92 mM external Ca2+, increased oocyte maturation rate in a dose-dependent manner and its stimulatory effect was attenuated by pre-treatment with the CASR antagonist NPS 2390. NPS R-467 had no effect in suboptimal external Ca2+ (0.5 mM), indicating that it requires higher external Ca2+ to promote oocyte maturation. In oocytes treated with NPS R-467, CASR staining increased at the plasmalemma and was reduced in the cytosol. Moreover, NPS R-467 increased the activity of MAPK, also called ERK, in cumulus cells and oocytes. These results provide evidence of a novel signal transduction pathway modulating oocyte meiotic maturation in mammals in addition to the well-known systemic hormones. PMID:19494043

De Santis, Teresa; Casavola, Valeria; Reshkin, Stephan Joel; Guerra, Lorenzo; Ambruosi, Barbara; Fiandanese, Nadia; Dalbies-Tran, Rozenn; Goudet, Ghylene; Dell'Aquila, Maria Elena

2009-09-01

404

Female-biased anorexia and anxiety in the Syrian hamster.  

PubMed

Anorexia and anxiety cause significant mortality and disability with female biases and frequent comorbidity after puberty, but the scarcity of suitable animal models impedes understanding of their biological underpinnings. It is reported here that in adult or weanling Syrian hamsters, relative to social housing (SH), social separation (SS) induced anorexia characterized as hypophagia, weight loss, reduced adiposity, and hypermetabolism. Following anorexia, SS increased reluctance to feed, and thigmotaxis, in anxiogenic environments. Importantly, anorexia and anxiety were induced post-puberty with female biases. SS also reduced hypothalamic corticotrophin-releasing factor mRNA and serum corticosteroid levels assessed by RT-PCR and RIA, respectively. Consistent with the view that sex differences in adrenal suppression contributed to female biases in anorexia and anxiety by disinhibiting neuroimmune activity, SS elevated hypothalamic interleukin-6 and toll-like receptor 4 mRNA levels. Although corticosteroids were highest during SH, they were within the physiological range and associated with juvenile-like growth of white adipose, bone, and skeletal muscle. These results suggest that hamsters exhibit plasticity in bioenergetic and emotional phenotypes across puberty without an increase in stress responsiveness. Thus, social separation of hamsters provides a model of sex differences in anorexia and anxiety during adulthood and their pathogeneses during adolescence. PMID:24866911

Shannonhouse, John L; Fong, Li An; Clossen, Bryan L; Hairgrove, Ross E; York, Daniel C; Walker, Benjamin B; Hercules, Gregory W; Mertesdorf, Lauren M; Patel, Margi; Morgan, Caurnel

2014-06-22

405

Histogenesis of pancreatic carcinogenesis in the hamster: ultrastructural evidence  

SciTech Connect

Pancreatic carcinogenesis in the Syrian hamster, induced by ..beta..-oxidized derivatives of N-nitroso-di-n-propylamine, constitutes a valuable model of human cancer of the exocrine pancreas. In both species the majority of tumors are adenocarcinomas: superficially, on the basis of their histological appearance, these appear to be ductal in origin. However, sequential analysis, by electron microscopy, of the development of pancreatic neoplasia in the hamster model indicates that acinar cells may participate in the histogenesis of ductal adenomas and carcinomas. Acinar cells appear to undergo changes in differentiation, including pseudoductular transformation, giving rise to a new population of cells that resemble ductular or centroacinar types. This new population may then proliferate to form, first, cystic foci and subsequently cytadenomas and adenocarcinomas. Mucous metaplasia appears to develop at late stages of tumor development. Although the participation of ductular and centroacinar cells in pancreatic carcinogenesis cannot be excluded, very few tumors arise from the ductal epithelium. It is possible that some human pancreatic adenocarcinomas may also have their origin from dysplastic acinar cells, by analogy with the hamster model: focal acinar dyplasia being common in human pancreatic cancer patients. 90 references, 18 figures.

Flaks, B.

1984-06-01

406

Melanin content of hamster tissues, human tissues, and various melanomas  

SciTech Connect

Melanin content (percentage by weight) was determined in both pigmented and nonpigmented tissues of Syrian golden hamsters bearing Greene melanoma. Melanin content was also measured in various other melanoma models (B-16 in C57 mice, Harding-Passey in BALB/c mice, and KHDD in C3H mice) and in nine human melanomas, as well as in selected normal tissues. The purpose was to evaluate the possible efficacy of chlorpromazine, which is known to bind to melanin, as a vehicle for boron transport in neutron capture therapy. Successful therapy would depend upon selective uptake and absolute concentration of borated compounds in tumors; these parameters will in turn depend upon melanin concentration in melanomas and nonpigmented ''background'' tissues. Hamster whole eyes, hamster melanomas, and other well-pigmented animal melanomas were found to contain 0.3 to 0.8% melanin by weight, whereas human melanomas varied from 0.1 to 0.9% (average, 0.35%). Other tissues, with the exception of skin, were lower in content by a factor of greater than or equal to30. Melanin pigment was extracted from tissues, and the melanin content was determined spectrophotometrically. Measurements were found to be sensitive to the presence of other proteins. Previous procedures for isolating and quantifying melanin often neglected the importance of removing proteins and other interfering nonmelanic substances.

Watts, K.P.; Fairchild, R.G.; Slatkin, D.N.; Greenberg, D.; Packer, S.; Atkins, H.L.; Hannon, S.J.

1981-02-01

407

Learned magnetic compass orientation by the Siberian hamster, Phodopus sungorus  

SciTech Connect

Magnetic orientation has been demonstrated in Siberian hamsters, Phodopus sungorus. The behavior, using a nest building assay, shows a directional preference in nest position and appears in this animal to be a learned behavior. Hamsters were housed prior to testing in rectangular cages aligned along perpendicular axes. When subsequently tested in a radially-symmetrical arena, the hamsters positioned their nests in a bimodal distribution that coincided with the magnetic direction of the long-axis of the holding cages. In addition, results are presented that illustrate some of the factors that can influence behavioral responses to the magnetic field. In particular for P. sungorus, holding conditions prior to testing and the presence of non-magnetic cues may influence the strength and expression of magnetic orientation. Failure to consider these and other factors may help to explain why previous attempts to demonstrate magnetic orientation in a number of rodent species have failed or, when positive results have been obtained, have been difficult to replicate in other laboratories.

Deutschlander, Mark E.; Freake, Michael J.; Borland, Christopher; Phillips, John B.; Madden, R C.; Anderson, Larry E.; Wilson, B W.

2003-04-01

408

Asynchronism in sterlet oocyte development under industrial conditions  

Microsoft Academic Search

Under natural conditions in the course of gametogenesis, the asynchronism in oocyte development in fish ovaries is observed\\u000a on early stages of gonad maturation and mostly in species with batch spawning. When investigating gametogenesis in sterlet\\u000a under conditions of a warm-water farm (Konakovsky sturgeon farm), rather uniform development of gametes is noted; asynchronism\\u000a occurs in separate individuals and also in

O. P. Melekhova; E. A. Chertikhina

2009-01-01

409

Oocyte-somatic cells interactions, lessons from evolution  

PubMed Central

Background Despite the known importance of somatic cells for oocyte developmental competence acquisition, the overall mechanisms underlying the acquisition of full developmental competence are far from being understood, especially in non-mammalian species. The present work aimed at identifying key molecular signals from somatic origin that would be shared by vertebrates. Results Using a parallel transcriptomic analysis in 4 vertebrate species - a teleost fish, an amphibian, and two mammals - at similar key steps of developmental competence acquisition, we identified a large number of species-specific differentially expressed genes and a surprisingly high number of orthologous genes exhibiting similar expression profiles in the 3 tetrapods and in the 4 vertebrates. Among the evolutionary conserved players participating in developmental competence acquisition are genes involved in key processes such as cellular energy metabolism, cell-to-cell communications, and meiosis control. In addition, we report many novel molecular actors from somatic origin that have never been studied in the vertebrate ovary. Interestingly, a significant number of these new players actively participate in Drosophila oogenesis. Conclusions Our study provides a comprehensive overview of evolutionary-conserved mechanisms from somatic origin participating in oocyte developmental competence acquisition in 4 vertebrates. Together our results indicate that despite major differences in ovarian follicular structure, some of the key players from somatic origin involved in oocyte developmental competence acquisition would be shared, not only by vertebrates, but also by metazoans. The conservation of these mechanisms during vertebrate evolution further emphasizes the important contribution of the somatic compartment to oocyte quality and paves the way for future investigations aiming at better understanding what makes a good egg. PMID:23083410

2012-01-01

410

Physiological and molecular basis of fish oocyte hydration  

Microsoft Academic Search

As in other lower vertebrates, teleost oocytes growing within the ovary pass through a series of developmental stages that\\u000a eventually culminate in the production of a mature female gamete or egg. During most of its time they are temporarily arrested\\u000a in meiotic prophase I, and energy expenditures are concentrated on the synthesis and uptake of various substances (e.g. vitellogenin\\u000a (Vg)

Joan Cerdà; Mercedes Fabra; Demetrio Raldúa

411

Chemical structure of sterols that activate oocyte meiosis  

Microsoft Academic Search

GONADOTROPHINS and various growth factors, but not sex steroids, can induce resumption of meiosis in vitro, but only in oocytes enclosed by cumulus-granulosa cells1. Follicular purines prevent resumption of meiosis2,3. This process can be overcome, in vitro, by a transient elevation of cyclic AMP resulting in the production of a diffusible meiosis-inducing substance secreted by the cumulus cells4. A meiosis-inducing

Anne Grete Byskov; Claus Yding Andersen; Lars Nordholm; Henning Thogersen; Xia Guoliang; Ole Wassmann; Jan Vanggaard Andersen; Erling Guddal; Tiny Roed

1995-01-01

412

A hyperpolarization-activated ion current of amphibian oocytes.  

PubMed

A comparative analysis of a hyperpolarization-activated ion current present in amphibian oocytes was performed using the two-electrode voltage-clamp technique in Xenopus laevis, Xenopus tropicalis, and Ambystoma mexicanum. This current appears to be driven mainly by Cl(-) ions, is independent of Ca(2+), and is made evident by applying extremely negative voltage pulses; it shows a slow activating phase and little or no desensitization. The pharmacological profile of the current is complex. The different channel blocker used for Cl(-), K(+), Na(+) and Ca(2+) conductances, exhibited various degrees of inhibition depending of the species. The profiles illustrate the intricacy of the components that give rise to this current. During X. laevis oogenesis, the hyperpolarization-activated current is present at all stages of oocytes tested (II-VI), and the amplitude of the current increases from about 50 nA in stage I to more than 1 ?A in stage VI; nevertheless, there was no apparent modification of the kinetics. Our results suggest that the hyperpolarization-activated current is present both in order Anura and Urodela oocytes. However, the electrophysiological and pharmacological characteristics are quite perplexing and seem to suggest a mixture of ionic conductances that includes the activation of both anionic and cationic channels, most probably transiently opened due to the extreme hyperpolarizion of the plasma membrane. As a possible mechanism for the generation of the current, a kinetic model which fits the data suggests the opening of pores in the plasma membrane whose ion selectivity is dependent on the extracellular Cl(-) concentration. The extreme voltage conditions could induce the opening of otherwise latent pores in plasma membrane proteins (i.e., carriers), resembling the ´slippage´ events already described for some carriers. These observations should be valuable for other groups trying to express cloned, voltage-dependent ion channels in oocytes of amphibian in which hyperpolarizing voltage pulses are applied to activate the channels. PMID:23440457

Ochoa-de la Paz, L D; Salazar-Soto, D B; Reyes, J P; Miledi, R; Martinez-Torres, A

2013-08-01

413

Purification and characterization of RNase from Rana catesbiana (bullfrog) oocytes  

SciTech Connect

In vitro transcription study under conditions where 5S ribosomal (rRNA) synthesis was highly active in oocyte extracts of X. laevis, the transcriptional activity was not detected in oocyte extracts of cold treated R. catesbiana. The lack of 5S rRNA transcription was not due to the absence of RNA polymerase III, since this enzyme was still active when poly d(A-T) was used as a template. It was found that R. catesbiana extracts could cleave exogenously added 5S rRNA, tRNA and VA-RNA while the X. laevis extract could not. The presence of this RNase activity was not a result of oocyte destruction because eggs derived from R. catesbiana oocytes, which contained this RNase activity, developed into tadpoles after artificial fertilization. In order to elucidate the relationship between the RNase activity and the regulation of gene transcription of cold treated R. catesbiana, this enzyme was purified to homogeneity and characterized. This enzyme could be inactivated by heating to 80C for 15 min, but was resistant to acid and alkaline conditions. The optimal temperature for activity was 55C-65C, while the optimal pH was 4 in 50 mM acetate buffer and 8 in 50 mM Tris-buffer. The optimal cation concentration for the enzyme activity was 4 mM and 0.5 mM for Mg{sup ++} and Zn{sup ++} respectively. The specific cleavage site of this enzyme was located at the phosphodiester bond to the 3{prime} side of the pyrimidine in the pyrimidine-p-G segment. The antiserum against the purified RNase was prepared and used for quantitating this enzyme under different condition.

Liao, Y.D.; Lin, C.T. (Academia Sinica, Taipei (Taiwan))

1991-03-11

414

Extra- and intracellular ice formation in mouse oocytes.  

PubMed

The occurrence of intracellular ice formation (IIF) during freezing, or the lack there of, is the single most important factor determining whether or not cells survive cryopreservation. One important determinant of IIF is the temperature at which a supercooled cell nucleates. To avoid intracellular ice formation, the cell must be cooled slowly enough so that osmotic dehydration eliminates nearly all cell supercooling before reaching that temperature. This report is concerned with factors that determine the nucleation temperature in mouse oocytes. Chief among these is the concentration of cryoprotective additive (here, glycerol or ethylene glycol). The temperature for IIF decreases from -14 degrees C in buffered isotonic saline (PBS) to -41 degrees C in 1M glycerol/PBS and 1.5M ethylene glycol/PBS. The latter rapidly permeates the oocyte; the former does not. The initial extracellular freezing at -3.9 to -7.8 degrees C, depending on the CPA concentration, deforms the cell. In PBS that deformation often leads to IIF; in CPA it does not. The oocytes are surrounded by a zona pellucida. That structure appears to impede the growth of external ice through it, but not to block it. In most cases, IIF is characterized by an abrupt blackening or flashing during cooling. But in some cases, especially with dezonated oocytes, a pale brown veil abruptly forms during cooling followed by slower blackening during warming. Above -30 degrees C, flashing occurs in a fraction of a second. Below -30 degrees C, it commonly occurs much more slowly. We have observed instances where flashing is accompanied by the abrupt ejection of cytoplasm. During freezing, cells lie in unfrozen channels between the growing external ice. From phase diagram data, we have computed the fraction of water and solution that remains unfrozen at the observed flash temperatures and the concentrations of salt and CPA in those channels. The results are somewhat ambiguous as to which of these characteristics best correlates with IIF. PMID:15975568

Mazur, Peter; Seki, Shinsuke; Pinn, Irina L; Kleinhans, F W; Edashige, Keisuke

2005-08-01

415

Multiple Requirements of PLK1 during Mouse Oocyte Maturation  

PubMed Central

Polo-like kinase 1 (PLK1) orchestrates multiple events of cell division. Although PLK1 function has been intensively studied in centriole-containing and rapidly cycling somatic cells, much less is known about its function in the meiotic divisions of mammalian oocytes, which arrest for a long period of time in prophase before meiotic resumption and lack centrioles for spindle assembly. Here, using specific small molecule inhibition combined with live mouse oocyte imaging, we comprehensively characterize meiotic PLK1’s functions. We show that PLK1 becomes activated at meiotic resumption on microtubule organizing centers (MTOCs) and later at kinetochores. PLK1 is required for efficient meiotic resumption by promoting nuclear envelope breakdown. PLK1 is also needed to recruit centrosomal proteins to acentriolar MTOCs to promote normal spindle formation, as well as for stable kinetochore-microtubule attachment. Consequently, PLK1 inhibition leads to metaphase I arrest with misaligned chromosomes activating the spindle assembly checkpoint (SAC). Unlike in mitosis, the metaphase I arrest is not bypassed by the inactivation of the SAC. We show that PLK1 is required for the full activation of the anaphase promoting complex/cyclosome (APC/C) by promoting the degradation of the APC/C inhibitor EMI1 and is therefore essential for entry into anaphase I. Moreover, our data suggest that PLK1 is required for proper chromosome segregation and the maintenance of chromosome condensation during the meiosis I-II transition, independently of the APC/C. Thus, our results define the meiotic roles of PLK1 in oocytes and reveal interesting differential requirements of PLK1 between mitosis and oocyte meiosis in mammals. PMID:25658810

Solc, Petr; Kitajima, Tomoya S.; Yoshida, Shuhei; Brzakova, Adela; Kaido, Masako; Baran, Vladimir; Mayer, Alexandra; Samalova, Pavlina; Motlik, Jan; Ellenberg, Jan

2015-01-01

416

Developmental Control of Oocyte Maturation and Egg Activation in Metazoan Models  

PubMed Central

Production of functional eggs requires meiosis to be coordinated with developmental signals. Oocytes arrest in prophase I to permit oocyte differentiation, and in most animals, a second meiotic arrest links completion of meiosis to fertilization. Comparison of oocyte maturation and egg activation between mammals, Caenorhabditis elegans, and Drosophila reveal conserved signaling pathways and regulatory mechanisms as well as unique adaptations for reproductive strategies. Recent studies in mammals and C. elegans show the role of signaling between surrounding somatic cells and the oocyte in maintaining the prophase I arrest and controlling maturation. Proteins that regulate levels of active Cdk1/cyclin B during prophase I arrest have been identified in Drosophila. Protein kinases play crucial roles in the transition from meiosis in the oocyte to mitotic embryonic divisions in C. elegans and Drosophila. Here we will contrast the regulation of key meiotic events in oocytes. PMID:21709181

Von Stetina, Jessica R.; Orr-Weaver, Terry L.

2011-01-01

417

A role for retrotransposon LINE-1 in fetal oocyte attrition in mice.  

PubMed

Fetal oocyte attrition (FOA) is a conserved but poorly understood process of elimination of more than two-thirds of meiotic prophase I (MPI) oocytes before birth. We now implicate retrotransposons LINE-1 (L1), activated during epigenetic reprogramming of the embryonic germline, in FOA in mice. We show that wild-type fetal oocytes possess differential nuclear levels of L1ORF1p, an L1-encoded protein essential for L1 ribonucleoprotein particle (L1RNP) formation and L1 retrotransposition. We demonstrate that experimental elevation of L1 expression correlates with increased MPI defects, FOA, oocyte aneuploidy, and embryonic lethality. Conversely, reverse transcriptase (RT) inhibitor AZT has a profound effect on the FOA dynamics and meiotic recombination, and it implicates an RT-dependent trigger in oocyte elimination in early MPI. We propose that FOA serves to select oocytes with limited L1 activity that are therefore best suited for the next generation. PMID:24882376

Malki, Safia; van der Heijden, Godfried W; O'Donnell, Kathryn A; Martin, Sandra L; Bortvin, Alex

2014-06-01

418

Ovarian Germline Stem Cells: An Unlimited Source of Oocytes?  

PubMed Central

While there has been progress in directing the development of embryonic stem cells and induced pluripotent stem cells toward a germ cell state, their ability to serve as a source of functional oocytes in a clinically relevant model or situation has yet to be established. Recent studies suggest the adult mammalian ovary is not endowed with a finite number of oocytes, but instead possesses stem cells that contribute to their renewal. The ability to isolate and promote the growth and development of such ovarian germline stem cells (GSCs) would provide a novel means to treat infertility in women. While such ovarian GSCs are well-characterized in non-mammalian model organisms, the findings that support the existence of adult ovarian GSCs in mammals have been met with considerable evidence that disputes their existence. Thus, this review details the lessons provided by model organisms that successfully utilize ovarian GSCs to allow for a continual and high level of female germ cell production throughout their life, with a specific focus on the cellular mechanisms involved in GSC self-renewal and oocyte development. Such an overview of the role oogonial stem cells play in maintaining fertility in non-mammalian species serves as a backdrop for the data generated to-date that supports or disputes the existence of GSCs in mammals as well as the future of this area of research in terms of its potential for any application in reproductive medicine. PMID:24382341

Hanna, Carol; Hennebold, Jon

2014-01-01

419

Rearranged mitochondrial genomes are present in human oocytes  

SciTech Connect

Using quantitative PCR, we have determined that a human oocyte contains {approximately}100,000 mitochondrial genomes (mtDNAs). We have also found that some oocytes harbor measurable levels (up to 0.1%) of the so-called common deletion, an mtDNA molecule containing a 4,977-bp rearrangement that is present in high amounts in many patients with {open_quotes}sporadic{close_quotes} Kearns-Sayre syndrome (KSS) and progressive external ophthalmoplegia (PEO). This is the first demonstration that rearranged mtDNAs are present in human oocytes, and it provides experimental support for the supposition that pathogenic deletions associated with the ontogeny of sporadic KSS and PEO can be transmitted in the female germ line, from mother to child. The relevance of these findings to the accumulation of extremely low levels of deleted mtDNAs in both somatic and germ-line tissues during normal human aging is also discussed. 42 refs., 6 figs., 1 tab.

Xi, Chen; Prosser, R.; Simonetti, S. [Columbia Univ., New York, NY (United States)] [and others

1995-08-01

420

The gametic synapse: RNA transfer to the bovine oocyte.  

PubMed

Even after several decades of quiescent storage in the ovary, the female germ cell is capable of reinitiating transcription to build the reserves that are essential to support early embryonic development. In the current model of mammalian oogenesis, there exists bilateral communication between the gamete and the surrounding cells that is limited to paracrine signaling and direct transfer of small molecules via gap junctions existing at the end of the somatic cells' projections that are in contact with the oolemma. The purpose of this work was to explore the role of cumulus cell projections as a means of conductance of large molecules, including RNA, to the mammalian oocyte. By studying nascent RNA with confocal and transmission electron microscopy in combination with transcript detection, we show that the somatic cells surrounding the fully grown bovine oocyte contribute to the maternal reserves by actively transferring large cargo, including mRNA and long noncoding RNA. This occurrence was further demonstrated by the reconstruction of cumulus-oocyte complexes with transfected cumulus cells transferring a synthetic transcript. We propose selective transfer of transcripts occurs, the delivery of which is supported by a remarkable synapselike vesicular trafficking connection between the cumulus cells and the gamete. This unexpected exogenous contribution to the maternal stores offers a new perspective on the determinants of female fertility. PMID:25143353

Macaulay, Angus D; Gilbert, Isabelle; Caballero, Julieta; Barreto, Rodrigo; Fournier, Eric; Tossou, Prudencio; Sirard, Marc-André; Clarke, Hugh J; Khandjian, Édouard W; Richard, Francois J; Hyttel, Poul; Robert, Claude

2014-10-01

421

Pharmaceutical Options for Triggering of Final Oocyte Maturation in ART  

PubMed Central

Since the pioneering days of in vitro fertilization, hCG has been the gold standard to induce final follicular maturation. We herein reviewed different pharmaceutical options for triggering of final oocyte maturation in ART. The new upcoming agent seems to be GnRHa with its potential advantages over hCG trigger. GnRHa triggering elicits a surge of gonadotropins resembling the natural midcycle surge of gonadotropins, without the prolonged action of hCG, resulting in the retrieval of more mature oocytes and a significant reduction in or elimination of OHSS as compared to hCG triggering. The induction of final follicular maturation using GnRHa represents a paradigm shift in the ovulation triggering concept in ART and, thus, a way to develop a safer IVF procedure. Kisspeptins are key central regulators of the neuroendocrine mechanisms of human reproduction, who have been shown to effectively elicit an LH surge and to induce final oocyte maturation in IVF cycles. This new trigger concept may, therefore, offer a completely new, “natural” pharmacological option for ovulation induction. Whether kisspeptins will be the future agent to trigger ovulation remains to be further explored. PMID:25133168

Humaidan, Peter; Bernabéu, Rafael

2014-01-01

422

Dieldrin modifies the hydrolysis of PIP 2 and decreases the fertilization rate in Bufo arenarum oocytes  

Microsoft Academic Search

Carbachol treatment in Bufo arenarum oocytes decreases the radioactivity in [32P]PIP2 in the following 20 min after stimulation and increases the [3H]glycerol labeling of 1,2-DAG at 1 min of stimulation. On the contrary, in Dieldrin treated oocytes carbachol stimulation produces an increase in [32P]PIP2 labeling without changes in [3H]1,2-DAG radioactivity. The sustained hydrolysis of PIP2 observed in Control oocytes is

Teresa M. Fonovich de Schroeder; Ana M. Pechén de D'Angelo

1995-01-01

423

?-endosulfine (ENSA) regulates exit from prophase I arrest in mouse oocytes.  

PubMed

Mammalian oocytes in ovarian follicles are arrested in meiosis at prophase I. This arrest is maintained until ovulation, upon which the oocyte exits from this arrest, progresses through meiosis I and to metaphase of meiosis II. The progression from prophase I to metaphase II, known as meiotic maturation, is mediated by signals that coordinate these transitions in the life of the oocyte. ENSA (?-endosulfine) and ARPP19 (cAMP-regulated phosphoprotein-19) have emerged as regulators of M-phase, with function in inhibition of protein phosphatase 2A (PP2A) activity. Inhibition of PP2A maintains the phosphorylated state of CDK1 substrates, thus allowing progression into and/or maintenance of an M-phase state. We show here ENSA in mouse oocytes plays a key role in the progression from prophase I arrest into M-phase of meiosis I. The majority of ENSA-deficient oocytes fail to exit from prophase I arrest. This function of ENSA in oocytes is dependent on PP2A, and specifically on the regulatory subunit PPP2R2D (also known as B55?). Treatment of ENSA-deficient oocytes with Okadaic acid to inhibit PP2A rescues the defect in meiotic progression, with Okadaic acid-treated, ENSA-deficient oocytes being able to exit from prophase I arrest. Similarly, oocytes deficient in both ENSA and PPP2R2D are able to exit from prophase I arrest to an extent similar to wild-type oocytes. These data are evidence of a role for ENSA in regulating meiotic maturation in mammalian oocytes, and also have potential relevance to human oocyte biology, as mouse and human have genes encoding both Arpp19 and Ensa. PMID:24675883

Matthews, Lauren M; Evans, Janice P

2014-01-01

424

Non-invasive method to assess genotoxicity of nocodazole interfering with spindle formation in mammalian oocytes  

Microsoft Academic Search

Trisomies due to nondisjunction in oogenesis are still a major cause of genetic diseases in humans. In this study, we analysed spindle morphology of in vitro matured nocodazole-exposed mouse oocytes by novel non-invasive Polscope-microscopy, and compared images to those obtained by anti-tubulin immunofluorescence of fixed oocytes. Polscope revealed a reduction in the numbers of oocytes expressing a birefringent spindle, and

Ying Shen; Ilse Betzendahl; Fengyun Sun; Hans-Rudolf Tinneberg; Ursula Eichenlaub-Ritter

2005-01-01

425

In Vitro Ovine Embryo Production: the Study of Seasonal and Oocyte Recovery Method Effects  

PubMed Central

Background: To current knowledge, different oocyte's recovery method and various seasons have profound impact on in vitro embryo production (IVEP). Objectives: The aim of this study was to define an efficient recovery method for oocytes harvesting from slaughterhouse material in different seasons, and their effects on IVEP yield. Materials and Methods: Ovaries from slaughtered ewes in breeding season (BS) and non-breeding season (NBS) were collected from a local abattoir. The oocytes were recovered through aspiration, centrifugation (ORC), puncture and slicing, and categorized into three classes (I, oocytes with more than three layers of cumulus cells; II, less than three layers with damaged cumulus cells; III, denuded oocytes). After cultivation in TCM 199 for 24 hours, matured oocytes were subjected to in vitro fertilization (IVF) and in vitro culture (IVC). The oocyte recovery using ORC in BS and NBS was significantly higher (P < 0.05) compared with other recovery methods. Results: No significant dissimilarities in the proportion of oocytes reaching M-II stage were recorded when using different oocyte recovery methods in different seasons. Aspiration resulted in lower (P < 0.05) proportion of class I (BS, 60.0 ± 2.1; NBS, 51.1 ± 2.1) compared to ORC (BS, 82.0 ± 1.2; NBS, 70.0 ± 1.2), slicing (BS, 80.0 ± 2.1; NBS, 71.0 ± 1.4) and puncture (BS, 80.0 ± 1.5; NBS, 72.0 ± 2.0). Monospermy and blastocyst development rates were significantly higher using ORC than other recovery techniques in both BS and NBS. More oocytes with high quality, greater blastocyst development and oocyte recovery rates were achieved in BS. Conclusions: The results revealed that oocytes harvesting technique and season are effective in the rate of cleavage and blastocysts’ development, and suggest that despite same meiotic resumption rate in all treatments, it would be better to use ORC. PMID:25593733

Dadashpour Davachi, Navid; Zare Shahneh, Ahmad; Kohram, Hamid; Zhandi, Mahdi; Dashti, Saeed; Shamsi, Helia; Moghadam, Razieh

2014-01-01

426

Ultrastructure of previtellogene oocytes in the neotenic cave salamander Proteus anguinus anguinus (Amphibia, Urodela, Proteidae)  

Microsoft Academic Search

Oogenesis in the neotenic, cave dwelling salamander Proteus anguinus anguinus has not been studied yet, and this study provides a detailed description of the early growth of the oocytes. Early previtellogene\\u000a oocytes ranging from 100 to 600?µm in diameter were examined by light and transmission electron microscopy. The oocytes were\\u000a divided into two stages based on size, color, and histology.

Lilijana Bizjak Mali; Boris Bulog

2010-01-01

427

Morphological and cytogenetic analysis of human giant oocytes and giant embryos  

Microsoft Academic Search

BACKGROUND: Giant binuclear oocytes occur with considerable frequency in human ovaries, but their ultimate fate remains unknown. We report the morphology, cytogenetics and developmental potential of human giant oocytes from patients undergoing assisted reproductive technologies. METHODS AND RESULTS: A total of 44 giant oocytes was collected from patients aged 22-44 years old, with an overall frequency of 0.3% (44\\/14 272

Hanna Balakier; Derek Bouman; Agata Sojecki; Clifford Librach; Jeremy A. Squire

428

Penetrative calcretes and their stratigraphic implications  

SciTech Connect

Multiple calcrete horizons previously considered representative of true subaerial exposures may, in fact, be false penetrative calcretes related to only one subaerial exposure event. Individual subaerial exposure periods can produce a true surficial calcrete plus a series of penetrative calcretes. Although penetrative calcrete horizons are not laterally traceable over long distances, they are very similar to surficial calcretes, are commonly found at sequence and/or lithologic boundaries (permeability anomalies), and can be easily misinterpreted in core borings as representing individual exposure events. Recent penetrative calcretes are common on elevated limestone ridges of the Caicos Islands where roots penetrate downward, seeking ground-water moisture. In glacial times, penetrative calcretes may have been present throughout carbonate platforms, as ground-water levels were much lower. Penetrative and surficial calcretes can be differentiated by the lower concentrations of insoluble Al and Fe in penetrative calcretes.

Rossinsky, V. Jr; Wanless, H.R.; Swart, P.K. (Univ. of Miami, FL (United States))

1992-04-01

429

Bovine cumulus-oocyte-complex-quality is reflected in sensitivity for ?-amanitin, oocyte-diameter and developmental capacity  

Microsoft Academic Search

The aim of the present study was to find more parameters to define developmental competence of cumulus-oocyte-complexes (COCs). Bovine COCs were divided into five groups based on their morphology. In order of increasing level of atresia: COC-A had a bright, compact cumulus investment; COC-B1 also had a compact cumulus investment, but was darker than COC-A; the color of COC-B2 was

A. A. C. de Wit; Th. A. M. Kruip

2001-01-01

430

Long-term radioimmunotherapy studies of Cu-64 anti-colon carcinoma monoclonal antibody (MAb)-1A3, intact and F(ab{prime}){sub 2} singly and in combination, in the GW39-hamster model  

SciTech Connect

In previous studies we have shown that Cu-64 has potential for use in radioimmunotherapy (RIT). The present study was undertaken to examine the therapeutic potential of Cu-64-benzyl-TETA-MAb 1A3, intact and F(ab{prime}){sub 2} fragments, injected single or in combination. Using the model of hamsters carrying the GW39 human colon carcinoma in their thighs, we were interested in whether injecting Cu-64-MAb 1A3 intact and F(ab{prime}){sub 2} fragments together would give improved RIT results compared to either agent alone due to the better tumor penetrating properties of F(ab{prime}){sub 2} fragments and the higher uptake and long tumor residence time of intact MAbs. Hamsters were injected with either 1.5 mCi Cu-64-1A3, 1.5 mCi Cu-64-1A3 F(ab{prime}){sub 2} or a combination of 0.75 mCi Cu-64-1A3 intact and 0.75 mCi Cu-64-1A3 F(ab{prime}){sub 2}. These suboptimal doses of Cu-64 were administered in order to detect any enhanced RIT effects with the combination of Cu-64-labeled MAb and fragments. Control groups received saline along. Hamsters were sacrificed when tumors were > 10 g or after surviving for 6 months. Mean lifespans for hamsters treated with Cu-64-1A3 intact, F(ab{prime}){sub 2}, and the combination were 92 {plus_minus} 44 days, 104 {plus_minus} 54 days and 129 {plus_minus} 48 days respectively, compared to 32 {plus_minus} 5 days for the saline controls (p,0.001). 6 months following treatment 43% of the hamsters (3/7) treated with 1.5 mCi Cu-64 1A3 F(ab{prime}){sub 2}, and 50% of hamsters (5/10) treated with 0.75 mCi Cu-64-1A3 and 0.75 mCi Cu-64-1A3 F(ab{prime}){sub 2} in combination were alive and tumor free. Although tumor grown inhibition was also seen in the group receiving 1.5 mCi Cu-64 1A3 intact, only one hamster (1/7) survived tumor free to 6 months. Results show that Cu-64-1A3 F(ab{prime}){sub 2} as well as intact Cu-64-1A3 can increase survival and effect long term tumor inhibition.

Connett, J.M.; Anderson, C.J.; Guo, L.W. [Washington Univ. School of Medicine, St. Louis, MO (United States)] [and others

1996-05-01

431

Mouse Oocyte Control of Granulosa Cell Development and Function: Paracrine Regulation of Cumulus Cell Metabolism  

PubMed Central

Bi-directional communication between oocytes and the companion granulosa cells is essential for the development and functions of both compartments. Oocytes are deficient in their ability to transport certain amino acids and in carrying out glycolysis and cholesterol biosynthesis, and require that cumulus cells provide them with the specific amino acids and the products in these metabolic pathways. Oocytes control metabolic activities in cumulus cells by promoting the expression of genes in cumulus cells encoding specific amino acid transporters and enzymes essential for the oocyte-deficient metabolic processes. Hence, oocytes outsource metabolic functions to cumulus cells to compensate for oocyte metabolic deficiencies. Oocyte control of granulosa cell metabolism may also participate in regulating the rate of follicular development in coordination with endocrine, paracrine and autocrine signals. Oocytes influence granulosa cell development mainly by secretion of paracrine factors although juxtacrine signals probably also participate. Key oocyte-derived paracine factors include growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15) 15, and fibroblast growth factor 8B (FGF8B). PMID:19197803

Su, You-Qiang; Sugiura, Koji; Eppig, John J.

2009-01-01

432

In vitro maturation of oocytes from non-stimulated common wombats.  

PubMed

Assisted reproductive techniques, such as in vitro oocyte maturation in conjunction with in vitro fertilisation, may be used as a tool to manipulate reproduction. Using the common wombat as a model for the critically endangered northern hairy-nosed wombat, the present study examined whether oocyte maturation could be achieved under field conditions. At the time of collection, no oocytes were at the metaphase II (MII) stage (0/42). After 60 h culture using the submarine incubation system, 34% of oocytes (24/70) matured to telophase/MII, as indicated by the presence of a polar body. The proportion of oocytes that reached MII was higher for oocytes collected from follicles >2 mm in diameter compared with follicles <2 mm (40% v. 22%, respectively). The presence of cumulus cells alone did not influence the maturation potential. Oocytes without cumulus cells collected from follicles >2 mm in diameter had the highest maturation rate (58%). Maturation was not affected by the reproductive status of the common wombat or a delay of up to 5 h before oocyte collection. In conclusion, the present study demonstrated that oocytes collected from non-stimulated common wombats can mature to MII in culture. PMID:14588188

Cleary, M; West, M; Shaw, J; Jenkin, G; Trounson, A

2003-01-01

433

Ultrastructure of oocytes of the Urostreptus atrobrunneus (Diplopoda, Spirostreptida, Spirostreptidae): a potential urban centers plague.  

PubMed

The knowledge of the process of egg formation is indispensable for understanding the mechanisms involved in the reproduction of different species. In this context, the objective of this work was to describe the ultrastructure of the oocytes of Urostreptus atrobrunneus (Spirostreptida), a potential plague of urban centers in different locations of São Paulo State. The lack of knowledge about the morphology, physiology, and the reproductive behavior of the species have hindered an effective control of it. The oocytes of U. atrobrunneus presented three development stages: young oocyte or type I; intermediary oocyte or type II; and mature oocyte or type III. During the oocyte development, the cytoplasm become filled with several globules of protein, drops of lipids, and sphaerocrystals, and it was not observed in many organelles in the oocytes with exception of mitochondria, abundant, principally in young oocytes. The vitelline membrane is also deposited in a discontinuous form and the chorion does not present differentiation of layers. The follicular epithelium alters its shape according to the development phase of the oocyte. Part of the vitellus is from exogenous origin and part is endogenous. Before this, only two studies about the ultrastructural analysis of the female germ cells of diplopods were published. PMID:22791626

Fontanetti, Carmem S; Calligaris, Izabela Braggião; Souza, Tatiana Da Silva; Iamonte, Mônika

2012-11-01

434

Live Birth from Slow-Frozen Rabbit Oocytes after In Vivo Fertilisation  

PubMed Central

In vivo fertilisation techniques such as intraoviductal oocyte transfer have been considered as alternatives to bypass the inadequacy of conventional in vitro fertilisation in rabbit. There is only one study in the literature, published in 1989, that reports live offspring from cryopreserved rabbit oocytes. The aim of the present study was to establish the in vivo fertilisation procedure to generate live offspring with frozen oocytes. First, the effect of two recipient models (i) ovariectomised or (ii) oviduct ligated immediately after transfer on the ability of fresh oocytes to fertilise were compared. Second, generation of live offspring from slow-frozen oocytes was carried out using the ligated oviduct recipient model. Throughout the experiment, recipients were artificially inseminated 9 hours prior to oocyte transfer. In the first experiment, two days after unilateral transfer of fresh oocytes, oviducts and uterine horns were flushed to assess embryo recovery rates. The embryo recovery rates were low compared to control in both ovariectomised and ligated oviduct groups. However, ligated oviduct recipient showed significantly (P<0.05) higher embryo recovery rates compared to ovariectomised and control-transferred. In the second experiment, using bilateral oviduct ligation model, all females that received slow-frozen oocytes became pregnant and delivered a total of 4 live young naturally. Thus, in vivo fertilisation is an effective technique to generate live offspring using slow-frozen oocytes in rabbits. PMID:24358281

Jiménez-Trigos, Estrella; Vicente, José S.; Marco-Jiménez, Francisco

2013-01-01

435

SRC-family tyrosine kinases in oogenesis, oocyte maturation and fertilization: an evolutionary perspective.  

PubMed

The oocyte is a highly specialized cell poised to respond to fertilization with a unique set of actions needed to recognize and incorporate a single sperm, complete meiosis, reprogram maternal and paternal genomes and assemble them into a unique zygotic genome, and finally initiate the mitotic cell cycle. Oocytes accomplish this diverse series of events through an array of signal transduction pathway components that include a characteristic collection of protein tyrosine kinases. The src-family protein kinases (SFKs) figure importantly in this signaling array and oocytes characteristically express certain SFKs at high levels to provide for the unique actions that the oocyte must perform. The SFKs typically exhibit a distinct pattern of subcellular localization in oocytes and perform critical functions in different subcellular compartments at different steps during oocyte maturation and fertilization. While many aspects of SFK signaling are conserved among oocytes from different species, significant differences exist in the extent to which src-family-mediated pathways are used by oocytes from species that fertilize externally vs those which are fertilized internally. The observation that several oocyte functions which require SFK signaling appear to represent common points of failure during assisted reproductive techniques in humans, highlights the importance of these signaling pathways for human reproductive health. PMID:25030759

Kinsey, William H

2014-01-01

436

Resveratrol improves the quality of pig oocytes derived from early antral follicles through sirtuin 1 activation.  

PubMed

During oocyte growth, the number of mitochondria drastically increases and mitochondrial function profoundly affects the oocyte competence. Resveratrol is a well-known activator of sirtuin 1 (SIRT1), which has a role in cellular energy homeostasis and mitochondrial biogenesis. The main aim of the present study was to examine the effect of supplementation of culture media with resveratrol on oocyte development and mitochondrial number and functions. Lipid contents and developmental ability of the oocytes grown in vitro were also examined. Oocyte-granulosa cell complexes were collected from early antral follicles of gilt ovaries and were cultured in medium containing 0 or 2 ?M resveratrol for 14 days. Immunostaining revealed that resveratrol enhanced SIRT1 expression in oocytes. Antrum formation during the culture period and survivability of the granulosa cells surrounding the developed oocytes did not differ between the two concentrations of resveratrol. In addition, the ability of oocytes to complete meiotic maturation did not differ between the two concentrations of resveratrol, whereas the ability of oocytes to develop to the blastocyst stage was improved significantly by resveratrol (7.4% vs. 1.6%; P < 0.05). Resveratrol upregulated the ATP content in oocytes grown in vitro, and the addition of 2 ?M of the SIRT1 inhibitor 6-Chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (EX527) diminished this effect although EX527 alone had no effect on ATP content. The mitochondrial DNA copy number in oocytes determined by quantitative real-time polymerase chain reaction increased during in vitro oocyte development, but resveratrol did not affect the kinetics of the mitochondrial DNA copy number. We found that resveratrol also increased the expression level of phospho-5'-adenosine monophosphate-activated protein kinase in oocytes but decreased the lipid content in oocytes grown in vitro. These results suggest that resveratrol increased the ATP content in oocytes via energy homeostasis and improved the developmental ability of oocytes grown in vitro. PMID:25724287

Itami, N; Shirasuna, K; Kuwayama, T; Iwata, H

2015-05-01

437

Association between nondisjunction and maternal age in meiosis-II human oocytes  

SciTech Connect

The relationship between advanced maternal age and increased risk of trisomic offspring is well know clinically but not clearly understood at the level of the oocyte. A total of 383 oocytes that failed fertilization from 107 patients undergoing in vitro fertilization were analyzed by FISH using X-, 18-, and 13/21-chromosome probes simultaneously. The corresponding polar bodies were also analyzed in 188 of these oocytes. The chromosomes in the oocyte and first polar body complement each other and provide an internal control to differentiate between aneuploidy and technical errors. Two mechanisms of nondisjunction were determined. First, nondisjunction of bivalent chromosomes resulting in two univalents going to the same pole and, second, nondisjunction by premature chromatid separation (predivision) of univalent chromsomes producing either a balanced (2 + 2) or unbalanced (3 + 1) distribution of chromatids into the first polar body and M-II oocytes. Balanced predivision of chromatids, previously proposed as a major mechanism of aneuploidy, was found to increase significantly with time in culture (P < .005), which suggests that this phenomenon should be interpreted carefully. Unbalanced predivision and classical nondisjunction were unaffected by oocyte aging. In comparing oocytes from women <35 years of age with oocytes from women {ge}40 years of age, a significant increase (P < .001) in nondisjunction of full dyads was found in the oocytes with analyzable polar bodies and no FISH errors. Premature predivision of chromatids was also found to cause nondisjunction, but it did not increase with maternal age. 44 refs., 3 figs., 2 tabs.

Dailey, T.; Cohen, J.; Munne, S.; Dale, B.

1996-07-01

438

Direct Real-Time Measurement of Intra-Oocyte Nitric Oxide Concentration In Vivo  

PubMed Central

Nitric oxide (NO) is reported to play significant a role in oocyte activation and maturation, implantation, and early embryonic development. Previously we have shown that NO forms an important component of the oocyte microenvironment, and functions effectively to delay oocyte aging. Thus, precise information about intra-oocyte NO concentrations [NO] will result in designing more accurate treatment plans in assisted reproduction. In this work, the direct, real-time and quantitative intra-oocyte [NO] was measured utilizing an L-shaped amperometric integrated NO-selective electrode. This method not only provides an elegant and convenient approach to real-time the measurement of NO in physiological environments, but also mimics the loss of NO caused by rapid NO diffusion combined with its reactivity in the biological milieu. This experiment suggests that the NO levels of oocytes obtained from young animals are significantly higher than those of oocytes obtained from old animals. Additionally the NO levels stay constant during the measurements; however, the intra-oocyte [NO] is reduced significantly (70–75% reduction) in response to L-NAME incubation, suggesting that NO measurements are truly NOS based rather than caused by an unknown interfering substance in our system. We believe this first demonstration of the direct quantitative measurement of [NO] in situ in an intact cellular complex should be useful in tracking real-time and rapid changes at nanomolar levels. Moreover, this finding confirms and extends our previous work showing that supplementation with NO delays the oocyte aging process. PMID:24887331

Goud, Pravin T.; Goud, Anuradha P.; Najafi, Tohid; Gonik, Bernard; Diamond, Michael P.; Saed, Ghassan M.; Zhang, Xueji; Abu-Soud, Husam M.

2014-01-01

439

Src-family Tyrosine Kinases in Oogenesis, Oocyte Maturation, and Fertilization: An Evolutionary Perspective  

PubMed Central

The oocyte is a highly specialized cell poised to respond to fertilization with a unique set of actions needed to recognize and incorporate a single sperm, complete meiosis, reprogram maternal and paternal genomes and assemble them into a unique zygotic genome, and finally initiate the mitotic cell cycle. Oocytes accomplish this diverse series of events through an array of signal transduction pathway components that include a characteristic collection of protein tyrosine kinases. The src-family protein kinases figure importantly in this signaling array and oocytes characteristically express certain SFKs at high levels to provide for the unique actions that the oocyte must perform. The SFKs typically exhibit a distinct pattern of subcellular localization in oocytes and perform critical functions in different subcellular compartments at different steps during oocyte maturation and fertilization. While many aspects of SFK signaling are conserved among oocytes from different species, significant differences exist in the extent to which src-family -mediated pathways are used by oocytes from species that fertilize externally vs those which are fertilized internally. The observation that several oocyte functions which require SFK signaling appear to represent common points of failure during assisted reproductive techniques in humans, highlights the importance of these signaling pathways for human reproductive health. PMID:25030759

Kinsey, William H.

2015-01-01

440