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Sample records for functional sperm tests

  1. Sperm function test

    PubMed Central

    Talwar, Pankaj; Hayatnagarkar, Suryakant

    2015-01-01

    With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give insight on the events in fertilization of the oocyte. The sperms need to get nutrition from the seminal plasma in the form of fructose and citrate (this can be assessed by fructose qualitative and quantitative estimation, citrate estimation). They should be protected from the bad effects of pus cells and reactive oxygen species (ROS) (leukocyte detection test, ROS estimation). Their number should be in sufficient in terms of (count), structure normal to be able to fertilize eggs (semen morphology). Sperms should have intact and functioning membrane to survive harsh environment of vagina and uterine fluids (vitality and hypo-osmotic swelling test), should have good mitochondrial function to be able to provide energy (mitochondrial activity index test). They should also have satisfactory acrosome function to be able to burrow a hole in zona pellucida (acrosome intactness test, zona penetration test). Finally, they should have properly packed DNA in the nucleus to be able to transfer the male genes (nuclear chromatic decondensation test) to the oocyte during fertilization. PMID:26157295

  2. Sperm function test.

    PubMed

    Talwar, Pankaj; Hayatnagarkar, Suryakant

    2015-01-01

    With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give insight on the events in fertilization of the oocyte. The sperms need to get nutrition from the seminal plasma in the form of fructose and citrate (this can be assessed by fructose qualitative and quantitative estimation, citrate estimation). They should be protected from the bad effects of pus cells and reactive oxygen species (ROS) (leukocyte detection test, ROS estimation). Their number should be in sufficient in terms of (count), structure normal to be able to fertilize eggs (semen morphology). Sperms should have intact and functioning membrane to survive harsh environment of vagina and uterine fluids (vitality and hypo-osmotic swelling test), should have good mitochondrial function to be able to provide energy (mitochondrial activity index test). They should also have satisfactory acrosome function to be able to burrow a hole in zona pellucida (acrosome intactness test, zona penetration test). Finally, they should have properly packed DNA in the nucleus to be able to transfer the male genes (nuclear chromatic decondensation test) to the oocyte during fertilization. PMID:26157295

  3. [Eosin Y-water test for sperm function examination].

    PubMed

    Zha, Shu-wei; Lü, Nian-qing; Xu, Hao-qin

    2015-06-01

    Based on the principles of the in vitro staining technique, hypotonic swelling test, and water test, the Eosin Y-water test method was developed to simultaneously detect the integrity of the sperm head and tail and sperm membrane structure and function. As a widely used method in clinical laboratories in China, the Eosin Y-water test is methodologically characterized by three advantages. Firstly, both the sperm head and tail can be detected at the same time, which allows easy and comprehensive assessment of membrane damage in different parts of sperm. Secondly, distilled water is used instead of the usual formula solution to simplify and standardize the test by eliminating any potential effects on the water molecules through the sperm membrane due to different osmotic pressure or different sugar proportions and electrolyte solutions. Thirdly, the test takes less time and thus can be repeated before and after treatment. This article focuses on the fundamental principles and modification of the Eosin Y-water test and its application in sperm function examination and routine semen analysis for male infertility, assessment of the quality of sperm retrieved by testicular fine needle aspiration, semen cryopreservation program development, and evaluation of sperm membrane integrity after microwave radiation. PMID:26242051

  4. Generation of functional eggs and sperm from cryopreserved whole testes

    PubMed Central

    Lee, Seungki; Iwasaki, Yoshiko; Shikina, Shinya; Yoshizaki, Goro

    2013-01-01

    The conservation of endangered fish is of critical importance. Cryobanking could provide an effective backup measure for use in conjunction with the conservation of natural populations; however, methodology for cryopreservation of fish eggs and embryos has not yet been developed. The present study established a methodology capable of deriving functional eggs and sperm from frozen type A spermatogonia (ASGs). Whole testes taken from rainbow trout were slowly frozen in a cryomedium, and the viability of ASGs within these testes did not decrease over a 728-d freezing period. Frozen-thawed ASGs that were intraperitoneally transplanted into sterile triploid hatchlings migrated toward, and were incorporated into, recipient genital ridges. Transplantability of ASGs did not decrease after as much as 939 d of cryopreservation. Nearly half of triploid recipients produced functional eggs or sperm derived from the frozen ASGs and displayed high fecundity. Fertilization of resultant gametes resulted in the successful production of normal, frozen ASG-derived offspring. Feasibility and simplicity of this methodology will call for an immediate application for real conservation of endangered wild salmonids. PMID:23319620

  5. Generation of functional eggs and sperm from cryopreserved whole testes.

    PubMed

    Lee, Seungki; Iwasaki, Yoshiko; Shikina, Shinya; Yoshizaki, Goro

    2013-01-29

    The conservation of endangered fish is of critical importance. Cryobanking could provide an effective backup measure for use in conjunction with the conservation of natural populations; however, methodology for cryopreservation of fish eggs and embryos has not yet been developed. The present study established a methodology capable of deriving functional eggs and sperm from frozen type A spermatogonia (ASGs). Whole testes taken from rainbow trout were slowly frozen in a cryomedium, and the viability of ASGs within these testes did not decrease over a 728-d freezing period. Frozen-thawed ASGs that were intraperitoneally transplanted into sterile triploid hatchlings migrated toward, and were incorporated into, recipient genital ridges. Transplantability of ASGs did not decrease after as much as 939 d of cryopreservation. Nearly half of triploid recipients produced functional eggs or sperm derived from the frozen ASGs and displayed high fecundity. Fertilization of resultant gametes resulted in the successful production of normal, frozen ASG-derived offspring. Feasibility and simplicity of this methodology will call for an immediate application for real conservation of endangered wild salmonids. PMID:23319620

  6. Implications of diversity in sperm size and function for sperm competition and fertility

    E-print Network

    Museo Nacional de Ciencias Naturales

    Implications of diversity in sperm size and function for sperm competition and fertility MONTSERRAT, University of London, London, U.K. ABSTRACT Sperm competition is now recognised as a potent selective force shaping many male reproductive traits. While the influence of sperm competition on sperm number is widely

  7. Functional distribution of synapsin I in human sperm

    PubMed Central

    Coleman, William L.; Kulp, Adam C.; Venditti, Jennifer J.

    2015-01-01

    Proteins known to function during cell–cell communication and exocytosis in neurons and other secretory cells have recently been reported in human sperm. Synapsins are a group of proteins that have been very well characterized in neurons, but little is known about synapsin function in other cell types. Based upon previous findings and the known function of synapsin, we tested the hypothesis that synapsin I was present in human sperm. Washed, capacitated, and acrosome induced sperm preparations were used to evaluate the functional distribution of synapsin I using immunocytochemistry. Protein extracts from mouse brain, mouse testis/epididymis, and human semen were used for protein blotting techniques. Immunolocalization revealed synapsin I was enriched in the sperm equatorial segment. Protein extracts from mouse brain, mouse testis/epididymis, and human semen were positive for synapsin I using several different antibodies, and dot blot results were confirmed by Western blot analyses. Finally, treatment of capacitated and acrosome reaction induced samples with anti-synapsin antibodies significantly reduced sperm motility. Localization of synapsin I in human sperm is a novel finding. The association of synapsin I with the sperm equatorial segment and effects on motility are suggestive of a role associated with capacitation and/or acrosome reaction, processes that render sperm capable of fertilizing an oocyte. PMID:26566474

  8. The hypo-osmotic swelling test for evaluation of sperm membrane integrity.

    PubMed

    Ramu, Sivakumar; Jeyendran, Rajasingam S

    2013-01-01

    A functional membrane is requisite for the fertilizing ability of spermatozoa, as it plays an integral role in sperm capacitation, acrosome reaction, and binding of the spermatozoon to the egg surface. The hypo-osmotic swelling (HOS) test evaluates the functional integrity of the sperm's plasma membrane and also serves as a useful indicator of fertility potential of sperm. The HOS test predicts membrane integrity by determining the ability of the sperm membrane to maintain equilibrium between the sperm cell and its environment. Influx of the fluid due to hypo-osmotic stress causes the sperm tail to coil and balloon or "swell." A higher percentage of swollen sperm indicates the presence of sperm having a functional and intact plasma membrane. Here, we present the detailed protocol for performing the HOS test and explain the results for interpretation. PMID:22992900

  9. Intramale variation in sperm size: functional significance in a polygynous mammal

    PubMed Central

    Pintus, Eliana; Garde, José Julián

    2015-01-01

    Studies concerning the relationships between sperm size and velocity at the intraspecific level are quite limited and often yielded contradictory results across the animal kingdom. Intramale variation in sperm size may represent a meaningful factor to predict sperm velocity, due to its relationship with the level of sperm competition among related taxa. Because sperm phenotype is under post-copulatory sexual selection, we hypothesized that a reduced intramale variation in sperm size is associated with sperm competitiveness in red deer. Our results show that low variation in sperm size is strongly related to high sperm velocity and normal sperm morphology, which in turn are good predictors of male fertility in this species. Furthermore, it is well known that the red deer show high variability in testicular mass but there is limited knowledge concerning the significance of this phenomenon at intraspecific level, even though it may reveal interesting processes of sexual selection. Thereby, as a preliminary result, we found that absolute testes mass is negatively associated with intramale variation in sperm size. Our findings suggest that sperm size variation in red deer is under a strong selective force leading to increase sperm function efficiency, and reveal new insights into sexual selection mechanisms. PMID:26664807

  10. Investigation of the cause of low sperm motility in asthenozoospermic patients by multiple quantitative tests.

    PubMed

    Yeung, C H; Bals-Pratsch, M; Knuth, U A; Nieschlag, E

    1988-08-01

    Multiple tests were done on the ejaculates of 10 asthenozoospermic patients and nine healthy normozoospermic volunteers in an attempt to identify individually the cause of low sperm motility in these patients. Possible defects in the sperm plasma membrane and the motility apparatus of sperm, and in epididymal function affecting the development of motility, were investigated. The presence of seminal sperm antibodies or any motility-inhibiting factors in the seminal plasma that could be removed by washing were also tested. Each test was positive in only one or two patients but axonemal dysfunction was identified in nine patients. Removal of seminal plasma from asthenozoospermic samples did not improve sperm motility to any greater extent than with donor ejaculates, and the motile sperm of these patients exhibited characteristics mostly similar to those of donors under various incubation conditions. Selection procedures are, therefore, required to obtain samples of good quality sperm from such asthenozoospermic ejaculates. PMID:3170019

  11. Impact of kudzu and puerarin on sperm function.

    PubMed

    Gray, Sandra L; Lackey, Brett R; Boone, William R

    2015-06-01

    The goal of this study was to investigate the impact of kudzu (Pueraria mirifica) and the isoflavone puerarin in functional toxicological tests on spermatozoa and to assess the affinity of extracts and pure isoflavones for estrogen receptor (ER)-alpha and -beta (ER?, ER?) in receptor binding assays. Capacitation, acrosome reaction and chromatin decondensation in spermatozoa were analyzed using microscopic analysis. Kudzu, but not puerarin, reduced motility of sperm. Puerarin reduced the percent spontaneous acrosome reaction in spermatozoa. The pathways used by kudzu that affect sperm function are not fully mirrored by puerarin. Puerarin, kudzu and its other phytoestrogenic components displayed preferential affinity for ER?, however the diverse effects of kudzu and puerarin on sperm function implicate the involvement of multiple signaling systems. PMID:25828059

  12. Functional Amyloids in the Mouse Sperm Acrosome

    PubMed Central

    Guyonnet, Benoit; Egge, Nathan

    2014-01-01

    The acrosomal matrix (AM) is an insoluble structure within the sperm acrosome that serves as a scaffold controlling the release of AM-associated proteins during the sperm acrosome reaction. The AM also interacts with the zona pellucida (ZP) that surrounds the oocyte, suggesting a remarkable stability that allows its survival despite being surrounded by proteolytic and hydrolytic enzymes released during the acrosome reaction. To date, the mechanism responsible for the stability of the AM is not known. Our studies demonstrate that amyloids are present within the sperm AM and contribute to the formation of an SDS- and formic-acid-resistant core. The AM core contained several known amyloidogenic proteins, as well as many proteins predicted to form amyloid, including several ZP binding proteins, suggesting a functional role for the amyloid core in sperm-ZP interactions. While stable at pH 3, at pH 7, the sperm AM rapidly destabilized. The pH-dependent dispersion of the AM correlated with a change in amyloid structure leading to a loss of mature forms and a gain of immature forms, suggesting that the reversal of amyloid is integral to AM dispersion. PMID:24797071

  13. Trigonellae Semen Enhances Sperm Motility and the Expression of the Cation Sperm Channel Proteins in Mouse Testes

    PubMed Central

    Kim, Do Rim; Kim, Hyu Young; Kim, Ha Young; Chang, Mun Seog; Park, Seong Kyu

    2015-01-01

    Genetic defects during spermatogenesis can lead to a reduction in sperm motility and cause male infertility. The cation channels of sperm (CatSper) play a role in the regulation of hyperactivated sperm motility in mouse testes. The effect of Trigonellae Semen (TS) on the male reproductive system and CatSper protein in mouse testes during spermatogenesis was examined. C57BL/c mice were divided into the following five groups: normal, cyclophosphamide- (CP-) only treated (control group), and three groups treated with varying concentrations of TS with CP (100, 500, and 1000?mg/kg TS and 100?mg/kg CP). Real-time PCR, western blot analysis, and a testosterone immunoassay were performed to assess CatSper protein levels in the five groups. Additionally, sperm cell counts and motility were examined. Results indicate that sperm motility and sperm counts increased in the TS treated groups in a dose-dependent manner (p < 0.01). CatSper levels were also significantly higher in the TS treated groups compared to that of the control group (p < 0.001). Therefore, TS treatment could enhance sperm function by promoting spermatogenesis and the expression of CatSper proteins in mouse testes. PMID:26539234

  14. Variability in sperm form and function in the context of sperm competition risk in two Tupinambis lizards

    PubMed Central

    Blengini, Cecilia S; Sergio, Naretto; Gabriela, Cardozo; Giojalas, Laura C; Margarita, Chiaraviglio

    2014-01-01

    In polyandrous species, sperm morphometry and sperm velocity are under strong sexual selection. Although several hypotheses have been proposed to explain the role of sperm competition in sperm trait variation, this aspect is still poorly understood. It has been suggested that an increase in sperm competition pressure could reduce sperm size variation or produce a diversity of sperm to maximize male fertilization success. We aim at elucidating the variability of sperm morphometric traits and velocity in two Tupinambis lizards in the context of sperm competition risk. Sperm traits showed substantial variation at all levels examined: between species, among males within species, and within the ejaculate of individual males. Sperm velocity was found to be positively correlated with flagellum: midpiece ratio, with relatively longer flagella associated with faster sperm. Our results document high variability in sperm form and function in lizards. PMID:25505535

  15. Ejaculate Economics: Testing the Effects of Male Sexual History on the Trade-Off between Sperm and Immune Function in Australian Crickets

    PubMed Central

    Dowling, Damian K.; Simmons, Leigh W.

    2012-01-01

    Trade-offs between investment into male sexual traits and immune function provide the foundation for some of the most prominent models of sexual selection. Post-copulatory sexual selection on the male ejaculate is intense, and therefore trade-offs should occur between investment into the ejaculate and the immune system. Examples of such trade-offs exist, including that between sperm quality and immunity in the Australian cricket, Teleogryllus oceanicus. Here, we explore the dynamics of this trade-off, examining the effects that increased levels of sexual interaction have on the viability of a male's sperm across time, and the concomitant effects on immune function. Males were assigned to a treatment, whereby they cohabited with females that were sexually immature, sexually mature but incapable of copulation, or sexually mature and capable of copulation. Sperm viability of each male was then assessed at two time points: six and 13 days into the treatment, and immune function at day 13. Sperm viability decreased across the time points, but only for males exposed to treatment classes involving sexually mature females. This decrease was similar in magnitude across both sexually mature classes, indicating that costs to the expression of high sperm viability are incurred largely through levels of pre-copulatory investment. Males exposed to immature females produced sperm of low viability at both time points. Although we confirmed a weak negative association between sperm viability and lytic activity (a measure of immune response to bacterial infection) at day 13, this relationship was not altered across the mating treatment. Our results highlight that sperm viability is a labile trait, costly to produce, and subject to strategic allocation in these crickets. PMID:22253916

  16. DEVELOPMENT OF A SPERM CELL TOXICITY TEST FOR MARINE WATERS

    EPA Science Inventory

    Preliminary methods for conducting a quick and sensitive sperm cell toxicity test for marine waters have been developed. This paper presents a simple static test in which sea urchin or sand dollar sperm cells are exposed to test or control solutions for short periods of time (typ...

  17. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...2013-04-01 false Seminal fluid (sperm) immunological test system. 866...Systems § 866.5800 Seminal fluid (sperm) immunological test system. (a) Identification. A seminal fluid (sperm) immunological test system...

  18. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...2011-04-01 false Seminal fluid (sperm) immunological test system. 866...Systems § 866.5800 Seminal fluid (sperm) immunological test system. (a) Identification. A seminal fluid (sperm) immunological test system...

  19. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 false Seminal fluid (sperm) immunological test system. 866...Systems § 866.5800 Seminal fluid (sperm) immunological test system. (a) Identification. A seminal fluid (sperm) immunological test system...

  20. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...2014-04-01 false Seminal fluid (sperm) immunological test system. 866...Systems § 866.5800 Seminal fluid (sperm) immunological test system. (a) Identification. A seminal fluid (sperm) immunological test system...

  1. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...2012-04-01 false Seminal fluid (sperm) immunological test system. 866...Systems § 866.5800 Seminal fluid (sperm) immunological test system. (a) Identification. A seminal fluid (sperm) immunological test system...

  2. The regulation of spermatogenesis and sperm function in nematodes

    PubMed Central

    Ellis, Ronald E.; Stanfield, Gillian M.

    2014-01-01

    In the nematode C. elegans, both males and self-fertile hermaphrodites produce sperm. As a result, researchers have been able to use a broad range of genetic and genomic techniques to dissect all aspects of sperm development and function. Their results show that the early stages of spermatogenesis are controlled by transcriptional and translational processes, but later stages are dominated by protein kinases and phosphatases. Once spermatids are produced, they participate in many interactions with other cells — signals from the somatic gonad determine when sperm activate and begin to crawl, signals from the female reproductive tissues guide the sperm, and signals from sperm stimulate oocytes to mature and be ovulated. The sperm also show strong competitive interactions with other sperm and oocytes. Some of the molecules that mediate these processes have conserved functions in animal sperm, others are conserved proteins that have been adapted for new roles in nematode sperm, and some are novel proteins that provide insights into evolutionary change. The advent of new techniques should keep this system on the cutting edge of research in cellular and reproductive biology. PMID:24718317

  3. High Quality RNA in Semen and Sperm: Isolation, Analysis and Potential Application in Clinical Testing

    PubMed Central

    Georgiadis, Andrew P.; Kishore, Archana; Zorrilla, Michelle; Jaffe, Thomas M.; Sanfilippo, Joseph S.; Volk, Etta; Rajkovic, Aleksandar; Yatsenko, Alexander N.

    2015-01-01

    Purpose Male infertility is a complex health condition. To our knowledge there are no molecular biomarkers of male infertility. Sperm RNA is a potential biomarker for detecting sperm abnormalities and viability at infertility clinics. However, RNA use is hindered by its inconsistent quantity, quality, multiple cell types in semen and condensed sperm structure. Materials and Methods We tested the usefulness of high quality RNA isolated from mature sperm and whole semen by our protocol, which reduces RNA degradation by maintaining semen and protocol components at 37C and decreasing processing time. We isolated RNA from 83 whole semen samples, 18 samples of motile sperm prepared by the swim-up protocol and 18 of sperm prepared by the standard Percoll gradient method. Results Electrophoretic and spectral analysis of RNA revealed high quality 18S and 28S rRNAs in 71 of 83 whole semen samples (86%) and 15 of 18 mature sperm swim-up samples (83%). However, high quality RNA was isolated from only 7 of 18 Percoll gradient sperm samples (39%). Interestingly quantitative reverse transcriptase-polymerase chain reaction analysis of 4 somatic and 10 germ cell markers showed that whole semen and swim-up samples had similar RNA profiles. RNA sequencing revealed that most encoded proteins were involved in mature sperm function, regulation of DNA replication, transcription, translation, cell cycle and embryo development. Conclusions We believe that semen and sperm specific RNAs are highly informative biomarkers for germ cell stages and somatic cell contribution. Therefore, these RNAs could be valuable diagnostic indicators of sperm survival, fertilization and early embryogenesis, and could serve as a predictor of the in vitro fertilization prognosis. PMID:25088949

  4. Stability, delivery and functions of human sperm RNAs at fertilization

    PubMed Central

    Sendler, Edward; Johnson, Graham D.; Mao, Shihong; Goodrich, Robert J.; Diamond, Michael P.; Hauser, Russ; Krawetz, Stephen A.

    2013-01-01

    Increasing attention has focused on the significance of RNA in sperm, in light of its contribution to the birth and long-term health of a child, role in sperm function and diagnostic potential. As the composition of sperm RNA is in flux, assigning specific roles to individual RNAs presents a significant challenge. For the first time RNA-seq was used to characterize the population of coding and non-coding transcripts in human sperm. Examining RNA representation as a function of multiple methods of library preparation revealed unique features indicative of very specific and stage-dependent maturation and regulation of sperm RNA, illuminating their various transitional roles. Correlation of sperm transcript abundance with epigenetic marks suggested roles for these elements in the pre- and post-fertilization genome. Several classes of non-coding RNAs including lncRNAs, CARs, pri-miRNAs, novel elements and mRNAs have been identified which, based on factors including relative abundance, integrity in sperm, available knockout data of embryonic effect and presence or absence in the unfertilized human oocyte, are likely to be essential male factors critical to early post-fertilization development. The diverse and unique attributes of sperm transcripts that were revealed provides the first detailed analysis of the biology and anticipated clinical significance of spermatozoal RNAs. PMID:23471003

  5. Presence and Function of Dopamine Transporter (DAT) in Stallion Sperm: Dopamine Modulates Sperm Motility and Acrosomal Integrity

    PubMed Central

    Covarrubias, Alejandra A.; Rodríguez-Gil, Joan Enric; Ramírez-Reveco, Alfredo; Concha, Ilona I.

    2014-01-01

    Dopamine is a catecholamine with multiple physiological functions, playing a key role in nervous system; however its participation in reproductive processes and sperm physiology is controversial. High dopamine concentrations have been reported in different portions of the feminine and masculine reproductive tract, although the role fulfilled by this catecholamine in reproductive physiology is as yet unknown. We have previously shown that dopamine type 2 receptor is functional in boar sperm, suggesting that dopamine acts as a physiological modulator of sperm viability, capacitation and motility. In the present study, using immunodetection methods, we revealed the presence of several proteins important for the dopamine uptake and signalling in mammalian sperm, specifically monoamine transporters as dopamine (DAT), serotonin (SERT) and norepinephrine (NET) transporters in equine sperm. We also demonstrated for the first time in equine sperm a functional dopamine transporter using 4-[4-(Dimethylamino)styryl]-N-methylpyridinium iodide (ASP+), as substrate. In addition, we also showed that dopamine (1 mM) treatment in vitro, does not affect sperm viability but decreases total and progressive sperm motility. This effect is reversed by blocking the dopamine transporter with the selective inhibitor vanoxerine (GBR12909) and non-selective inhibitors of dopamine reuptake such as nomifensine and bupropion. The effect of dopamine in sperm physiology was evaluated and we demonstrated that acrosome integrity and thyrosine phosphorylation in equine sperm is significantly reduced at high concentrations of this catecholamine. In summary, our results revealed the presence of monoamine transporter DAT, NET and SERT in equine sperm, and that the dopamine uptake by DAT can regulate sperm function, specifically acrosomal integrity and sperm motility. PMID:25402186

  6. Presence and function of dopamine transporter (DAT) in stallion sperm: dopamine modulates sperm motility and acrosomal integrity.

    PubMed

    Urra, Javier A; Villaroel-Espíndola, Franz; Covarrubias, Alejandra A; Rodríguez-Gil, Joan Enric; Ramírez-Reveco, Alfredo; Concha, Ilona I

    2014-01-01

    Dopamine is a catecholamine with multiple physiological functions, playing a key role in nervous system; however its participation in reproductive processes and sperm physiology is controversial. High dopamine concentrations have been reported in different portions of the feminine and masculine reproductive tract, although the role fulfilled by this catecholamine in reproductive physiology is as yet unknown. We have previously shown that dopamine type 2 receptor is functional in boar sperm, suggesting that dopamine acts as a physiological modulator of sperm viability, capacitation and motility. In the present study, using immunodetection methods, we revealed the presence of several proteins important for the dopamine uptake and signalling in mammalian sperm, specifically monoamine transporters as dopamine (DAT), serotonin (SERT) and norepinephrine (NET) transporters in equine sperm. We also demonstrated for the first time in equine sperm a functional dopamine transporter using 4-[4-(Dimethylamino)styryl]-N-methylpyridinium iodide (ASP(+)), as substrate. In addition, we also showed that dopamine (1 mM) treatment in vitro, does not affect sperm viability but decreases total and progressive sperm motility. This effect is reversed by blocking the dopamine transporter with the selective inhibitor vanoxerine (GBR12909) and non-selective inhibitors of dopamine reuptake such as nomifensine and bupropion. The effect of dopamine in sperm physiology was evaluated and we demonstrated that acrosome integrity and thyrosine phosphorylation in equine sperm is significantly reduced at high concentrations of this catecholamine. In summary, our results revealed the presence of monoamine transporter DAT, NET and SERT in equine sperm, and that the dopamine uptake by DAT can regulate sperm function, specifically acrosomal integrity and sperm motility. PMID:25402186

  7. New insights into transduction pathways that regulate boar sperm function.

    PubMed

    Hurtado de Llera, A; Martin-Hidalgo, D; Gil, M C; Garcia-Marin, L J; Bragado, M J

    2016-01-01

    Detailed molecular mechanisms mediating signal transduction cascades that regulate boar sperm function involving Ser/Thr and tyrosine phosphorylation of proteins have been reviewed previously. Therefore, this review will focus in those kinase pathways identified recently (<10 years) in boar spermatozoa that regulate different functional spermatozoa processes. AMP-activated protein kinase (AMPK) is a cell energy sensor kinase that was first identified in mammalian spermatozoa in 2012, and since then it has emerged as an essential regulator of boar sperm function. Signaling pathways leading to AMPK activation in boar sperm are highlighted in this review (PKA, CaMKK?/?, and PKC as well as Ca(2+) and cAMP messengers as upstream regulators). Interestingly, stimuli considered as cell stress (hyperosmotic stress, inhibition of mitochondrial activity, absence of intracellular Ca(2+)) markedly activate AMPK in boar spermatozoa. Moreover, AMPK plays a remarkable and necessary regulatory role in mammalian sperm function, controlling essential boar sperm functional processes such as motility, viability, mitochondrial membrane potential, organization and fluidity of plasma membrane, and outer acrosome membrane integrity. These mentioned processes are all required under fluctuating environment of spermatozoa when transiting through the female reproductive tract to achieve fertilization. An applied role of AMPK in artificial insemination techniques is also suggested as during boar seminal doses preservation at 17 °C, physiological levels of AMPK activity markedly increase (maximum on Day 7) and result essential to maintain the aforementioned fundamental sperm processes. Moreover, regulation of sperm function exerted by the glycogen synthase kinase 3 and Src family kinase pathways is summarized. PMID:26074068

  8. Ubiquitination Regulates the Morphogenesis and Function of Sperm Organelles

    PubMed Central

    Nakamura, Nobuhiro

    2013-01-01

    It is now understood that protein ubiquitination has diverse cellular functions in eukaryotes. The molecular mechanism and physiological significance of ubiquitin-mediated processes have been extensively studied in yeast, Drosophila and mammalian somatic cells. Moreover, an increasing number of studies have emphasized the importance of ubiquitination in spermatogenesis and fertilization. The dysfunction of various ubiquitin systems results in impaired sperm development with abnormal organelle morphology and function, which in turn is highly associated with male infertility. This review will focus on the emerging roles of ubiquitination in biogenesis, function and stability of sperm organelles in mammals. PMID:24709878

  9. Effect of varicocelectomy on sperm functional characteristics and DNA methylation.

    PubMed

    Tavalaee, M; Bahreinian, M; Barekat, F; Abbasi, H; Nasr-Esfahani, M H

    2015-10-01

    In individuals with varicocele, DNA is damaged due to high level of oxidative stress, and varicocelectomy can overcome this effect. Damaged DNA is less liable to DNA methylation, and antioxidant therapy appears to have the potential to reduce sperm oxidative stress and DNA damage and thereby maintain DNA methylation, while effect of varicocelectomy on DNA methylation patterns has remained unclear. In the light of these considerations, we aimed to examine the effect of varicocelectomy on sperm DNA methylation and functional characteristics. Fifty-two men with left-sided varicocele (grade II &III) were included. Sperm parameters, DNA fragmentation, protamine deficiency, oxidative stress and global DNA methylation were evaluated before and 3 months after surgery. Our data show that sperm concentration, percentages of spermatozoon with abnormal morphology, DNA fragmentation, protamine deficiency and oxidative stress significantly improved after surgery. Percentage of sperm motility, global DNA methylation and intensity of DNA methylation also improved after surgery, although the differences were not significant when compared with before surgery. Categorisation of individuals to subgroups revealed that improvement of DNA methylation appears to take place in oligozoospermic individuals, which are more severely affected by state of varicocele. However, this is a preliminary study, and further studies are required to solidify this conclusion. PMID:25234073

  10. Effects of the Czech Propolis on Sperm Mitochondrial Function

    PubMed Central

    Cedikova, Miroslava; Miklikova, Michaela; Stachova, Lenka; Grundmanova, Martina; Tuma, Zdenek; Vetvicka, Vaclav; Zech, Nicolas; Kralickova, Milena; Kuncova, Jitka

    2014-01-01

    Propolis is a natural product that honeybees collect from various plants. It is known for its beneficial pharmacological effects. The aim of our study was to evaluate the impact of propolis on human sperm motility, mitochondrial respiratory activity, and membrane potential. Semen samples from 10 normozoospermic donors were processed according to the World Health Organization criteria. Propolis effects on the sperm motility and mitochondrial activity parameters were tested in the fresh ejaculate and purified spermatozoa. Propolis preserved progressive motility of spermatozoa in the native semen samples. Oxygen consumption determined in purified permeabilized spermatozoa by high-resolution respirometry in the presence of adenosine diphosphate and substrates of complex I and complex II (state OXPHOSI+II) was significantly increased in the propolis-treated samples. Propolis also increased uncoupled respiration in the presence of rotenone (state ETSII) and complex IV activity, but it did not influence state LEAK induced by oligomycin. Mitochondrial membrane potential was not affected by propolis. This study demonstrates that propolis maintains sperm motility in the native ejaculates and increases activities of mitochondrial respiratory complexes II and IV without affecting mitochondrial membrane potential. The data suggest that propolis improves the total mitochondrial respiratory efficiency in the human spermatozoa in vitro thereby having potential to improve sperm motility. PMID:25104965

  11. Effects of the czech propolis on sperm mitochondrial function.

    PubMed

    Cedikova, Miroslava; Miklikova, Michaela; Stachova, Lenka; Grundmanova, Martina; Tuma, Zdenek; Vetvicka, Vaclav; Zech, Nicolas; Kralickova, Milena; Kuncova, Jitka

    2014-01-01

    Propolis is a natural product that honeybees collect from various plants. It is known for its beneficial pharmacological effects. The aim of our study was to evaluate the impact of propolis on human sperm motility, mitochondrial respiratory activity, and membrane potential. Semen samples from 10 normozoospermic donors were processed according to the World Health Organization criteria. Propolis effects on the sperm motility and mitochondrial activity parameters were tested in the fresh ejaculate and purified spermatozoa. Propolis preserved progressive motility of spermatozoa in the native semen samples. Oxygen consumption determined in purified permeabilized spermatozoa by high-resolution respirometry in the presence of adenosine diphosphate and substrates of complex I and complex II (state OXPHOSI+II) was significantly increased in the propolis-treated samples. Propolis also increased uncoupled respiration in the presence of rotenone (state ETSII) and complex IV activity, but it did not influence state LEAK induced by oligomycin. Mitochondrial membrane potential was not affected by propolis. This study demonstrates that propolis maintains sperm motility in the native ejaculates and increases activities of mitochondrial respiratory complexes II and IV without affecting mitochondrial membrane potential. The data suggest that propolis improves the total mitochondrial respiratory efficiency in the human spermatozoa in vitro thereby having potential to improve sperm motility. PMID:25104965

  12. PATHS OF MOTION AND VELOCITY DISTRIBUTION FUNCTION OF LIVING SPERM CELL STUDIED BY OPTICAL METHODS

    E-print Network

    PATHS OF MOTION AND VELOCITY DISTRIBUTION FUNCTION OF LIVING SPERM CELL STUDIED BY OPTICAL METHODS Academy of Sciencies, Al. Lotników 32/46, 02-668,Warszawa ABSTRACT The method of sperm motility analysis of the movie images illustrating the dynamics of the sperm cells movement. An appropriate analysis of a grey

  13. Bisphenol A reduces fertilizing ability and motility by compromising mitochondrial function of sperm.

    PubMed

    Singh, Ram P; Shafeeque, Chathathayil M; Sharma, Sanjeev K; Pandey, Nitin K; Singh, Renu; Mohan, Jag; Kolluri, Gautham; Saxena, Meeta; Sharma, Bhaskar; Sastry, Kochiganti V H; Kataria, Jag M; Azeez, Parappurath A

    2015-07-01

    Bisphenol A (BPA) acts as an endocrine disruptor, affects animal reproductive success in vivo and affects sperm functions in vitro at environmentally relevant concentrations, leading to reduction in sperm motility and fertilizing ability in fish. The effect of in vitro BPA on avian sperm functions has not been explored. The present study examined the effect of environmentally relevant concentrations of BPA (0?mM, 0.18?mM, 0.37?mM, and 0.74?mM) on sperm functions in chicken in vitro. Sperm were exposed to concentrations of BPA for 30 min and analyzed for motility, fertilizing ability, live sperm percentage, and mitochondrial membrane potential (??m). Results showed that BPA at a concentration of 0.74 mM significantly decreased motility, fertilizing ability, live sperm count percentage, and sperm ??m. Sperm motility was positively correlated with fertility (r =?0.73, p?? 0.01), live sperm percentage (r?=?0.64, p???0.01), and high ??m (r?=?0.44, p???0.01). A dose-dependent and time-dependent effect of BPA was observed on sperm motility at all BPA concentrations. However, sperm's fertilizing ability was unaffected in low BPA concentration (0.18?mM and 0.37?mM). A significantly higher percentage of moribund sperm was observed at 0.37?mM and 0.74?mM BPA compared with at 0.18?mM BPA, in the negative control, and in the vehicle control. The present study confirms that environmentally relevant concentrations of BPA are capable of compromising sperm functions, leading to reduction in fertilizing ability of chicken sperm. PMID:25728985

  14. Systematic mapping and functional analysis of a family of human epididymal secretory sperm-located proteins.

    PubMed

    Li, JianYuan; Liu, FuJun; Wang, HaiYan; Liu, Xin; Liu, Juan; Li, Ning; Wan, FengChun; Wang, WenTing; Zhang, ChengLin; Jin, ShaoHua; Liu, Jie; Zhu, Peng; Liu, YunXiang

    2010-11-01

    The mammalian spermatozoon has many cellular compartments, such as head and tail, permitting it to interact with the female reproductive tract and fertilize the egg. It acquires this fertilizing potential during transit through the epididymis, which secretes proteins that coat different sperm domains. Optimal levels of these proteins provide the spermatozoon with its ability to move to, bind to, fuse with, and penetrate the egg; otherwise male infertility results. As few human epididymal proteins have been characterized, this work was performed to generate a database of human epididymal sperm-located proteins involved in maturation. Two-dimensional gel electrophoresis of epididymal tissue and luminal fluid proteins, followed by identification using MALDI-TOF/MS or MALDI-TOF/TOF, revealed over a thousand spots in gels comprising 745 abundant nonstructural proteins, 408 in luminal fluids, of which 207 were present on spermatozoa. Antibodies raised to 619 recombinant or synthetic peptides, used in Western blots, histological sections, and washed sperm preparations to confirm antibody quality and protein expression, indicated their regional location in the epididymal epithelium and highly specific locations on washed functional spermatozoa. Sperm function tests suggested the role of some proteins in motility and protection against oxidative attack. A large database of these proteins, characterized by size, pI, chromosomal location, and function, was given a unified terminology reflecting their sperm domain location. These novel, secreted human epididymal proteins are potential targets for a posttesticular contraceptive acting to provide rapid, reversible, functional sterility in men and they are also biomarkers that could be used in noninvasive assessments of male fertility. PMID:20736409

  15. Relative testis size and sperm morphometry across mammals: no evidence for an association between sperm competition and sperm length.

    PubMed Central

    Gage, Matthew J G; Freckleton, Robert P

    2003-01-01

    Understanding why there is extensive variation in sperm form and function across taxa has been a challenge because sperm are specialized cells operating at a microscopic level in a complex environment. This comparative study collates published data to determine whether the evolution of sperm morphometry (sperm total length and separate component dimensions) is associated with sperm competition (when different males' sperm mix and compete for a female's ova) across 83 mammalian species. We use relative testes mass as an indicator of the intensity of sperm competition across taxa: relative investment into testes is widely accepted to predict the level of sperm competition that a species or population endures. Although we found evidence for positive associations between relative testes mass (controlling for allometry) and sperm morphometry across 83 mammalian species, these relationships were phylogenetically dependent. When we appropriately controlled for phylogenetic association using multiple regression within a phylogenetic framework, there was no relationship between relative testes mass and sperm length across mammals. Furthermore, we found no evidence for associations between relative testes mass and sperm head, mid-piece or flagellar lengths, nor was there a relationship with mid-piece or mitochondrial volumes. Results, therefore, indicate that sperm competition does not select for longer or shorter sperm across mammals, and alternative forces selecting on sperm form and function are discussed. PMID:12769463

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

    PubMed Central

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

    2015-01-01

    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

  17. Morphology-function relationships and repeatability in the sperm of Passer sparrows.

    PubMed

    Cramer, Emily R A; Laskemoen, Terje; Stensrud, Even; Rowe, Melissah; Haas, Fredrik; Lifjeld, Jan T; Saetre, Glenn-Peter; Johnsen, Arild

    2015-04-01

    Sperm performance is likely to be an important determinant of male reproductive success, especially when females copulate with multiple males. Understanding sperm performance is therefore crucial to fully understand the evolution of male reproductive strategies. In this study, we examined the repeatability of sperm morphology and motility measures over three breeding seasons, and we studied relationships between sperm morphology and function. We conducted this study in wild-derived captive house sparrows (Passer domesticus) and Spanish sparrows (P. hispaniolensis). Results for the two species were similar. As predicted from results in other passerine species, total sperm length was highly repeatable across ejaculates, and repeatability for the length of other components was moderate. The repeatability of sperm swimming speed across ejaculates was lower, but statistically significant, suggesting that sperm velocity may be a relatively dynamic trait. Surprisingly, swimming speed did not correlate with the relative length of the midpiece, and it correlated negatively with the relative length of the flagellum and with total sperm length. This pattern is the opposite of what theory predicts and differs from what has been found in house sparrows before. Also contrary to previous work, we found no evidence that total sperm length correlates with sperm longevity. These results therefore highlight the need for a better understanding of relationships between sperm morphology and function in passerine birds. PMID:25427840

  18. Impact of glycosylation on the unimpaired functions of the sperm.

    PubMed

    Cheon, Yong-Pil; Kim, Chung-Hoon

    2015-09-01

    One of the key factors of early development is the specification of competence between the oocyte and the sperm, which occurs during gametogenesis. However, the starting point, growth, and maturation for acquiring competence during spermatogenesis and oogenesis in mammals are very different. Spermatogenesis includes spermiogenesis, but such a metamorphosis is not observed during oogenesis. Glycosylation, a ubiquitous modification, is a preliminary requisite for distribution of the structural and functional components of spermatids for metamorphosis. In addition, glycosylation using epididymal or female genital secretory glycans is an important process for the sperm maturation, the acquisition of the potential for fertilization, and the acceleration of early embryo development. However, nonemzymatic unexpected covalent bonding of a carbohydrate and malglycosylation can result in falling fertility rates as shown in the diabetic male. So far, glycosylation during spermatogenesis and the dynamics of the plasma membrane in the process of capacitation and fertilization have been evaluated, and a powerful role of glycosylation in spermatogenesis and early development is also suggested by structural bioinformatics, functional genomics, and functional proteomics. Further understanding of glycosylation is needed to provide a better understanding of fertilization and embryo development and for the development of new diagnostic and therapeutic tools for infertility. PMID:26473106

  19. Impact of glycosylation on the unimpaired functions of the sperm

    PubMed Central

    2015-01-01

    One of the key factors of early development is the specification of competence between the oocyte and the sperm, which occurs during gametogenesis. However, the starting point, growth, and maturation for acquiring competence during spermatogenesis and oogenesis in mammals are very different. Spermatogenesis includes spermiogenesis, but such a metamorphosis is not observed during oogenesis. Glycosylation, a ubiquitous modification, is a preliminary requisite for distribution of the structural and functional components of spermatids for metamorphosis. In addition, glycosylation using epididymal or female genital secretory glycans is an important process for the sperm maturation, the acquisition of the potential for fertilization, and the acceleration of early embryo development. However, nonemzymatic unexpected covalent bonding of a carbohydrate and malglycosylation can result in falling fertility rates as shown in the diabetic male. So far, glycosylation during spermatogenesis and the dynamics of the plasma membrane in the process of capacitation and fertilization have been evaluated, and a powerful role of glycosylation in spermatogenesis and early development is also suggested by structural bioinformatics, functional genomics, and functional proteomics. Further understanding of glycosylation is needed to provide a better understanding of fertilization and embryo development and for the development of new diagnostic and therapeutic tools for infertility. PMID:26473106

  20. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Seminal fluid (sperm) immunological test system. 866.5800 Section 866.5800 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....5800 Seminal fluid (sperm) immunological test system. (a) Identification. A seminal fluid...

  1. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Seminal fluid (sperm) immunological test system. 866.5800 Section 866.5800 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....5800 Seminal fluid (sperm) immunological test system. (a) Identification. A seminal fluid...

  2. Carbonic anhydrases and their functional differences in human and mouse sperm physiology.

    PubMed

    José, O; Torres-Rodríguez, P; Forero-Quintero, L S; Chávez, J C; De la Vega-Beltrán, J L; Carta, F; Supuran, C T; Deitmer, J W; Treviño, C L

    2015-12-25

    Fertilization is a key reproductive event in which sperm and egg fuse to generate a new individual. Proper regulation of certain parameters (such as intracellular pH) is crucial for this process. Carbonic anhydrases (CAs) are among the molecular entities that control intracellular pH dynamics in most cells. Unfortunately, little is known about the function of CAs in mammalian sperm physiology. For this reason, we re-explored the expression of CAI, II, IV and XIII in human and mouse sperm. We also measured the level of CA activity, determined by mass spectrometry, and found that it is similar in non-capacitated and capacitated mouse sperm. Importantly, we found that CAII activity accounts for half of the total CA activity in capacitated mouse sperm. Using the general CA inhibitor ethoxyzolamide, we studied how CAs participate in fundamental sperm physiological processes such as motility and acrosome reaction in both species. We found that capacitated human sperm depend strongly on CA activity to support normal motility, while capacitated mouse sperm do not. Finally, we found that CA inhibition increases the acrosome reaction in capacitated human sperm, but not in capacitated mouse sperm. PMID:26551457

  3. Second messengers, steroids and signaling cascades: Crosstalk in sperm development and function.

    PubMed

    Lackey, B R; Gray, S L

    2015-12-01

    Signaling cascades control numerous aspects of sperm physiology, ranging from creation to fertilization. Novel aspects of several kinases and their influence on sperm development will be discussed in the first section and cover proliferation, chromatin remodeling and morphology. In the second section, protein kinases (A, B and C) that affect sperm function and their regulation by second messengers, cyclic-AMP and phosphoinositides, as well as steroids will be featured. Key areas of integration will be presented on the topics of sperm motility, capacitation, acrosome reaction and fertilization. PMID:26188217

  4. Fibroblast Growth Factor Receptors (FGFRs) in Human Sperm: Expression, Functionality and Involvement in Motility Regulation

    PubMed Central

    Saucedo, Lucía; Buffa, Gabriela N.; Rosso, Marina; Guillardoy, Tomás; Góngora, Adrian; Munuce, María J.

    2015-01-01

    Fibroblast growth factors receptors (FGFRs) have been widely characterized in somatic cells, but there is scarce evidence of their expression and function in mammalian gametes. The objective of the present study was to evaluate the expression of FGFRs in human male germ cells, to determine sperm FGFR activation by the FGF2 ligand and their participation in the regulation of sperm motility. The expression of FGFR1, 2, 3 and 4 mRNAs and proteins in human testis and localization of these receptors in germ cells of the seminiferous epithelium was demonstrated. In ejaculated sperm, FGFRs were localized to the acrosomal region and flagellum. Sperm exposure to FGF2 caused an increase in flagellar FGFR phosphorylation and activation of extracellular signal-regulated kinase (ERK) and protein kinase B (PKB or Akt) signaling pathways. Incubation with FGF2 led to a significant increase in the percentage of total and progressive sperm motility, as well as in sperm kinematics. All responses were prevented by sperm preincubation with BGJ398, a specific inhibitor of FGFR tyrosine kinase activity. In addition to confirming the expression of FGFRs in germ cells of the human testis, our study describes for the first time the presence, localization and functionality of human sperm FGFRs, and provides evidence of the beneficial effect of FGF2 upon sperm motility. PMID:25970615

  5. Functional deficit of sperm and fertility impairment in men with antisperm antibodies.

    PubMed

    Bozhedomov, V A; Nikolaeva, M A; Ushakova, I V; Lipatova, N A; Bozhedomova, G E; Sukhikh, G T

    2015-11-01

    Autoimmune reactions against the sperm cells play an ambiguous role in fertility impairment. The objective of this study was to characterize functional deficit of sperm conditioned by antisperm immune response in normozoospermic men. This was a multi-centric, cross-sectional, ?ase-control study. The study subjects were 1060 infertile normozoospermic men and 107 fertile men. The main outcome measures were clinical examination, semen analysis including MAR test for antisperm antibodies (ASA), computer-aided sperm analysis, acrosome reaction (AR) detected with flow cytometry, DNA fragmentation measured with sperm chromatin dispersion, reactive oxygen species (ROS) assessed using the luminol-dependent chemiluminescence method. 2% of the fertile men had MAR-IgG?50%, but all subjects with MAR-IgG>12% were outliers; 16% infertile men had MAR-IgG?50% (p<0.0001). There was a direct correlation between the infertility duration and MAR-IgG (R=0.3; ?<0.0001). The ASA-positive infertile men had AR disorders 2.1 times more frequently (?<0.02), predominantly inductivity disorders. We found signs of hyperactivation proportionate to the ASA level (p<0.001). DNA fragmentation was more highly expressed and was 1.6 and 1.3 times more frequent compared with the fertile and the ASA-negative patients, respectively (p<0.001 and p<0.05). We found signs of oxidative stress (OS): ROS generation by washed ASA-positive spermatozoa was 3.7 times higher than in the fertile men (p<0.00001) and depended on the ASA levels (R=0.5; p<0.0001). The ASA correlation with ROS generation in native sperm was weak (R=0.2; ?<0.001). We concluded that autoimmune reactions against spermatozoa are accompanied by a fertility decrease in normozoospermia. This results from AR and capacitation disorders and DNA fragmentation. The pathogenesis of sperm abnormalities in immune infertility is associated with the OS of spermatozoa. PMID:26409252

  6. Choline Dehydrogenase Polymorphism rs12676 Is a Functional Variation and Is Associated with Changes in Human Sperm Cell Function

    PubMed Central

    Johnson, Amy R.; Lao, Sai; Wang, Tongwen; Galanko, Joseph A.; Zeisel, Steven H.

    2012-01-01

    Approximately 15% of couples are affected by infertility and up to half of these cases arise from male factor infertility. Unidentified genetic aberrations such as chromosomal deletions, translocations and single nucleotide polymorphisms (SNPs) may be the underlying cause of many cases of idiopathic male infertility. Deletion of the choline dehydrogenase (Chdh) gene in mice results in decreased male fertility due to diminished sperm motility; sperm from Chdh?/? males have decreased ATP concentrations likely stemming from abnormal sperm mitochondrial morphology and function in these cells. Several SNPs have been identified in the human CHDH gene that may result in altered CHDH enzymatic activity. rs12676 (G233T), a non-synonymous SNP located in the CHDH coding region, is associated with increased susceptibility to dietary choline deficiency and risk of breast cancer. We now report evidence that this SNP is also associated with altered sperm motility patterns and dysmorphic mitochondrial structure in sperm. Sperm produced by men who are GT or TT for rs12676 have 40% and 73% lower ATP concentrations, respectively, in their sperm. rs12676 is associated with decreased CHDH protein in sperm and hepatocytes. A second SNP located in the coding region of IL17BR, rs1025689, is linked to altered sperm motility characteristics and changes in choline metabolite concentrations in sperm. PMID:22558321

  7. METHOD OF SPERM COLLECTION SIGNIFICANTLY INFLUENCES SPERM MOTION PARAMETERS FOLLOWING ETHANE DIMETHANESULPHONATE ADMINISTRATION IN THE RAT

    EPA Science Inventory

    Sperm motion analysis following exposure to a reproductive toxicant is one means of evaluating the functional integrity of the testes and epididymis. n this study we sought to determine whether the method used to collect sperm from the proximal cauda epididymidis, where sperm are...

  8. MEASUREMENT OF EPIDIDYMAL SPERM MOTILITY AS A TEST VARIABLE IN THE RAT

    EPA Science Inventory

    Several environmental contaminants, notably dibromochloropropane (Whorton et al. 1977) and kepone (Taylor et al. 1978; Cannon et al, 1978) have been implicated in sperm deficiencies among occupationally exposed males. These incidents emphasize the need for adequate testing of che...

  9. Heat Shock Protein 90 Has Roles in Intracellular Calcium Homeostasis, Protein Tyrosine Phosphorylation Regulation, and Progesterone-Responsive Sperm Function in Human Sperm

    PubMed Central

    Chen, Aijun; Jiang, Youfang; Xie, Haifeng; Shi, Qixian; Zhang, Songying; Ni, Ya

    2014-01-01

    Heat shock protein 90 plays critical roles in client protein maturation, signal transduction, protein folding and degradation, and morphological evolution; however, its function in human sperm is not fully understood. Therefore, our objective in this study was to elucidate the mechanism by which heat shock protein 90 exerts its effects on human sperm function. By performing indirect immunofluorescence staining, we found that heat shock protein 90 was localized primarily in the neck, midpiece, and tail regions of human sperm, and that its expression increased with increasing incubation time under capacitation conditions. Geldanamycin, a specific inhibitor of heat shock protein 90, was shown to inhibit this increase in heat shock protein 90 expression in western blotting analyses. Using a multifunctional microplate reader to examine Fluo-3 AM-loaded sperm, we observed for the first time that inhibition of heat shock protein 90 by using geldanamycin significantly decreased intracellular calcium concentrations during capacitation. Moreover, western blot analysis showed that geldanamycin enhanced tyrosine phosphorylation of several proteins, including heat shock protein 90, in a dose-dependent manner. The effects of geldanamycin on human sperm function in the absence or presence of progesterone was evaluated by performing chlortetracycline staining and by using a computer-assisted sperm analyzer. We found that geldanamycin alone did not affect sperm capacitation, hyperactivation, and motility, but did so in the presence of progesterone. Taken together, these data suggest that heat shock protein 90, which increases in expression in human sperm during capacitation, has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-stimulated sperm function. In this study, we provide new insights into the roles of heat shock protein 90 in sperm function. PMID:25541943

  10. Evaluation of sperm tests as indicators of germ-cell damage in men exposed to chemical or physical agents

    SciTech Connect

    Wyrobek, A.J.; Watchmaker, G.; Gordon, L.

    1983-06-15

    As reviewed here, at least 89 chemical exposures have been studied for their effects on human spermatogenesis using sperm tests, with the majority showing some effect on sperm count, motility, or morphology. Approximately 85% of these exposures were to experimental or therapeutic drugs, 10% to occupational or environmental agents, and 5% to recreational drugs. This paper briefly describes the more common sperm-based methods and reviews some of their applications. It also includes guidelines for undertaking a human sperm study, as well as a discussion of the predictive value of induced sperm changes, an evaluation of the role of animal sperm tests, and a summary of the advantages and disadvantages of the sperm tests.

  11. Sperm competition enhances functional capacity of mammalian spermatozoa

    E-print Network

    Museo Nacional de Ciencias Naturales

    there are differences in the proportion of spermatozoa that become ready to interact with the ovum (``capacitated to progesterone, an ovum-associated signal. Sperm competition thus favors a larger population of spermatozoa that are competent to fertilize, and spermatozoa that are more sensitive to the signals emitted by the ovum

  12. SIGNIFICANCE OF INCORPORATING MEASURES OF SPERM PRODUCTION AND FUNCTION INTO RAT TOXICOLOGY STUDIES

    EPA Science Inventory

    The rat is the preferred species for reproductive toxicity testing. The inclusion of measures of rat sperm quality, such as motility and morphology, into reproductive test protocols often increases the sensitivity of the test to detect effects, and provides the toxicologist and ...

  13. Role of C-type natriuretic peptide in the function of normal human sperm.

    PubMed

    Xia, Hui; Chen, Yao; Wu, Ke-Jia; Zhao, Hu; Xiong, Cheng-Liang; Huang, Dong-Hui

    2016-01-01

    C-type natriuretic peptide (CNP) is a newly discovered type of local regulatory factor that mediates its biological effects through the speci?c, membrane-bound natriuretic peptide receptor-B (NPR-B). Recent studies have established that CNP is closely related to male reproductive function. The aims of this study were to determine the distribution of CNP/NPR-B in human ejaculated spermatozoa through different methods (such as immunolocalization, real time polymerase chain reaction and Western Blot), and then to evaluate the influence of CNP on sperm function i n vitro, such as motility and acrosome reaction. Human semen samples were collected from consenting donors who met the criteria of the World Health Organization for normozoospermia. Our results show that the speci?c receptor NPR-B of CNP is localized in the acrosomal region of the head and the membrane of the front-end tail of the sperm, and there is no signal of CNP in human sperm. Compared with the control, CNP can induce a signi?cant dose-dependent increase in spermatozoa motility and acrosome reaction. In summary, CNP/NPR-B can affect sperm motility and acrosome reaction, thus regulating the reproductive function of males. CNP may be a new key factor in regulating sperm function. PMID:25926602

  14. Testes Mass, but Not Sperm Length, Increases with Higher Levels of Polyandry in an Ancient Sex Model

    PubMed Central

    Vrech, David E.; Olivero, Paola A.; Mattoni, Camilo I.; Peretti, Alfredo V.

    2014-01-01

    There is strong evidence that polyandrous taxa have evolved relatively larger testes than monogamous relatives. Sperm size may either increase or decrease across species with the risk or intensity of sperm competition. Scorpions represent an ancient direct mode with spermatophore-mediated sperm transfer and are particularly well suited for studies in sperm competition. This work aims to analyze for the first time the variables affecting testes mass, ejaculate volume and sperm length, according with their levels of polyandry, in species belonging to the Neotropical family Bothriuridae. Variables influencing testes mass and sperm length were obtained by model selection analysis using corrected Akaike Information Criterion. Testes mass varied greatly among the seven species analyzed, ranging from 1.6±1.1 mg in Timogenes dorbignyi to 16.3±4.5 mg in Brachistosternus pentheri with an average of 8.4±5.0 mg in all the species. The relationship between testes mass and body mass was not significant. Body allocation in testes mass, taken as Gonadosomatic Index, was high in Bothriurus cordubensis and Brachistosternus ferrugineus and low in Timogenes species. The best-fitting model for testes mass considered only polyandry as predictor with a positive influence. Model selection showed that body mass influenced sperm length negatively but after correcting for body mass, none of the variables analyzed explained sperm length. Both body mass and testes mass influenced spermatophore volume positively. There was a strong phylogenetic effect on the model containing testes mass. As predicted by the sperm competition theory and according to what happens in other arthropods, testes mass increased in species with higher levels of sperm competition, and influenced positively spermatophore volume, but data was not conclusive for sperm length. PMID:24736525

  15. Sperm functional parameters and erythrocytes oxidant-antioxidant imbalance during municipal landfill leachate treatment withdrawal in rats.

    PubMed

    Adedara, Isaac A; Lawal, Tajudeen A; Adesina, Adebayo A; Oyebiyi, Oluwatosin O; Ebokaiwe, Azubike P; Farombi, Ebenezer O

    2014-01-01

    Adequate information on how leachates affect hematological and reproductive functions is necessary to help in linking causality with predictable response. The present study investigated the effects of Olushosun municipal landfill leachate (OMLL) exposure and withdrawal on sperm characteristics and erythrocytes oxidant-antioxidant balance in rats. Adult male Wistar rats were exposed to 0%, 12.5% and 25% OMLL in drinking water for 28 days. One-half of the rats in each group were sacrificed on day 29 while the remaining one-half stayed an additional 28 days without treatment. OMLL exposure significantly decreased sperm functional parameters, disrupted antioxidant systems with concomitant elevation in hydrogen peroxide and malondialdehyde levels in erythrocytes and sperm. Following withdrawal of treatment, OMLL-mediated decrease in sperm count and daily sperm production were reversed to near control. However, erythrocytes and sperm oxidative damage, increased sperm abnormalities, decreased epididymis weight, sperm progressive motility and testicular sperm number persisted and were consistent with results obtained from rats sacrificed immediately after OMLL treatment. Collectively, OMLL-induced irreversible oxidative damage to erythrocytes and sperm in rats within the time course of investigation. These findings highlight potential adverse effects of OMLL on individuals unduly exposed to leachates contaminated substances. PMID:24486716

  16. Do highly ornamented and less parasitized males have high quality sperm? - an experimental test for parasite-induced reproductive trade-offs in European minnow (Phoxinus phoxinus).

    PubMed

    Kekäläinen, Jukka; Pirhonen, Juhani; Taskinen, Jouni

    2014-11-01

    Parasites take their resources from hosts and thus directly reduce available resources for hosts' own body functions, such as growth and reproduction. Furthermore, parasite infections cause significant indirect costs to their hosts in terms of increased investments on immune defense. In this study, we investigated the impact of parasite infection on the sperm quality and expression of secondary sexual ornamentation (saturation of the red abdominal colouration and number of breeding tubercles) in the Eurasian minnow (Phoxinus phoxinus). We exposed minnows to a high and low dose of common nonspecific fish ectoparasite, the glochidia larvae of duck mussel (Anodonta anatina) and tested whether parasite infection leads to trade-off in sperm quality and/or ornamental expression. We found that glochidia infection reduces the curvature of the sperm swimming trajectory, number of breeding tubercles, and possibly male competitive ability, but does not affect expression of male color ornamentation. Furthermore, glochidia infection was found to reduce sperm motility, but only when all the noninfected individuals were excluded from the model. Supporting one of the predictions by phenotype-linked fertility hypothesis both in high-infection and low-infection group male breeding colouration was positively associated with sperm quality. Our results suggest that although glochidia infection may have negative impact on male reproductive success, parasite-induced costs may not create strong trade-off between breeding colouration and sperm quality or that such trade-off become detectable only in resource-limited conditions. PMID:25540686

  17. Do highly ornamented and less parasitized males have high quality sperm? – an experimental test for parasite-induced reproductive trade-offs in European minnow (Phoxinus phoxinus)

    PubMed Central

    Kekäläinen, Jukka; Pirhonen, Juhani; Taskinen, Jouni

    2014-01-01

    Parasites take their resources from hosts and thus directly reduce available resources for hosts’ own body functions, such as growth and reproduction. Furthermore, parasite infections cause significant indirect costs to their hosts in terms of increased investments on immune defense. In this study, we investigated the impact of parasite infection on the sperm quality and expression of secondary sexual ornamentation (saturation of the red abdominal colouration and number of breeding tubercles) in the Eurasian minnow (Phoxinus phoxinus). We exposed minnows to a high and low dose of common nonspecific fish ectoparasite, the glochidia larvae of duck mussel (Anodonta anatina) and tested whether parasite infection leads to trade-off in sperm quality and/or ornamental expression. We found that glochidia infection reduces the curvature of the sperm swimming trajectory, number of breeding tubercles, and possibly male competitive ability, but does not affect expression of male color ornamentation. Furthermore, glochidia infection was found to reduce sperm motility, but only when all the noninfected individuals were excluded from the model. Supporting one of the predictions by phenotype-linked fertility hypothesis both in high-infection and low-infection group male breeding colouration was positively associated with sperm quality. Our results suggest that although glochidia infection may have negative impact on male reproductive success, parasite-induced costs may not create strong trade-off between breeding colouration and sperm quality or that such trade-off become detectable only in resource-limited conditions. PMID:25540686

  18. 1Sperm fate and function in reproductive isolation in Drosophila Corresponding author E-mail: tmarkow@arl.arizona.edu

    E-print Network

    Markow, Therese

    of the genus Drosophila provide an unusually attractive model system for studies of the reproductive isolating1Sperm fate and function in reproductive isolation in Drosophila Corresponding author E-mail: tmarkow@arl.arizona.edu Sperm fate and function in reproductive isolation in Drosophila Therese Ann Markow

  19. Impact of obesity on male fertility, sperm function and molecular composition

    PubMed Central

    Palmer, Nicole O.; Bakos, Hassan W.; Fullston, Tod; Lane, Michelle

    2012-01-01

    Male obesity in reproductive-age men has nearly tripled in the past 30 y and coincides with an increase in male infertility worldwide. There is now emerging evidence that male obesity impacts negatively on male reproductive potential not only reducing sperm quality, but in particular altering the physical and molecular structure of germ cells in the testes and ultimately mature sperm. Recent data has shown that male obesity also impairs offspring metabolic and reproductive health suggesting that paternal health cues are transmitted to the next generation with the mediator mostly likely occurring via the sperm. Interestingly the molecular profile of germ cells in the testes and sperm from obese males is altered with changes to epigenetic modifiers. The increasing prevalence of male obesity calls for better public health awareness at the time of conception, with a better understanding of the molecular mechanism involved during spermatogenesis required along with the potential of interventions in reversing these deleterious effects. This review will focus on how male obesity affects fertility and sperm quality with a focus on proposed mechanisms and the potential reversibility of these adverse effects. PMID:23248766

  20. Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

    PubMed Central

    Hossain, Md. Sharoare; Johannisson, Anders; Wallgren, Margareta; Nagy, Szabolcs; Siqueira, Amanda Pimenta; Rodriguez-Martinez, Heriberto

    2011-01-01

    Flow cytometry is now a recognized methodology within animal spermatology, and has moved from being a research tool to become routine in the assessment of animal semen destined to breeding. The availability of ‘bench-top' flow cytometers and of newer and versatile markers for cell structure and function had allowed the instrumentation to measure more sperm parameters, from viability to reactiveness when exposed to exogenous stimuli, and to increase our capabilities to sort spermatozoa for potential fertilizing capacity, or chromosomal sex. The present review summarizes the state of the art regarding flow cytometry applied to animal andrology, albeit keeping an open comparative intent. It critically evaluates the present and future capabilities of flow cytometry for the diagnostics of potential fertility and for the development of current reproductive technologies such as sperm freezing, sperm selection and sperm sorting. The flow cytometry methods will probably further revolutionize our understanding of the sperm physiology and their functionality, and will undoubtedly extend its application in isolating many uncharacterized features of spermatozoa. However, continuous follow-up of the methods is a necessity owing to technical developments and the complexity of mapping spermatozoa. PMID:21478895

  1. Metabolic Substrates Exhibit Differential Effects on Functional Parameters of Mouse Sperm Capacitation1

    PubMed Central

    Goodson, Summer G.; Qiu, Yunping; Sutton, Keith A.; Xie, Guoxiang; Jia, Wei; O'Brien, Deborah A.

    2012-01-01

    ABSTRACT Although substantial evidence exists that sperm ATP production via glycolysis is required for mammalian sperm function and male fertility, conflicting reports involving multiple species have appeared regarding the ability of individual glycolytic or mitochondrial substrates to support the physiological changes that occur during capacitation. Several mouse models with defects in the signaling pathways required for capacitation exhibit reductions in sperm ATP levels, suggesting regulatory interactions between sperm metabolism and signal transduction cascades. To better understand these interactions, we conducted quantitative studies of mouse sperm throughout a 2-h in vitro capacitation period and compared the effects of single substrates assayed under identical conditions. Multiple glycolytic and nonglycolytic substrates maintained sperm ATP levels and comparable percentages of motility, but only glucose and mannose supported hyperactivation. These monosaccharides and fructose supported the full pattern of tyrosine phosphorylation, whereas nonglycolytic substrates supported at least partial tyrosine phosphorylation. Inhibition of glycolysis impaired motility in the presence of glucose, fructose, or pyruvate but not in the presence of hydroxybutyrate. Addition of an uncoupler of oxidative phosphorylation reduced motility with pyruvate or hydroxybutyrate as substrates but unexpectedly stimulated hyperactivation with fructose. Investigating differences between glucose and fructose in more detail, we demonstrated that hyperactivation results from the active metabolism of glucose. Differences between glucose and fructose appeared to be downstream of changes in intracellular pH, which rose to comparable levels during incubation with either substrate. Sperm redox pathways were differentially affected, with higher levels of associated metabolites and reactive oxygen species generated during incubations with fructose than during incubations with glucose. PMID:22837480

  2. Methyl-parathion decreases sperm function and fertilization capacity after targeting spermatocytes and maturing spermatozoa

    SciTech Connect

    Pina-Guzman, Belem; Sanchez-Gutierrez, M.; Marchetti, Francesco; Hernandez-Ochoa, I.; Solis-Heredia, M.J .; Quintanilla-Vega, B.

    2009-05-03

    Paternal germline exposure to organophosphorous pesticides (OP) has been associated with reproductive failures and adverse effects in the offspring. Methyl parathion (Me-Pa), a worldwide-used OP, has reproductive adverse effects and is genotoxic to sperm. Oxidative damage has been involved in the genotoxic and reproductive effects of OP. The purpose of this study was to determine the effects of Me-Pa on spermatozoa function and ability to fertilize. Male mice were exposed to Me-Pa (20 mg/kg bw, i.p.) and spermatozoa from epididymis-vas deferens were collected at 7 or 28 days post-treatment (dpt) to assess the effects on maturing spermatozoa and spermatocytes, respectively. DNA damage was evaluated by nick translation (NT-positive cells) and SCSA (percentDFI); lipoperoxidation (LPO) by malondialdehyde production; sperm function by spontaneous- and induced-acrosome reactions (AR); mitochondrial membrane potential (MMP) by using the JC-1 flurochrome; and, fertilization ability by an in vitro assay and in vivo mating. Results showed alterations in DNA integrity (percentDFI and NT-positive cells) at 7 and 28 dpt, in addition to decreased sperm quality and a decrease in induced-AR; reduced MMP and LPO was observed only at 7 dpt. We found negative correlations between LPO and all sperm alterations. Altered sperm functional parameters were associated with reduced fertilization rates at both times, evaluated either in vitro or in vivo. These results show that Me-Pa exposure of maturing spermatozoa and spermatocytes affects many sperm functional parameters that result in a decreased fertilizing capacity. Oxidative stress seems to be a likely mechanism ofthe detrimental effects of Me-Pa in male germ cells.

  3. Environmental osmolality influences sperm motility activation in an anuran amphibian.

    PubMed

    Byrne, P G; Dunne, C; Munn, A J; Silla, A J

    2015-03-01

    Evolutionary theory predicts that selection will favour sperm traits that maximize fertilization success in local fertilization environments. In externally fertilizing species, osmolality of the fertilization medium is known to play a critical role in activating sperm motility, but there remains limited evidence for adaptive responses to local osmotic environments. In this study, we used a split-sample experimental design and computer-assisted sperm analysis to (i) determine the optimal medium osmolality for sperm activation (% sperm motility and sperm velocity) in male common eastern froglets (Crinia signifera), (ii) test for among-population variation in percentage sperm motility and sperm velocity at various activation-medium osmolalities and (iii) test for among-population covariation between sperm performance and environmental osmolality. Frogs were obtained from nine populations that differed in environmental osmolality, and sperm samples of males from different populations were subjected to a range of activation-medium osmolalities. Percentage sperm motility was optimal between 10 and 50 mOsm kg(-1) , and sperm velocity was optimal between 10 and 100 mOsm kg(-1) , indicating that C. signifera has evolved sperm that can function across a broad range of osmolalities. As predicted, there was significant among-population variation in sperm performance. Furthermore, there was a significant interaction between activation-medium osmolality and environmental osmolality, indicating that frogs from populations with higher environmental osmolality produced sperm that performed better at higher osmolalities in vitro. This finding may reflect phenotypic plasticity in sperm functioning, or genetic divergence resulting from spatial variation in the strength of directional selection. Both of these explanations are consistent with evolutionary theory, providing some of the first empirical evidence that local osmotic environments can favour adaptive sperm motility responses in species that use an external mode of fertilization. PMID:25586700

  4. Semen Quality and Sperm Function Loss by Hypercholesterolemic Diet Was Recovered by Addition of Olive Oil to Diet in Rabbit

    PubMed Central

    Romero, Aida A.; Funes, Abi K.; Cid-Barria, Macarena; Cabrillana, María E.; Monclus, María A.; Simón, Layla; Vicenti, Amanda E.; Fornés, Miguel W.

    2013-01-01

    Fat increment (0.05% cholesterol, chol) in standard diet promoted a significant increase in serum and sperm membrane chol, which ultimately altered membrane-coupled sperm specific functions: osmotic resistance, acrosomal reaction, and sperm capacitation in White New Zealand rabbits. These changes were also associated with a reduction in motility percentage and appearance of abnormal sperm morphology. The present study was carried out to evaluate the effect of dietary olive oil (OO, 7% v/w) administration to several male hypercholesterolemic rabbits (hypercholesterolemic rabbits, HCR) with altered fertility parameters. These HCR males were achieved by feeding normal rabbits with a high-fat diet (0.05% chol). HCR were associated with a modest non-significant increase in body weight (standard diet, 4.08±0.17 Kg, versus high-fat diet, 4.37±0.24 Kg). Hypercholesterolemic rabbits presented a marked decrease in semen volume, sperm cell count, and percentage of sperm motility, associated with a significant increase in sperm cell abnormalities. Moreover, sperm capacitation measured by the characteristic phosphorylated protein pattern in and induced acrosomal reaction were also altered suggesting sperm dysfunction. However, the administration of OO (for 16 weeks) to rabbits that were fed with 50% of the high-fat diet normalized serum chol. Curiously, OO supply succeeded to attenuate the seminal and sperm alterations observed in HCR group. Administration of OO alone did not cause any significant changes in above mentioned parameters. These data suggest that OO administration to HCR male rabbits recovers the loss of semen quality and sperm functionality. PMID:23326331

  5. Differences in the fatty-acid composition of rodent spermatozoa are associated to levels of sperm competition

    PubMed Central

    delBarco-Trillo, Javier; Mateo, Rafael; Roldan, Eduardo R. S.

    2015-01-01

    Sperm competition is a prevalent phenomenon that drives the evolution of sperm function. High levels of sperm competition lead to increased metabolism to fuel higher sperm velocities. This enhanced metabolism can result in oxidative damage (including lipid peroxidation) and damage to the membrane. We hypothesized that in those species experiencing high levels of sperm competition there are changes in the fatty-acid composition of the sperm membrane that makes the membrane more resistant to oxidative damage. Given that polyunsaturated fatty acids (PUFAs) are the most prone to lipid peroxidation, we predicted that higher sperm competition leads to a reduction in the proportion of sperm PUFAs. In contrast, we predicted that levels of sperm competition should not affect the proportion of PUFAs in somatic cells. To test these predictions, we quantified the fatty-acid composition of sperm, testis and liver cells in four mouse species (genus Mus) that differ in their levels of sperm competition. Fatty-acid composition in testis and liver cells was not associated to sperm competition levels. However, in sperm cells, as predicted, an increase in sperm competition levels was associated with an increase in the proportion of saturated fatty-acids (the most resistant to lipid peroxidation) and by a concomitant decrease in the proportion of PUFAs. Two particular fatty acids were most responsible for this pattern (arachidonic acid and palmitic acid). Our findings thus indicate that sperm competition has a pervasive influence in the composition of sperm cells that ultimately may have important effects in sperm function. PMID:25795911

  6. Insect sperm motility.

    PubMed

    Werner, Michael; Simmons, Leigh W

    2008-05-01

    The flagellosperm of insects, although following a general ground plan, exhibit considerable variation in morphology and ultrastructure across taxa, consistent with a history of rapid and divergent evolution. Sperm competition, which occurs when sperm of two or more males compete for the fertilization of a female's ova, has been recognized as a significant driving force in the evolution of insect sperm structure. Despite a considerable volume of data on sperm morphology, little is known about the motility of insect sperm. Understanding insect sperm motility would help to refine models of sexual selection on insect sperm, and would throw light on the selective mechanisms that shape insect sperm structure and function. This review updates our present knowledge of the proximate and ultimate aspects of insect sperm motility. PMID:18397179

  7. The porcine sperm reservoir in relation to the function of hyaluronan

    PubMed Central

    TIENTHAI, Paisan

    2015-01-01

    The oviduct plays a role in successful animal reproduction not only in spermatozoa and ova transport to the fertilization site but also by affording a microenvironment for fertilization and early embryonic development. The sperm reservoir (SR) is restricted in the uterotubal junction (UTJ) and caudal isthmus. Billions of porcine spermatozoa are distributed to the female reproductive tract during/after insemination, and small amounts of them are stored for about 36–40 hours in the SR, which maintains sperm viability in the pre-ovulation period through its surface epithelium and production of fluid. The SR regulates the release of spermatozoa so that only a small population moves towards the fertilization site (ampulla) to decrease polyspermy. This review attempts to provide information about the structure and function of the porcine SR, its intraluminal content (hyaluronan, HA), and the influences of HA on porcine spermatozoa in vivo. In pigs, the spermatozoa are stored in a mucous-like fluid within the UTJ and caudal isthmus in the pre-ovulation period. The oviduct fluid contains sulfated glycosaminoglycans (GAGs) and non-sulfated GAGs, i.e., HA. It is interesting to note that HA is synthesized by hyaluronan synthase-3 (HAS-3), and its receptor, CD44, is found in the epithelium of the porcine SR site. Additionally, sperm capacitation does not occur in vivo in the SR during the pre- and peri-ovulation periods, but spermatozoa in the SR will attempt to capacitate if exposed to bicarbonate. However, capacitation in the SR will rise in the post-ovulation period, indicating the role of HA in modulating sperm capacitation after ovulation. All data support the understanding that the porcine SR ensures the viability of fertile spermatozoa and maintains the non-capacitated status during the pre-ovulation period. This basic knowledge about the SR is believed to be useful to advance sperm preparation procedures for in vitro fertilization (IVF) and improve the preservation process of porcine semen. PMID:26311759

  8. Postnatal exposure of the male mouse to 2,2',3,3',4,4',5,5',6,6'-decabrominated diphenyl ether: decreased epididymal sperm functions without alterations in DNA content and histology in testis.

    PubMed

    Tseng, Li-Ho; Lee, Chia-Wei; Pan, Min-Hsiung; Tsai, Shinn-Shong; Li, Mei-Hui; Chen, Jenq-Renn; Lay, Jiunn-Jyi; Hsu, Ping-Chi

    2006-07-01

    2,2',3,3',4,4',5,5',6,6'-Decabrominated diphenyl ether (PBDE 209) is the second most used brominated flame retardant (BFRs) in constructed materials because it is considered less toxic than others, though other fire retardants, some congeners of PBDE 209, are reported to be toxic. This combined the fact that PBDE 209 has been found in high levels in human milk, blood, indoor environments as well as in foodstuffs has led us in this study attempt to find out whether PBDE 209, also known as decaBDE and decabrominated diphenyl oxide (DBDPO), has an adverse effect on this histology of testes and sperm in CD-1 male mice. The mice we studied were divided into groups and gavaged with 10, 100, 500 and 1500 mg/kg PBDE 209 in corn oil per day between postnatal Days 21 and 70. On Day 71, the mice were anesthetized and sperm function, testis DNA content, and histopathology were studied. We found in the 500- and 1500-mg/kg/day groups that neonatal exposure to PBDE 209 reduced sperm epididymal sperm mitochondrial membrane potential (MMP), reduced amplitude of the lateral head displacement (ALH) and induced the generation of hydrogen peroxide (H2O2) in the sperm of sexually mature male mice, without affecting the sperm count, motility, morphology, curvilinear velocity (VCL), angular progressive velocity (VAP), straight-line velocity (VSL), beat-cross frequency (BCF), sperm chromatin structure assay (SCSA), superoxide anion (O2-*) generation, DNA content in testis cells, or testicular histopathology. ALH was positively associated with an increase in MMP and negatively associated with generation of sperm H2O2. The reduction of MMP was negatively associated with an increase in generation of sperm H2O2. The presence of the relationships between sperm ALH, MMP, and generation of H2O2 indicate toxic action possibly resulting from PBDE 209-induced oxidative stress. In conclusion, this is the first study to report the lowest-observed-adverse-effect level (LOAEL) for sperm function to be 500 mg/kg of PBDE 209 in male mice. Decreased epididymal sperm MMP and ALH as well as induced generation of sperm H2O2 were some of the most serious effects of postnatal PBDE 209 exposure. Future investigations should be performed to study the effects of prenatal exposure of PBDE 209 and the mechanism behind PBDE 209-related oxidative stress in the fetal and pubertal stages of development. PMID:16713668

  9. Hyaluronidase 2: a novel germ cell hyaluronidase with epididymal expression and functional roles in mammalian sperm.

    PubMed

    Modelski, Mark J; Menlah, Gladys; Wang, Yipei; Dash, Soma; Wu, Kathie; Galileo, Deni S; Martin-DeLeon, Patricia A

    2014-11-01

    To initiate the crucial cell adhesion events necessary for fertilization, sperm must penetrate extracellular matrix barriers containing hyaluronic acid (HA), a task thought to be accomplished by neutral-active hyaluronidases. Here we report that the ~57 kDa hyaluronidase 2 (HYAL2) that in somatic tissues has been highly characterized to be acid-active is present in mouse and human sperm, as detected by Western blot, flow cytometric, and immunoprecipitation assays. Immunofluorescence revealed its presence on the plasma membrane over the acrosome, the midpiece, and proximal principal piece in mice where protein fractionation demonstrated a differential distribution in subcellular compartments. It is significantly more abundant in the acrosome-reacted (P = 0.04) and soluble acrosomal fractions (P = 0.006) (microenvironments where acid-active hyaluronidases function) compared to that of the plasma membrane where neutral hyaluronidases mediate cumulus penetration. Using HA substrate gel electrophoresis, immunoprecipitated HYAL 2 was shown to have catalytic activity at pH 4.0. Colocalization and coimmunoprecipitation assays reveal that HYAL2 is associated with its cofactor, CD44, consistent with CD44-dependent HYAL2 activity. HYAL2 is also present throughout the epididymis, where Hyal2 transcripts were detected, and in the epididymal luminal fluids. In vitro assays demonstrated that HYAL2 can be acquired on the sperm membrane from epididymal luminal fluids, suggesting that it plays a role in epididymal maturation. Because similar biphasic kinetics are seen for HYAL2 and SPAM1 (Sperm adhesion molecule 1), it is likely that HYAL2 plays a redundant role in the catalysis of megadalton HA to its 20 kDa intermediate during fertilization. PMID:25232017

  10. Selection of functional human sperm with higher DNA integrity and fewer reactive oxygen species

    PubMed Central

    Asghar, Waseem; Velasco, Vanessa; Kingsley, James L.; Shoukat, Muhammad S.; Shafiee, Hadi; Anchan, Raymond M.; Mutter, George L.; Tüzel, Erkan; Demirci, Utkan

    2014-01-01

    Fertilization and reproduction are central to the survival and propagation of a species. Couples who cannot reproduce naturally have to undergo in vitro clinical procedures. An integral part of these clinical procedures includes isolation of healthy sperm from raw semen. Existing sperm sorting methods are not efficient and isolate sperm having high DNA fragmentation and reactive oxygen species, and suffer from multiple manual steps and variations between embryologists. Inspired by in vivo natural sperm sorting mechanisms where vaginal mucus becomes less viscous to form microchannels to guide sperm towards egg, we present a chip that efficiently sorts healthy, motile and morphologically normal sperm without centrifugation. Higher percentage of sorted sperm show significantly lesser reactive oxygen species and DNA fragmentation than the conventional swim-up method. The presented chip is an easy-to-use high throughput sperm sorter that provides standardized sperm sorting assay with less reliance on embryologist’s skills, facilitating reliable operational steps. PMID:24753434

  11. Supplementation of IVF medium with melatonin: effect on sperm functionality and in vitro produced bovine embryos.

    PubMed

    Cheuquemán, C; Arias, M E; Risopatrón, J; Felmer, R; Álvarez, J; Mogas, T; Sánchez, R

    2015-08-01

    Gamete co-incubation generates high free radical levels surrounding growing zygotes which may impair subsequent embryo viability. Melatonin eliminates a wide variety of free radicals; hence, we tried to improve in vitro embryo production by adding melatonin to in vitro fertilisation (IVF) media in high (Exp. 1) and low concentrations (Exp. 2), and we evaluated its effect on bull sperm function during IVF co-incubation time (Exp. 3). In Experiment 1, we supplemented IVF media culture with 0.01, 0.1 and 1 mmol of melatonin, along with a no melatonin control group. In Experiment 2, melatonin levels were reduced to 10, 100 and 1000 nmol, with a no melatonin control group. In Experiment 3, spermatozoa were incubated in IVF media with melatonin (as Exp. 2) and functional parameters were analysed at 0, 4 and 18 h. In Experiment 1, only 1 mmol melatonin showed lesser blastocyst rates than control (C: 23.2 ± 6.7% versus 1 mmol: 2.0 ± 1.7%). In Experiment 2, no statistical differences were found in cleavage percentage, blastocyst percentage and total cell count for any melatonin treatment. In Experiment 3, sperm samples with 1000 nmol melatonin had a significantly higher wobbler (WOB) coefficient, a lower percentage of intact acrosomes, a lower percentage of viable spermatozoa with ROS, greater DNA fragmentation and higher DNA oxidation than controls. Total fluorescence intensity for ROS at 10 nmol melatonin was significantly greater than controls (P < 0.05). IVF media with 1 mmol melatonin is deleterious for embryo development, and in lower concentrations, it modulated sperm functionality, but had no effects on embryo production. PMID:25059349

  12. Toxic effects of anatoxin-a on testes and sperm counts of male mice.

    PubMed

    Yavasoglu, Altug; Karaaslan, M Ali; Uyanikgil, Yigit; Sayim, Ferah; Ates, Utku; Yavasoglu, N Ulku Karabay

    2008-08-01

    Anatoxin-a, a potent neurotoxin, is one of a number of toxins produced by cyanobacteria especially some strains of Anabaena. Toxic cyanobacteria are found worldwide in inland and coastal water environments. The present study was performed to evaluate the toxicity of anatoxin-a on testes and sperm counts of male mice. The animals of the treatment groups were administered with 50, 100 and 150microg/kg/day anatoxin-a for seven consecutive days by intraperitoneal (i.p.) route. Although there were no significant changes in body weight gain, and absolute and relative testes weights, absolute and relative weights of cauda epididymis reduced significantly in the 100 and 150microg/kg groups when compared with control group. The number of sperm count in cauda epididymis was reduced dose dependently in all treatment groups compared with control animals. Anatoxin-a caused dose-dependent histopathological changes in the testes of mice such as degenerations in seminiferous tubules, intercellular disassociation of spermatogenetic cell lines, sloughing of germ cells into tubular lumen, vacuolisation in Sertoli cells and loss of germ cells. The epithelial thickness of seminiferous tubules decreased significantly in all treatment groups in a dose-dependent manner. PMID:18514498

  13. A Systematic Analysis of a Deep Mouse Epididymal Sperm Proteome

    SciTech Connect

    Chauvin, Theodore; Xie, Fang; Liu, Tao; Nicora, Carrie D.; Yang, Feng; Camp, David G.; Smith, Richard D.; Roberts, Kenneth P.

    2012-12-21

    Spermatozoa are highly specialized cells that, when mature, are capable of navigating the female reproductive tract and fertilizing an oocyte. The sperm cell is thought to be largely quiescent in terms of transcriptional and translational activity. As a result, once it has left the male reproductive tract, the sperm cell is essentially operating with a static population of proteins. It is therefore theoretically possible to understand the protein networks contained in a sperm cell and to deduce its cellular function capabilities. To this end we have performed a proteomic analysis of mouse sperm isolated from the cauda epididymis and have confidently identified 2,850 proteins, which is the most comprehensive sperm proteome for any species reported to date. These proteins comprise many complete cellular pathways, including those for energy production via glycolysis, ?-oxidation and oxidative phosphorylation, protein folding and transport, and cell signaling systems. This proteome should prove a useful tool for assembly and testing of protein networks important for sperm function.

  14. Leukocytes and oxidative stress: dilemma for sperm function and male fertility

    PubMed Central

    Henkel, Ralf R

    2011-01-01

    Spermatozoa are constantly exposed to the interphase between oxidation through high amounts of reactive oxygen species (ROS) and leukocytes, and reduction by means of scavengers and antioxidants. Considering the very special functions as being the only cells with such high polarization and exerting their functions outside the body, even in a different individual, the female genital tract, the membranes of these cells are chemically composed of an extraordinary high amount of polyunsaturated fatty acids. This in turn, renders them very susceptible to oxidative stress, which is defined as an imbalance between oxidation and reduction towards the oxidative status. As a result, ROS deriving from both leukocytes and the male germ cells themselves cause a process called ‘lipid peroxidation' and other damages to the sperm cell. On the other hand, a certain limited amount of ROS is essential in order to trigger vital physiological reactions in cells, including capacitation or the acrosome reaction in sperm. The treatment of patients with antioxidants to compensate the oxidative status caused by oxidative stress is highly debated as uncontrolled antioxidative treatment might derail the system towards the reduced status, which is also unphysiological and can even induce cancer. This paradox is called the ‘antioxidant paradox'. Therefore, a proper andrological diagnostic work-up, including the evaluation of ROS levels and the antioxidant capacity of the semen, has to be carried out beforehand, aimed at keeping the fine balance between oxidation and scavenging of vital amounts of ROS. PMID:21076433

  15. Melatonin improve the sperm quality in forced swimming test induced oxidative stress in nandrolone treated Wistar rats.

    PubMed

    Minaii, Bagher; Moayeri, Ardeshir; Shokri, Saeed; Habibi Roudkenar, Mehryar; Golmohammadi, Taghi; Malek, Fatemeh; Barbarestani, Mohammad

    2014-01-01

    This study investigates the effects of melatonin on the sperm quality and testis weight after the combination of swimming exercise and nandrolone decanoate (DECA). Two groups of male Wistar rats were treated for eight weeks as follows; group A consist of CO (control), Sham, N (DECA), S (swimming) and NS (DECA plus swimming); and group B: Sham M (sham melatonin), M (melatonin), MN (melatonin plus DECA), MS (melatonin plus swimming), MNS (melatonin, DECA plus swimming). The motility of sperm was significantly improved in melatonin groups in comparison to N, S and NS groups (P?0.05).  The left testes weight was decreased in N, NS and MNS groups, and the right testes weight was decreased in N,S,NS, MS and MNS groups in compare with the control group. This study concluded that melatonin probably could improve the sperm motility and sex organs weight after the combination of DECA and exercise. PMID:25135257

  16. Glass wool filtration of bull cryopreserved semen: a rapid and effective method to obtain a high percentage of functional sperm.

    PubMed

    Arzondo, M M; Caballero, J N; Marín-Briggiler, C I; Dalvit, G; Cetica, P D; Vazquez-Levin, M H

    2012-07-01

    Frozen-thawed bull sperm are widely used in assisted reproductive technologies, but cryopreservation negatively affects semen quality. Several sperm selection techniques have been developed to separate motile sperm from non-motile cells. The aim of the present study was to evaluate the effectiveness of the glass wool column filtration to select functional sperm from frozen-thawed bull semen samples. Frozen semen from six Holstein bulls was thawed and filtered through a glass wool column, followed by assessment of routine and functional sperm parameters. In a set of experiments, sperm aliquots were also processed by swim up to compare both selection methods. Samples recovered in the glass wool filtrate had high percentages of viable (94 ± 3%, mean ± SD), progressively motile (89 ± 4%), acrosome-intact (98 ± 1%), and non-capacitated (80 ± 10%) sperm; these values were higher (P < 0.05) than those obtained after performing the swim up procedure. Moreover, the glass wool filtration yielded 67 ± 19% motile cells, in comparison with 18 ± 8% obtained with swim up (P < 0.05), calculated as the concentration of progressively motile cells selected relative to their concentration in the sample before the selection procedure. Glass wool-filtered sperm were able to undergo capacitation-related events, based on the increase in the percentage of cells classified as capacitated by CTC staining (B-pattern) after incubation with heparin (50 ± 5%) in comparison with control conditions with no heparin (17 ± 4%) or heparin + glucose (16 ± 2%; P < 0.05). Moreover, they underwent acrosomal exocytosis in response to pharmacologic (calcium ionophore A23187 and lysophosphatidylcholine) and physiological (follicular fluid) stimuli, and they fertilized in vitro matured cumulus-oocyte complexes and denuded oocytes (two-cell embryos: 72 ± 4% and 52 ± 6%, respectively). We conclude that glass wool filtration is a low-cost, simple, and highly effective procedure to select functionally competent sperm for reproductive technologies in the bull, which may be useful for other domestic and farm animals, as well as for endangered species. PMID:22537998

  17. The sperm motility pattern in ecotoxicological tests. The CRYO-Ecotest as a case study.

    PubMed

    Fabbrocini, Adele; D'Adamo, Raffaele; Del Prete, Francesco; Maurizio, Daniela; Specchiulli, Antonietta; Oliveira, Luis F J; Silvestri, Fausto; Sansone, Giovanni

    2016-01-01

    Changes in environmental stressors inevitably lead to an increasing need for innovative and more flexible monitoring tools. The aim of this work has been the characterization of the motility pattern of the cryopreserved sea bream semen after exposure to a dumpsite leachate sample, for the identification of the best representative parameters to be used as endpoints in an ecotoxicological bioassay. Sperm motility has been evaluated either by visual and by computer-assisted analysis; parameters concerning motility on activation and those describing it in the times after activation (duration parameters) have been assessed, discerning them in terms of sensitivity, reliability and methodology of assessment by means of multivariate analyses. The EC50 values of the evaluated endpoints ranged between 2.3 and 4.5ml/L, except for the total motile percentage (aTM, 7.0ml/L), which proved to be the less sensitive among all the tested parameters. According to the multivariate analyses, a difference in sensitivity among "activation" endpoints in respect of "duration" ones can be inferred; on the contrary, endpoints seem to be equally informative either describing total motile sperm or the rapid sub-population, as well as the assessment methodology seems to be not discriminating. In conclusion, the CRYO-Ecotest is a multi-endpoint bioassay that can be considered a promising innovative ecotoxicological tool, characterized by a high plasticity, as its endpoints can be easy tailored each time according to the different needs of the environmental quality assessment programs. PMID:26318919

  18. Extraocular muscle function testing

    MedlinePLUS

    Extraocular muscle function testing examines the function of the eye muscles. A health care provider observes the movement of ... evaluate weakness or other problem in the extraocular muscles. These problems may result in double vision or ...

  19. Functional Task Test (FTT)

    NASA Technical Reports Server (NTRS)

    Bloomberg, Jacob J.; Mulavara, Ajitkumar; Peters, Brian T.; Rescheke, Millard F.; Wood, Scott; Lawrence, Emily; Koffman, Igor; Ploutz-Snyder, Lori; Spiering, Barry A.; Feeback, Daniel L.; Platts, Steven H.; Stenger, Michael B.; Lee, Stuart M.C.; Arzeno, Natalia; Feiveson, Alan H.; Ryder, Jeffrey; Garcia, Yamil; Guilliams, Mark E.

    2009-01-01

    This slide presentation reviews the Functional Task Test (FTT), an interdisciplinary testing regimen that has been developed to evaluate astronaut postflight functional performance and related physiological changes. The objectives of the project are: (1) to develop a set of functional tasks that represent critical mission tasks for the Constellation Program, (2) determine the ability to perform these tasks after space flight, (3) Identify the key physiological factors that contribute to functional decrements and (4) Use this information to develop targeted countermeasures.

  20. Patch clamp studies of human sperm under physiological ionic conditions reveal three functionally and pharmacologically distinct cation channels

    PubMed Central

    Mansell, S.A.; Publicover, S.J.; Barratt, C.L.R.; Wilson, S.M.

    2014-01-01

    Whilst fertilizing capacity depends upon a K+ conductance (GK) that allows the spermatozoon membrane potential (Vm) to be held at a negative value, the characteristics of this conductance in human sperm are virtually unknown. We therefore studied the biophysical/pharmacological properties of the K+ conductance in spermatozoa from normal donors held under voltage/current clamp in the whole cell recording configuration. Our standard recording conditions were designed to maintain quasi-physiological, Na+, K+ and Cl? gradients. Experiments that explored the effects of ionic substitution/ion channel blockers upon membrane current/potential showed that resting Vm was dependent upon a hyperpolarizing K+ current that flowed via channels that displayed only weak voltage dependence and limited (?7-fold) K+ versus Na+ selectivity. This conductance was blocked by quinidine (0.3 mM), bupivacaine (3 mM) and clofilium (50 µM), NNC55-0396 (2 µM) and mibefradil (30 µM), but not by 4-aminopyridine (2 mM, 4-AP). Progesterone had no effect upon the hyperpolarizing K+ current. Repolarization after a test depolarization consistently evoked a transient inward ‘tail current’ (ITail) that flowed via a second population of ion channels with poor (?3-fold) K+ versus Na+ selectivity. The activity of these channels was increased by quinidine, 4-AP and progesterone. Vm in human sperm is therefore dependent upon a hyperpolarizing K+ current that flows via channels that most closely resemble those encoded by Slo3. Although 0.5 µM progesterone had no effect upon these channels, this hormone did activate the pharmacologically distinct channels that mediate ITail. In conclusion, this study reveals three functionally and pharmacologically distinct cation channels: Ik, ITail, ICatSper. PMID:24442342

  1. Evolution of sperm morphology in anurans: insights into the roles of mating system and spawning location

    PubMed Central

    2014-01-01

    Background The degree of postcopulatory sexual selection, comprising variable degrees of sperm competition and cryptic female choice, is an important evolutionary force to influence sperm form and function. Here we investigated the effects of mating system and spawning location on the evolution of sperm morphology in 67 species of Chinese anurans. We also examined how relative testes size as an indicator of the level of sperm competition affected variation in sperm morphology across a subset of 29 species. Results We found a significant association of mating system and spawning location with sperm morphology. However, when removing the effects of body mass or absolute testes mass for species for which such data were available, this effect became non-significant. Consistent with predictions from sperm competition theory, we found a positive correlation between sperm morphology and relative testes size after taking phylogeny into account. Conclusions Our findings suggest that sexual selection in Chinese anurans favors longer sperm when the level of sperm competition is high. Pre-copulatory male-male competition and spawning location, on the other hand, do not affect the evolution of sperm morphology after taking body mass and absolute testes mass into account. PMID:24884745

  2. Pancreatic exocrine function testing

    SciTech Connect

    Goff, J.S.

    1981-11-01

    It is important to understand which pancreatic function tests are available and how to interpret them when evaluating patients with malabsorption. Available direct tests are the secretin stimulation test, the Lundh test meal, and measurement of serum or fecal enzymes. Indirect tests assess pancreatic exocrine function by measuring the effect of pancreatic secretion on various nutrients. These include triglycerides labeled with carbon 14, cobalamin labeled with cobalt 57 and cobalt 58, and para-aminobenzoic acid bound to a dipeptide. Of all these tests the secretin stimulation test is the most accurate and reliable if done by experienced personnel. However, the indirect tests are simpler to do and appear to be comparable to the secretin test at detecting pancreatic exocrine insufficiency. These indirect tests are becoming clinically available and clinicians should familiarize themselves with the strengths and weaknesses of each.

  3. Transcripts encoding the sperm surface protein tMDC II are non-functional in the human.

    PubMed Central

    Frayne, J; Dimsey, E A; Jury, J A; Hall, L

    1999-01-01

    Five members of the MDC (metalloproteinase-like,disintegrin-like cysteine-rich domain) family of proteins (fertilin alpha, fertilin beta, tMDC I, tMDC II and tMDC III) are expressed on the surface of macaque (Macaca fascicularis) sperm, where they have been proposed to play a role in sperm-egg binding via an interaction between their disintegrin-like domain and one or more integrins on the egg plasma membrane. Of these, two (fertilin alpha and tMDC I) have recently been shown to be non-functional in the human. Here we report the existence of multiple isoforms of human tMDC II transcripts in the human, all of which are also non-functional owing to the presence of deletions and in-frame termination codons, when compared with the macaque orthologue, a finding which is further supported by the lack of immunoreactivity on Western blots of human testis and sperm extracts probed with a macaque anti-tMDC II polyclonal antiserum. These results are discussed in the context of our proposed model for multiple proteins implicated in sperm-egg interactions. PMID:10417343

  4. Immune Activation Reduces Sperm Quality in the Great Tit

    PubMed Central

    Losdat, Sylvain; Richner, Heinz; Blount, Jonathan D.; Helfenstein, Fabrice

    2011-01-01

    Mounting an immune response against pathogens incurs costs to organisms by its effects on important life-history traits, such as reproductive investment and survival. As shown recently, immune activation produces large amounts of reactive species and is suggested to induce oxidative stress. Sperm are highly susceptible to oxidative stress, which can negatively impact sperm function and ultimately male fertilizing efficiency. Here we address the question as to whether mounting an immune response affects sperm quality through the damaging effects of oxidative stress. It has been demonstrated recently in birds that carotenoid-based ornaments can be reliable signals of a male's ability to protect sperm from oxidative damage. In a full-factorial design, we immune-challenged great tit males while simultaneously increasing their vitamin E availability, and assessed the effect on sperm quality and oxidative damage. We conducted this experiment in a natural population and tested the males' response to the experimental treatment in relation to their carotenoid-based breast coloration, a condition-dependent trait. Immune activation induced a steeper decline in sperm swimming velocity, thus highlighting the potential costs of an induced immune response on sperm competitive ability and fertilizing efficiency. We found sperm oxidative damage to be negatively correlated with sperm swimming velocity. However, blood resistance to a free-radical attack (a measure of somatic antioxidant capacity) as well as plasma and sperm levels of oxidative damage (lipid peroxidation) remained unaffected, thus suggesting that the observed effect did not arise through oxidative stress. Towards the end of their breeding cycle, swimming velocity of sperm of more intensely colored males was higher, which has important implications for the evolution of mate choice and multiple mating in females because females may accrue both direct and indirect benefits by mating with males having better quality sperm. PMID:21765955

  5. Methyl glycol, methanol and DMSO effects on post-thaw motility, velocities, membrane integrity and mitochondrial function of Brycon orbignyanus and Prochilodus lineatus (Characiformes) sperm.

    PubMed

    Viveiros, Ana T M; Nascimento, Ariane F; Leal, Marcelo C; Gonçalves, Antônio C S; Orfão, Laura H; Cosson, Jacky

    2015-02-01

    The aim of this study was to use more accurate techniques to investigate the effects of cryoprotectants (CPAs) and extenders on post-thaw sperm quality of Brycon orbignyanus and Prochilodus lineatus. Six freezing media comprising the combination of three CPAs (DMSO, methanol and methyl glycol) and two extenders (BTS and glucose) were used. Sperm was diluted in each medium, loaded into 0.5-mL straws, frozen in a nitrogen vapor vessel (dry-shipper), and stored in liquid nitrogen at -196 °C. Post-thaw sperm motility rate and velocities (curvilinear = VCL; straight line = VSL; average path = VAP) were evaluated using a computer-assisted sperm analyzer. Membrane integrity and mitochondrial function were determined using fluorochromes. Post-thaw quality was considered high when samples presented the following minimum values: 60 % motile sperm, 140 µm/s of VCL, 50 % intact sperm membrane and 50 % mitochondrial function integrity. High post-thaw quality was observed in B. orbignyanus sperm frozen in BTS-methyl glycol and in P. lineatus sperm frozen in BTS-methyl glycol, glucose-methyl glycol and glucose-methanol. All samples frozen in DMSO yielded low quality. The presence of ions in the BTS extender affected post-thaw sperm quality positively in B. orbignyanus and negatively in P. lineatus. Methyl glycol was the most suitable CPA for both fish species, leading to a good protection of cell membrane, mitochondrial function and motility apparatus during the cryopreservation process. For an improved protection, B. orbignyanus sperm should be frozen in an ionic freezing medium. PMID:25433690

  6. DNA-binding sperm proteins with oligo-arginine clusters function as potent activators for egg CK-II.

    PubMed

    Ohtsuki, K; Nishikawa, Y; Saito, H; Munakata, H; Kato, T

    1996-01-01

    The stimulatory effect of DNA-binding sperm proteins (histone and protamine) on the phosphorylation of p98 (ERp99/GRp94, one of the Hsp-90 family of proteins) by egg casein kinase II (CK-II) was investigated in vitro. It was found that (i) phosphorylation of p98 by egg CK-II in vitro is greatly stimulated by poly-Arg, but not by poly-Lys; and (ii) similar stimulation is observed with sperm histones H2B2 and H2B3 (sea urchin) and fish protamines, such as salmine A1 (salmon) and protamine 3a (rainbow trout). These findings suggest that these DNA-binding sperm proteins function as potent activators for CK-II in fertilized eggs. All of these DNA-binding sperm proteins contain at least an oligo-Arg cluster as a common feature, which can interact with an acidic amino acid cluster of the regulatory beta-subunit CK-II. PMID:8549815

  7. Pulmonary function tests

    MedlinePLUS

    ... measured to estimate the lung volume. To measure diffusion capacity , you breathe a harmless gas, called a ... on your report after pulmonary function tests include: Diffusion capacity to carbon monoxide (DLCO) Expiratory reserve volume ( ...

  8. Testing an egg yolk supplemented diet on boars to aid in sperm adaptation at 5°C.

    PubMed

    Casas, Isabel; Miller-Lux, Yvonne; Osborne, Betty; Bonet, Sergi; Althouse, Gary C

    2015-01-01

    In many species, extended semen can be stored at low temperatures to slow bacterial growth. However, boar semen performs poorly at temperatures below 15 °C and this poses unique challenges, as it is not easy to maintain a constant 15-19 °C during shipment. Some extenders have been formulated with egg yolk for storage at 5 °C but the addition of egg yolk is not applicable in the majority of commercial operations. The purpose of this study was to evaluate if boar dietary supplementation with powdered egg yolk imparts any protective effects on sperm quality when stored at 15 °C and 5 °C for up to 11 days in a conventional extender. Ten boars were fed a commercial diet with the addition of 0.11 Kg of powdered egg yolk for 10 weeks. Ejaculates collected on weeks 4, 6, 8, and 10 were processed for storage at both 15 °C and 5 °C and compared with ejaculates from boars fed a standard diet. Throughout an 11-day storage period, sperm quality was assessed including several motility and morphologic parameters and select plasma membrane properties (fluidity, integrity, and triacylglycerol content). Linear regression models were used to describe effects of treatment, storage day, week and temperature on all sperm parameters. Overall, there were minimal beneficial effects of egg yolk treatment on sperm quality parameters. Sperm from egg yolk supplemented boars did have a slower decline in viability and plasma membrane fluidity than that observed in the control sperm when stored at 5 °C (p < 0.001). Additionally, there was an increase in total morphologic abnormalities in sperm from egg yolk fed boars compared to controls at week 10 (p sperm quality or resistance to cold storage when feeding a 10-week dietary supplementation of 0.11 Kg powdered egg yolk to crossbred boars. PMID:25966000

  9. Comparison on the Effects and Safety of Tualang Honey and Tribestan in Sperm Parameters, Erectile Function, and Hormonal Profiles among Oligospermic Males

    PubMed Central

    Ismail, Shaiful Bahari; Bakar, Mohd. Bustamanizan; Nik Hussain, Nik Hazlina; Sulaiman, Siti Amrah; Jaafar, Hasnan; Draman, Samsul; Ramli, Roszaman; Wan Yusoff, Wan Zahanim

    2014-01-01

    Introduction. This study aims to evaluate the effectiveness of Tualang honey on sperm parameters, erectile function, and hormonal and safety profiles. Methodology. A randomized control trial was done using Tualang honey (20 grams) and Tribestan (750?mg) over a period of 12 weeks. Sperm parameters including sperm concentration, motility, and morphology were analyzed and erectile function was assessed using IIEF-5 questionnaire. Hormonal profiles of testosterone, FSH, and LH were studied. The volunteers were randomized into two groups and the outcomes were analyzed using SPSS version 18. Results. A total of 66 participants were involved. A significant increment of mean sperm concentration (P < 0.001), motility (P = 0.015) and morphology (P = 0.008) was seen in Tualang honey group. In Tribestan group, a significant increment of mean sperm concentration (P = 0.007), and morphology (P = 0.009) was seen. No significant differences of sperm concentration, motility, and morphology were seen between Tualang honey and Tribestan group and similar results were also seen in erectile function and hormonal profile. All safety profiles were normal and no adverse event was reported. Conclusion. Tualang honey effect among oligospermic males was comparable with Tribestan in improving sperm concentration, motility, and morphology. The usage of Tualang honey was also safe with no reported adverse event. PMID:25505918

  10. Sperm assays in man and other mammals as indicators of chemically induced testicular dysfunction

    SciTech Connect

    Wyrobek, A.J.

    1980-07-31

    Human sperm assays can be effective in identifying chemical agents that alter testicular function. Four human sperm assays are available - sperm density, motility, morphology, and the YFF test. Sperm assays have practical advantages over other approaches for assessing chemically induced changes in human testicular function, and they have been used in more than 60 different occupational, environmental, and drug-related chemical exposures. Studies of chemically induced sperm anomalies in other mammals have provided data on several hundred agents and encompass numerous species. The most widely used animal sperm assay is sperm morphology of mice. It is simple, quantitative, and sensitive to carcinogens, mutagens, and teratogens. The availability of both human and animal sperm assays provides a valuable link between human and animal studies which may be of potential benefit in assessing the heritable genetic risk associated with chemically induced sperm defects. Results from sperm assays may be used in conjunction with results from short term mutagenicity assays (those with definitive genetic endpoints) to identify those mutagens that may be active in the mammalian testes.

  11. Effect of short-term semen storage in salmon (Oncorhynchus mykiss) on sperm functional parameters evaluated by flow cytometry.

    PubMed

    Trigo, P; Merino, O; Figueroa, E; Valdebenito, I; Sánchez, R; Risopatrón, J

    2015-05-01

    The short-term storage of salmonid semen is a viable method for in vitro fertilisation. Previous studies have found that short-term storage affects sperm motility, compromising quality and fertilising capacity. However, the functional characteristics of the spermatozoa of O. mykiss during storage time and its relation to the spawning period are little known. This study was designed to evaluate the effects of in vitro short-term storage on sperm functional parameters in O. mykiss, determined by flow cytometry. Semen samples of the first spawning - undiluted (SSD) and diluted (SD) (Storfish(®) 1 : 2v/v; IMV AI solutions, France) - were stored at 4 °C for 14 days. Motility, viability (PMI: plasma membrane integrity) and mitochondrial membrane potential (??M) were assessed. On the fifth day of storage, spermatozoa showed a motility >70% (SSD: 78.3% versus SD 85.0%), PMI (81.5% SSD/87.2% SD) and ??M (72.5% SSD/SD 80.0%) (P < 0.05). However, a significant decline in the percentage of all functional parameters (P < 0.05) was observed after 5 days of storage for all samples of both undiluted (SSD) and diluted semen. In conclusion, the results here provide new data on O. mykiss sperm quality with respect to in vitro short-term storage evaluated by flow cytometry. PMID:24717099

  12. Role of the epididymis in sperm competition.

    PubMed

    Jones, Russell C; Dacheux, Jean-Louis; Nixon, Brett; Ecroyd, Heath W

    2007-07-01

    Although it is generally understood that the testes recruited kidney ducts for reproductive function during the evolution of vertebrates, little is understood of the biological significance of the adaptation. In the context of the evolution of the mammalian epididymis, this report provides evidence that a major role of the epididymis is to enhance a male's chance of achieving paternity in a competitive mating system. A unique example of sperm cooperation in monotremes is used as evidence that the epididymis produces sperm competition proteins to form groups of 100 sperm into bundles that have a forward motility nearly thrice that of individual spermatozoa. As it required 3-h incubation in vitro under capacitation conditions to release motile sperm from the bundles, it is suggested that the monotremes provide an example of capacitation that is quite different from capacitation in higher mammals. It is suggested that variation between species in the intensity of sperm competition could explain the variation that occurs between species in the amount of post-testicular sperm maturation and storage in the epididymis, an explanation of why the human epididymis does not play as important a role in reproduction as the epididymis of most mammals. PMID:17589786

  13. Oxidative phosphorylation is essential for felid sperm function, but is substantially lower in cheetah (Acinonyx jubatus) compared to domestic cat (Felis catus) ejaculate.

    PubMed

    Terrell, Kimberly A; Wildt, David E; Anthony, Nicola M; Bavister, Barry D; Leibo, S P; Penfold, Linda M; Marker, Laurie L; Crosier, Adrienne E

    2011-09-01

    Compared with the normospermic domestic cat, sperm metabolic function is compromised in the teratospermic cat and cheetah, but the pathway(s) involved in this deficiency are unknown. Glycolysis is essential for sperm motility, yet it appears to function normally in spermatozoa of either species regardless of structural morphology. We conducted a comparative study to further understand the mechanisms of energy production in felid spermatozoa, with the hypothesis that oxidative phosphorylation is required for normal sperm function and is impaired in teratospermic ejaculates. Electroejaculates from both species were stained with MitoTracker to quantify mitochondrial membrane potential (MMP) or were incubated to assess changes in sperm function (motility, acrosomal integrity, and lactate production) after mitochondrial inhibition with myxothiazol. Sperm midpiece dimensions also were quantified. Sperm mitochondrial fluorescence (directly proportional to MMP) was ~95% lower in the cheetah compared with the normospermic and teratospermic cat, despite the cheetah having a 10% longer midpiece. In both species, MMP was increased 5-fold in spermatozoa with retained cytoplasm compared with structurally normal cells. Inhibition of oxidative phosphorylation impaired sperm function in both species, but a 100-fold higher inhibitor concentration was required in the cat compared with the cheetah. Collectively, findings revealed that oxidative phosphorylation was required for sperm function in the domestic cat and cheetah. This pathway of energy production appeared markedly less active in the cheetah, indicating a species-specific vulnerability to mitochondrial dysfunction. The unexpected, cross-species linkage between retained cytoplasmic droplets and elevated MMP may reflect increased concentrations of metabolic enzymes or substrates in these structures. PMID:21593479

  14. Modern vestibular function testing.

    PubMed Central

    Baloh, R W; Furman, J M

    1989-01-01

    Current tests of vestibular function concentrate on the horizontal semicircular canal-ocular reflex because it is the easiest reflex to stimulate (calorically and rotationally) and record (using electro-oculography). Tests of the other vestibulo-ocular reflexes (vertical semicircular canal and otolith) and of the vestibulospinal reflexes have yet to be shown useful in the clinical setting. Digital video recording of eye movements and vestibular-evoked responses are promising new technologies that may affect clinical testing in the near future. PMID:2660408

  15. Platelet Function Tests

    MedlinePLUS

    ... services. Advertising & Sponsorship: Policy | Opportunities PLEASE NOTE: Your web browser does not have JavaScript enabled. Unless you enable Javascript , your ability to navigate and access the features of this website will be ... Platelet Function Tests Share this page: Was this page helpful? Also ...

  16. Natural Variants of C. elegans Demonstrate Defects in Both Sperm Function and Oogenesis at Elevated Temperatures

    PubMed Central

    Petrella, Lisa N.

    2014-01-01

    The temperature sensitivity of the germ line is conserved from nematodes to mammals. Previous studies in C. briggsae and Drosophila showed that isolates originating from temperate latitudes lose fertility at a lower temperature than strains originating from tropical latitudes. In order to investigate these relationships in C. elegans, analysis of the fertility of 22 different wild-type isolates of C. elegans isolated from equatorial, tropical and temperate regions was undertaken. It was found that there are significant temperature, genotype and temperature × genotype effects on fertility but region of isolation showed no significant effect on differences in fertility. For most isolates 100% of the population maintained fertility from 20°C to 26°C, but there was a precipitous drop in the percentage of fertile hermaphrodites at 27°C. In contrast, all isolates show a progressive decrease in brood size as temperature increases from 20°C to 26°C, followed by a brood size near zero at 27°C. Temperature shift experiments were performed to better understand the causes of high temperature loss of fertility. Males up-shifted to high temperature maintained fertility, while males raised at high temperature lost fertility. Down-shifting males raised at high temperature generally did not restore fertility. This result differs from that observed in Drosophila and suggested that in C. elegans spermatogenesis or sperm function is irreversibly impaired in males that develop at high temperature. Mating and down-shifting experiments with hermaphrodites were performed to investigate the relative contributions of spermatogenic and oogenic defects to high temperature loss of fertility. It was found that the hermaphrodites of all isolates demonstrated loss in both spermatogenic and oogenic germ lines that differed in their relative contribution by isolate. These studies uncovered unexpectedly high variation in both the loss of fertility and problems with oocyte function in natural variants of C. elegans at high temperature. PMID:25380048

  17. The small RNA content of human sperm reveals pseudogene-derived piRNAs complementary to protein-coding genes.

    PubMed

    Pantano, Lorena; Jodar, Meritxell; Bak, Mads; Ballescà, Josep Lluís; Tommerup, Niels; Oliva, Rafael; Vavouri, Tanya

    2015-06-01

    At the end of mammalian sperm development, sperm cells expel most of their cytoplasm and dispose of the majority of their RNA. Yet, hundreds of RNA molecules remain in mature sperm. The biological significance of the vast majority of these molecules is unclear. To better understand the processes that generate sperm small RNAs and what roles they may have, we sequenced and characterized the small RNA content of sperm samples from two human fertile individuals. We detected 182 microRNAs, some of which are highly abundant. The most abundant microRNA in sperm is miR-1246 with predicted targets among sperm-specific genes. The most abundant class of small noncoding RNAs in sperm are PIWI-interacting RNAs (piRNAs). Surprisingly, we found that human sperm cells contain piRNAs processed from pseudogenes. Clusters of piRNAs from human testes contain pseudogenes transcribed in the antisense strand and processed into small RNAs. Several human protein-coding genes contain antisense predicted targets of pseudogene-derived piRNAs in the male germline and these piRNAs are still found in mature sperm. Our study provides the most extensive data set and annotation of human sperm small RNAs to date and is a resource for further functional studies on the roles of sperm small RNAs. In addition, we propose that some of the pseudogene-derived human piRNAs may regulate expression of their parent gene in the male germline. PMID:25904136

  18. Automated detection of sperm whale sounds as a function of abrupt changes in sound intensity

    NASA Astrophysics Data System (ADS)

    Walker, Christopher D.; Rayborn, Grayson H.; Brack, Benjamin A.; Kuczaj, Stan A.; Paulos, Robin L.

    2003-04-01

    An algorithm designed to detect abrupt changes in sound intensity was developed and used to identify and count sperm whale vocalizations and to measure boat noise. The algorithm is a MATLAB routine that counts the number of occurrences for which the change in intensity level exceeds a threshold. The algorithm also permits the setting of a ``dead time'' interval to prevent the counting of multiple pulses within a single sperm whale click. This algorithm was used to analyze digitally sampled recordings of ambient noise obtained from the Gulf of Mexico using near bottom mounted EARS buoys deployed as part of the Littoral Acoustic Demonstration Center experiment. Because the background in these data varied slowly, the result of the application of the algorithm was automated detection of sperm whale clicks and creaks with results that agreed well with those obtained by trained human listeners. [Research supported by ONR.

  19. Hyaluronidase 2: A Novel Germ Cell Hyaluronidase with Epididymal Expression and Functional Roles in Mammalian Sperm1

    PubMed Central

    Modelski, Mark J.; Menlah, Gladys; Wang, Yipei; Dash, Soma; Wu, Kathie; Galileo, Deni S.; Martin-DeLeon, Patricia A.

    2014-01-01

    ABSTRACT To initiate the crucial cell adhesion events necessary for fertilization, sperm must penetrate extracellular matrix barriers containing hyaluronic acid (HA), a task thought to be accomplished by neutral-active hyaluronidases. Here we report that the ?57 kDa hyaluronidase 2 (HYAL2) that in somatic tissues has been highly characterized to be acid-active is present in mouse and human sperm, as detected by Western blot, flow cytometric, and immunoprecipitation assays. Immunofluorescence revealed its presence on the plasma membrane over the acrosome, the midpiece, and proximal principal piece in mice where protein fractionation demonstrated a differential distribution in subcellular compartments. It is significantly more abundant in the acrosome-reacted (P = 0.04) and soluble acrosomal fractions (P = 0.006) (microenvironments where acid-active hyaluronidases function) compared to that of the plasma membrane where neutral hyaluronidases mediate cumulus penetration. Using HA substrate gel electrophoresis, immunoprecipitated HYAL 2 was shown to have catalytic activity at pH 4.0. Colocalization and coimmunoprecipitation assays reveal that HYAL2 is associated with its cofactor, CD44, consistent with CD44-dependent HYAL2 activity. HYAL2 is also present throughout the epididymis, where Hyal2 transcripts were detected, and in the epididymal luminal fluids. In vitro assays demonstrated that HYAL2 can be acquired on the sperm membrane from epididymal luminal fluids, suggesting that it plays a role in epididymal maturation. Because similar biphasic kinetics are seen for HYAL2 and SPAM1 (Sperm adhesion molecule 1), it is likely that HYAL2 plays a redundant role in the catalysis of megadalton HA to its 20 kDa intermediate during fertilization. PMID:25232017

  20. Mitochondrial fusion protein MFN2 interacts with the mitostatin-related protein MNS1 required for mouse sperm flagellar structure and function

    PubMed Central

    2014-01-01

    Background Cilia and the sperm flagellum share many structural properties. Meiosis-specific nuclear structural 1 (MNS1) is a recently characterized protein that is abundantly expressed in post-meiotic spermatids and is required for proper flagellar and motile cilia formation. To explore the possible functions of MNS1, we performed a BLAST search and determined it is homologous to the conserved domain pfam13868, exemplified by mitostatin. This protein interacts with mitofusin 2 (MFN2), a protein that participates in regulating mitochondrial associations to subcellular organelles. We hypothesized that an association between MFN2 and MNS1 in the sperm is involved in flagellar biogenesis and function. Results In the studies reported here, MFN2 was found in murine reproductive and somatic tissues high in ciliary content while MNS1 was present as two closely migrating bands in reproductive tissues. Interestingly, mitostatin was also present in reproductive tissues. Similar to Mns1 and mitostatin, Mfn2 was expressed in the testis as detected by RT-PCR. In addition, Mfn2 and Mns1 decreased in expression from pachytene spermatocytes to condensing spermatids as assessed by quantitative RT-PCR. Co-immunoprecipitation demonstrated an association between MFN2 and MNS1 in spermatogenic cells. Indirect immunofluorescence indicated that MFN2 and MNS1 co-localized to the sperm flagellum in freshly collected cauda epididymal sperm. MFN2 associated with the midpiece while MNS1 was present throughout the sperm tail in caput and cauda epididymal sperm. In spermatogenic cells, MFN2 was seen in the mitochondria, and MNS1 was present throughout the cell cytoplasm. MFN2 and MNS1 were present in detergent-resistant flagellar structures of the sperm. Conclusions These results demonstrate that MFN2 and MNS1 are present in spermatogenic cells and are an integral part of the sperm flagellum, indicating they play a role in flagellar biogenesis and/or function. PMID:24876927

  1. Aneuploidy in human sperm: The use of multicolor FISH to test various theories of nondisjunction

    SciTech Connect

    Spriggs, E.L.; Martin, R.H.

    1996-02-01

    While it is known that all chromosomes are susceptible to meiotic nondisjunction, it is not clear whether all chromosomes display the same frequency of nondisjunction. By use of multicolor FISH and chromosome-specific probes, the frequency of disomy in human sperm was determined for chromosomes 1, 2, 4, 9, 12, 15, 16, 18, 20, and 21, and the sex chromosomes. A minimum of 10,000 sperm nuclei were scored from each of five healthy, chromosomally normal donors for every chromosome studied, giving a total of 418,931 sperm nuclei. The mean frequencies of disomy obtained were 0.09% for chromosome 1; 0.08% for chromosome 2; 0.11% for chromosome 4; 0.14% for chromosome 9; 0.16% for chromosome 12; 0.11% for chromosomes 15, 16, and 18; 0.12% for chromosome 20; 0.29% for chromosome 21; and 0.43% for the sex chromosomes. Data for chromosomes 1, 12, 15, and 18, and the sex chromosomes have been published elsewhere. When the mean frequencies of disomy were compared, the sex chromosomes and chromosome 21 had significantly higher frequencies of disomy than that of any other autosome studied. These results corroborate the pooled data obtained from human sperm karyotypes and suggest that the sex chromosome bivalent and the chromosome 21 bivalent are more susceptible to nondisjunction during spermatogenesis. From these findings, theories proposed to explain the variable incidence of nondisjunction can be supported or discarded as improbable. 33 refs., 4 tabs.

  2. Effects of reactive oxygen species action on sperm function in spermatozoa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Reactive oxygen species (ROS) formation and lipid peroxidation have been recognized as problems for sperm survival and fertility. The precise roles and detection of superoxide (SO), hydrogen peroxide (HP), and membrane lipid peroxidation have been problematic because of the low specificity and sens...

  3. Sperm Proteome Maturation in the Mouse Epididymis

    PubMed Central

    Skerget, Sheri; Rosenow, Matthew A.; Petritis, Konstantinos; Karr, Timothy L.

    2015-01-01

    In mammals, transit through the epididymis, which involves the acquisition, loss and modification of proteins, is required to confer motility and fertilization competency to sperm. The overall dynamics of maturation is poorly understood, and a systems level understanding of the complex maturation process will provide valuable new information about changes occurring during epididymal transport. We report the proteomes of sperm collected from the caput, corpus and cauda segments of the mouse epididymis, identifying 1536, 1720 and 1234 proteins respectively. This study identified 765 proteins that are present in sperm obtained from all three segments. We identified 1766 proteins that are potentially added (732) or removed (1034) from sperm during epididymal transit. Phenotypic analyses of the caput, corpus and cauda sperm proteomes identified 60 proteins that have known sperm phenotypes when mutated, or absent from sperm. Our analysis indicates that as much as one-third of proteins with known sperm phenotypes are added to sperm during epididymal transit. GO analyses revealed that cauda sperm are enriched for specific functions including sperm-egg recognition and motility, consistent with the observation that sperm acquire motility and fertilization competency during transit through the epididymis. In addition, GO analyses revealed that the immunity protein profile of sperm changes during sperm maturation. Finally, we identified components of the 26S proteasome, the immunoproteasome, and a proteasome activator in mature sperm. PMID:26556802

  4. How is a giant sperm ejaculated? Anatomy and function of the sperm pump, or "Zenker organ," in Pseudocandona marchica (Crustacea, Ostracoda, Candonidae)

    NASA Astrophysics Data System (ADS)

    Yamada, Shinnosuke; Matzke-Karasz, Renate

    2012-07-01

    `Giant sperm', in terms of exceptionally long spermatozoa, occur in a variety of taxa in the animal kingdom, predominantly in arthropod groups, but also in flatworms, mollusks, and others. In some freshwater ostracods (Cypridoidea), filamentous sperm cells reach up to ten times the animal's body length; nonetheless, during a single copulation several dozen sperm cells can be transferred to the female's seminal receptacle. This highly effective ejaculation has traditionally been credited to a chitinous-muscular structure within the seminal duct, which has been interpreted as a sperm pump. We investigated this organ, also known as the Zenker organ, of a cypridoid ostracod, Pseudocandona marchica, utilizing light and electron microscope techniques and produced a three-dimensional reconstruction based on serial semi-thin histological sections. This paper shows that numerous muscle fibers surround the central tube of the Zenker organ, running in parallel with the central tube and that a thin cellular layer underlies the muscular layer. A cellular inner tube exists inside the central tube. A chitinous-cellular structure at the entrance of the organ has been recognized as an ejaculatory valve. In male specimens during copulation, we confirmed a small hole derived from the passage of a single spermatozoon through the valve. The new data allowed for proposing a detailed course of operation of the Zenker organ during giant sperm ejaculation.

  5. Di-(2-ethylhexyl)-phthalate disrupts pituitary and testicular hormonal functions to reduce sperm quality in mature goldfish.

    PubMed

    Golshan, Mahdi; Hatef, Azadeh; Socha, Magdalena; Milla, Sylvain; Butts, Ian A E; Carnevali, Oliana; Rodina, Marek; Soko?owska-Miko?ajczyk, Miros?awa; Fontaine, Pascal; Linhart, Otomar; Alavi, Sayyed Mohammad Hadi

    2015-06-01

    Di-(2-ethylhexyl) phthalate (DEHP) interferes with male reproductive endocrine system in mammals, however its effects on fish reproduction are largely unknown. We evaluated sperm quality and investigated reproductive endocrine system in mature goldfish (Carassius auratus) exposed to nominal 1, 10, and 100?g/L DEHP. To examine DEHP estrogenic activity, one group of goldfish was exposed to 17?-estradiol (5?g/L E2) for comparison. Following 30d of exposure, sperm production was decreased and suppressed in DEHP and E2 treated goldfish, respectively. Sperm motility and velocity were decreased in goldfish exposed to 100 and 10?g/L DEHP at 15s post-sperm activation, respectively. Compared to control, 11-ketotestosterone (11-KT) levels were decreased at 10 and 1?g/L DEHP at day 15 and 30, respectively. In E2 treated goldfish, 11-KT levels were decreased compared to control during the period of exposure. E2 levels were increased in goldfish exposed to E2, but remained unchanged in DEHP treated goldfish during the period of exposure. StAR mRNA levels encoding regulator of cholesterol transfer to steroidogenesis were decreased in DEHP and E2 treated goldfish following 15 and 30d of exposure, respectively. Luteinizing hormone (LH) levels were decreased in DEHP and E2 treated goldfish following 15 and 30d of exposure, respectively. In DEHP treated goldfish, gnrh3, kiss1 and its receptor (gpr54) mRNA levels did not change during the experimental period. In E2 treated goldfish, gnrh3 mRNA levels were decreased at day 7, but kiss1 and gpr54 mRNA levels were increased at day 30 of exposure. The mRNA levels of genes encoding testicular LH and androgen receptors remained unchanged in DEHP and E2 treated goldfish. In contrast to E2 treated goldfish, vitellogenin production was not induced in DEHP treated goldfish and mRNA levels of genes with products mediating estrogenic effects remained unchanged or decreased. In conclusion, DEHP interferes with testis and pituitary hormonal functions to reduce sperm quality in goldfish and does not exhibit estrogenic activity. PMID:25827748

  6. Defective Human Sperm Cells Are Associated with Mitochondrial Dysfunction and Oxidant Production.

    PubMed

    Cassina, Adriana; Silveira, Patricia; Cantu, Lidia; Montes, Jose Maria; Radi, Rafael; Sapiro, Rossana

    2015-11-01

    Infertility affects about 15% of couples of reproductive age. The male factor is involved in nearly 50% of infertility cases. Defective human sperm function has been associated with evidence of high levels of reactive oxygen species (ROS) and a resultant loss of fertilizing potential in vivo and in vitro. Analogous to what has been observed in somatic cells, mitochondria are likely the major sources of ROS in sperm cells. In this study, we analyzed mitochondrial function using high-resolution respirometry, ROS production, and footprints of oxidative and nitrative stress processes in intact human sperm cells. We showed that mitochondrial dysfunction (measured through the respiratory control ratio) was correlated with a decrease in human sperm motility. The samples analyzed presented nitro-oxidative modifications of proteins, such as protein 3-nitrotyrosine, that were observed mainly in the mid-piece (where mitochondria are localized) and in the sperm head. Semen samples presenting lower percentage of motile sperm showed higher amounts of nitro-oxidative protein modifications than those with larger quantities of motile sperm. When spermatozoa were exposed to inhibitors of the respiratory mitochondrial function, in the presence of a nitric oxide flux, sperm produced potent nitro-oxidative species (i.e., peroxynitrite). This effect was observed in more than 90% of intact living sperm cells and in sperm mitochondrial fractions. These data suggest that dysfunctional mitochondria in sperm cells produce oxidants that may contribute to male infertility. These data provide the rationale for testing the potential of compounds that improve sperm mitochondrial function to treat male infertility. PMID:26447142

  7. The function of male sperm whale slow clicks in a high latitude habitat: communication, echolocation, or prey debilitation?

    PubMed

    Oliveira, Cláudia; Wahlberg, Magnus; Johnson, Mark; Miller, Patrick J O; Madsen, Peter T

    2013-05-01

    Sperm whales produce different click types for echolocation and communication. Usual clicks and buzzes appear to be used primarily in foraging while codas are thought to function in social communication. The function of slow clicks is less clear, but they appear to be produced by males at higher latitudes, where they primarily forage solitarily, and on the breeding grounds, where they roam between groups of females. Here the behavioral context in which these vocalizations are produced and the function they may serve was investigated. Ninety-nine hours of acoustic and diving data were analyzed from sound recording tags on six male sperm whales in Northern Norway. The 755 slow clicks detected were produced by tagged animals at the surface (52%), ascending from a dive (37%), and during the bottom phase (11%), but never during the descent. Slow clicks were not associated with the production of buzzes, other echolocation clicks, or fast maneuvering that would indicate foraging. Some slow clicks were emitted in seemingly repetitive temporal patterns supporting the hypothesis that the function for slow clicks on the feeding grounds is long range communication between males, possibly relaying information about individual identity or behavioral states. PMID:23654416

  8. Protein-Tyrosine Kinase Signaling in the Biological Functions Associated with Sperm

    PubMed Central

    Ijiri, Takashi W.; Mahbub Hasan, A. K. M.; Sato, Ken-ichi

    2012-01-01

    In sexual reproduction, two gamete cells (i.e., egg and sperm) fuse (fertilization) to create a newborn with a genetic identity distinct from those of the parents. In the course of these developmental processes, a variety of signal transduction events occur simultaneously in each of the two gametes, as well as in the fertilized egg/zygote/early embryo. In particular, a growing body of knowledge suggests that the tyrosine kinase Src and/or other protein-tyrosine kinases are important elements that facilitate successful implementation of the aforementioned processes in many animal species. In this paper, we summarize recent findings on the roles of protein-tyrosine phosphorylation in many sperm-related processes (from spermatogenesis to epididymal maturation, capacitation, acrosomal exocytosis, and fertilization). PMID:23209895

  9. Methamidophos alters sperm function and DNA at different stages of spermatogenesis in mice.

    PubMed

    Urióstegui-Acosta, Mayrut; Hernández-Ochoa, Isabel; Sánchez-Gutiérrez, Manuel; Piña-Guzmán, Belem; Rafael-Vázquez, Leticia; Solís-Heredia, M J; Martínez-Aguilar, Gerardo; Quintanilla-Vega, Betzabet

    2014-09-15

    Methamidophos (MET) is a highly toxic organophosphate (OP) pesticide that is widely used in developing countries. MET has male reproductive effects, including decreased fertility. We evaluated MET effects on sperm quality, fertilization and DNA integrity, exploring the sensitivity of different stages of spermatogenesis. Adult male mice received MET (3.75 or 5mg/kg-bw/ip/day/4 days) and were euthanized 1, 28 or 45 days post-treatment (dpt) to evaluate MET's effects on epididymal maturation, meiosis or mitosis, respectively. Spermatozoa were obtained from the cauda epididymis-vas deferens and were evaluated for sperm quality, acrosome reaction (AR; Coomassie staining), mitochondrial membrane potential (by JC-1), DNA damage (comet assay), oxidative damage (malondialdehyde (MDA) production), in vitro fertilization and protein phosphorylation (immunodetection), and erythrocyte acetylcholinesterase (AChE) activity. At 1-dpt, MET inhibited AChE (43-57%) and increased abnormal cells (6%). While at 28- and 45-dpt, sperm motility and viability were significantly reduced with an increasing MET dose, and abnormal morphology increased at 5mg/kg/day/4 days. MDA and mitochondrial activity were not affected at any dose or time. DNA damage (OTM and %DNA) was observed at 5mg/kg/day/4 days in a time-dependent manner, whereas both parameters were altered in cells from mice exposed to 3.75 mg/kg/day/4 days only at 28-dpt. Depending on the time of collection, initial-, spontaneous- and induced-AR were altered at 5mg/kg/day/4 days, and the fertilization capacity also decreased. Sperm phosphorylation (at serine and tyrosine residues) was observed at all time points. Data suggest that meiosis and mitosis are the more sensitive stages of spermatogenesis for MET reproductive toxicity compared to epididymal maturation. PMID:24998973

  10. Mammalian Sperm Fertility Related Proteins

    PubMed Central

    Ashrafzadeh, Ali; Karsani, Saiful Anuar; Nathan, Sheila

    2013-01-01

    Infertility is an important aspect of human and animal reproduction and still presents with much etiological ambiguity. As fifty percent of infertility is related to the male partner, molecular investigations on sperm and seminal plasma can lead to new knowledge on male infertility. Several comparisons between fertile and infertile human and other species sperm proteome have shown the existence of potential fertility markers. These proteins have been categorized into energy related, structural and other functional proteins which play a major role in sperm motility, capacitation and sperm-oocyte binding. The data from these studies show the impact of sperm proteome studies on identifying different valuable markers for fertility screening. In this article, we review recent development in unraveling sperm fertility related proteins. PMID:24151436

  11. Pulmonary Function Tests

    MedlinePLUS

    ... test. 4 If you have difficulty with closed spaces (claustrophobia), let the test center know in case one of the tests involves being enclosed 4 Ask if there are any medicines you should stop taking before being tested and ...

  12. Mammalian sperm morphometry Matthew J. G. Gage

    E-print Network

    Cambridge, University of

    Mammalian sperm morphometry Matthew J. G. Gage Population Biology Research Group, School.ac.uk) Understanding the adaptive signi¢cance of sperm form and function has been a challenge to biologists because sperm are highly specialized cells operating at a microscopic level in a complex environment. A fruitful

  13. Cytometric analysis of shape and DNA content in mammalian sperm

    SciTech Connect

    Gledhill, B.L.

    1983-10-10

    Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. Sperm were analyzed by flow cytometry and slit-scan flow analysis for injury following the exposure of testes to mutagens. The utility of flow cytometry in genotoxin screening and monitoring of occupational exposure was evaluated. The technique proved valuable in separation of X- and Y-chromosome bearing sperm and the potential applicability of this technique in artificial insemination and a solution, of accurately assessing the DNA content of sperm were evaluated-with reference to determination of X- and Y-chromosome bearing sperm.

  14. Subcellular preservation in giant ostracod sperm from an early Miocene cave deposit in Australia.

    PubMed

    Matzke-Karasz, Renate; Neil, John V; Smith, Robin J; Symonová, Radka; Mo?kovský, Libor; Archer, Michael; Hand, Suzanne J; Cloetens, Peter; Tafforeau, Paul

    2014-07-01

    Cypridoidean ostracods are one of a number of animal taxa that reproduce with giant sperm, up to 10 000 µm in length, but they are the only group to have aflagellate, filamentous giant sperm. The evolution and function of this highly unusual feature of reproduction with giant sperm are currently unknown. The hypothesis of long-term evolutionary persistence of this kind of reproduction has never been tested. We here report giant sperm discovered by propagation phase contrast X-ray synchrotron micro- and nanotomography, preserved in five Miocene ostracod specimens from Queensland, Australia. The specimens belong to the species Heterocypris collaris Matzke-Karasz et al. 2013 (one male and three females) and Newnhamia mckenziana Matzke-Karasz et al. 2013 (one female). The sperm are not only the oldest petrified gametes on record, but include three-dimensional subcellular preservation. We provide direct evidence that giant sperm have been a feature of this taxon for at least 16 Myr and provide an additional criterion (i.e. longevity) to test hypotheses relating to origin and function of giant sperm in the animal kingdom. We further argue that the highly resistant, most probably chitinous coats of giant ostracod sperm may play a role in delaying decay processes, favouring early mineralization of soft tissue. PMID:24827442

  15. Subcellular preservation in giant ostracod sperm from an early Miocene cave deposit in Australia

    PubMed Central

    Matzke-Karasz, Renate; Neil, John V.; Smith, Robin J.; Symonová, Radka; Mo?kovský, Libor; Archer, Michael; Hand, Suzanne J.; Cloetens, Peter; Tafforeau, Paul

    2014-01-01

    Cypridoidean ostracods are one of a number of animal taxa that reproduce with giant sperm, up to 10 000 µm in length, but they are the only group to have aflagellate, filamentous giant sperm. The evolution and function of this highly unusual feature of reproduction with giant sperm are currently unknown. The hypothesis of long-term evolutionary persistence of this kind of reproduction has never been tested. We here report giant sperm discovered by propagation phase contrast X-ray synchrotron micro- and nanotomography, preserved in five Miocene ostracod specimens from Queensland, Australia. The specimens belong to the species Heterocypris collaris Matzke-Karasz et al. 2013 (one male and three females) and Newnhamia mckenziana Matzke-Karasz et al. 2013 (one female). The sperm are not only the oldest petrified gametes on record, but include three-dimensional subcellular preservation. We provide direct evidence that giant sperm have been a feature of this taxon for at least 16 Myr and provide an additional criterion (i.e. longevity) to test hypotheses relating to origin and function of giant sperm in the animal kingdom. We further argue that the highly resistant, most probably chitinous coats of giant ostracod sperm may play a role in delaying decay processes, favouring early mineralization of soft tissue. PMID:24827442

  16. Effect of male age on sperm traits and sperm competition success in the guppy (Poecilia reticulata).

    PubMed

    Gasparini, C; Marino, I A M; Boschetto, C; Pilastro, A

    2010-01-01

    Deleterious mutations can accumulate in the germline with age, decreasing the genetic quality of sperm and imposing a cost on female fitness. If these mutations also affect sperm competition ability or sperm production, then females will benefit from polyandry as it incites sperm competition and, consequently, minimizes the mutational load in the offspring. We tested this hypothesis in the guppy (Poecilia reticulata), a species characterized by polyandry and intense sperm competition, by investigating whether age affects post-copulatory male traits and sperm competition success. Females did not discriminate between old and young males in a mate choice experiment. While old males produced longer and slower sperm with larger reserves of strippable sperm, compared to young males, artificial insemination did not reveal any effect of age on sperm competition success. Altogether, these results do not support the hypothesis that polyandry evolved in response to costs associated with mating with old males in the guppy. PMID:19912453

  17. Thyroid Function Tests.

    ERIC Educational Resources Information Center

    Glover, Irving T.

    1979-01-01

    Describes two tests, T-4 and T-3, for hypothyroid based on the binding of the hormones by proteins. The tests were performed in courses for physicians, clinical chemists, laboratory technicians, and undergraduate science students by the individuals involved and on their own sera. These tests are commercially available in kit form. (GA)

  18. No evidence for sperm priming responses under varying sperm competition risk or intensity in guppies

    NASA Astrophysics Data System (ADS)

    Evans, Jonathan P.

    2009-07-01

    Sperm competition theory predicts that males should tailor their investment in ejaculates according to the number of rival males competing to fertilize a female’s eggs. Research spanning several taxa supports this prediction by showing that males are often sensitive to the level of sperm competition and adjust their investment in sperm numbers accordingly. More recent work has revealed that males may also tailor the quality of sperm according to the number of males competing for fertilization. Here I test for both effects in guppies ( Poecilia reticulata) in an experiment that simultaneously evaluates the risk and intensity models of sperm competition. The experiment determined whether male guppies adjust the number (stripped ejaculate size) and quality (sperm velocity and viability) of sperm that are primed over a 3-day period according to experimental changes in the perceived level of sperm competition. A total of 136 focal males were initially stripped of all retrievable sperm and assayed for these sperm traits before being allocated at random to one of four treatments simulating different levels of sperm competition risk and intensity. During the 3-day treatment phase, focal males had visual and olfactory access to a sexually receptive (initially virgin) female maintained with different numbers of stimulus males to simulate variation in the risk and intensity of sperm competition. Following this, males were assayed again for the sperm traits. Contrary to predictions, there was no significant change in any of the measured variables among treatments, although qualitatively the patterns for sperm velocity and viability did conform to expectation. The lack of any trend for the number of sperm primed was unequivocal and future work examining the effects of sperm competition on sperm production should focus on whether males differentially allocate sperm numbers among matings that differ in the level of sperm competition.

  19. Osmotic tolerance of avian spermatozoa: Influence of time, temperature, cryoprotectant and membrane ion pump function on sperm viability

    USGS Publications Warehouse

    Blanco, J.M.; Long, J.A.; Gee, G.; Donoghue, A.M.; Wildt, D.E.

    2008-01-01

    Potential factors influencing sperm survival under hypertonic conditions were evaluated in the Sandhill crane (Grus canadensis) and turkey (Meleagridis gallopavo). Sperm osmotolerance (300-3000 mOsm/kg) was evaluated after: (1) equilibration times of 2, 10, 45 and 60 min at 4 ?C versus 21 ?C; (2) pre-equilibrating with dimethylacetamide (DMA) or dimethylsulfoxide (Me2SO) at either 4 ?C or 21 ?C; and (3) inhibition of the Na+/K+ and the Na+/H+ antiporter membrane ionic pumps. Sperm viability was assessed using the eosin-nigrosin live/dead stain. Species-specific differences occurred in response to hypertonic conditions with crane sperm remaining viable under extreme hypertonicity (3000 mOsm/kg), whereas turkey sperm viability was compromised with only slightly hypertonic (500 mOsm/kg) conditions. The timing of spermolysis under hypertonic conditions was also species-specific, with a shorter interval for turkey (2 min) than crane (10 min) sperm. Turkey sperm osmotolerance was slightly improved by lowering the incubation temperature from 21 to 4 ?C. Pre-equilibrating sperm with DMA reduced the incidence of hypertonic spermolysis only in the crane, at both room and refrigeration temperature. Inhibiting the Na+/K+ and the Na+/H+ antiporter membrane ion pumps did not impair resistance of crane and turkey spermatozoa to hypertonic stress; pump inhibition actually increased turkey sperm survival compared to control sperm. Results demonstrate marked species specificity in osmotolerance between crane and turkey sperm, as well as in the way temperature and time of exposure affect sperm survival under hypertonic conditions. Differences are independent of the role of osmotic pumps in these species.

  20. The use of a hypo-osmotic swelling (HOS) test on sperm of the pig (Sus scrofa domesticus), emu (Dromaius novaehollandiae), Asian elephant (Elephas maximus), hamadryas baboon (Papio hamadryas hamadryas), and central rock rat (Zyzomys pedunculatus).

    PubMed

    Matson, Phillip; Kappelle, Wendy; Malecki, Irek

    2009-07-01

    A hypo-osmotic swelling test using TALP-HEPES medium over a range of 50 to 300 mOsm/kg was applied to sperm from domestic and endangered species. Maximal responses of curling of the sperm tails were seen over a range of osmolalities for epididymal sperm from the pig (100 mOsm/kg), hamadryas baboon (range 50-125 mOsm/kg), and central rock rat (range 50-100 mOsm/kg), and the ejaculated sperm from the emu (50 mOsm/kg) and the Asian elephant (range 75-150 mOsm/kg). A solution of TALP-HEPES medium at 100 mOsm/kg would be suitable to obtain the maximal response in this range of mammals tested, though it would need to be diluted to at least 50 mOsm/kg when testing the viability of the emu sperm. PMID:19734955

  1. Drosophila pericentrin requires interaction with calmodulin for its function at centrosomes and neuronal basal bodies but not at sperm basal bodies

    PubMed Central

    Galletta, Brian J.; Guillen, Rodrigo X.; Fagerstrom, Carey J.; Brownlee, Chris W.; Lerit, Dorothy A.; Megraw, Timothy L.; Rogers, Gregory C.; Rusan, Nasser M.

    2014-01-01

    Pericentrin is a critical centrosomal protein required for organizing pericentriolar material (PCM) in mitosis. Mutations in pericentrin cause the human genetic disorder Majewski/microcephalic osteodysplastic primordial dwarfism type II, making a detailed understanding of its regulation extremely important. Germaine to pericentrin's function in organizing PCM is its ability to localize to the centrosome through the conserved C-terminal PACT domain. Here we use Drosophila pericentrin-like-protein (PLP) to understand how the PACT domain is regulated. We show that the interaction of PLP with calmodulin (CaM) at two highly conserved CaM-binding sites in the PACT domain controls the proper targeting of PLP to the centrosome. Disrupting the PLP-CaM interaction with single point mutations renders PLP inefficient in localizing to centrioles in cultured S2 cells and Drosophila neuroblasts. Although levels of PCM are unaffected, it is highly disorganized. We also demonstrate that basal body formation in the male testes and the production of functional sperm does not rely on the PLP-CaM interaction, whereas production of functional mechanosensory neurons does. PMID:25031429

  2. Functional Sperm of the Yellowtail (Seriola quinqueradiata) Were Produced in the Small-Bodied Surrogate, Jack Mackerel (Trachurus japonicus).

    PubMed

    Morita, Tetsuro; Morishima, Kagayaki; Miwa, Misako; Kumakura, Naoki; Kudo, Satomi; Ichida, Kensuke; Mitsuboshi, Toru; Takeuchi, Yutaka; Yoshizaki, Goro

    2015-10-01

    Production of xenogeneic gametes from large-bodied, commercially important marine species in closely related smaller surrogates with short generation times may enable rapid domestication of the targeted species. In this study, we aimed to produce gametes of Japanese yellowtail (Seriola quinqueradiata) using jack mackerel (Trachurus japonicus) as a surrogate with a smaller body size and shorter maturation period. Donor spermatogonia were collected from the testes of yellowtail males and transferred into the peritoneal cavity of 10- and 12-day-old jack mackerel larvae. Twenty days later, 59.5% of the recipients survived of which 88.2% had donor-derived germ cells in their gonads. One year later, genomic DNA templates were prepared from the semen of 96 male recipients and subjected to polymerase chain reaction (PCR) analyses using primers specific for the yellowtail vasa sequence, resulting in the detection of positive signals in semen from two recipients. The milt collected from the recipients was used for fertilization with yellowtail eggs. Of eight hatchlings obtained from the crosses, two were confirmed to be derived from donor yellowtail by DNA markers, although the others were gynogenetic diploids. These findings indicate that it is possible to produce donor-derived sperm in xenogeneic recipients with a smaller body size and shorter generation time by transplanting spermatogonia. Thus, the xenogeneic transplantation of spermatogonia might be a potential tool to produce gametes of large-bodied, commercially important fish, although the efficiency of the method requires further improvement. This is the first report demonstrating that donor-derived sperm could be produced in xenogeneic recipient via spermatogonial transplantation in carangid fishes. PMID:26239188

  3. Decoding mechanisms of loss of fertilization ability of cryopreserved mouse sperm

    NASA Astrophysics Data System (ADS)

    Gray, Jeffrey Earl

    Cryopreservation of mouse sperm is an important technology for management of biomedical research resources. Dramatic progress has been made recently in the development of protocols that combat mouse-strain specific reduction of IVF after cryopreservation. Equal emphasis, however, has not been placed on investigating the biological mechanisms underlying these improvements to IVF. This dissertation broadly investigates the basic question of how mouse-strain specific reduction of IVF occurs after cryopreservation, and how recently developed protocols prevent this process. My research investigated the effects of antioxidants, the cholesterol-acceptor CD, reduced calcium media, and TYH capacitation media on sperm function and oxidative stress after cryopreservation in a variety of mouse strains. I found that reduced IVF was associated with loss of capacitation-dependent sperm function in three strains, B6/J, B6/N, and 129X1, and CD improved sperm function and IVF in all three strains. These findings suggest that cryopreservation inhibits cholesterol efflux resulting in reduced IVF of many mouse strains. I also found that cryopreservation induces uniquely high production of mitochondrial H2O2 by B6/J sperm. H2O2 present in other cellular compartments of B6/J sperm was not elevated compared to other strains. High levels of mitochondrial H2O2 were associated with lipid peroxidation of the sperm head and inability to acrosome react. Antioxidants reduced mitochondrial H2O2 production, decreased sperm head lipid peroxidation, and improved acrosome reaction. The cryopreservation-induced increase in mitochondrial H2O2 production of B6/J and B6129XF1 sperm was associated with elevation of intracellular calcium after cryopreservation and dependent on mitochondrial metabolic substrates. Reducing intracellular calcium levels or removing mitochondrial metabolic substrates decreased mitochondrial H2O2 production and increased IVF rates of cryopreserved B6/J sperm. Many of the strains I tested exhibited increased H2O2 production after cryopreservation, but cryopreservation-induced H2O2 only interfered with IVF of B6/J sperm. This dissertation describes two means to improve IVF of cryopreserved sperm, mitigation of oxidative stress in B6/J sperm and improvement of capacitation-dependent sperm function for several mouse strains.

  4. Structure, material characteristics and function of the upper respiratory tract of the pygmy sperm whale.

    PubMed

    Davenport, John; Cotter, Liz; Rogan, Emer; Kelliher, Denis; Murphy, Colm

    2013-12-15

    Cetaceans are neckless, so the trachea is very short. The upper respiratory tract is separate from the mouth and pharynx, and the dorsal blowhole connects, via the vestibular and nasopalatine cavities, directly to the larynx. Toothed cetaceans (Odontoceti) are capable of producing sounds at depth, either for locating prey or for communication. It has been suggested that during dives, air from the lungs and upper respiratory tract can be moved to the vestibular and nasal cavities to permit sound generation to continue when air volume within these cavities decreases as ambient pressure rises. The pygmy sperm whale, Kogia breviceps, is a deep diver (500-1000 m) that is known to produce hunting clicks. Our study of an immature female shows that the upper respiratory tract is highly asymmetrical: the trachea and bronchi are extremely compressible, whereas the larynx is much more rigid. Laryngeal and tracheal volumes were established. Calculations based on Boyle's Law imply that all air from the lungs and bronchi would be transferred to the larynx and trachea by a depth of 270 m and that the larynx itself could not accommodate all respiratory air mass at a depth of 1000 m. This suggests that no respiratory air would be available for vocalisation. However, the bronchi, trachea and part of the larynx have a thick vascular lining featuring large, thin-walled vessels. We propose that these vessels may become dilated during dives to reduce the volume of the upper respiratory tract, permitting forward transfer of air through the larynx. PMID:24072789

  5. Sperm DNA fragmentation is related to sperm morphological staining patterns.

    PubMed

    Sá, Rosália; Cunha, Mariana; Rocha, Eduardo; Barros, Alberto; Sousa, Mário

    2015-10-01

    In this prospective comparative study, sperm DNA fragmentation (sDNAfrag) was compared at each step of a sequential semen preparation, with semen parameters according to their degree of severity. At each step (fractions) of the sequential procedure, sDNAfrag was determined: fresh (Raw), after gradient centrifugation, washing, and swim-up (SU) for 70 infertile men enrolled in intracytoplasmic sperm injection cycles. sDNAfrag significantly (P = 0.04; P < 0.0001) decreased throughout the steps of semen preparation, with centrifugation and washing not increasing it. A negative correlation to sperm motility was observed in Raw and SU fractions, and a higher sDNAfrag was observed in samples with lower semen quality. Our results confirm that the steps of the sequential procedure do not compromise sperm DNA integrity and progressively decreased sDNAfrag regardless of the sperm abnormality and that semen parameters with lower quality present higher sDNAfrag. Four distinct patterns were observed, of which the entire sperm head staining was the pattern most expressed in all studied fractions. Additionally, the sperm head gene-rich region staining pattern was reduced by the procedure. This suggests that pattern quantification might be a useful adjunct when performing sDNAfrag testing for male infertility. PMID:26278809

  6. Anatase titanium dioxide nanoparticles in mice: evidence for induced structural and functional sperm defects after short-, but not long-, term exposure

    PubMed Central

    Smith, Michelle A; Michael, Rowan; Aravindan, Rolands G; Dash, Soma; Shah, Syed I; Galileo, Deni S; Martin-DeLeon, Patricia A

    2015-01-01

    Titanium dioxide (TiO2) nanoparticles (TNPs) are widely used commercially and exist in a variety of products. To determine if anatase TNPs (ATNPs) in doses smaller than previously used reach the scrotum after entry in the body at a distant location and induce sperm defects, 100% ATNP (2.5 or 5 mg kg?1 body weight) was administered intraperitoneally to adult males for three consecutive days, followed by sacrifice 1, 2, 3, or 5 weeks later (long-) or 24, 48 or 120 h (short-term exposure). Transmission electron microscopy revealed the presence of ANTP in scrotal adipose tissues collected 120 h postinjection when cytokine evaluation showed an inflammatory response in epididymal tissues and fluid. At 120 h and up to 3 weeks postinjection, testicular histology revealed enlarged interstitial spaces. Significantly increased numbers of terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling-positive (apoptotic) germ (P = 0.002) and interstitial space cells (P = 0.04) were detected in treated males. Caudal epididymal sperm from the short-term, but not a long-term, arm showed significantly (P < 0.001) increased frequencies of flagellar abnormalities, excess residual cytoplasm (ERC), and unreacted acrosomes in treated versus controls (dose-response relationship). A novel correlation between ERC and unreacted acrosomes was uncovered. At 120 h, there were significant decreases in hyperactivated motility (P < 0.001) and mitochondrial membrane potential (P < 0.05), and increased reactive oxygen species levels (P < 0.00001) in treated versus control sperm. These results indicate that at 4–8 days postinjection, ANTP induce structural and functional sperm defects associated with infertility, and DNA damage via oxidative stress. Sperm defects were transient as they were not detected 10 days to 5 weeks postinjection. PMID:25370207

  7. Anatase titanium dioxide nanoparticles in mice: evidence for induced structural and functional sperm defects after short-, but not long-, term exposure.

    PubMed

    Smith, Michelle A; Michael, Rowan; Aravindan, Rolands G; Dash, Soma; Shah, Syed I; Galileo, Deni S; Martin-DeLeon, Patricia A

    2015-01-01

    Titanium dioxide (TiO 2 ) nanoparticles (TNPs) are widely used commercially and exist in a variety of products. To determine if anatase TNPs (ATNPs) in doses smaller than previously used reach the scrotum after entry in the body at a distant location and induce sperm defects, 100% ATNP (2.5 or 5 mg kg-1 body weight) was administered intraperitoneally to adult males for three consecutive days, followed by sacrifice 1, 2, 3, or 5 weeks later (long-) or 24, 48 or 120 h (short-term exposure). Transmission electron microscopy revealed the presence of ANTP in scrotal adipose tissues collected 120 h postinjection when cytokine evaluation showed an inflammatory response in epididymal tissues and fluid. At 120 h and up to 3 weeks postinjection, testicular histology revealed enlarged interstitial spaces. Significantly increased numbers of terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling-positive (apoptotic) germ (P = 0.002) and interstitial space cells (P = 0.04) were detected in treated males. Caudal epididymal sperm from the short-term, but not a long-term, arm showed significantly (P < 0.001) increased frequencies of flagellar abnormalities, excess residual cytoplasm (ERC), and unreacted acrosomes in treated versus controls (dose-response relationship). A novel correlation between ERC and unreacted acrosomes was uncovered. At 120 h, there were significant decreases in hyperactivated motility (P < 0.001) and mitochondrial membrane potential (P < 0.05), and increased reactive oxygen species levels (P < 0.00001) in treated versus control sperm. These results indicate that at 4-8 days postinjection, ANTP induce structural and functional sperm defects associated with infertility, and DNA damage via oxidative stress. Sperm defects were transient as they were not detected 10 days to 5 weeks postinjection. PMID:25370207

  8. Exposure in utero to 2,2',3,3',4,6'-hexachlorobiphenyl (PCB 132) impairs sperm function and alters testicular apoptosis-related gene expression in rat offspring

    SciTech Connect

    Hsu, P.-C.; Pan, M.-H.; Li, L.-A.; Chen, C.-J.; Tsai, S.-S.; Guo, Y.L. . E-mail: leonguo@ha.mc.ntu.edu.tw

    2007-05-15

    Toxicity of the polychlorinated biphenyls (PCBs) depends on their molecular structure. Mechanisms by prenatal exposure to a non-dioxin-like PCB, 2,2',3,4',5',6-hexachlorobiphenyl (PCB 132) that may act on reproductive pathways in male offspring are relatively unknown. The purpose was to determine whether epididymal sperm function and expression of apoptosis-related genes were induced or inhibited by prenatal exposure to PCB 132. Pregnant rats were treated with a single dose of PCB 132 at 1 or 10 mg/kg on gestational day 15. Male offspring were killed and the epididymal sperm counts, motility, velocity, reactive oxygen species (ROS) generation, sperm-oocyte penetration rate (SOPR), testicular histopathology, apoptosis-related gene expression and caspase activation were assessed on postnatal day 84. Prenatal exposure to PCB 132 with a single dose of 1 or 10 mg/kg decreased cauda epididymal weight, epididymal sperm count and motile epididymal sperm count in adult offspring. The spermatozoa of PCB 132-exposed offspring produced significantly higher levels of ROS than the controls; ROS induction and SOPR reduction were dose-related. In the low-dose PCB 132 group, p53 was significantly induced and caspase-3 was inhibited. In the high-dose group, activation of caspase-3 and -9 was significantly increased, while the expressions of Fas, Bax, bcl-2, and p53 genes were significantly decreased. Gene expression and caspase activation data may provide insight into the mechanisms by which exposure to low-dose or high-dose PCB 132 affects reproduction in male offspring in rats. Because the doses of PCB 132 administered to the dams were approximately 625-fold in low-dose group and 6250-fold higher in high-dose group than the concentration in human tissue levels, the concentrations are not biologically or environmentally relevant. Further studies using environmentally relevant doses are needed for hazard identification.

  9. Methods and functions: Breath tests.

    PubMed

    Braden, Barbara

    2009-01-01

    Breath tests provide a valuable non-invasive diagnostic strategy to in vivo assess a variety of enzyme activities, organ functions or transport processes. Both the hydrogen breath tests and the (13)C-breath tests using the stable isotope (13)C as tracer are non-radioactive and safe, also in children and pregnancy. Hydrogen breath tests are widely used in clinical practice to explore gastrointestinal disorders. They are applied for diagnosing carbohydrate malassimilation, small intestinal bacterial overgrowth and for measuring the orocecal transit time. (13)C-breath tests non-invasively monitor the metabolisation of a (13)C-labelled substrate. Depending on the choice of the substrate they enable the assessment of gastric bacterial Helicobacter pylori infection, gastric emptying, liver and pancreatic function as well as measurements of many other enzyme activities. The knowledge of potential pitfalls and influencing factors are important for correct interpretation of breath test results before drawing clinical conclusions. PMID:19505663

  10. Structural and Functional Studies of the Protamine 2-Zinc Complex from Syrian Gold Hamster (Mesocricetus Auratus) Spermatids and Sperm

    SciTech Connect

    Dolan, C E

    2004-08-30

    The research described in this dissertation consists of four major areas: (1) sequence analysis of protamine 2 from Muroid rodents to identify potential zinc-binding domain(s) of protamine 2; (2) structural studies of the protamine 2-zinc complex from Syrian Gold hamster sperm and spermatids to elucidate the role of zinc during spermiogenesis; (3) structural studies of an unique protamine 2-zinc complex from chinchilla sperm; and (4) Nuclear Magnetic Resonance (NMR) studies of soluble complexes of hairpin oligonucleotides with synthetic arginine-rich peptides or protamine 1 isolated from bull sperm. First, zinc was quantitated in spermatids and sperm by Proton-Induced X-ray Emission (PIXE) to determine whether zinc is present in the early stages of spermiogenesis. The PIXE results revealed the zinc content varies proportionately with the amount of protamine 2 in both spermatid and sperm nuclei. An exception was chinchilla sperm containing twice the amount of protamine 2 than zinc. Further analyses by PIXE and X-ray Absorption Spectroscopy (XAS) of zinc bound to protamines isolated from hamster sperm confirmed the majority of the zinc is bound to protamine and identified the zinc ligands of protamine 2 in hamster spermatids and sperm in vivo. These studies established that zinc is bound to the protamine 2 precursor in hamster spermatids and the coordination of zinc by protamine 2 changes during spermiogenesis. Finally, the sequence analysis combined with the XAS results suggest that the zinc-binding domain in protamine 2 resides in the amino-terminus. Similar analyses of chinchilla sperm by XAS were performed to clarify the unusual PIXE results and revealed that chinchilla has an atypical protamine 2-zinc structure. Two protamine 2 molecules coordinate one zinc atom, forming homodimers that facilitate the binding of protamine 2 to DNA and provide an organizational scheme that would accommodate the observed species-specific protamine stoichiometry in mammalian sperm. Based on these results, we propose the binding of zinc to protamine 2 molecules stabilizes a dimerization domain in other mammalian sperm. Future experiments will use the knowledge we gained of the interactions between protamine 1 and DNA from the NMR studies to obtain structural data for the DNA-protamine 2-zinc complex.

  11. Ejaculated Mouse Sperm Enter Cumulus-Oocyte Complexes More Efficiently In Vitro than Epididymal Sperm

    PubMed Central

    Suarez, Susan S.

    2015-01-01

    The mouse is an established and popular animal model for studying reproductive biology. Epididymal mouse sperm, which lack exposure to secretions of male accessory glands and do not precisely represent ejaculated sperm for the study of sperm functions, have been almost exclusively used in studies. We compared ejaculated and epididymal sperm in an in vitro fertilization setting to examine whether ejaculated sperm enter cumulus-oocyte complexes more efficiently. In order to prepare sperm for fertilization, they were incubated under capacitating conditions. At the outset of incubation, ejaculated sperm stuck to the glass surfaces of slides and the incidences of sticking decreased with time; whereas, very few epididymal sperm stuck to glass at any time point, indicating differences in surface charge. At the end of the capacitating incubation, when sperm were added to cumulus-oocyte complexes, the form of flagellar movement differed dramatically; specifically, ejaculated sperm predominantly exhibited increased bending on one side of the flagellum (a process termed pro-hook hyperactivation), while epididymal sperm equally exhibited increased bending on one or the other side of the flagellum (pro-hook or anti-hook hyperactivation). This indicates that accessory sex gland secretions might have modified Ca2+ signaling activities in sperm, because the two forms of hyperactivation are reported to be triggered by different Ca2+ signaling patterns. Lastly, over time, more ejaculated than epididymal sperm entered the cumulus oocyte complexes. We concluded that modification of sperm by male accessory gland secretions affects the behavior of ejaculated sperm, possibly providing them with an advantage over epididymal sperm for reaching the eggs in vivo. PMID:25996155

  12. Sperm cryopreservation update: Cryodamage, markers, and factors affecting the sperm freezability in pigs.

    PubMed

    Yeste, Marc

    2016-01-01

    Cryopreservation is the most efficient method for long-term preservation of mammalian sperm. However, freeze-thawing procedures may strongly impair the sperm function and survival and thus decrease the reproductive performance. In addition, the sperm resilience to withstand cryopreservation, also known as freezability, presents a high individual variability. The present work summarizes the principles of cryoinjury and the relevance of permeating and nonpermeating cryoprotective agents. Descriptions about sperm cryodamage are mainly focused on boar sperm, but reference to other mammalian species is also made when relevant. Main cryoinjuries not only regard to sperm motility and membrane integrity, but also to the degradation effect exerted by freeze-thawing on other important components for sperm fertilizing ability, such as mRNAs. After delving into the main differences between good and poor freezability boar ejaculates, those protein markers predicting the sperm ability to sustain cryopreservation are also mentioned. Moreover, factors that may influence sperm freezability, such as season, diet, breed, or ejaculate fractions are discussed, together with the effects of different additives, like seminal plasma and antioxidants. After briefly referring to the effects of long-term sperm preservation in frozen state and the reproductive performance of frozen-thawed boar sperm, this work speculates with new research horizons on the preservation of boar sperm, such as vitrification and freeze-drying. PMID:26506124

  13. Epigallocatechin-3-Gallate (EGCG) Reduces Rotenone Effect on Stallion Sperm-Zona Pellucida Heterologous Binding.

    PubMed

    Plaza Dávila, M; Bucci, D; Galeati, G; Peña, F J; Mari, G; Giaretta, E; Tamanini, C; Spinaci, M

    2015-12-01

    Stallion spermatozoa are highly dependent on oxidative phosphorylation for ATP production to achieve normal sperm function and to fuel the motility. The aim of this study was to evaluate the response of equine sperm under capacitating conditions to the inhibition of mitochondrial complex I by rotenone and to test whether epigallocatechin-3-gallate (EGCG), a natural polyphenol component of green tea, could counteract this effect. After 2-h incubation of stallion spermatozoa in modified Tyrode's medium, rotenone (100 nm, 500 nm and 5 ?m) and EGCG (10, 20 and 60 ?m), alone or in combination, did not induce any significant difference on the percentage of viable cells, live sperm with active mitochondria and spermatozoa with intact acrosome. The inhibition of complex I of mitochondrial respiratory chain of stallion sperm with rotenone exerted a negative effect on heterologous ZP binding ability. EGCG at the concentrations of 10 and 20 ?m (but not of 60 ?m) induced a significant increase in the number of sperm bound to the ZP compared with that for control. Moreover, when stallion sperm were treated with rotenone 100 nm, the presence of EGCG at all the concentrations tested (10, 20 and 60 ?m) significantly increased the number of sperm bound to the ZP up to control levels, suggesting that this green tea polyphenol is able to reduce the toxicity of rotenone. PMID:26482419

  14. Female presence influences sperm velocity in the guppy.

    PubMed

    Gasparini, Clelia; Peretti, Alfredo V; Pilastro, Andrea

    2009-12-23

    As sperm production is costly, males are expected to strategically allocate resources to sperm production according to mating opportunities. While sperm number adjustments have been reported in several taxa, only a few studies investigated whether sperm quality shows adaptive plasticity as well. We tested this prediction in the guppy, Poecilia reticulata. A total of 46 males were initially stripped of all retrievable sperm before being randomly allocated to one of two treatments simulating different levels of mating opportunities (visual contact with females or female deprived). After 3 days, males were stripped and sperm velocity was assayed using Computer Assisted Sperm Analysis. Males in the presence of females produced significantly faster sperm than their counterparts. Implications for the evolution of this ejaculate plasticity in the light of results of sperm competition studies are discussed. PMID:19656863

  15. Sperm viability - Determination of sperm viability using fluorescence microscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To determine the percentage of viable sperm in a semen sample using stains that differentiates viable (live) sperm from nonviable (dead) sperm. Viable sperm are detected by SYBR-14, which stains the sperm nuclei green. Nonviable sperm are detected by propidium iodide (PI), which stains the sperm red...

  16. Testing and learning Boolean functions

    E-print Network

    Matulef, Kevin Michael

    2009-01-01

    Given a function f on n inputs, we consider the problem of testing whether f belongs to a concept class C, or is far from every member of C. An algorithm that achieves this goal for a particular C is called a property ...

  17. Cytometry of mammalian sperm

    SciTech Connect

    Gledhill, B.L.

    1983-10-11

    Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. The accessibility of male cells makes them well suited for analytical cytology. We might automate the process of determining sperm morphology but should not do so solely for increased speed. Rather, richer tangible benefits will derive from cytometric evaluation through increased sensitivity, reduced subjectivity, standardization between investigators and laboratories, enhanced archival systems, and the benefits of easily exchanged standardized data. Inroads on the standardization of assays for motility and functional integrity are being made. Flow cytometric analysis of total DNA content of individual sperm is an insensitive means to detect exposure to reproductive toxins because of the small size and low frequency of the DNA content errors. Flow cytometry can be applied to determine the proportions of X- and Y-sperm in semen samples.

  18. Ameliorative Effect of Zinc Oxide Nanoparticles on Antioxidants and Sperm Characteristics in Streptozotocin-Induced Diabetic Rat Testes

    PubMed Central

    Afifi, Mohamed; Almaghrabi, Omar A.; Kadasa, Naif Mohammed

    2015-01-01

    The present study investigated the impact of zinc oxide nanoparticles (ZnONPs) on the oxidative status and sperm characteristics in diabetic rat testicular tissue. Forty male albino rats were used in this study; 10 of them served as a control and 30 rats were injected with a single dose (100?mg/kg) of streptozotocin intraperitoneally. They were subdivided into diabetic, diabetic + ZnONPs (10?mg/kg B.W.), and diabetic and cotreated with ZnONPs + insulin groups. The sperm count and motility were assessed. The activity and mRNA expression of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GRD), and Glutathion-S-Transferase (GST) were determined in the testicular tissue. Malondialdehyde (MDA) and reduced glutathione (GSH) levels were estimated in the testicular tissue. Sperm count and motility increased in ZnONPs treated diabetic rats. A significant increase in the activity and mRNA expression of SOD, CAT, GPx, GRD, and GST was shown in ZnONPs treated diabetic rats. MDA significantly decreased, while GSH increased in testicular tissue of ZnONPs treated diabetic rats. It was concluded that ZnONPs either alone or in combination with insulin have the ability to increase the sperm count and motility and protect the testicular tissue against the oxidative stress induced by diabetes in rats. PMID:26581756

  19. OSMOTIC TOLERANCE OF AVIAN SPERMATOZOA: INFLUENCE OF TIME, TEMPERATURE, CRYOPROTECTANT AND MEMBRANE ION PUMP FUNCTION ON SPERM VIABILITY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An experiment was conducted to evaluate the sensitivity of crane and turkey spermatozoa to hyperosmotic conditions similar to those encountered during a cryogenic cycle. Sperm from both species were exposed to hypertonic media ranging from 500-3000 mOsm (300 mOsm = isotonic) for differing lengths o...

  20. LOCALIZATION OF SP22 ON HUMAN SPERM OF DIFFERING QUALITY

    EPA Science Inventory

    LOCALIZATION OF SP22 ON HUMAN SPERM OF DIFFERING QUALITY. AE Lavers*1, GR Klinefelter2, DW Hamilton1, KP Roberts1, 1University of Minnesota, Minneapolis, MN and 2US EPA, Research Triangle Park, NC.
    SP22 is a sperm membrane protein that has been implicated in sperm function d...

  1. GALACSI integration and functional tests

    NASA Astrophysics Data System (ADS)

    La Penna, P.; Ströbele, S.; Aller Carpentier, E.; Argomedo, J.; Arsenault, R.; Conzelmann, R. D.; Delabre, B.; Donaldson, R.; Duchateau, M.; Fedrigo, E.; Gago, F.; Hubin, N.; Quentin, J.; Jolley, P.; Kiekebusch, M.; Kirchbauer, J. P.; Klein, B.; Kolb, J.; Kuntschner, H.; Le Louarn, M.; Lizon, J. L.; Madec, P.-.; Manescau, A.; Mehrgan, L.; Sedghi, B.; Suarez Valles, M.; Soenke, C.; Tordo, S.; Vernet, J.; Zampieri, S.

    2014-07-01

    GALACSI is the Adaptive Optics (AO) modules of the ESO Adaptive Optics Facility (AOF) that will correct the wavefront delivered to the MUSE Integral Field Spectrograph. It will sense with four 40×40 subapertures Shack-Hartmann wavefront sensors the AOF 4 Laser Guide Stars (LGS), acting on the 1170 voice-coils actuators of the Deformable Secondary Mirror (DSM). GALACSI has two operating modes: in Wide Field Mode (WFM), with the four LGS at 64" off axis, the collected energy in a 0.2"×0.2" pixel will be enhanced by a factor 2 at 750 nm over a Field of View (FoV) of 1'×1' using the Ground Layer AO (GLAO) technique. The other mode, the Narrow Field Mode (NFM), provides an enhanced wavefront correction (Strehl Ratio (SR) of 5% (goal 10%) at 650 nm) but in a smaller FoV (7.5"×7.5"), using Laser Tomography AO (LTAO), with the 4 LGS located closer, at 10" off axis. Before being shipped to Paranal, GALACSI will be first integrated and fully tested in stand-alone, and then moved to a dedicated AOF facility to be tested with the DSM in Europe. At present the module is fully assembled, its main functionalities have been implemented and verified, and AO system tests with the DSM are starting. We present here the main system features and the results of the internal functional tests of GALACSI.

  2. Turbidity as a method of preparing sperm dilutions in the echinoid sperm/egg bioassay

    SciTech Connect

    Hall, T.J.; Haley, R.K.; Battan, K.J. )

    1993-11-01

    The use of turbidimeter for preparing sperm dilutions used in the echinoid sperm/egg bioassay was evaluated. Regression analyses of the relationship between sperm density and turbidity for the sea urchins Strongylocentrotus purpuratus and Strongylocentrotus droebachiensis and the sand dollar Dendraster excentricus indicated that although there were slope differences for each species, each coefficient of determination was highly significant. For Dendraster excentricus, triplicate hemacytometer counts over a range of turbidities as well as repeated preparations of a single sperm turbidity indicated similar variability for each. The use of the turbidimeter has time-saving advantages over conventional hemacytometer methods without sacrificing precision. Sperm dilutions can be prepared rapidly, minimizing seawater sperm preactivation before test initiation, and may therefore contribute to increased test precision.

  3. Hybridization of modified-heme reconstitution and distal histidine mutation to functionalize sperm whale myoglobin.

    PubMed

    Sato, Hideaki; Hayashi, Takashi; Ando, Tsutomu; Hisaeda, Yoshio; Ueno, Takafumi; Watanabe, Yoshihito

    2004-01-21

    To modulate the physiological function of a hemoprotein, most approaches have been demonstrated by site-directed mutagenesis. Replacement of the native heme with an artificial prosthetic group is another way to modify a hemoprotein. However, an alternate method, mutation or heme reconstitution, does not always demonstrate sufficient improvement compared with the native heme enzyme. In the present study, to convert a simple oxygen storage hemoprotein, myoglobin, into an active peroxidase, we applied both methods at the same time. The native heme of myoglobin was replaced with a chemically modified heme 2 having two aromatic rings at the heme-propionate termini. The constructed myoglobins were examined for 2-methoxyphenol (guaiacol) oxidation in the presence of H2O2. Compared with native myoglobin, rMb(H64D.2) showed a 430-fold higher kcat/Km value, which is significantly higher than that of cytochrome c peroxidase and only 3-fold less than that of horseradish peroxidase. In addition, myoglobin-catalyzed degradation of bisphenol A was examined by HPLC analysis. The rMb(H64D.2) showed drastic acceleration (>35-fold) of bisphenol A degradation compared with the native myoglobin. In this system, a highly oxidized heme reactive species is smoothly generated and a substrate is effectively bound in the heme pocket, while native myoglobin only reversibly binds dioxygen. The present results indicate that the combination of a modified-heme reconstitution and an amino acid mutation should offer interesting perspectives toward developing a useful biomolecule catalyst from a hemoprotein. PMID:14719919

  4. The sperm structure of Cryptocercus punctulatus Scudder (Blattodea) and sperm evolution in Dictyoptera.

    PubMed

    Dallai, Romano; Thipaksorn, Apisit; Gottardo, Marco; Mercati, David; Machida, Ryuichiro; Beutel, Rolf Georg

    2015-04-01

    Sperm of the dictyopteran key taxon Cryptocercus punctulatus was examined. It has largely maintained a blattodean groundplan condition, with a three-layered acrosome, an elongate nucleus, a single centriole, a conspicuous centriole adjunct material, two connecting bands (=accessory bodies), and a long functional flagellum with a 9+9+2 axoneme provided with accessory tubules with 16 protofilaments and intertubular material. These sperm characters are shared with several other polyneopterans. The sperm of C. punctulatus is very similar to what is found in Periplaneta americana and species of other groups of roaches, including the sperm of Loboptera decipiens described here for the first time. The general sperm organization here described can be assumed for the groundplan of Insecta and Pterygota. The following evolutionary path can be suggested: after the split between Cryptocercidae and the common ancestor of Isoptera, the typical pattern of sperm formation was altered very distinctly, resulting in a duplication or multiplication (Mastotermitidae) of the centrioles. Mastotermes has maintained a certain sperm motility, but with a very unusual apparatus of multiple flagella with a 9+0 axoneme pattern. After the split into Mastotermitidae and the remaining Isoptera, sperm motility was completely abandoned, and different modifications of sperm components occurred, and even the loss of the sperm flagellum. PMID:25503102

  5. The FASEB Journal Research Communication The galectin-1glycan axis controls sperm fertilizing

    E-print Network

    Gleeson, Joseph G.

    The FASEB Journal · Research Communication The galectin-1­glycan axis controls sperm fertilizing capacity by regulating sperm motility and membrane hyperpolarization Gustavo Vasen,* Maria Agustina, in the control of sperm function. We found that Gal-1 is expressed in the epi- didymis and associates with sperm

  6. vol. 169, no. 6 the american naturalist june 2007 Sperm Competition and the Evolution of

    E-print Network

    Day, Troy

    vol. 169, no. 6 the american naturalist june 2007 E-Article Sperm Competition and the Evolution abstract: We present a model of sperm competition that incor- porates both sperm and nonsperm parts of the effects of seminal fluid on male reproductive success and as a function of asymmetry in sperm usage

  7. The development of cat testicular sperm cryopreservation protocols: Effects of tissue fragments or sperm cell suspension.

    PubMed

    Chatdarong, Kaywalee; Thuwanut, Paweena; Morrell, Jane M

    2016-01-15

    In endangered animals that have been found dead or sterilized for medical reasons, testis is the ultimate source of haploid DNA or sperm. Thus, preservation of testicular sperm may be performed to rescue their genetics. The aim of this study was to evaluate protocols for testicular sperm freezing: as tissue fragments or cell suspension in domestic cats as a model. A pair of testes from each cat (n = 9) were cut into eight equal pieces. Four randomly selected pieces were cryopreserved as: (1) tissue pieces using two-step freezing; (2) tissue pieces using a slow passive cooling device (CoolCell); (3) sperm suspension after single-layer centrifugation (SLC) through colloids; and (4) sperm suspension without being processed through SLC. A testicular piece from each cat served as fresh control. Testicular sperm membrane and DNA integrity were evaluated before, and after, the cryopreservation process. In addition, spermatogenic cell types (testicular sperm, spermatogonia, spermatocyte, and spermatid) present in the suspension samples were counted before and after SLC. The results found that testicular sperm membrane integrity in the suspension after SLC process was higher than that in the fragment form neither using the two-step nor CoolCell freezing, both before and after freezing (before freezing: 92.3 ± 3.4 vs. 81 ± 4.5 and 80.0 ± 7.0; after freezing: 84.5 ± 4.6 vs. 71.2 ± 12 and 76.2 ± 4.6; P ? 0.05). Testicular sperm DNA integrity was, however, not different among groups. Furthermore, the samples processed through the SLC had higher ration of sperm cells: other spermatogenic cells than those were not processed through the SLC (88.9 ± 3.8 vs. 30 ± 7.9; P ? 0.05). In summary, testicular sperm cryopreserved as a minced suspension is considered suitable in terms of preventing sperm membrane integrity, and SLC is considered a selection tool for enriching haploid sperm cells from castrated or postmortem cats. PMID:26498389

  8. Two Types of Assays for Detecting Frog Sperm Chemoattraction

    PubMed Central

    Burnett, Lindsey A.; Tholl, Nathan; Chandler, Douglas E.

    2011-01-01

    Sperm chemoattraction in invertebrates can be sufficiently robust that one can place a pipette containing the attractive peptide into a sperm suspension and microscopically visualize sperm accumulation around the pipette1. Sperm chemoattraction in vertebrates such as frogs, rodents and humans is more difficult to detect and requires quantitative assays. Such assays are of two major types - assays that quantitate sperm movement to a source of chemoattractant, so-called sperm accumulation assays, and assays that actually track the swimming trajectories of individual sperm. Sperm accumulation assays are relatively rapid allowing tens or hundreds of assays to be done in a single day, thereby allowing dose response curves and time courses to be carried out relatively rapidly. These types of assays have been used extensively to characterize many well established chemoattraction systems - for example, neutrophil chemotaxis to bacterial peptides and sperm chemotaxis to follicular fluid. Sperm tracking assays can be more labor intensive but offer additional data on how chemoattractancts actually alter the swimming paths that sperm take. This type of assay is needed to demonstrate the orientation of sperm movement relative to the chemoattrractant gradient axis and to visualize characteristic turns or changes in orientation that bring the sperm closer to the egg. Here we describe methods used for each of these two types of assays. The sperm accumulation assay utilized is called a "two-chamber" assay. Amphibian sperm are placed in a tissue culture plate insert with a polycarbonate filter floor having 12 ?m diameter pores. Inserts with sperm are placed into tissue culture plate wells containing buffer and a chemoatttractant carefully pipetted into the bottom well where the floor meets the wall (see Fig. 1). After incubation, the top insert containing the sperm reservoir is carefully removed, and sperm in the bottom chamber that have passed through the membrane are removed, pelleted and then counted by hemocytometer or flow cytometer. The sperm tracking assay utilizes a Zigmond chamber originally developed for observing neutrophil chemotaxis and modified for observation of sperm by Giojalas and coworkers2,3. The chamber consists of a thick glass slide into which two vertical troughs have been machined. These are separated by a 1 mm wide observation platform. After application of a cover glass, sperm are loaded into one trough, the chemoattractant agent into the other and movement of individual sperm visualized by video microscopy. Video footage is then analyzed using software to identify two-dimensional cell movements in the x-y plane as a function of time (xyt data sets) that form the trajectory of each sperm. PMID:22231741

  9. 14 CFR 35.40 - Functional test.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Functional test. 35.40 Section 35.40... STANDARDS: PROPELLERS Tests and Inspections § 35.40 Functional test. The variable-pitch propeller system must be subjected to the applicable functional tests of this section. The same propeller system used...

  10. 14 CFR 35.40 - Functional test.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Functional test. 35.40 Section 35.40... STANDARDS: PROPELLERS Tests and Inspections § 35.40 Functional test. The variable-pitch propeller system must be subjected to the applicable functional tests of this section. The same propeller system used...

  11. Functional Task Test: Data Review

    NASA Technical Reports Server (NTRS)

    Cromwell, Ronita

    2014-01-01

    After space flight there are changes in multiple physiological systems including: Cardiovascular function; Sensorimotor function; and Muscle function. How do changes in these physiological system impact astronaut functional performance?

  12. Flow cytometry of sperm

    SciTech Connect

    Gledhill, B.L.

    1987-09-21

    This brief paper summarizes automated flow cytometric determination of sperm morphology and flow cytometry/sorting of sperm with application to sex preselection. In the latter context, mention is made of results of karyotypic determination of sex chromosome ratios in albumin-processed human sperm. 23 refs., 1 fig., 1 tab.

  13. Predictive capacity of sperm quality parameters and sperm subpopulations on field fertility after artificial insemination in sheep.

    PubMed

    Santolaria, P; Vicente-Fiel, S; Palacín, I; Fantova, E; Blasco, M E; Silvestre, M A; Yániz, J L

    2015-12-01

    This study was designed to evaluate the relevance of several sperm quality parameters and sperm population structure on the reproductive performance after cervical artificial insemination (AI) in sheep. One hundred and thirty-nine ejaculates from 56 adult rams were collected using an artificial vagina, processed for sperm quality assessment and used to perform 1319 AI. Analyses of sperm motility by computer-assisted sperm analysis (CASA), sperm nuclear morphometry by computer-assisted sperm morphometry analysis (CASMA), membrane integrity by acridine orange-propidium iodide combination and sperm DNA fragmentation using the sperm chromatin dispersion test (SCD) were performed. Clustering procedures using the sperm kinematic and morphometric data resulted in the classification of spermatozoa into three kinematic and three morphometric sperm subpopulations. Logistic regression procedures were used, including fertility at AI as the dependent variable (measured by lambing, 0 or 1) and farm, year, month of AI, female parity, female lambing-treatment interval, ram, AI technician and sperm quality parameters (including sperm subpopulations) as independent factors. Sperm quality variables remaining in the logistic regression model were viability and VCL. Fertility increased for each one-unit increase in viability (by a factor of 1.01) and in VCL (by a factor of 1.02). Multiple linear regression analyses were also performed to analyze the factors possibly influencing ejaculate fertility (N=139). The analysis yielded a significant (P<0.05) relationship between sperm viability and ejaculate fertility. The discriminant ability of the different semen variables to predict field fertility was analyzed using receiver operating characteristic (ROC) curve analysis. Sperm viability and VCL showed significant, albeit limited, predictive capacity on field fertility (0.57 and 0.54 Area Under Curve, respectively). The distribution of spermatozoa in the different subpopulations was not related to fertility. PMID:26507945

  14. Lectin staining and flow cytometry reveals female-induced sperm acrosome reaction and surface carbohydrate reorganization.

    PubMed

    Kekäläinen, Jukka; Larma, Irma; Linden, Matthew; Evans, Jonathan P

    2015-01-01

    All cells are covered by glycans, an individually unique layer of oligo- and polysaccharides that are critical moderators of self-recognition and other cellular-level interactions (e.g. fertilization). The functional similarity between these processes suggests that gamete surface glycans may also have an important, but currently overlooked, role in sexual selection. Here we develop a user-friendly methodological approach designed to facilitate future tests of this possibility. Our proposed method is based on flow cytometric quantification of female-induced sperm acrosome reaction and sperm surface glycan modifications in the Mediterranean mussel Mytilus galloprovincialis. In this species, as with many other taxa, eggs release water-soluble factors that attract conspecific sperm (chemoattraction) and promote potentially measurable changes in sperm behavior and physiology. We demonstrate that flow cytometry is able to identify sperm from other seawater particles as well as accurately measure both acrosome reaction and structural modifications in sperm glycans. This methodological approach can increase our understanding of chemically-moderated gamete-level interactions and individual-specific gamete recognition in Mytilus sp. and other taxa with similar, easily identifiable acrosome structure. Our approach is also likely to be applicable to several other species, since carbohydrate-mediated cellular-level interactions between gametes are universal among externally and internally fertilizing species. PMID:26470849

  15. Lectin staining and flow cytometry reveals female-induced sperm acrosome reaction and surface carbohydrate reorganization

    PubMed Central

    Kekäläinen, Jukka; Larma, Irma; Linden, Matthew; Evans, Jonathan P.

    2015-01-01

    All cells are covered by glycans, an individually unique layer of oligo- and polysaccharides that are critical moderators of self-recognition and other cellular-level interactions (e.g. fertilization). The functional similarity between these processes suggests that gamete surface glycans may also have an important, but currently overlooked, role in sexual selection. Here we develop a user-friendly methodological approach designed to facilitate future tests of this possibility. Our proposed method is based on flow cytometric quantification of female-induced sperm acrosome reaction and sperm surface glycan modifications in the Mediterranean mussel Mytilus galloprovincialis. In this species, as with many other taxa, eggs release water-soluble factors that attract conspecific sperm (chemoattraction) and promote potentially measurable changes in sperm behavior and physiology. We demonstrate that flow cytometry is able to identify sperm from other seawater particles as well as accurately measure both acrosome reaction and structural modifications in sperm glycans. This methodological approach can increase our understanding of chemically-moderated gamete-level interactions and individual-specific gamete recognition in Mytilus sp. and other taxa with similar, easily identifiable acrosome structure. Our approach is also likely to be applicable to several other species, since carbohydrate-mediated cellular-level interactions between gametes are universal among externally and internally fertilizing species. PMID:26470849

  16. Snail sperm production characteristics vary with sperm competition risk

    PubMed Central

    Oppliger, A.; Hosken, D. J.; Ribi, G.

    1998-01-01

    Sperm competition is widespread and influences both male investment in spermatogenic tissue and ejaculate characteristics. Sperm competition models assume trade-offs between sperm size and number, although such trade-offs may be difficult to detect. This study examines the effects of sperm competition risk on the sperm production characteristics of the freshwater snail Viviparus ater. In this prosobranch, females mate frequently and store sperm, generating sperm competition. Males produce two sperm morphs, fertile eupyrene sperm and non-fertilizing oligopyrene sperm. Non-fertilizing sperm may play a role in sperm competition and therefore, like fertilizing sperm, the number produced could vary relative to sperm competition risk. In addition, trade-offs between sperm number and sperm size may be expected. We manipulated the sex ratio of sexually mature snails and found the presence of rivals affected the ratio of oligopyrene/eupyrene sperm males produced. In experimental and natural populations, the number of oligopyrene sperm produced, but not the number of eupyrene sperm, was significantly higher when the sex ratio was male biased. Testis mass did not vary between experimental treatments. We also found a negative relationship between the number and size of oligopyrene sperm produced, which is consistent with evolutionary models of sperm competition, and is, to our knowledge, the first intraspecific demonstration of a trade-off between these traits.

  17. Sperm competition and sperm midpiece size: no consistent pattern in passerine birds.

    PubMed

    Immler, Simone; Birkhead, Tim R

    2007-02-22

    Sperm competition is thought to be a major force driving the evolution of sperm shape and function. However, previous studies investigating the relationship between the risk of sperm competition and sperm morphometry revealed inconclusive results and marked differences between taxonomic groups. In a comparative study of two families of passerines (Fringillidae and Sylviidae) and also across species belonging to different passerine families, we investigated the relative importance of the phylogenetic background on the relationship between sperm morphometry and the risk of sperm competition. The risk of sperm competition was inferred from relative testis mass as an indicator of investment in sperm production. We found: (i) a significant positive association between both midpiece length and flagellum length and relative testis mass in the Fringillidae, (ii) a significant negative association between sperm trait dimensions and relative testis mass in the Sylviidae, and (iii) no association across all species. Despite the striking difference in the patterns shown by the Sylviidae and the Fringillidae, the relationship between midpiece length and flagellum length was positive in both families and across all species with positive allometry. Reasons for the differences and similarities between passerine families are discussed. PMID:17476777

  18. Male size, sperm transfer, and colony fitness in the western harvester ant, Pogonomyrmex occidentalis.

    PubMed

    Wiernasz, D C; Sater, A K; Abell, A J; Cole, B J

    2001-02-01

    Mating success in the western harvester ant, Pogonomyrmex occidentalis, increases with male size. We tested the hypothesis that increased mating success increases male fitness and the fitness of colonies that make large males by comparing the sperm content of males prior to and at the conclusion of the mating swarm. The number of sperm a male initially possesses is a function of male size, and large males transfer a greater proportion of their sperm than do small males. For colonies, the payoff per unit of investment is an increasing function of male size, and investment in large males is not equivalent to investing in a larger number of small males. Allocation ratios in species that show size variation in reproductives may need to be modified by the individual fitness functions. PMID:11308090

  19. The future of computer-aided sperm analysis

    PubMed Central

    Mortimer, Sharon T; van der Horst, Gerhard; Mortimer, David

    2015-01-01

    Computer-aided sperm analysis (CASA) technology was developed in the late 1980s for analyzing sperm movement characteristics or kinematics and has been highly successful in enabling this field of research. CASA has also been used with great success for measuring semen characteristics such as sperm concentration and proportions of progressive motility in many animal species, including wide application in domesticated animal production laboratories and reproductive toxicology. However, attempts to use CASA for human clinical semen analysis have largely met with poor success due to the inherent difficulties presented by many human semen samples caused by sperm clumping and heavy background debris that, until now, have precluded accurate digital image analysis. The authors review the improved capabilities of two modern CASA platforms (Hamilton Thorne CASA-II and Microptic SCA6) and consider their current and future applications with particular reference to directing our focus towards using this technology to assess functional rather than simple descriptive characteristics of spermatozoa. Specific requirements for validating CASA technology as a semi-automated system for human semen analysis are also provided, with particular reference to the accuracy and uncertainty of measurement expected of a robust medical laboratory test for implementation in clinical laboratories operating according to modern accreditation standards. PMID:25926614

  20. The Rhesus Macaque (Macaca mulatta) Sperm Proteome*

    PubMed Central

    Skerget, Sheri; Rosenow, Matthew; Polpitiya, Ashoka; Petritis, Konstantinos; Dorus, Steve; Karr, Timothy L.

    2013-01-01

    Mass spectrometry based proteomics has facilitated sperm composition studies in several mammalian species but no studies have been undertaken in non-human primate species. Here we report the analysis of the 1247 proteins that comprise the Rhesus macaque (Macaca mulatta) sperm proteome (termed the MacSP). Comparative analysis with previously characterized mouse and human sperm proteomes reveals substantial levels of orthology (47% and 40% respectively) and widespread overlap of functional categories based on Gene Ontology analyses. Approximately 10% of macaque sperm genes (113/1247) are significantly under-expressed in the testis as compared with other tissues, which may reflect proteins specifically acquired during epididymal maturation. Phylogenetic and genomic analyses of three MacSP ADAMs (A-Disintegrin and Metalloprotease proteins), ADAM18-, 20- and 21-like, provides empirical support for sperm genes functioning in non-human primate taxa which have been subsequently lost in the lineages leading to humans. The MacSP contains proteasome proteins of the 20S core subunit, the 19S proteasome activator complex and an alternate proteasome activator PA200, raising the possibility that proteasome activity is present in mature sperm. Robust empirical characterization of the Rhesus sperm proteome should greatly expand the possibility for targeted molecular studies of spermatogenesis and fertilization in a commonly used model species for human infertility. PMID:23816990

  1. Individual adjustment of sperm expenditure accords with sperm competition theory

    E-print Network

    Pilastro, Andrea

    Individual adjustment of sperm expenditure accords with sperm competition theory Andrea Pilastro Rica, and approved May 30, 2002 (received for review March 6, 2002) Sperm competition theory predicts that males should strategically allocate their sperm reserves according to the level of sperm competition

  2. ORIGINAL PAPER Sperm competition affects sex allocation but not sperm

    E-print Network

    Schärer, Lukas

    ORIGINAL PAPER Sperm competition affects sex allocation but not sperm morphology in a flatworm Tim /Published online: 16 April 2010 # Springer-Verlag 2010 Abstract Sperm competition has been shown. One morphological trait that is particularly likely to be affected by sperm competition is sperm size

  3. Track/train dynamics test procedure transfer function test

    NASA Technical Reports Server (NTRS)

    Vigil, R. A.

    1975-01-01

    A transfer function vibration test was made on an 80 ton open hopper freight car in an effort to obtain validation data on the car's nonlinear elastic model. Test configuration, handling, test facilities, test operations, and data acquisition/reduction activities necessary to meet the conditions of test requirements are given.

  4. 14 CFR 35.40 - Functional test.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...tests of this section. The same propeller system used in the endurance test (§ 35.39) must be used in the functional tests...failure or malfunction. This test may be combined with the endurance test for accumulation of cycles. (a)...

  5. Microscopic analysis of MTT stained boar sperm cells

    PubMed Central

    van den Berg, B.M.

    2015-01-01

    The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI) Stations is limited. PMID:26623368

  6. Role of sperm surface arylsulfatase A in mouse sperm-zona pellucida binding.

    PubMed

    Tantibhedhyangkul, Julierut; Weerachatyanukul, Wattana; Carmona, Euridice; Xu, Hongbin; Anupriwan, Araya; Michaud, Dominick; Tanphaichitr, Nongnuj

    2002-07-01

    We have previously described the zonae pellucidae (ZP) binding ability of a pig sperm surface protein, P68. Our recent results on peptide sequencing of 3 P68 tryptic peptides and molecular cloning of pig testis arylsulfatase A (AS-A) revealed the identity of P68 as AS-A. In this report, we demonstrate the presence of AS-A on the mouse sperm surface and its role in ZP binding. Using anti-AS-A antibody, we have shown by immunoblotting that AS-A was present in a Triton X-100 extract of mouse sperm. The presence of AS-A on the sperm plasma membrane was conclusively demonstrated by indirect immunofluorescence, immunogold electron microscopy, and AS-A's desulfation activity on live mouse sperm. The AS-A remained on the head surface of in vivo capacitated sperm, as revealed by positive immunofluorescent staining of oviductal/uterine sperm. Significantly, the role of mouse sperm surface AS-A on ZP binding was demonstrated by dose-dependent decreases of sperm-ZP binding on sperm pretreatment with anti-AS-A IgG/Fab. Furthermore, Alexa-430 conjugated AS-A bound to mouse ZP of unfertilized eggs but not to fertilized ones, and this level of binding increased and approached saturation with increasing Alexa-430 AS-A concentrations. Moreover, in vivo fertilization was markedly decreased when mouse sperm pretreated with anti-AS-A IgG were artificially inseminated into females. All of these results designated a new function for AS-A in mouse gamete interaction. PMID:12080020

  7. Defining the mechanisms by which the reactive oxygen species by-product, 4-hydroxynonenal, affects human sperm cell function.

    PubMed

    Baker, Mark A; Weinberg, Anita; Hetherington, Louise; Villaverde, Ana-Izabel; Velkov, Tony; Baell, Jonathan; Gordon, Christopher P

    2015-04-01

    Lipid peroxidation products such as the naturally occurring aldehyde 4-hydroxynonenal (4-HNE) are known to be cytotoxic toward different cell types, including spermatozoa. In order to understand this at the molecular level, we have employed a proteomic approach to characterize direct 4-HNE adducts on human spermatozoa. Several proteins were identified to be of particular interest, including aldehyde labeling of histone methyltransferase and dynein heavy chain. In addition, we found that 4-HNE bound to part of the activation segment, cysteine residue 199, of protein kinase A (PKA). Interestingly, at low levels, addition of 4-HNE had a stimulatory effect on PKA. However, this did not correlate to increased phosphotyrosine levels during capacitation. This data explains the link between reactive oxygen species and sperm toxicity. Given that epigenetic regulation is likely affected in oxidative-stressed spermatozoa, this data show that spermatozoa appear to shut down under these conditions before reaching the egg. PMID:25673561

  8. Regulation and roles of Ca2+ stores in human sperm

    PubMed Central

    Correia, Joao; Michelangeli, Francesco; Publicover, Stephen

    2015-01-01

    [Ca2 +]i signalling is a key regulatory mechanism in sperm function. In mammalian sperm the Ca2 +-permeable plasma membrane ion channel CatSper is central to [Ca2 +]i signalling, but there is good evidence that Ca2 + stored in intracellular organelles is also functionally important. Here we briefly review the current understanding of the diversity of Ca2 + stores and the mechanisms for the regulation of their activity. We then consider the evidence for the involvement of these stores in [Ca2 +]i signalling in mammalian (primarily human) sperm, the agonists that may activate these stores and their role in control of sperm function. Finally we consider the evidence that membrane Ca2 + channels and stored Ca2 + may play discrete roles in the regulation of sperm activities and propose a mechanism by which these different components of the sperm Ca2 +-signalling apparatus may interact to generate complex and spatially diverse [Ca2 +]i signals. PMID:25964382

  9. Ocean acidification impacts on sperm mitochondrial membrane potential bring sperm swimming behaviour near its tipping point.

    PubMed

    Schlegel, Peter; Binet, Monique T; Havenhand, Jonathan N; Doyle, Christopher J; Williamson, Jane E

    2015-04-01

    Broadcast spawning marine invertebrates are susceptible to environmental stressors such as climate change, as their reproduction depends on the successful meeting and fertilization of gametes in the water column. Under near-future scenarios of ocean acidification, the swimming behaviour of marine invertebrate sperm is altered. We tested whether this was due to changes in sperm mitochondrial activity by investigating the effects of ocean acidification on sperm metabolism and swimming behaviour in the sea urchin Centrostephanus rodgersii. We used a fluorescent molecular probe (JC-1) and flow cytometry to visualize mitochondrial activity (measured as change in mitochondrial membrane potential, MMP). Sperm MMP was significantly reduced in ?pH -0.3 (35% reduction) and ?pH -0.5 (48% reduction) treatments, whereas sperm swimming behaviour was less sensitive with only slight changes (up to 11% decrease) observed overall. There was significant inter-individual variability in responses of sperm swimming behaviour and MMP to acidified seawater. We suggest it is likely that sperm exposed to these changes in pH are close to their tipping point in terms of physiological tolerance to acidity. Importantly, substantial inter-individual variation in responses of sperm swimming to ocean acidification may increase the scope for selection of resilient phenotypes, which, if heritable, could provide a basis for adaptation to future ocean acidification. PMID:25833135

  10. Polyspermy in birds: sperm numbers and embryo survival

    PubMed Central

    Hemmings, N.; Birkhead, T. R.

    2015-01-01

    Polyspermy is a major puzzle in reproductive biology. In some taxa, multiple sperm enter the ovum as part of the normal fertilization process, whereas in others, penetration of the ovum by more than one sperm is lethal. In birds, several sperm typically enter the germinal disc, yet only one fuses with the female pronucleus. It is unclear whether supernumerary sperm play an essential role in the avian fertilization process and, if they do, how females regulate the progression of sperm through the oviduct to ensure an appropriate number reach the ovum. Here, we show that when very few sperm penetrate the avian ovum, embryos are unlikely to survive beyond the earliest stages of development. We also show that when the number of inseminated sperm is limited, a greater proportion than expected reach and penetrate the ovum, indicating that females compensate for low sperm numbers in the oviduct. Our results suggest a functional role for supernumerary sperm in the processes of fertilization and early embryogenesis, providing an exciting expansion of our understanding of sperm function in birds. PMID:26511048

  11. Polyspermy in birds: sperm numbers and embryo survival.

    PubMed

    Hemmings, N; Birkhead, T R

    2015-11-01

    Polyspermy is a major puzzle in reproductive biology. In some taxa, multiple sperm enter the ovum as part of the normal fertilization process, whereas in others, penetration of the ovum by more than one sperm is lethal. In birds, several sperm typically enter the germinal disc, yet only one fuses with the female pronucleus. It is unclear whether supernumerary sperm play an essential role in the avian fertilization process and, if they do, how females regulate the progression of sperm through the oviduct to ensure an appropriate number reach the ovum. Here, we show that when very few sperm penetrate the avian ovum, embryos are unlikely to survive beyond the earliest stages of development. We also show that when the number of inseminated sperm is limited, a greater proportion than expected reach and penetrate the ovum, indicating that females compensate for low sperm numbers in the oviduct. Our results suggest a functional role for supernumerary sperm in the processes of fertilization and early embryogenesis, providing an exciting expansion of our understanding of sperm function in birds. PMID:26511048

  12. Chemical composition of seminal and ovarian fluids of chinook salmon (Oncorhynchus tshawytscha) and their effects on sperm motility traits

    E-print Network

    Montgomerie, Bob

    ) and their effects on sperm motility traits Patrice Rosengrave a, , Harry Taylor a, , Robert Montgomerie b , Victoria: Computer-assisted sperm analysis (CASA) Chinook salmon Ionic composition Ovarian fluid Seminal fluid Sperm, seminal fluids and sperm function are not well known in teleostean fish species. The objective

  13. Seminal characteristics and cryopreservation of sperm from the squirrel monkey, Saimiri collinsi.

    PubMed

    Oliveira, K G; Leão, D L; Almeida, D V C; Santos, R R; Domingues, S F S

    2015-09-15

    The Neotropical nonhuman primate squirrel monkey (Saimiri sp.) is one of the most commonly used species in research in several areas of knowledge. However, little progress has been reported in respect to techniques for preservation of their gametes. Thus, the main objectives of this study were (1) to describe testicular and seminal aspects of a new species, Saimiri collinsi, (2) to preserve semen of this species by cooling or freezing using ACP-118 (powdered coconut water), and (3) to test two glycerol (GLY) concentrations (1.5% or 3%) for semen freezing in the presence of ACP-118. The experimental group started with 14 captive males, but only 11 were suitable to collect ejaculates containing sperm. After anesthesia, both testes were evaluated: length, width, height, and testicular circumference. Semen was collected by electroejaculation and evaluated, followed by dilution, cooling, and freezing. Seminal parameters and sperm motility, vigor, plasma membrane integrity, and normal morphology were evaluated after each step; functionality was also checked in fresh and frozen-thawed sperm. Sperm motility, plasma membrane integrity, and normal sperm in cooled semen (n = 11) were 44.1 ± 34.0, 63.1 ± 15.6, and 73.8 ± 19.8, respectively, with vigor ranging of 2 to 3. Sperm motility, plasma membrane integrity, normal and functional sperm in frozen semen (n = 5) were 0.6 ± 1.3 (1.5% and 3% GLY); 4.4 ± 4.9 (1.5% GLY) and 6.6 ± 7.2 (3% GLY); 86.8 ± 3.0 (1.5% GLY) and 88.8 ± 5.1 (3% GLY); 13.3 ± 11.9 (1.5% GLY) and 14.3 ± 13.5 (3% GLY), respectively, and vigor 0 for both 1.5% and 3% GLY. No significant difference between GLY concentrations was observed. We concluded that electroejaculation was efficient for semen collection of S collinsi and tested the cooling protocol that allowed to recover a satisfactory percentage (63%) of membrane intact sperm. However, the freezing protocol was not appropriate to sperm preservation. PMID:26047706

  14. Rheotaxis guides mammalian sperm

    PubMed Central

    Miki, Kiyoshi; Clapham, David E

    2013-01-01

    Background In sea urchins, spermatozoan motility is altered by chemotactic peptides, giving rise to the assumption that mammalian eggs also emit chemotactic agents that guide spermatozoa through the female reproductive tract to the mature oocyte. Mammalian spermatozoa indeed undergo complex adaptations within the female (the process of capacitation) that are initiated by agents ranging from pH to progesterone, but these factors are not necessarily taxic. Currently, chemotaxis, thermotaxis, and rheotaxis have not been definitively established in mammals. Results Here, we show that positive rheotaxis, the ability of organisms to orient and swim against the flow of surrounding fluid, is a major taxic factor for mouse and human sperm. This flow is generated within 4 hours of sexual stimulation and coitus in female mice; prolactin-triggered oviductal fluid secretion clears the oviduct of debris, lowers viscosity, and generates the stream that guides sperm migration in the oviduct. Rheotaxic movement is demonstrated in capacitated and uncapacitated spermatozoa in low and high viscosity medium. Finally, we show that a unique sperm motion we quantify using the sperm head's rolling rate reflects sperm rotation that generates essential force for positioning the sperm in the stream. Rotation requires CatSper channels, presumably by enabling Ca2+ influx. Conclusions We propose that rheotaxis is a major determinant of sperm guidance over long distances in the mammalian female reproductive tract. Coitus induces fluid flow to guide sperm in the oviduct. Sperm rheotaxis requires rotational motion during CatSper channel-dependent hyperactivated motility. PMID:23453951

  15. Functional Assays for Neurotoxicity Testing

    EPA Science Inventory

    Neurobehavioral and pathological evaluations of the nervous system are complementary components of basic research and toxicity testing of pharmaceutical and environmental chemicals. While neuropathological assessments provide insight as to cellular changes in neurons, behavioral ...

  16. Functional Assays for Neurotoxicity Testing*

    EPA Science Inventory

    Neurobehavioral and pathological evaluations of the nervous system are complementary components of basic research and toxicity testing of pharmaceutical and environmental chemicals. While neuropathological assessments provide insight as to cellular changes in neurons, behavioral ...

  17. What Are Lung Function Tests?

    MedlinePLUS

    ... COPD How the Lungs Work Idiopathic Pulmonary Fibrosis Sarcoidosis Send a link to NHLBI to someone by ... caused by conditions such as pulmonary fibrosis and sarcoidosis (sar-koy-DOE-sis). Also, these tests might ...

  18. A new platelet function test.

    PubMed

    Ryan, W L; Hakenkamp, K; Sullivan, E

    1990-04-15

    A method for screening EDTA(K3) blood samples for platelet function is described. The general availability of whole blood platelet counters in the clinical laboratory suggested their use to measure platelet aggregation. To simplify the collection and storage of blood samples, a method utilizing EDTA collected blood is described. Addition of calcium and citrate restores platelet function to blood samples even when stored at room temperature for several hours. Thus, the blood sample used to assay RBC, WBC and platelet parameters may also be employed in platelet aggregation. A comparison of this method with aggregometry on 120 subjects indicated a similar response to arachidonate, collagen, ristocetin and ADP. PMID:2112272

  19. The in vitro effects of melatonin on human sperm function and its scavenging activities on NO and ROS.

    PubMed

    du Plessis, S S; Hagenaar, K; Lampiao, F

    2010-04-01

    Various systems of antioxidants exist endogenously in the body to help protect it against free radical damage by scavenging excessive ROS and RNS. Melatonin, a hormone secreted by the pineal gland, and responsible for controlling the circadian rhythm, is one such endogenous antioxidant. Melatonin has been reported to be present in human seminal fluid, but its antioxidant activities in semen are rather contradictory. This study aimed at establishing the effects of melatonin treatment on human spermatozoa. Spermatozoa were incubated with 2 mm melatonin (120 min, 37 degrees C, 5% CO(2)) after which motility parameters were measured by computer aided motility analysis, while cell viability (PI), intracellular NO (DAF-2/DA) and ROS (DCFH-DA) were assessed using flow cytometry. In vitro melatonin treated samples (n = 12) showed a significantly higher percentage of motile, progressive motile and rapid cells, while simultaneously reducing the number of nonviable spermatozoa when compared with the control. Endogenous NO was significantly decreased, but no effect was observed on ROS levels. From these results, it can be concluded that melatonin was able to directly or indirectly scavenge NO, as indicated by the reduction in 4,5-diaminofluorescein-2/diacetate fluorescence. Future studies will indicate whether melatonin treatment during sperm preparation techniques could protect spermatozoa from excessive NO production. PMID:20384801

  20. Pulmonary Function Testing in Children

    MedlinePLUS

    ... can be compared to the results of other children of the same sex, age, and height, like the standard ranges on a growth chart. Two common PFT’s done in children are s pirometry and airway resistance tests . What is ...

  1. Dynamics of sperm transfer in the ant Leptothorax gredleri

    NASA Astrophysics Data System (ADS)

    Oppelt, Angelika; Heinze, Jürgen

    2007-09-01

    Mating tactics differ remarkably between and within species of social Hymenoptera (bees, wasps, ants) concerning, e.g., mating frequencies, sperm competition, and the degree of male sperm limitation. Although social Hymenoptera might, therefore, potentially be ideal model systems for testing sexual selection theory, the dynamics of mating and sperm transfer have rarely been studied in species other than social bees, and basic information needed to draw conclusions about possible sperm competition and female choice is lacking. We investigated sperm transfer in the ant Leptothorax gredleri, a species in which female sexuals attract males by “female calling.” The analysis of 38 female sexuals fixed immediately or up to 7 days after copulation with a single male each revealed that the sperm is transferred into the female bursa copulatrix embedded in a gelatinous mass, presumably a spermatophore. Sperm cells rapidly start to migrate from the tip of the spermatophore towards the spermatheca, but transfer is drastically slowed down by an extreme constriction of the spermathecal duct, through which sperm cells have to pass virtually one by one. This results in the spermatheca being filled only between one and several hours after mating. During this time, the posterior part of the spermatophore seals the junction between bursa copulatrix and spermathecal duct and prevents sperm loss. The prolonged duration of sperm transfer might allow female sexuals to chose between ejaculates and explain previously reported patterns of single paternity of the offspring of multiply mated queens.

  2. Suprazero Cooling Rate, Rather Than Freezing Rate, Determines Post Thaw Quality Of Rhesus Macaque Sperm

    PubMed Central

    Martorana, Kelly; Klooster, Katie; Meyers, Stuart

    2013-01-01

    Sperm become most sensitive to cold shock when cooled from 37ºC to 5ºC at rates that are too fast or too slow; cold shock increases the susceptibility to oxidative damage due to its influence on reactive oxygen species (ROS) production ([1]. ROS are significant stress factors that are generated during cooling and low temperature storage, and may be a main cause of decreased motility and fertility upon warming. ROS have been shown to change cellular function through the disruption of the sperm plasma membrane and through damage to proteins and DNA. The objective of this study was to determine which cryopreservation rates result in the lowest degree of oxidative damage and greatest sperm quality. In the rhesus model it has not been determined whether suprazero cooling or subzero freezing rates causes a significant amount of ROS damage to sperm. Semen samples were collected from male rhesus macaques, washed, and resuspended in TEST-yolk cryopreservation buffer to 100 x 106 sperm/mL. Sperm were frozen in 0.5mL straws at four different combinations of suprazero and subzero rates. Three different suprazero rates were used between 22ºC and 0ºC: 0.5ºC/min (Slow), 45ºC/min (Medium), and 93ºC/min (Fast). These suprazero rates were used in combination with two different subzero rates for temperatures 0ºC to ?110ºC: 42ºC/min (Medium) and 87ºC/min (Fast). The different freezing groups were as follows: Slow-Med (SM), Slow-Fast (SF), Med-Med (MM), and Fast-Fast (FF). Flow cytometry was used to detect lipid peroxidation (LPO), a result of ROS generation. Motility was evaluated using a computer assisted sperm motion analyzer. The MM and FF treated sperm had less viable (P < 0.0001) and motile sperm (P < 0.001) than the SM, SF, or fresh sperm. Sperm exposed to MM and FF treatments demonstrated significantly higher oxidative damage than SM, SF, or fresh sperm (P < 0.05). The SM and SF treated sperm showed decreased motility, membrane integrity, and LPO compared to fresh semen (P<0.001). Slow cooling from room temperature promotes higher membrane integrity and motility post thaw, compared to medium or fast cooling rates. Cells exposed to similar cooling rates with differing freezing rates were not different in motility and membrane integrity, whereas comparison of cells exposed to differing cooling rates with similar freezing rates indicated significant differences in motility, membrane integrity, and LPO. These data suggest that sperm quality appears to be more sensitive to the cooling, rather than freezing rate and highlight the role of the suprazero cooling rate in post thaw sperm quality. PMID:24239181

  3. Viability and fertilizing capacity of cryopreserved sperm from three North American acipenseriform species: A retrospective study

    USGS Publications Warehouse

    Horvath, A.; Wayman, W.R.; Dean, J.C.; Urbanyi, B.; Tiersch, T.R.; Mims, S.D.; Johnson, D.; Jenkins, J.A.

    2008-01-01

    Populations of sturgeon across the globe are threatened due to unregulated harvest and habitat loss, and the status varies among species across North America. Ready access to viable and functional sperm would contribute to recovery programmes for these species. In this study, we examined the motility, viability (cell membrane integrity) of cryopreserved sperm from three North American acipenseriform species and fertilizing capacity. Milt samples were collected from captive shortnose sturgeon (Acipenser brevirostrum), wild paddlefish (Polyodon spathula) and pallid sturgeon (Scaphirhynchus albus) and cryopreserved using combinations of Modified Tsvetkova's (MT) extender, Original Tsvetkova's extender, and modified Hanks' balanced salt solution, along with the cryoprotectants methanol (MeOH) or dimethyl sulfoxide (DMSO). A dual-staining technique using the fluorescent stains SYBR-14 and propidium iodide was employed with flow cytometry to determine the percentages of spermatozoa that were viable by virtue of having intact membranes. The percentage of viable spermatozoa ranged from 5% to 12% in shortnose sturgeon, 30-59% in paddlefish, and 44-58% in pallid sturgeon. In the first experiment with shortnose sturgeon sperm, methanol allowed for higher values for dependent variables than did DMSO, and sperm viability generally correlated with post-thaw motility. However, fertilization rate, neurulation, or hatching rates were independent from these factors. In the second experiment with shortnose sturgeon, 5% MeOH combined with MT yielded higher values for all parameters tested than the other combinations: viability was correlated with motility, fertilization rate, and hatching rate. Overall, viability and post-thaw motility was not affected by the use of hyperosmotic extenders (OT) or cryoprotectants (DMSO), but their use decreased fertilization percentages. For paddlefish sperm (experiment 3), MT combined with 10% MeOH was clearly a good choice for cryopreservation; viability and motility results were correlated, but independent of fertilization. For pallid sturgeon sperm (experiment 4), MT with 5-10% MeOH showed significantly higher sperm quality and fertilization parameters. Membrane integrity can be used as a predictor of fertilization by cryopreserved sperm, however additional sperm quality parameters, supplementary to motility and membrane integrity, would be useful in the refining and optimizing cryopreservation protocols with acipenseriform sperm. ?? 2008 Blackwell Verlag, Berlin.

  4. Sperm head phenotype and male fertility in ram semen.

    PubMed

    Maroto-Morales, A; Ramón, M; García-Álvarez, O; Montoro, V; Soler, A J; Fernández-Santos, M R; Roldan, E R S; Pérez-Guzmán, M D; Garde, J J

    2015-12-01

    Although there is ample evidence for the effects of sperm head shape on sperm function, its impact on fertility has not been explored in detail at the intraspecific level in mammals. Here, we assess the relationship between sperm head shape and male fertility in a large-scale study in Manchega sheep (Ovis aries), which have not undergone any selection for fertility. Semen was collected from 83 mature rams, and before insemination, head shapes were measured for five parameters: area, perimeter, length, width, and p2a (perimeter(2)/2×?×area) using a computer-assisted sperm morphometric analysis. In addition, a cluster analysis using sperm head length and p2a factor was performed to determine sperm subpopulations (SPs) structure. Our results show the existence of four sperm SPs, which present different sperm head phenotype: SP1 (large and round), SP2 (short and elongated), SP3 (shortest and round), and SP4 (large and the most elongated). No relationships were found between males' fertility rates and average values of sperm head dimensions. However, differences in fertility rates between rams were strongly associated to the proportion of spermatozoa in an ejaculate SP with short and elongated heads (P < 0.001). These findings show how the heterogeneity in sperm head shape of the ejaculate has an effect on reproductive success, and highlight the important role of modulation of the ejaculate at the intraspecific level. PMID:26318229

  5. Sperm Patch-Clamp

    PubMed Central

    Lishko, Polina; Clapham, David E.; Navarro, Betsy; Kirichok, Yuriy

    2014-01-01

    Sperm intracellular pH and calcium concentration ([Ca2+]i) are two central factors that control sperm activity within the female reproductive tract. As such, the ion channels of the sperm plasma membrane that alter intracellular sperm [Ca2+] and pH play important roles in sperm physiology and the process of fertilization. Indeed, sperm ion channels regulate sperm motility, control sperm chemotaxis toward the egg in some species, and may trigger the acrosome reaction. Until recently, our understanding of these important molecules was rudimentary due to the inability to patch-clamp spermatozoa and directly record the activity of these ion channels under voltage clamp. Recently, we overcame this technical barrier and developed a method for reproducible application of the patch-clamp technique to mouse and human spermatozoa. This chapter covers important aspects of application of the patch-clamp technique to spermatozoa, such as selection of the electrophysiological equipment, isolation of spermatozoa for patch-clamp experiments, formation of the gigaohm seal with spermatozoa, and transition into the whole-cell mode of recording. We also discuss potential pitfalls in application of the patch-clamp technique to flagellar ion channels. PMID:23522465

  6. Detection of dilute sperm samples using photoacoustic flowmetry

    NASA Astrophysics Data System (ADS)

    Viator, J. A.; Sutovsky, P.; Weight, R. M.

    2008-02-01

    Detection of sperm cells in dilute samples may have application in forensic testing and diagnosis of male reproductive health. Due to the optically dense subcellular structures in sperm cells, irradiation by nanosecond laser pulses induces a photoacoustic response detectable using a custom flow cytometer. We determined the detection threshold of bull sperm using various concentrations, from 200 to 1,000,000 sperm cells per milliliter. Using a tunable laser system set to 450nm with a 5 ns pulse duration and 11-12 mJ/pulse, we obtained a detection threshold of 3 sperm cells. The flow rate was 4 ml/minute through the flow chamber. The acoustic sensor was a 100 ?m PVDF film attached to the glass flow chamber. The acoustic signal was preamplified and sent to an oscilloscope. The threshold signal indicated a signal to noise ratio of approximately 6 to 1. Improved system design may decrease the threshold to single sperm cells.

  7. Elemental composition of human semen is associated with motility and genomic sperm defects among older men

    PubMed Central

    Schmid, Thomas E.; Grant, Patrick G.; Marchetti, Francesco; Weldon, Rosana H.; Eskenazi, Brenda; Wyrobek, Andrew J.

    2013-01-01

    BACKGROUND Older men tend to have poorer semen quality and are generally at higher risks for infertility and abnormal reproductive outcomes. METHODS We employed proton-induced X-ray emission (PIXE, 3 MeV proton beam) to investigate the concentrations of zinc, copper, calcium, sulfur, chlorine, potassium, titanium, iron and nickel in washed sperm and seminal plasma from non-smoking groups of 10 older men (65–80 years old) and 10 younger men (22–28 years old) who were concurrently assayed for sperm function and genomicly defective sperm. RESULTS The older group showed elevated zinc, copper and calcium in sperm and elevated sulfur in seminal plasma compared with the younger men. The older group also showed reduced motility as well as increased sperm DNA fragmentation, achondroplasia mutations, DNA strand breaks and chromosomal aberrations. Sperm calcium and copper were positively associated with sperm DNA fragmentation (P < 0.03). Seminal sulfur was positively associated with sperm DNA fragmentation and chromosomal aberrations (P < 0.04), and negatively associated with sperm motility (P < 0.05). Sperm calcium was negatively associated with sperm motility, independent of male age (P = 0.01). CONCLUSIONS We identified major differences in elemental concentrations between sperm and seminal plasma and that higher sperm copper, sulfur and calcium are quantitatively associated with poorer semen quality and increased frequencies of genomic sperm defects. PMID:23042799

  8. An update on sperm retrieval techniques for azoospermic males

    PubMed Central

    Esteves, Sandro C; Miyaoka, Ricardo; Orosz, José Eduardo; Agarwal, Ashok

    2013-01-01

    The use of non-ejaculated sperm coupled with intracytoplasmic sperm injection has become a globally established procedure for couples with azoospermic male partners who wish to have biological offspring. Surgical methods have been developed to retrieve spermatozoa from the epididymides and the testes of such patients. This article reviews the methods currently available for sperm acquisition in azoospermia, with a particular focus on the perioperative, anesthetic and technical aspects of these procedures. A critical analysis of the advantages and disadvantages of these sperm retrieval methods is provided, including the authors' methods of choice and anesthesia preferences. PMID:23503959

  9. Activity of the Na,K-ATPase ?4 isoform is regulated during sperm capacitation to support sperm motility.

    PubMed

    Jimenez, Tamara; Sánchez, Gladis; Blanco, Gustavo

    2012-01-01

    The ?4 polypeptide is a testis-specific isoform of the catalytic subunit of the Na,K-ATPase, which is essential for sperm motility and fertility. In the present study, we have investigated the regulation of activity of the ?4 isoform and the relevance of this event for sperm capacitation. We have performed this by taking advantage of the selective high affinity of ?4 for the inhibitor ouabain. Our results show that ouabain-sensitive hydrolysis of ATP and uptake of (86)Rb, corresponding to the enzymatic and ion transport activities of ?4, respectively, increased during sperm capacitation in a time-dependent manner. Specific labeling of ?4 with the fluorescent indicator bodipy-ouabain and immunoblot analysis of biotinylated and streptavidin-precipitated sperm plasma membrane proteins indicated a capacitation- and time-dependent rise in levels of active ?4 isoform at the sperm surface. Ouabain inhibition of ?4 blocked the increase in total sperm motility and the hyperactive motility pattern characteristic of sperm capacitation. Moreover, interference of ?4 activity with ouabain partially prevented the intracellular decrease in Na(+) and the plasma membrane hyperpolarization that typically accompany sperm capacitation. In contrast, ouabain inhibition of ?4 did not affect the spontaneous sperm acrosomal reaction following capacitation. Together, these results demonstrate that Na,K-ATPase ?4 activity is up-regulated during sperm capacitation through mechanisms that involve both increases in molecular activity and levels of ?4 at the sperm plasma membrane. This increase in ?4 activity helps maintain the changes in motility that are associated with sperm capacitation, emphasizing the biologic relevance of the Na,K-ATPase ?4 isoform in sustaining sperm function. PMID:22441762

  10. Sweat testing to evaluate autonomic function.

    PubMed

    Illigens, Ben M W; Gibbons, Christopher H

    2009-04-01

    Sudomotor dysfunction is common in many subtypes of neuropathy but is one of the earliest detectable neurophysiologic abnormalities in distal small fiber neuropathy. Clinical assessments of sudomotor function include thermoregulatory sweat testing (TST), quantitative sudomotor axon reflex testing (QSART), silicone impressions, the sympathetic skin response (SSR), the acetylcholine sweat-spot test and quantitative direct and indirect axon reflex testing (QDIRT). These testing techniques, when used in combination, can detect and localize pre- and postganglionic lesions, can provide early diagnosis of sudomotor dysfunction and can monitor disease progression or disease recovery. In this article, we describe many of the common clinical tests available for evaluation of sudomotor function with focus on the testing methodology and limitations while providing concrete examples of test results. PMID:18989618

  11. Assessing risks of invasion through gamete performance: farm Atlantic salmon sperm and eggs show equivalence in function, fertility, compatibility and competitiveness to wild Atlantic salmon

    PubMed Central

    Yeates, Sarah E; Einum, Sigurd; Fleming, Ian A; Holt, William V; Gage, Matthew JG

    2014-01-01

    Adaptations at the gamete level (a) evolve quickly, (b) appear sensitive to inbreeding and outbreeding and (c) have important influences on potential to reproduce. We apply this understanding to problems posed by escaped farm salmon and measure their potential to reproduce in the wild. Farm Atlantic salmon (Salmo salar) are a threat to biodiversity, because they escape in large numbers and can introgress, dilute or disrupt locally adapted wild gene pools. Experiments at the whole fish level have found farm reproductive potential to be significant, but inferior compared to wild adults, especially for males. Here, we assess reproductive performance at the gamete level through detailed in vitro comparisons of the form, function, fertility, compatibility and competitiveness of farm versus wild Atlantic salmon sperm and eggs, in conditions mimicking the natural gametic microenvironment, using fish raised under similar environmental conditions. Despite selective domestication and reduced genetic diversity, we find functional equivalence in all farm fish gamete traits compared with their wild ancestral strain. Our results identify a clear threat of farm salmon reproduction with wild fish and therefore encourage further consideration of using triploid farm strains with optimized traits for aquaculture and fish welfare, as triploid fish remain reproductively sterile following escape. PMID:24822083

  12. Sperm competition and the evolution of sperm design in mammals

    PubMed Central

    2011-01-01

    Background The influence of sperm competition upon sperm size has been a controversial issue during the last 20 years which remains unresolved for mammals. The hypothesis that, when ejaculates compete with rival males, an increase in sperm size would make sperm more competitive because it would increase sperm swimming speed, has generated contradictory results from both theoretical and empirical studies. In addition, the debate has extended to which sperm components should increase in size: the midpiece to accommodate more mitochondria and produce more energy to fuel motility, or the principal piece to generate greater propulsion forces. Results In this study we examined the influence of sperm competition upon sperm design in mammals using a much larger data set (226 species) than in previous analyses, and we corrected for phylogenetic effects by using a more complete and resolved phylogeny, and more robust phylogenetic control methods. Our results show that, as sperm competition increases, all sperm components increase in an integrated manner and sperm heads become more elongated. The increase in sperm length was found to be associated with enhanced swimming velocity, an adaptive trait under sperm competition. Conclusions We conclude that sperm competition has played an important role in the evolution of sperm design in mammals, and discuss why previous studies have failed to detect it. PMID:21232104

  13. Supporters of sperm

    PubMed Central

    Løvlie, Hanne

    2014-01-01

    The Biology of Spermatozoa (BoS) meetings have run on a biannual basis since the early 1990s. They are dedicated to the fascinating research topic of sperm and their complicated route to fertilization. The BoS meetings focus on sperm, but they also explore additional supporting factors important in fertilization, such as those present in seminal and ovarian fluid, as well as the genomic bases of sperm biology. Here, I present a report of the recent BoS meeting, and showcase some of the highlights of this year’s meeting. PMID:25225623

  14. Eliminating the effect of pathomorphologically formed sperm on resulting gravidity using the intracytoplasmic sperm injection method

    PubMed Central

    BLAHOVÁ, EVA; MÁCHAL, JAN; MÁCHAL, LADISLAV; MILAKOVI?, IRENA; HANULÁKOVÁ, ŠÁRKA

    2014-01-01

    The aim of the present study was to test whether it is possible to eliminate a high percentage of morphologically abnormal sperm in male ejaculate by assisted reproduction using the intracytoplasmic sperm injection (ICSI) method. Treatment success was evaluated by comparing fertilization, clinical pregnancy and reproduction rates between males with heavy teratospermia (?1% morphologically normal spermatozoa) and males with a higher percentage (>1%) of normal sperm. In total, 174 patients who had previously undergone 174 ICSI cycles (1 per each pair) were evaluated retrospectively. In the group of patients with heavily impaired sperm morphology (n=37), the percentage of normal spermatozoa was ?1%. In the second group, males with >1% normal spermatozoa (n=137) were considered as patients with mildly impaired sperm morphology. The results of partner fertilization in these two groups were compared and a lower number of fertilized oocytes was identified in the patients with heavily impaired sperm morphology (P=0.038). However, neither the gravidity nor the take-home baby rates of the partners differed between the patients with mildly and heavily impaired sperm morphology. Trends opposite to that for fertilization were observed for gravidity and delivery [odds ratio (OR), 0.62; 95% confidence interval (CI), 0.29–1.30; OR, 0.55; 95% CI, 0.26–1.24, respectively]. This indicates that the lower number of fertilized oocytes was not associated with the overall outcome of fertilization and that patients with heavily impaired sperm morphology experience the same benefit from ICSI as patients with mildly impaired sperm morphology. PMID:24669266

  15. Sperm survival and heterogeneity are correlated with fertility after intrauterine insemination

    E-print Network

    Zaragoza, Universidad de

    Sperm survival and heterogeneity are correlated with fertility after intrauterine insemination estimations of fertilizing ability. One key factor is the plasma membrane of the sperm cell, which is actively to analyze sperm surface changes accounting for loss of viability and different functional states

  16. Adjustment of copula duration and ejaculate size according to the risk of sperm

    E-print Network

    Museo Nacional de Ciencias Naturales

    Adjustment of copula duration and ejaculate size according to the risk of sperm competition of prolonged copulations in insects, which include mate guarding and sperm loading functions. We have explored) and the effect of an increased risk of sperm competition on ejaculate investment. Our data support predictions

  17. Human Sperm Tail Proteome Suggests New Endogenous Metabolic Pathways*

    PubMed Central

    Amaral, Alexandra; Castillo, Judit; Estanyol, Josep Maria; Ballescà, José Luís; Ramalho-Santos, João; Oliva, Rafael

    2013-01-01

    Proteomic studies are contributing greatly to our understanding of the sperm cell, and more detailed descriptions are expected to clarify additional cellular and molecular sperm attributes. The aim of this study was to characterize the subcellular proteome of the human sperm tail and, hopefully, identify less concentrated proteins (not found in whole cell proteome studies). Specifically, we were interested in characterizing the sperm metabolic proteome and gaining new insights into the sperm metabolism issue. Sperm were isolated from normozoospermic semen samples and depleted of any contaminating leukocytes. Tail fractions were obtained by means of sonication followed by sucrose-gradient ultracentrifugation, and their purity was confirmed via various techniques. Liquid chromatography and tandem mass spectrometry of isolated sperm tail peptides resulted in the identification of 1049 proteins, more than half of which had not been previously described in human sperm. The categorization of proteins according to their function revealed two main groups: proteins related to metabolism and energy production (26%), and proteins related to sperm tail structure and motility (11%). Interestingly, a great proportion of the metabolic proteome (24%) comprised enzymes involved in lipid metabolism, including enzymes for mitochondrial beta-oxidation. Unexpectedly, we also identified various peroxisomal proteins, some of which are known to be involved in the oxidation of very long chain fatty acids. Analysis of our data using Reactome suggests that both mitochondrial and peroxisomal pathways might indeed be active in sperm, and that the use of fatty acids as fuel might be more preponderant than previously thought. In addition, incubation of sperm with the fatty acid oxidation inhibitor etomoxir resulted in a significant decrease in sperm motility. Contradicting a common concept in the literature, we suggest that the male gamete might have the capacity to obtain energy from endogenous pools, and thus to adapt to putative exogenous fluctuations. PMID:23161514

  18. PULMONARY FUNCTION TESTING IN SMALL LABORATORY MAMMALS

    EPA Science Inventory

    The lung is the primary organ likely to be exposed by inhalation studies and, therefore, measurement of changes in lung function are of particular interest to the pulmonary physiologist and toxicologist. Tests of pulmonary function have been developed which can be used with small...

  19. Long-lived sperm in the geothermal bryophyte Pohlia nutans

    PubMed Central

    Rosenstiel, Todd N.; Eppley, Sarah M.

    2009-01-01

    Non-vascular plants rely on sperm to cross the distance between male and female reproductive organs for fertilization and sexual reproduction to occur. The majority of non-vascular plants have separate sexes, and thus, this distance may be a few millimetres to many metres. Because sperm need water for transport, it has been assumed that sperm lifespans are short and that this type of sexual reproduction limits the expansion of non-vascular plants in terrestrial environments. However, little data is available on the lifespan of sperm in non-vascular plants, and none is available for bryophytes, the group thought to have first colonized terrestrial habitats. Here, we documented the lifespan of sperm of Pohlia nutans, collected from a geothermal spring's area, and tested the effects of variation under environmental conditions on this lifespan. Surprisingly, 20 per cent of the sperm were still motile after 100 h, and sperm lifespan was not significantly affected by temperature variation between 22 and 60°C. Lifespan was significantly affected by sperm dilution and temperatures above 75°C. These results suggest the need to reconsider the importance of sperm motility in bryophyte fertilization. PMID:19640871

  20. The Full Function Test Explosive Generator

    SciTech Connect

    Reisman, D B; Javedani, J B; Griffith, L V; Ellsworth, G F; Kuklo, R M; Goerz, D A; White, A D; Tallerico, L J; Gidding, D A; Murphy, M J; Chase, J B

    2009-12-13

    We have conducted three tests of a new pulsed power device called the Full Function Test (FFT). These tests represented the culmination of an effort to establish a high energy pulsed power capability based on high explosive pulsed power (HEPP) technology. This involved an extensive computational modeling, engineering, fabrication, and fielding effort. The experiments were highly successful and a new US record for magnetic energy was obtained.

  1. Adaptation to long sperm in Drosophila: correlated development of the sperm roller and sperm packaging.

    PubMed

    Joly, Dominique; Luck, Nathalie; Dejonghe, Béatrice

    2008-03-15

    Sperm are generally small and produced in huge numbers, but some species combine exaggerated sperm length with extremely limited numbers of sperm, an evolutionary trend that deviates from the theory of anisogamy. Sperm gigantism has arisen recurrently in various species, but insects exhibit the longest sperm, with some species of the Drosophilidae family producing sperm up to 6 cm in length. The anatomical, cytological, and physiological requirements for males to cope with these giant sperm were hitherto poorly understood. In this paper, we investigate the internal morphology of the male reproductive tract, and highlight specific features that may be linked to this increase in sperm size. We focus on species in the repleta group, within which sperm length varies by a factor of 35. An associated development of the sperm roller, a special twisting device inserted between the testis and the seminal vesicle, is demonstrated. Its length and the number of coils involved increase with sperm size, and it allows individual sperm to swell and roll into a spermatic pellet before reaching the seminal vesicle. This process occurs independently of and in addition to the sperm bundle coiling that takes place at the base of the testis. It is suggested that the emergence and development of the sperm roller may be a male adaptation to sperm gigantism. PMID:17377954

  2. Intracytoplasmic Sperm Injection (ICSI)

    MedlinePLUS

    ... fertilization occurs, the fertilized egg (now called an embryo) grows in a laboratory for 1 to 5 ... damaged. • The egg might not grow into an embryo even after it is injected with sperm. • The ...

  3. Effect of postthaw storage time and sperm-to-egg ratio on fertility of cryopreserved brook trout sperm.

    PubMed

    Nynca, J; Dietrich, G J; Dobosz, S; Zalewski, T; Ciereszko, A

    2015-01-15

    The aim of this study was to test the influence of postthaw storage time on sperm motility parameters of brook trout (n = 9). Furthermore, we examined the effect of sperm-to-egg ratios of 300,000:1 and 600,000:1 on fertility of postthaw, cryopreserved, brook trout sperm. The application of a cryopreservation procedure produced very high postthaw sperm motility (56.8 ± 4.0%). The cryopreserved sperm of brook trout could be stored up to 60 minutes without loss of the percentage of sperm motility (52.0 ± 9.0%). The fertilization capacity of brook trout postthaw sperm was comparable with the fertilization rate of fresh semen at a sperm-to-egg ratio as low as 300,000:1 (42.4 ± 14.0% and 36.5 ± 11.0% for eyed and hatched stages, respectively). The possibility of postthaw semen storage for the prolonged time and the obtainment of high fertilization rate at low sperm-to-egg ratio can lead to the significant improvement of brook trout semen cryopreservation procedure. PMID:25308053

  4. Quantification of mammalian sperm morphology by slit-scan flow cytometry

    SciTech Connect

    Benaron, D.A.; Gray, J.W.; Gledhill, B.L.; Lake, S.; Wyrobek, A.J.; Young, I.T.

    1982-03-01

    The head shapes of mammalian sperm have been measured by slit-scan flow cytometry (SSFCM). In this approach, the distribution of fluorescence along acriflavine stained mammalian sperm is recorded and used as a measure of head shape. Fluorescence profiles were measured for sperm from mice, rabbits, hamsters, and bulls, and for sperm from mice exposed to testicular x-irradiation from 0 to 900 rads. The profiles for sperm from nonirradiated animals were characteristic of each species and were reproducible from sperm to sperm. Some of the fluorescence profiles for sperm from the irradiated mice differed significantly from the profiles usually measured for sperm from exposed mice. An algorithm was developed to determine the frequency of these sperm. The estimated frequencies of atypical profiles correlated well (r . 0.99) with the frequencies of abnormally shaped sperm determined by microscopic scoring. The maximum SSFCM sensitivity (minimum detectable dose . 199 rad) was not as high as that for the visual assay (minimum detectable dose . 116 rad). However, only 100 profiles were measured by SSFCM at each dose while at least 500 sperm were scored visually at each dose. The sensitivity of the SSFCM assay should be increased substantially by measuring more profiles. The objective nature of SSFCM couple with the high correlation with results from the visually based assay of morphology suggests the use of SSFCM to measure frequencies of misshapen sperm when testing for mutagens or monitoring for effects of environmental contaminants.

  5. Chemotactic Maneuverability of Sperm

    E-print Network

    Guasto, Jeffrey S; Zimmer, Richard K; Stocker, Roman

    2011-01-01

    In this fluid mechanics video, we explore the kinematics of chemotaxing sperm cells (sea urchin, \\textit{Arbacia punctulata}) swimming in a chemoattractant gradient. We demonstrate that the complex swimming trajectories resulting in chemotactic behavior (`turn-and-run' motility) are comprised of several distinct flagellar maneuvers. These motility patterns likely play an important role optimizing chemotaxic motility and navigation, when the sperm cells are subjected external fluid flows.

  6. Male genital tract antioxidant enzymes: their source, function in the female, and ability to preserve sperm DNA integrity in the golden hamster.

    PubMed

    Chen, Hong; Chow, Pak Ham; Cheng, So Kwan; Cheung, Annie L M; Cheng, Lydia Y L; O, Wai-Sum

    2003-01-01

    Recently, we reported that male accessory sex gland (ASG) secretions protect sperm genomic integrity by demonstrating that DNA damage was more extensive in sperm not exposed to the secretions. The present study was conducted to find out if ASGs secrete the main antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPx or GSH-Px), and catalase (CAT) and if the most abundant one, SOD, can protect those sperm that were not exposed to ASG secretions against NADPH-induced oxidative stress. Four experimental groups of male golden hamsters were used: intact animals with proven fertility, animals with all major ASGs removed (TX), animals that were bilaterally vasectomized, and sham-operated controls. SOD, CAT, and GPx activities were measured in secretions from all 5 ASGs and sperm-free uterine flushing from virgin females and those mated with the experimental males. The alkaline comet assay was used to analyze DNA integrity of the TX group sperm after incubation in a medium containing 50 U/mL of SOD along with 0 to 20 mmol/L NADPH. The main antioxidant enzyme in ASGs was SOD from coagulating glands (P <.05) and GPx together with CAT from ampullary glands (P <.05). Uterine flushing of ejaculates that contained ASG secretions had more SOD and CAT activities than those with epididymal secretions alone (P <.05 and P <.001, respectively), whereas activity of GPx was the same (P >.05). Addition of SOD in vitro dose dependently decreased the incidence of single-strand DNA damage in sperm not exposed to ASG secretions incubated in the presence of 0 to 20 mmol/L NADPH (P <.001). These results indicated that, in terms of abundance, SOD was the main antioxidant enzyme secreted by male ASGs, whereas CAT was the second one. The GPx activity came from both epididymis and ASGs. We conclude that ASG secretions play a significant role in protecting sperm against oxidative stress. PMID:12954661

  7. Sperm Cells of a Primitive Strepsipteran.

    PubMed

    Nardi, James B; Delgado, Juan A; Collantes, Francisco; Miller, Lou Ann; Bee, Charles M; Kathirithamby, Jeyaraney

    2013-01-01

    The unusual life style of Strepsiptera has presented a long-standing puzzle in establishing its affinity to other insects. Although Strepsiptera share few structural similarities with other insect orders, all members of this order share a parasitic life style with members of two distinctive families in the Coleoptera-the order now considered the most closely related to Strepsiptera based on recent genomic evidence. Among the structural features of several strepsipteran families and other insect families that have been surveyed are the organization of testes and ultrastructure of sperm cells. For comparison with existing information on insect sperm structure, this manuscript presents a description of testes and sperm of a representative of the most primitive extant strepsipteran family Mengenillidae, Eoxenos laboulbenei. We compare sperm structure of E. laboulbenei from this family with that of the three other families of Strepsiptera in the other strepsipteran suborder Stylopidia that have been studied as well as with members of the beetle families Meloidae and Rhipiphoridae that share similar life histories with Strepsiptera. Meloids, Rhipiphorids and Strepsipterans all begin larval life as active and viviparous first instar larvae. This study examines global features of these insects' sperm cells along with specific ultrastructural features of their organelles. PMID:26462430

  8. Sperm Cells of a Primitive Strepsipteran

    PubMed Central

    Nardi, James B.; Delgado, Juan A.; Collantes, Francisco; Miller, Lou Ann; Bee, Charles M.; Kathirithamby, Jeyaraney

    2013-01-01

    The unusual life style of Strepsiptera has presented a long-standing puzzle in establishing its affinity to other insects. Although Strepsiptera share few structural similarities with other insect orders, all members of this order share a parasitic life style with members of two distinctive families in the Coleoptera—the order now considered the most closely related to Strepsiptera based on recent genomic evidence. Among the structural features of several strepsipteran families and other insect families that have been surveyed are the organization of testes and ultrastructure of sperm cells. For comparison with existing information on insect sperm structure, this manuscript presents a description of testes and sperm of a representative of the most primitive extant strepsipteran family Mengenillidae, Eoxenos laboulbenei. We compare sperm structure of E. laboulbenei from this family with that of the three other families of Strepsiptera in the other strepsipteran suborder Stylopidia that have been studied as well as with members of the beetle families Meloidae and Rhipiphoridae that share similar life histories with Strepsiptera. Meloids, Rhipiphorids and Strepsipterans all begin larval life as active and viviparous first instar larvae. This study examines global features of these insects’ sperm cells along with specific ultrastructural features of their organelles. PMID:26462430

  9. Fertilizing characteristics of bovine sperm with flattened or indented acrosomes.

    PubMed

    Thundathil, J; Palomino, J; Barth, A; Mapletoft, R; Barros, C

    2001-09-15

    Frozen semen from a control bull (C: 89% morphologically normal sperm) and two bulls with acrosomal defects (K1: 92% flattened acrosomes; K2: 82% indented acrosomes) were used to investigate the fertilizing ability of bull sperm with flattened or indented acrosomes. In experiment 1, frozen-thawed sperm were evaluated for acrosomal integrity with fluorescent microscopy. In experiment 2, proteolytic activity of the acrosomal contents of sperm was evaluated through a gelatin digestion assay. In experiment 3, an IVF test system was used to determine the ability of sperm with flattened or indented acrosomes to bind to bovine oocytes and penetrate the zona pellucida. In experiment 4, IVM zona-free bovine oocytes (ZFO) were fertilized and examined to evaluate sperm chromatin decondensation. In experiment 1, bulls K1 and K2 had a lower proportion of sperm with intact acrosomes (0 and 13.6 +/- 4.5%, respectively) than bull C (30.2 +/- 5.6%) after 2h of incubation. In experiment 2, the proportion of sperm with proteolytic activity, as indicated by gelatin digestion around sperm heads, did not differ among bulls (C: 55%, n=410; K1: 43%, n=426; K2: 48%, n=324). In experiment 3, a lower proportion of sperm with flattened (K1) or indented acrosomes (K2) bound to oocytes than sperm from the control bull, C. The percentage of zona penetrated (55%, n=20; 13%, n=23; 4%, n=25) and the mean (+/- S.E.M.) number of sperm penetrating these zona pellucida (19.7 +/- 2.5; 6.9 +/- 1.0; and 2.6 +/- 0.5) was higher (P<0.05) for bull C than for bulls K1 or K2, respectively. In experiment 4, the percentage of ZFO penetrated (95%, n=20; 52%, n=30; 30%, n=33) and the mean (+/- S.E.M.) number of sperm with chromatin decondensation (7.8 +/- 1.6; 0.8 +/- 0.2; and 0.3 +/- 0.1) were also higher (P<0.05) for the control bull, C than for bulls K1 or K2, respectively. Results suggest that although sperm with the flattened or indented acrosomes had a tendency to undergo spontaneous acrosome reaction on incubation after thawing, the proteolytic activity of the acrosomal contents appeared to be normal. Sperm with the flattened or indented acrosomes also appeared to have a reduced ability to fuse with oolemma as demonstrated by confocal microscopy. This would impair the ability to penetrate ooplasm and undergo sperm chromatin decondensation. PMID:11530269

  10. Sperm studies in anesthesiologists

    SciTech Connect

    Wyrobek, A.J.; Brodsky, J.; Gordon, l.; Moore, D.H., II; Watchmaker, G.; Cohen, E.N.

    1981-11-01

    Semen samples were collected from 46 anesthesiologists each of whom had worked a minimum of one year in hospital operating rooms ventilated with modern gas-scavenging devices. Samples collected from 26 beginning residents in anesthesiology served as controls. Concentrations of sperm and percentage of sperm having abnormal head shapes were determined for each sample. No significant differences were found between anesthesiologists and beginning residents. Limiting the analyses to men having no confounding factors (varicocele, recent illness, medications, heavy smoking, frequent sauna use) did not change the results. The sperm concentration and morphology in 13 men did not change signficantly after one year of exposure to anesthetic gases. However, the group of men who had one or more confounding factors (excluding exposure to anesthetic gases) showed significantly higher percentages of sperm abnormalities than did the group of men without such factors. These results suggest that limited exposure to anesthetic gases does not significantly affect sperm production as judged by changes in sperm concentration and morphology. These data are reassuring, but since the hospitals surveyed used modern gas-scavenging devices, men who are occupationally exposed to anesthetic gases without this protection should be studied for fuller assessment of the possible human spermatotoxic effects.

  11. CIT photoheliograph functional verification unit test program

    NASA Technical Reports Server (NTRS)

    1973-01-01

    Tests of the 2/3-meter photoheliograph functional verification unit FVU were performed with the FVU installed in its Big Bear Solar Observatory vacuum chamber. Interferometric tests were run both in Newtonian (f/3.85) and Gregorian (f/50) configurations. Tests were run in both configurations with optical axis horizontal, vertical, and at 45 deg to attempt to determine any gravity effects on the system. Gravity effects, if present, were masked by scatter in the data associated with the system wavefront error of 0.16 lambda rms ( = 6328A) apparently due to problems in the primary mirror. Tests showed that the redesigned secondary mirror assembly works well.

  12. Track/train dynamics test report transfer function test. Volume 1: Test

    NASA Technical Reports Server (NTRS)

    Vigil, R. A.

    1975-01-01

    A description is presented of the transfer function test performed on an open hopper freight car loaded with 80 tons of coal. Test data and a post-test update of the requirements document and test procedure are presented. Included are a statement of the test objective, a description of the test configurations, test facilities, test methods, data acquisition/reduction operations, and a chronological test summary. An index to the data for the three test configurations (X, Y, and Z-axis tests) is presented along with test sequence, run number, test reference, and input parameters.

  13. Infertility in a dog due to proximal cytoplasmic droplets in the ejaculate: investigation of the significance for sperm functionality in vitro.

    PubMed

    Peña, A I; Barrio, M; Becerra, J J; Quintela, L A; Herradón, P G

    2007-10-01

    A 4-year-old Basque Shepherd male dog was presented for breeding soundness evaluation after the dog failed to impregnate the three bitches he had mated. Clinical examination showed no anomaly of the reproductive system. Semen evaluation showed normal sperm count (640 x 10(6)), 80% had progressively motile spermatozoa, and 96% had morphologically abnormal sperm of which 84% had proximal cytoplasmic droplet and 12% had proximal droplet plus other anomaly. A zona pellucida-binding assay, using canine oocytes derived from frozen-thawed ovaries, was performed in order to investigate the zona-binding ability of dog spermatozoa with proximal cytoplasmic droplets. For the zona pellucida-binding assay, ovaries were thawed and minced in phosphate-buffered saline + 0.4% bovine serum albumin, the oocytes recovered were divided into two groups of 35-40 oocytes to be, respectively, used with the infertile dog and with a control fertile dog. Spermatozoa were capacitated in Canine Capacitating Medium (CCM) at 38.5 degrees C and 5% CO(2) in air for 2 h before oocyte insemination. Groups of five to six oocytes placed in 45 microl droplets of CCM were incubated for 1 h. Afterwards, 5 microl of CCM containing 25,000 spermatozoa were added to each droplet and co-incubated for 2 h before fixation and evaluation of the complexes. After oocyte insemination, sperm motility and viability were evaluated: the sample from the infertile dog had 85% sperm motility with fast and linear progressive movement, and sperm viability of 92%. The sample from the control dog showed 40% sperm motility with fast and highly curvilinear and erratic movement, high degree of sperm agglutination and sperm viability of 32%. For the infertile dog the mean number of bound spermatozoa/oocyte was 0.33 whereas for the control dog it was 1.80. It was concluded that dog sperm with proximal cytoplasmic droplets seem to lack normal capacitating ability in vitro, and consequently, they may have reduced capacity to bind to the zona pellucida of canine oocytes. PMID:17845602

  14. Changes in Gene Expression of Goat Developing Testes and Sperm During Breeding and Non-breeding Season 

    E-print Network

    Faucette, Azure

    2012-07-16

    Testicular function is fundamental to male fertility, since testicular cells act in collaboration with each other to signal sex differentiation, the initiation of puberty and spermatogenesis. Complications that can be influenced by many factors...

  15. Myristoylated and Nonmyristoylated Pools of Sea Urchin Sperm Flagellar Creatine Kinase Exist Side-by-Side: Myristoylation Is Necessary for Efficient Lipid

    E-print Network

    Myristoylated and Nonmyristoylated Pools of Sea Urchin Sperm Flagellar Creatine Kinase Exist SideVed March 5, 1997X ABSTRACT: In sperm of the sea urchin Strongylocentrotus purpuratus, a functional in distinct locations, is essential for sperm motility. S. purpuratus sperm have an unusually large, 145 k

  16. Diisopropyl fluorophosphate labeling of sperm-associated proteinases

    SciTech Connect

    Odem, R.R.; Willand, J.L.; Polakoski, K.L. )

    1990-02-01

    Proteinase inhibitors have been shown to be capable of preventing various aspects of fertilization. Diisopropyl fluorophosphate (DFP) is an irreversible inhibitor of trypsin-like enzymes that is commercially available in a radiolabeled form. The experiments described herein were designed to determine if DFP would prevent sperm function in live, motile sperm and to identify the sperm proteins bound with DFP. DFP at 5 mM concentrations had no observable effect on sperm motility, but inhibited the penetration of zona-free hamster ova by human sperm (5.5%) compared to controls (33.5%). Acid extracts of motile sperm that had been incubated with radiolabeled DFP and collected by the swim-up procedure demonstrated the presence of radiolabeled DFP, and the autoradiography of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of these extracts localized the uptake of radiolabeled DFP to proteins in the molecular weight region of the proacrosin-acrosin system. Acid-extracted proteinases from semen samples incubated with DFP demonstrated a concentration-dependent inhibition of both esterolytic hydrolysis of benzoyl-arginine ethyl ester on spectrophotometric analysis and proteolytic activity on gelatin SDS-PAGE zymography. DFP-labeled proteins were precipitated by highly specific antibodies to proacrosin. These results demonstrated that DFP is capable of inhibiting sperm function, and that it associates with the proacrosin-acrosin system in live motile sperm.

  17. ACS CCD functional test for WFC

    NASA Astrophysics Data System (ADS)

    Mutchler, Max

    2009-07-01

    During the ACS Repair {ACS-R} conducted during Hubble Servicing Mission 4 {SM4}, astronauts will install a CCD Electronics Box Replacement {CEB-R} and Low-Voltage Power Supply Replacement {LVPS-R}. A rudimentary aliveness/functional test {AT/FT} will be conducted on-orbit during SM4. Shortly after SM4, the more comprehensive CCD function test {FT} defined in this program will be conducted as part of SMOV. This program is modeled after the original CCD functional test {HST program 9005, PI Mark Clampin} conducted following the initial installation of ACS during Hubble Servicing Mission 3B. This WFC-only proposal has a companion HRC proposal 11396.

  18. An automated system for pulmonary function testing

    NASA Technical Reports Server (NTRS)

    Mauldin, D. G.

    1974-01-01

    An experiment to quantitate pulmonary function was accepted for the space shuttle concept verification test. The single breath maneuver and the nitrogen washout are combined to reduce the test time. Parameters are defined from the forced vital capacity maneuvers. A spirometer measures the breath volume and a magnetic section mass spectrometer provides definition of gas composition. Mass spectrometer and spirometer data are analyzed by a PDP-81 digital computer.

  19. Air Pollution and Quality of Sperm: A Meta-Analysis

    PubMed Central

    Fathi Najafi, Tahereh; Latifnejad Roudsari, Robab; Namvar, Farideh; Ghavami Ghanbarabadi, Vahid; Hadizadeh Talasaz, Zahra; Esmaeli, Mahin

    2015-01-01

    Context: Air pollution is common in all countries and affects reproductive functions in men and women. It particularly impacts sperm parameters in men. This meta-analysis aimed to examine the impact of air pollution on the quality of sperm. Evidence Acquisition: The scientific databases of Medline, PubMed, Scopus, Google scholar, Cochrane Library, and Elsevier were searched to identify relevant articles published between 1978 to 2013. In the first step, 76 articles were selected. These studies were ecological correlation, cohort, retrospective, cross-sectional, and case control ones that were found through electronic and hand search of references about air pollution and male infertility. The outcome measurement was the change in sperm parameters. A total of 11 articles were ultimately included in a meta-analysis to examine the impact of air pollution on sperm parameters. The authors applied meta-analysis sheets from Cochrane library, then data extraction, including mean and standard deviation of sperm parameters were calculated and finally their confidence interval (CI) were compared to CI of standard parameters. Results: The CI for pooled means were as follows: 2.68 ± 0.32 for ejaculation volume (mL), 62.1 ± 15.88 for sperm concentration (million per milliliter), 39.4 ± 5.52 for sperm motility (%), 23.91 ± 13.43 for sperm morphology (%) and 49.53 ± 11.08 for sperm count. Conclusions: The results of this meta-analysis showed that air pollution reduces sperm motility, but has no impact on the other sperm parameters of spermogram. PMID:26023349

  20. Sperm Membrane Behaviour during Cooling and Cryopreservation.

    PubMed

    Sieme, H; Oldenhof, H; Wolkers, W F

    2015-09-01

    Native sperm is only marginally stable after collection. Cryopreservation of semen facilitates transport and storage for later use in artificial reproduction technologies, but cryopreservation processing may result in cellular damage compromising sperm function. Membranes are thought to be the primary site of cryopreservation injury. Therefore, insights into the effects of cooling, ice formation and protective agents on sperm membranes may help to rationally design cryopreservation protocols. In this review, we describe membrane phase behaviour of sperm at supra- and subzero temperatures. In addition, factors affecting membrane phase transitions and stability, sperm osmotic tolerance limits and mode of action of cryoprotective agents are discussed. It is shown how cooling only results in minor thermotropic non-cooperative phase transitions, whereas freezing causes sharp lyotropic fluid-to-gel phase transitions. Membrane cholesterol content affects suprazero membrane phase behaviour and osmotic tolerance. The rate and extent of cellular dehydration coinciding with freezing-induced membrane phase transitions are affected by the cooling rate and ice nucleation temperature and can be modulated by cryoprotective agents. Permeating agents such as glycerol can move across cellular membranes, whereas non-permeating agents such as sucrose cannot. Both, permeating and non-permeating protectants preserve biomolecular and cellular structures by forming a protective glassy state during freezing. PMID:26382025

  1. Effect of donkey seminal plasma on sperm movement and sperm-polymorphonuclear neutrophils attachment in vitro.

    PubMed

    Miró, Jordi; Vilés, Karina; García, Wilber; Jordana, Jordi; Yeste, Marc

    2013-08-01

    To evaluate the effect of seminal plasma in endometrial inflammation in donkeys, samples from fresh pure, fresh diluted and frozen-thawed semen of three different jackasses were co-incubated in water bath at 37°C with uterine Jennie's secretions collected 6h after artificial insemination with frozen-thawed donkey semen. Individual sperm movement parameters using the computerised sperm analysis system (CASA) and sperm-polymorphonuclear neutrophils (sperm-PMN) attachment observed in Diff-Quick stained smears were evaluated at 0, 1, 2, 3 and 4h of co-incubation. Controls consisted of incubating diluted or frozen-thawed sperm in the absence of uterine secretions. For data analyses, a repeated measures ANOVA was performed with incubation time as intra-subject factor and with treatment and donkey as inter-subject factor, followed by a post-hoc Bonferroni's test. Greater values (P<0.05) of sperm-PMN percentages and a loss of progressive motility were observed in frozen-thawed semen compared with pure and diluted fresh semen samples throughout the incubation time. In addition, the presence of seminal plasma in fresh and diluted semen samples reduced the inflammatory response of polymorphonuclear neutrophils produced after insemination by suppressing the sperm-PMN attachment in vitro. Motility sperm parameters analysed by CASA were also less affected than those in frozen-thawed semen samples. In conclusion, seminal plasma in jennies appears to have a modulation on the endometrial response after artificial insemination with frozen-thawed donkey semen. As a result, spermatozoa with the greater motility characteristics are selected. PMID:23891218

  2. Effect of sorafenib on sperm count and sperm motility in male Swiss albino mice

    PubMed Central

    Shetty, Surekha Devadasa; Bairy, Laxminarayana Kurady

    2015-01-01

    The issue of male germ line mutagenesis and the effects on developmental defects in the next generation has become increasingly high profile over recent years. Mutagenic substance affects germinal cells in the testis. Since the cells are undergoing different phases of cell division and maturation, it is an ideal system to study the effect of chemotherapeutic agents. There are lacunae in the literature on the effect of sorafenib on gonadal function. With background, a study was planned to evaluate the effects of sorafenib on sperm count and sperm motility in male Swiss albino mice. Male Swiss albino mice were used for the study. The animals were segregated into control, positive control (PC) and three treatment groups. PC received oral imatinib (100 mg/kg body weight) and treatment groups received 25, 50, and 100 mg/kg body weight of sorafenib orally for 7 consecutive days at intervals of 24 h between two administrations. The control group remained in the home cage for an equal duration of time to match their corresponding treatment groups. The animals were sacrificed at the end of 1st, 2nd, 4th, 5th, 7th, and 10th weeks after the last exposure to drug, respectively. Sperm suspensions were prepared and introduced into a counting chamber. Total sperm count and motility were recorded. There was a significant decrease in sperm count and sperm motility by sorafenib which was comparable with the effect of PC imatinib. Sorafenib adversely affects sperm count and sperm motility which are reversible after discontinuation of treatment. PMID:26605157

  3. Male sperm storage compromises sperm motility in guppies.

    PubMed

    Gasparini, Clelia; Kelley, Jennifer L; Evans, Jonathan P

    2014-11-01

    Sperm senescence can have important evolutionary implications due to its deleterious effects on sperm quality and offspring performance. Consequently, it has been argued that polyandry (female multiple mating) may facilitate the selection of younger, and therefore competitively superior, sperm when ejaculates from multiple males compete for fertilization. Surprisingly, however, unequivocal evidence that sperm ageing influences traits that underlie sperm competitiveness is lacking. Here, we used a paired experimental design that compares sperm quality between 'old' and 'young' ejaculates from individual male guppies (Poecilia reticulata). We show that older sperm exhibit significant reductions in sperm velocity compared with younger sperm from the same males. We found no evidence that the brightness of the male's orange (carotenoid) spots, which are thought to signal resistance to oxidative stress (and thus age-related declines in sperm fitness), signals a male's ability to withstand the deleterious effects of sperm ageing. Instead, polyandry may be a more effective strategy for females to minimize the likelihood of being fertilized by aged sperm. PMID:25392314

  4. Posttesticular sperm maturation, infertility, and hypercholesterolemia

    PubMed Central

    Whitfield, Marjorie; Pollet-Villard, Xavier; Levy, Rachel; Drevet, Joël R; Saez, Fabrice

    2015-01-01

    Cholesterol is a key molecule in the mammalian physiology of especial particular importance for the reproductive system as it is the common precursor for steroid hormone synthesis. Cholesterol is also a recognized modulator of sperm functions, not only at the level of gametogenesis. Cholesterol homeostasis regulation is crucial for posttesticular sperm maturation, and imbalanced cholesterol levels may particularly affect these posttesticular events. Metabolic lipid disorders (dyslipidemia) affect male fertility but are most of the time studied from the angle of endocrine/testicular consequences. This review will focus on the deleterious effects of a particular dyslipidemia, i.e., hypercholesterolemia, on posttesticular maturation of mammalian spermatozoa. PMID:26067871

  5. Posttesticular sperm maturation, infertility, and hypercholesterolemia.

    PubMed

    Whitfield, Marjorie; Pollet-Villard, Xavier; Levy, Rachel; Drevet, Joël R; Saez, Fabrice

    2015-01-01

    Cholesterol is a key molecule in the mammalian physiology of especial particular importance for the reproductive system as it is the common precursor for steroid hormone synthesis. Cholesterol is also a recognized modulator of sperm functions, not only at the level of gametogenesis. Cholesterol homeostasis regulation is crucial for posttesticular sperm maturation, and imbalanced cholesterol levels may particularly affect these posttesticular events. Metabolic lipid disorders (dyslipidemia) affect male fertility but are most of the time studied from the angle of endocrine/testicular consequences. This review will focus on the deleterious effects of a particular dyslipidemia, i.e., hypercholesterolemia, on posttesticular maturation of mammalian spermatozoa. PMID:26067871

  6. Sound production in neonate sperm whales (L)

    NASA Astrophysics Data System (ADS)

    Madsen, P. T.; Carder, D. A.; Au, W. W. L.; Nachtigall, P. E.; Møhl, B.; Ridgway, S. H.

    2003-06-01

    Acoustic data from two sperm whale neonates (Physeter macrocephalus) in rehabilitation are presented and implications for sound production and function are discussed. The clicks of neonate sperm whale are very different from usual clicks of adult specimens in that neonate clicks are of low directionality [SL anomaly (0°-90°) <8 dB], long duration (2-12 ms), and low frequency (centroid frequency between 300 and 1700 Hz) with estimated SLs between 140 and 162 dB//1 ?Pa (rms). Such neonate clicks are unsuited for biosonar, but can potentially convey homing information between calves and submerged conspecifics in open ocean waters at ranges of some 2 km. Moreover, it is demonstrated that sperm whale clicks are produced at the anterior placed monkey lips, thereby substantiating a key point in the modified Norris and Harvey theory and supporting the unifying theory of sound production in odontocetes.

  7. Chimeras of sperm PLC? reveal disparate protein domain functions in the generation of intracellular Ca2+ oscillations in mammalian eggs at fertilization

    PubMed Central

    Theodoridou, Maria; Nomikos, Michail; Parthimos, Dimitris; Gonzalez-Garcia, J. Raul; Elgmati, Khalil; Calver, Brian L.; Sideratou, Zili; Nounesis, George; Swann, Karl; Lai, F. Anthony

    2013-01-01

    Phospholipase C-zeta (PLC?) is a sperm-specific protein believed to cause Ca2+ oscillations and egg activation during mammalian fertilization. PLC? is very similar to the somatic PLC?1 isoform but is far more potent in mobilizing Ca2+ in eggs. To investigate how discrete protein domains contribute to Ca2+ release, we assessed the function of a series of PLC?/PLC?1 chimeras. We examined their ability to cause Ca2+ oscillations in mouse eggs, enzymatic properties using in vitro phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis and their binding to PIP2 and PI(3)P with a liposome interaction assay. Most chimeras hydrolyzed PIP2 with no major differences in Ca2+ sensitivity and enzyme kinetics. Insertion of a PH domain or replacement of the PLC? EF hands domain had no deleterious effect on Ca2+ oscillations. In contrast, replacement of either XY-linker or C2 domain of PLC? completely abolished Ca2+ releasing activity. Notably, chimeras containing the PLC? XY-linker bound to PIP2-containing liposomes, while chimeras containing the PLC? C2 domain exhibited PI(3)P binding. Our data suggest that the EF hands are not solely responsible for the nanomolar Ca2+ sensitivity of PLC? and that membrane PIP2 binding involves the C2 domain and XY-linker of PLC?. To investigate the relationship between PLC enzymatic properties and Ca2+ oscillations in eggs, we have developed a mathematical model that incorporates Ca2+-dependent InsP3 generation by the PLC chimeras and their levels of intracellular expression. These numerical simulations can for the first time predict the empirical variability in onset and frequency of Ca2+ oscillatory activity associated with specific PLC variants. PMID:24152875

  8. Sperm Motility in Flow

    NASA Astrophysics Data System (ADS)

    Guasto, Jeffrey; Juarez, Gabriel; Stocker, Roman

    2012-11-01

    A wide variety of plants and animals reproduce sexually by releasing motile sperm that seek out a conspecific egg, for example in the reproductive tract for mammals or in the water column for externally fertilizing organisms. Sperm are aided in their quest by chemical cues, but must also contend with hydrodynamic forces, resulting from laminar flows in reproductive tracts or turbulence in aquatic habitats. To understand how velocity gradients affect motility, we subjected swimming sperm to a range of highly-controlled straining flows using a cross-flow microfluidic device. The motion of the cell body and flagellum were captured through high-speed video microscopy. The effects of flow on swimming are twofold. For moderate velocity gradients, flow simply advects and reorients cells, quenching their ability to cross streamlines. For high velocity gradients, fluid stresses hinder the internal bending of the flagellum, directly inhibiting motility. The transition between the two regimes is governed by the Sperm number, which compares the external viscous stresses with the internal elastic stresses. Ultimately, unraveling the role of flow in sperm motility will lead to a better understanding of population dynamics among aquatic organisms and infertility problems in humans.

  9. Porcine sperm bind to specific 6-sialylated biantennary glycans to form the oviduct reservoir.

    PubMed

    Kadirvel, Govindasamy; Machado, Sergio A; Korneli, Claudia; Collins, Emily; Miller, Paul; Bess, Kelsey N; Aoki, Kazuhiro; Tiemeyer, Michael; Bovin, Nicolai; Miller, David J

    2012-06-01

    After mating, many female mammals store a subpopulation of sperm in the lower portion of the oviduct, forming a reservoir. The reservoir lengthens sperm lifespan, regulates sperm capacitation, controls polyspermy, and selects normal sperm. It is believed that sperm bind to glycans on the oviduct epithelium to form the reservoir, but the specific adhesion molecules that retain sperm are unclear. Herein, using a glycan array to test 377 glycans for their ability to bind porcine sperm, we found two glycan motifs in common among all glycans with sperm-binding ability: the Lewis X trisaccharide and biantennary structures containing a mannose core with 6-sialylated lactosamine at one or more termini. Binding to both motifs was specific; isomers of each motif did not bind sperm. Further work focused on sialylated lactosamine. Sialylated lactosamine was found abundantly on the apical side of epithelial cells collected from the oviduct isthmus, among N-linked and O-linked glycans. Sialylated lactosamine bound to the head of sperm, the region that interacts with the oviduct epithelium. After capacitation, sperm lost affinity for sialylated lactosamine. Receptor modification may contribute to release from the reservoir so that sperm can move to the site of fertilization. Sialylated lactosamine was required for sperm to bind oviduct cells. Simbucus nigra agglutinin or an antibody specific to sialylated lactosamine with a preference for Neu5Acalpha2-6Gal rather than Neu5Acalpha2-3Gal reduced sperm binding to oviduct isthmic cells, as did occupying putative receptors on sperm with sialylated biantennary glycans. These results demonstrate that sperm binding to oviduct 6-sialylated biantennary glycans is necessary for normal adhesion to the oviduct. PMID:23115267

  10. Freeze-dried dog sperm: Dynamics of DNA integrity.

    PubMed

    Olaciregui, M; Luño, V; Gonzalez, N; De Blas, I; Gil, L

    2015-10-01

    Freeze-drying (FD) has been proposed as an alternative method to preserve spermatozoa. During the FD procedure, sperm DNA might become damaged by both freezing and drying stresses caused by the endonucleases, the oxidative stress and the storage conditions. We examined the DNA integrity of dog sperm freeze-dried with two kinds of chelating agents in FD buffers and storage at two different temperatures. Ejaculated sperm from four dogs were suspended in basic medium (10mM Tris-HCl buffer+50mM NaCl) supplemented with 50mM EGTA or with 50mM EDTA and then freeze-dried. Sperm samples were stored at 4°C as room temperature, and the analysis of DNA damage was performed after a month and 5months of storage using a Sperm Chromatin Dispersion test. We found four different sperm populations according to the size of the halos around the sperm head: (1) absent halo, (2) <6?m, (3) 6-10?m, (4) >10?m. All of them coexisted in each freeze-dried dog semen samples and differed significantly among different treatments. The highest percentage of spermatozoa with halo >10?m was obtained when the semen samples were freeze-dried in EDTA medium and stored at room temperature for five months. Results suggested that both, the kind of chelating agent as well as storage temperature and period, influenced DNA integrity of freeze-dried dog sperm. PMID:26247315

  11. Epidemiological assessment of occupationally related, chemically induced sperm count suppression

    SciTech Connect

    Milby, T.H.; Whorton, D.

    1980-02-01

    Occupationally related, chemically induced sperm count suppression is a recently recognized problem, first brought to light in connection with the manufacture and formulation of dibromochloropropane (DBCP). The authors studied sperm count data from four occupational cohorts - two exposed to DBCP and two exposed to epichlorohydrin (ECH). In both DBCP cohorts there was a significant difference (alpha = 0.05) between sperm count distribution functions of the exposed group and of the non-exposed group. A much higher percentage of exposed men was oligospermic and the median sperm count for each exposed group was substantially lower than that for the respective non-exposed group. In the ECH cohorts there was no significant difference between sperm count data for the exposed group and for the non-exposed group. The authors concluded that exposure to DBCP, but not to ECH, was positively associated with detectable sperm count suppression. It is suggested that the key to identifying and assessing occupationally related sperm count suppression lies in the proper classification and interpretation of group sperm count data.

  12. Oviducal sperm storage in poultry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hens are capable of fertilizing a daily succession of ovulated ova due to their ability to store sperm in the oviduct for several weeks. However, the precise biological mechanisms describing how sperm are selected and survive in the oviduct, and which sperm actually reach the site of fertilization c...

  13. The Sperm Whale, Physeter macrocephalus

    E-print Network

    The Sperm Whale, Physeter macrocephalus MERRILL E. GOSHO, DALE W. RICE, and JEFFREY M. BREIWICK A sperm whale surfaces in the Southern Ocean. Note the characteristic wrinkl- ed appearance of the body surface and the obtusely rounded dorsal hump. Photo by G. Joyce. Introduction The sperm whale Physeter

  14. Role of human- and animal-sperm studies in the evaluation of male reproductive hazards

    SciTech Connect

    Wyrobek, A.J.; Gordon, L.; Watchmaker, G.

    1982-04-07

    Human sperm tests provide a direct means of assessing chemically induced spermatogenic dysfunction in man. Available tests include sperm count, motility, morphology (seminal cytology), and Y-body analyses. Over 70 different human exposures have been monitored in various groups of exposed men. The majority of exposures studied showed a significant change from control in one or more sperm tests. When carefully controlled, the sperm morphology test is statistically the most sensitive of these human sperm tests. Several sperm tests have been developed in nonhuman mammals for the study of chemical spermatotoxins. The sperm morphology test in mice has been the most widely used. Results with this test seem to be related to germ-cell mutagenicity. In general, animal sperm tests should play an important role in the identification and assessment of potential human reproductive hazards. Exposure to spermatotoxins may lead to infertility, and more importantly, to heritable genetic damage. While there are considerable animal and human data suggesting that sperm tests may be used to detect agents causing infertility, the extent to which these tests detect heritable genetic damage remains unclear. (ERB)

  15. 33 CFR 157.12f - Workshop functional test requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false Workshop functional test... CARRYING OIL IN BULK Design, Equipment, and Installation § 157.12f Workshop functional test requirements... functional test on a suitable test bench prior to delivery. The detailed program for a functional test...

  16. 33 CFR 157.12f - Workshop functional test requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 2 2011-07-01 2011-07-01 false Workshop functional test... CARRYING OIL IN BULK Design, Equipment, and Installation § 157.12f Workshop functional test requirements... functional test on a suitable test bench prior to delivery. The detailed program for a functional test...

  17. Effects of hypothermic storage of striped bass (Morone saxatilis) sperm on intracellular calcium, reactive oxygen species formation, mitochondrial function, motility, and viability

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Experiments were conducted to determine the effect of hypothermic 24 h storage of striped bass sperm cells on viability, intracellular Ca2+ ([Ca2+]i), mitochondrial membrane potential (D'm), and reactive oxygen species (ROS) formation (oxidation of hydroethidine to ethidium) as determined by flow cy...

  18. Effects of hypothermic storage of striped bass (Morone saxatilis) sperm on intracellular calcium, reactive oxygen species formation, mitochondrial function, motility, and viability

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Experiments were conducted to determine the effect of hypothermic 24 h storage of striped bass sperm cells (Morone saxatilis) on viability, intracellular Ca2+ [Ca2+]i, mitochondrial membrane potential (''m), and reactive oxygen species (ROS) formation as determined by flow cytometry; motion activati...

  19. Sperm proteases that may be involved in the initiation of sperm motility in the newt, Cynops pyrrhogaster.

    PubMed

    Yokoe, Misato; Sano, Makoto; Shibata, Honami; Shibata, Daisuke; Takayama-Watanabe, Eriko; Inaba, Kazuo; Watanabe, Akihiko

    2014-01-01

    A protease of sperm in the newt Cynops pyrrhogaster that is released after the acrosome reaction (AR) is proposed to lyse the sheet structure on the outer surface of egg jelly and release sperm motility-initiating substance (SMIS). Here, we found that protease activity in the sperm head was potent to widely digest substrates beneath the sperm. The protease activity measured by fluorescein thiocarbamoyl-casein digestion was detected in the supernatant of the sperm after the AR and the activity was inhibited by 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF), an inhibitor for serine or cysteine protease, suggesting the release of serine and/or cysteine proteases by AR. In an in silico analysis of the testes, acrosins and 20S proteasome were identified as possible candidates of the acrosomal proteases. We also detected another AEBSF-sensitive protease activity on the sperm surface. Fluorescence staining with AlexaFluor 488-labeled AEBSF revealed a cysteine protease in the principal piece; it is localized in the joint region between the axial rod and undulating membrane, which includes an axoneme and produces powerful undulation of the membrane for forward sperm motility. These results indicate that AEBSF-sensitive proteases in the acrosome and principal piece may participate in the initiation of sperm motility on the surface of egg jelly. PMID:25170808

  20. Identification of porcine sperm plasma membrane proteins that may play a role in sperm-egg fusion.

    PubMed

    Ash, K; Berger, T; Horner, C M; Calvert, C C

    1995-05-01

    Sperm plasma membrane (PM) proteins that demonstrate affinity for egg PM preparations have the potential to be biologically important during sperm-egg binding and/or fusion. In this study four such proteins have been identified. To provide quantitative evidence for possible biological function, the large natural variation among different porcine sperm populations with regard to their ability to interact with the egg was compared with the relative binding of egg PM material to individual proteins. An aliquot from each of 24 porcine ejaculates was evaluated by the zona-free hamster ova bioassay and the remainder processed to yield sperm PM vesicles. Aliquots of sperm PM were solubilised, separated by SDS-PAGE, western blotted and probed with partially purified, biotinylated egg PM protein. Bound egg PM proteins were visualised on western blots by an avidin/biotin/horseradish peroxidase system and analysed by scanning laser densitometry. Four sperm PM proteins (62, 39, 27 and 7 kDa estimated molecular mass) were the predominant binders of egg PM. The amount of egg PM bound to the 62 kDa protein was significantly correlated with the ability of sperm from the 24 ejaculates to penetrate zona-free hamster ova (percentage of ova penetrated, p = 0.01, R = 0.65; number of penetrated sperm per ovum, p = 0.02, R = 0.63). PMID:7582918

  1. A new scintigraphic test of neorectal function

    SciTech Connect

    O'Connell, P.R.; Omdahl, A.L.; Brown, M.L.; Kelly, K.A.

    1985-05-01

    Current tests of neorectal function after colectomy, mucosal rectectomy and ileal pouch-anal anastomosis are inadequate. The authors developed a new scintigraphic test that uses an artificial stool made of aluminium magnesium silicate gel (Veegum). Solutions of the gel were labeled with 1 mCi Tc-99m by stirring Veegum powder into water containing the isotope at 37/sup 0/C. Excellent binding was obtained with the sulfur colloid form of the isotope compared with the pertechnetate or ovalbumin forms (Table). The sulfur colloid isotope in 7.5% gel was used in subsequent tests. The volume given to the patients corresponded to the maximum capacity of the ileal pouch as measured manometrically; it averaged 300 ml. Ten patients have been studied to date with minimal patient discomfort. The gel was introduced over a 2 minute period via a 16Fr. transanal tube with the patient in the left lateral position. After 5 minutes AP standing and R lateral scans were taken. Then a dynamic scan was taken while the patient evacuated the gel into a commode. After evacuation AP and R lateral scans were repeated. Computer analysis was used to determine both the functional pouch capacity and the rates of emptying of the pouch and the more proximal ileum. The test has proven useful in the assessment of neorectal anatomy and function after ileal pouch-anal anastomosis.

  2. Platelet function tests: a comparative review

    PubMed Central

    Paniccia, Rita; Priora, Raffaella; Alessandrello Liotta, Agatina; Abbate, Rosanna

    2015-01-01

    In physiological hemostasis a prompt recruitment of platelets on the vessel damage prevents the bleeding by the rapid formation of a platelet plug. Qualitative and/or quantitative platelet defects promote bleeding, whereas the high residual reactivity of platelets in patients on antiplatelet therapies moves forward thromboembolic complications. The biochemical mechanisms of the different phases of platelet activation – adhesion, shape change, release reaction, and aggregation – have been well delineated, whereas their complete translation into laboratory assays has not been so fulfilled. Laboratory tests of platelet function, such as bleeding time, light transmission platelet aggregation, lumiaggregometry, impedance aggregometry on whole blood, and platelet activation investigated by flow cytometry, are traditionally utilized for diagnosing hemostatic disorders and managing patients with platelet and hemostatic defects, but their use is still limited to specialized laboratories. To date, a point-of-care testing (POCT) dedicated to platelet function, using pertinent devices much simpler to use, has now become available (ie, PFA-100, VerifyNow System, Multiplate Electrode Aggregometry [MEA]). POCT includes new methodologies which may be used in critical clinical settings and also in general laboratories because they are rapid and easy to use, employing whole blood without the necessity of sample processing. Actually, these different platelet methodologies for the evaluation of inherited and acquired bleeding disorders and/or for monitoring antiplatelet therapies are spreading and the study of platelet function is strengthening. In this review, well-tried and innovative platelet function tests and their methodological features and clinical applications are considered. PMID:25733843

  3. Correlation Testing in Nuclear Density Functional Theory

    E-print Network

    M. G. Bertolli

    2013-03-18

    Correlation testing provides a quick method of discriminating amongst potential terms to include in a nuclear mass formula or functional and is a necessary tool for further nuclear mass models; however a firm mathematical foundation of the method has not been previously set forth. Here, the necessary justification for correlation testing is developed and more detail of the motivation behind its use is give. Examples are provided to clarify the method analytically and for computational benchmarking. We provide a quantitative demonstration of the method's performance and short-comings, highlighting also potential issues a user may encounter. In concluding we suggest some possible future developments to improve the limitations of the method.

  4. The secretory products of Trichomonas vaginalis decrease fertilizing capacity of mice sperm in vitro

    PubMed Central

    Roh, Jaesook; Lim, Young-Su; Seo, Min-Young; Choi, Yuri; Ryu, Jae-Sook

    2015-01-01

    Trichomonas vaginalis infection is one of the most prevalent sexually transmitted infections in humans and is now recognized as an important cause of infertility in men. There is little information about the effect of extracellular polymeric substances (EPS) from T. vaginalis on sperm, but previous reports do not provide a conclusive description of the functional integrity of the sperm. To investigate the impact of EPS on the fertilizing capacity of sperm, we assessed sperm motility, acrosomal status, hypo-osmotic swelling, and in vitro fertilization rate after incubating the sperm with EPS in vitro using mice. The incubation of sperm with EPS significantly decreased sperm motility, viability, and functional integrity in a concentration and time-dependent manner. These effects on sperm quality also resulted in a decreased fertilization rate in vitro. This is the first report that demonstrates the direct negative impact of the EPS of T. vaginalis on the fertilization rate of sperm in vitro. However, further study should be performed using human sperm to determine if EPS has similar negative impact on human sperm fertilizing capacity in vitro. PMID:25578937

  5. Role of the oviduct in sperm capacitation.

    PubMed

    Rodriguez-Martinez, H

    2007-09-01

    Following insemination of spermatozoa pre-ovulation, the mammalian oviduct ensures, by the formation of a functional sperm reservoir (SR), that suitable (low) numbers of viable and potentially fertile spermatozoa are available for fertilization at the ampullary isthmic junction (AIJ). As ovulation approaches, a proportion of the SR-stored spermatozoa is continuously distributed towards the AIJ and individually activated leading to step-wise capacitation and the attainment of hyperactivated motility. This paper reviews in vivo changes in the intra-luminal milieu of the oviduct of pigs and cows, in particular the SR and the AIJ which relate to the modulation of sperm capacitation around spontaneous ovulation. In vivo, most viable spermatozoa in the pre-ovulatory SR are uncapacitated. Capacitation rates significantly increase after ovulation, apparently not massively but concurrent with the individual, continuous sperm dislocation from the SR. Bicarbonate, whose levels differ between the SR and the AIJ, appears as the common primary effector of the membrane destabilizing changes that encompasses the first stages of capacitation. Sperm activation can be delayed or even reversed by co-incubation with membrane proteins of the tubal lining, isthmic fluid or specific tubal glycosaminoglycans, such as hyaluronan. Although the pattern of response to in vitro induction of sperm activation - capacitation in particular - is similar for all spermatozoa, the capacity and speed of the response is very individual. Such diversity in responsiveness among spermatozoa insures full sperm viability before ovulation and the presence of spermatozoa at different stages of capacitation at the AIJ, thus maximizing the chances of normal fertilization. PMID:17452049

  6. Evaluation of sperm retrieval rate with bilateral testicular sperm extraction in infertile patients with azoospermia

    PubMed Central

    Moein, Mohammad Reza; Moein, Mahmoud Reza; Ghasemzadeh, Jalal; Pourmasoumi, Soheila

    2015-01-01

    Background: About 10% to 15% of infertile men have azoospermia, which could be obstructive or non-obstructive. Diagnostic biopsy from the testis and recently testicular sperm extraction (TESE) are the most precise investigations in these patients. Testicular biopsy can be done unilaterally or bilaterally. The worth of unilateral or bilateral testicular biopsy in men with azoospermia is controversial. Objective: To evaluate the necessity of bilateral diagnostic biopsy from the testis in new era of diagnosis and treatment of male infertility. Materials and Methods: In this retrospective study, we reviewed the results of testis biopsy in 419 azoospermic men, referred to Yazd Research and Clinical Center for Infertility from 2009-2013. Patients with known obstructive azoospermia were excluded from the study. Results: In totally, 254 infertile men (60.6%) were underwent unilateral TESE, which in 175 patients (88.4%) sperm were extracted from their testes successfully. Bilateral testis biopsy was done in 165 patients (39.4%) which in 37 patients (22.4%), sperm were found in their testes tissues. Conclusion: Due to the low probability of positive bilateral TESE results especially when we can’t found sperm in the first side, we recommend that physicians re-evaluate the risk and benefit of this procedure in era of newer and more precise technique of sperm retrieval like micro TESE.

  7. The flagellar beat of rat sperm is organized by the interaction of two functionally distinct populations of dynein bridges with a stable central axonemal partition.

    PubMed

    Lindemann, C B; Orlando, A; Kanous, K S

    1992-06-01

    Two distinct patterns of microtubular sliding were observed in rat sperm flagellar axonemes. The particular pattern of sliding was determined by the extraction conditions used to prepare the sperm for axoneme disintegration. Sperm prepared by incubating concentrated suspensions of Triton X-100-extracted sperm at pH 9.0 disintegrated by extruding the doublets and outer dense fibers numbered 4 through 7 in response to Mg-ATP. Sperm prepared by incubating motile Triton X-100-extracted models at 37 degrees C for 1 to 3 hours extruded doublets and outer dense fibers 9, 1 and 2. Axonemes disintegrated by both regimens tended to have doublets 3 and 8 (with their corresponding outer dense fibers), as well as the central pair, in place. In numerous instances, the 3-central-8 complex with outer dense fibers 3 and 8 could be found isolated in midpiece sections prepared from both methods. The 3-central-8 partition was also sometimes seen in isolation in cross-sections of the principal piece where it remained attached to the fibrous sheath. The flagellar remnant produced by extrusion of fibers 4 through 7 under high pH conditions was generally straight or randomly curved. In contrast, the flagellar remnant produced by extrusion of the 9-1-2 bundle of fibers was most often curved into a hook in the midpiece region. While the hook-like configuration was not Ca(2+)-dependent, it may be based on a related mechanism. The sliding of the 9-1-2 group of fibers is a consequence of dynein-tubulin sliding between the 2 and 3 doublets. This sliding pattern appears to be preferentially activated in the motile sperm models in EGTA, but seldom if ever produced sliding in the high-pH-extracted models. We conclude that the 3-central pair-8 complex and associated outer dense fibers form an I-beam-like partition that does not participate in sliding, but acts as a structural foundation for organizing a planar beat. In addition, it is clear that preferential activation of certain dynein arms can be evoked, depending on the treatment regimen employed. This shows definitively that the types of microtubule sliding in the two bend directions are not identical. PMID:1400632

  8. Patterns of sperm-specific histone variation in sea stars and sea urchins: primary structural homologies in the N-terminal region of spermatogenic H1.

    PubMed

    Massey, C B; Watts, S A

    1992-04-15

    An electrophoretic characterization of histones from pyloric caeca, testes, and sperm of Asterias vulgaris revealed a sperm/testes-specific variant of histone H1 significantly larger than its somatic counterpart from pyloric caeca. Additional proteins were observed in H1 regions of acetic acid-urea polyacrylamide gels in testicular extracts. Sperm or testis-specific variants of H2B observed in sea urchins were not found in the sea star. Evidence presented suggests that sperm- or testes-specific H1 species of intermediate mobility may arise from a single, slow-migrating H1 species (SpH1). Although an increase in nonspecific DNA binding by nuclear proteins must occur during the process of spermatogenesis, different organisms exhibit various patterns of sperm-specific protein mediating differential binding during the process. Sperm-specific variants of both H1 and H2B histones are observed in sea urchins, while the only variant observed in sea stars during spermatogenesis is SpH1. Sequencing of the N-terminus of SpH1 from A. vulgaris revealed a repeating tetrapeptide in residues 3-6 and 8-11 (Ser-Pro-Arg-Lys and Ser-Pro-Lys-Lys, respectively), homologous to repeats in the N-termini of sperm-specific H1s from sea urchins. Primary structure within critical, variable regions of molecules responsible for nonspecific DNA binding appear conserved in many organisms. The occurrence of repeating tetrapeptides in SpH1 and other DNA binding proteins suggests that such domains may function similarly in various chromatins undergoing regulated or reversible condensation. PMID:1583456

  9. Cryopreservation of transgenic Huntington's disease rhesus macaque sperm-A Case Report

    PubMed Central

    Putkhao, Kittiphong; Chan, Anthony W.S.; Agca, Yuksel; Parnpai, Rangsun

    2014-01-01

    The cryoprotective effects of glycerol in three different semen freezing extenders, Tris-citrate (TRIS), TEST, and Tes-Tris-Egg yolk (TTE), on wild-type (WT) rhesus monkey (Macaca mulatta) sperm cryopreservation have been tested. Sperm motility and viability were examined to evaluate the integrity of frozen-thawed sperm, and the best extender was selected to cryopreserve sperm from transgenic Huntington's disease (HD) rhesus monkey. The results showed no post-thaw motility difference among the freezing extender tested (P>0.05). However, sperm membrane integrity in TEST and TTE were significantly better than in TRIS extender (P<0.05). TEST was chosen for HD rhesus monkey sperm cryopreservation. The results showed that post-thawed HD sperm motility and viability was not different compared with WT control group (P>0.05). The present study demonstrates that TEST and TTE were excellent extenders and suitable for rhesus monkey sperm cryopreservation and no detectible differences of post-thaw sperm motility and viability between HD and WT rhesus monkeys resulted from TEST extender. PMID:25431746

  10. December 1998 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    54 December 1998 PYGMY SPERM WHALE (Kogia breviceps): Western North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale (Kogia breviceps) and the pygmy sperm whale (K. simus) appear. 1998). Pygmy sperm whales and dwarf sperm whales are difficult to distinguish and sightings of either

  11. Functional Task Test: 2. Spaceflight-Induced Cardiovascular Change and Recovery During NASA's Functional Task Test

    NASA Technical Reports Server (NTRS)

    Phillips, Tiffany; Arzeno, Natalia M.; Stenger, Michael; Lee, Stuart M. C.; Bloomberg, Jacob J.; Platts, Steven H.

    2011-01-01

    The overall objective of the functional task test (FTT) is to correlate spaceflight-induced physiological adaptations with changes in performance of high priority exploration mission-critical tasks. This presentation will focus on the recovery from fall/stand test (RFST), which measures the cardiovascular response to the transition from the prone posture (simulated fall) to standing in normal gravity, as well as heart rate (HR) during 11 functional tasks. As such, this test describes some aspects of spaceflight-induced cardiovascular deconditioning and the course of recovery in Space Shuttle and International Space Station (ISS) astronauts. The sensorimotor and neuromuscular components of the FTT are described in two separate abstracts: Functional Task Test 1 and 3.

  12. CRYPTIC CHOICE OF CONSPECIFIC SPERM CONTROLLED BY THE IMPACT OF OVARIAN FLUID ON SPERM SWIMMING BEHAVIOR

    PubMed Central

    Yeates, Sarah E; Diamond, Sian E; Einum, Sigurd; Emerson, Brent C; Holt, William V; Gage, Matthew J G

    2013-01-01

    Despite evidence that variation in male–female reproductive compatibility exists in many fertilization systems, identifying mechanisms of cryptic female choice at the gamete level has been a challenge. Here, under risks of genetic incompatibility through hybridization, we show how salmon and trout eggs promote fertilization by conspecific sperm. Using in vitro fertilization experiments that replicate the gametic microenvironment, we find complete interfertility between both species. However, if either species’ ova were presented with equivalent numbers of both sperm types, conspecific sperm gained fertilization precedence. Surprisingly, the species’ identity of the eggs did not explain this cryptic female choice, which instead was primarily controlled by conspecific ovarian fluid, a semiviscous, protein-rich solution that bathes the eggs and is released at spawning. Video analyses revealed that ovarian fluid doubled sperm motile life span and straightened swimming trajectory, behaviors allowing chemoattraction up a concentration gradient. To confirm chemoattraction, cell migration tests through membranes containing pores that approximated to the egg micropyle showed that conspecific ovarian fluid attracted many more spermatozoa through the membrane, compared with heterospecific fluid or water. These combined findings together identify how cryptic female choice can evolve at the gamete level and promote reproductive isolation, mediated by a specific chemoattractive influence of ovarian fluid on sperm swimming behavior. PMID:24299405

  13. Sperm storage mediated by cryptic female choice for nuptial gifts

    PubMed Central

    Albo, Maria J.; Bilde, Trine; Uhl, Gabriele

    2013-01-01

    Polyandrous females are expected to discriminate among males through postcopulatory cryptic mate choice. Yet, there is surprisingly little unequivocal evidence for female-mediated cryptic sperm choice. In species in which nuptial gifts facilitate mating, females may gain indirect benefits through preferential storage of sperm from gift-giving males if the gift signals male quality. We tested this hypothesis in the spider Pisaura mirabilis by quantifying the number of sperm stored in response to copulation with males with or without a nuptial gift, while experimentally controlling copulation duration. We further assessed the effect of gift presence and copulation duration on egg-hatching success in matings with uninterrupted copulations with gift-giving males. We show that females mated to gift-giving males stored more sperm and experienced 17% higher egg-hatching success, compared with those mated to no-gift males, despite matched copulation durations. Uninterrupted copulations resulted in both increased sperm storage and egg-hatching success. Our study confirms the prediction that the nuptial gift as a male signal is under positive sexual selection by females through cryptic sperm storage. In addition, the gift facilitates longer copulations and increased sperm transfer providing two different types of advantage to gift-giving in males. PMID:24266042

  14. Identification of Peroxiredoxin-5 in Bovine Cauda Epididymal Sperm

    PubMed Central

    Nagdas, Subir K; Buchanan, Teresa; Raychoudhury, Samir

    2013-01-01

    Developing spermatozoa require a series of post-testicular modifications within the luminal environment of the epididymis to achieve maturation; this involves several surface modifications including changes in plasma membrane lipids, proteins, carbohydrates, and alterations in the outer acrosomal membrane. Epididymal maturation can therefore allow sperm to gain forward motility and fertilization capabilities. The objective of this study was to identify maturation dependent protein(s) and to investigate their role with the production of functionally competent spermatozoa. Lectin blot analyses of caput and cauda sperm plasma membrane fractions identified a 17.5kDa Wheat Germ Agglutinin (WGA) binding polypeptide present in the cauda sperm plasma membrane not in the caput sperm plasma membrane. Among the several WGA stained bands, the presence of a 17.5kDa WGA binding polypeptide band was detected only in cauda epididymal fluid not in caput epididymal fluid suggesting that the 17.5kDa WGA-binding polypeptide is secreted from the cauda epididymis and binds to the cauda sperm plasma membrane during epididymal transit. Proteomic identification of the 17.5kDa polypeptide yielded 13 peptides that matched the sequence of peroxiredoxin-5 (PRDX5) protein (Bos Taurus). We propose that bovine cauda sperm PRDX5 acts as an antioxidant enzyme in the epididymal environment, which is crucial in protecting the viable sperm population against the damage caused by endogeneous or exogeneous peroxide. PMID:24186847

  15. Role of epididymal anti sticking factor in sperm capacitation.

    PubMed

    Roy, Debarun; Dey, Souvik; Majumder, Gopal C; Bhattacharyya, Debdas

    2015-08-01

    Sperm capacitation depends on several features like hormones, ions, intracellular signaling, sperm associated molecules, etc. Anti sticking factor (ASF) is a novel sperm surface associated glycoprotein isolated from epididymal plasma. Function of ASF in vivo has not been revealed yet. The current study is an attempt to highlight the surface localization of ASF and corresponding biochemical changes that occurs in sperm cells during in vitro capacitation. In the presence of 1 nM ASF, percentage of bicarbonate and BSA induced capacitated cells in modified Tyrode medium (7.2) decreased from 72.45% to 16.25% as per Merocyanine 540 (M540)/DAPI stained flowcytometric analysis. Indirect immunocytostaining and western blot analysis shows that the amount of sperm surface bound residual ASF decline during in vitro capacitation. ASF at its effective concentrations notably reduced the bicarbonate and BSA induced cholesterol efflux. These data help in concluding ASF as a majorly responsible molecule that maintains caprine sperm membrane integrity by inhibiting cholesterol efflux. As the capacitation process, progress at in vitro condition, ASF is found to be released from the sperm surface and cell moved from non-capacitated to the capacitated state. PMID:26100206

  16. The monopulsed nature of sperm whale clicks

    NASA Astrophysics Data System (ADS)

    Møhl, Bertel; Wahlberg, Magnus; Madsen, Peter T.; Heerfordt, Anders; Lund, Anders

    2003-08-01

    Traditionally, sperm whale clicks have been described as multipulsed, long duration, nondirectional signals of moderate intensity and with a spectrum peaking below 10 kHz. Such properties are counterindicative of a sonar function, and quite different from the properties of dolphin sonar clicks. Here, data are presented suggesting that the traditional view of sperm whale clicks is incomplete and derived from off-axis recordings of a highly directional source. A limited number of assumed on-axis clicks were recorded and found to be essentially monopulsed clicks, with durations of 100 ?s, with a composite directionality index of 27 dB, with source levels up to 236 dB re: 1 ?Pa (rms), and with centroid frequencies of 15 kHz. Such clicks meet the requirements for long-range biosonar purposes. Data were obtained with a large-aperture, GPS-synchronized array in July 2000 in the Bleik Canyon off Vestera?len, Norway (69°28' N, 15°40' E). A total of 14 h of sound recordings was collected from five to ten independent, simultaneously operating recording units. The sound levels measured make sperm whale clicks by far the loudest of sounds recorded from any biological source. On-axis click properties support previous work proposing the nose of sperm whales to operate as a generator of sound.

  17. Acceleration of sperm transit time and reduction of sperm reserves in the epididymis of rats exposed to sibutramine.

    PubMed

    Bellentani, Fernanda F; Fernandes, Glaura S A; Perobelli, Juliana E; Pacini, Enio S A; Kiguti, Luiz R A; Pupo, André S; Kempinas, Wilma D G

    2011-01-01

    Sibutramine is a drug globally used for the treatment of obesity. The aim of this study was to investigate male reproductive disorders caused by sibutramine in adult rats. Wistar rats were treated for 28 consecutive days (gavage) with 10 mg/kg of sibutramine. Control animals received only vehicle (dimethylsulfoxide and saline). The rats were sacrificed for evaluation of body and reproductive organ weights, sperm parameters, hormone levels (luteinizing hormone, follicle-stimulating hormone, and testosterone), testicular and epididymal histopathology, sexual behavior, fertility and in vitro contractility of the epididymal duct. Sibutramine decreased (P < .05) weights of the epididymis and ventral prostate, but not of other reproductive organs. The sperm number and transit time in the epididymal cauda were decreased (P < .001), but the daily sperm production was not altered. Moreover, morphology and sperm motility, histopathology of the testes and epididymis, sexual behavior, fertility, and serum hormone levels were not altered by the treatment. Sibutramine increased the potency of norepinephrine and, per se, increased the mechanical activity of the epididymal duct in vitro. Thus, although sibutramine in these experimental conditions did not interfere with the reproductive process of rats, it provoked acceleration of the sperm transit time and a decrease in the sperm reserves in the epididymal cauda. This alteration is probably related to the sympathomimetic effect of this drug, as shown by the in vitro assays. In humans, use of this drug might present a threat for male fertility because sperm reserves in men are naturally lower than those in rats. PMID:21764897

  18. High level of intracellular sperm oxidative stress negatively influences embryo pronuclear formation after intracytoplasmic sperm injection treatment.

    PubMed

    Ghaleno, L R; Valojerdi, M R; Hassani, F; Chehrazi, M; Janzamin, E

    2014-12-01

    This study evaluates the relationship between sperm intracellular reactive oxygen species (ROS; H2 O2 , O2 ), DNA fragmentation (DF), low mitochondria membrane potential (MMP) of sperm and normal pronuclear formation among intracytoplasmic sperm injection (ICSI) patients. Semen samples were obtained from 62 infertile male who were candidates for ICSI treatment. After sperm processing, metaphase II (MII) oocytes were injected, and the mean percentages of intracellular ROS, MMP and DF were evaluated using flow cytometry. The mean percentages of pronuclear formation and zygote score (Z) were also recorded, and Pearson, Spearman's rank correlation coefficient and Kruskal-Wallis tests were applied to analyse the data. The amounts of sperm intracellular H2 O2 and O2-? had significant positive correlation with low MMP (P < 0.01). The intracellular ROS had a negative correlation with pronuclear formation (P < 0.05), and its effect was higher than 66.66%. In addition, the mean percentages of neither H2 O2 nor O2-? affected the quality of pronuclear embryos (Z-score). This study shows that although high levels of both sperm intracellular H2 O2 and O2-? in ICSI patients have deleterious effect on sperm MMP, only H2 O2 may interfere in pronuclear formation. PMID:24313687

  19. Power functions and envelopes for unit root tests

    E-print Network

    Juhl, Ted P.; Xiao, Z. J.

    2003-04-01

    This paper studies power functions and envelopes for covariate augmented unit root tests. The power functions are calculated by integrating the characteristic function, allowing accurate evaluation of the power envelope and the power functions...

  20. Variation in sperm displacement and its association with accessory gland protein loci in Drosophila melanogaster

    SciTech Connect

    Clark, A.G.; Prout, T.; Harshman, L.G.

    1995-01-01

    Genes that influence mating and/or fertilization success may be targets for strong natural selection. If females remate frequently relative to the duration of sperm storage and rate of sperm use, sperm displacement may be an important component of male reproductive success. Although it has long been known that mutant laboratory stocks of Drosophila differ in sperm displacement, the magnitude of the naturally occurring genetic variation in this character has not been systematically quantified. Here we report the results of a screen for variation in sperm displacement among 152 lines of Drosophila melanogaster that were made homozygous for second and/or third chromosomes recovered from natural populations. Sperm displacement was assayed by scoring the progeny of cn;bw females that had been mated sequentially to cn;bw and tested males in either order. Highly significant differences were seen in both the ability to displace sperm that is resident in the female`s reproductive tract and in the ability to resist displacement by subsequent sperm. Most lines exhibited nearly complete displacement, having nearly all progeny sired by the second male, but several lines had as few as half the progeny fathered by the second male. Lines that were identified in the screen for naturally occurring variation in sperm displacement were also characterized for single-strand conformation polymorphisms (SSCP) at seven accessory gland protein (Acp) genes. Significant associations were found between particular Acp alleles at four different loci (Acp26Aa/Ab, Acp29B, Acp36DE and Acp53E) and the ability of males to resist displacement by subsequent sperm. There was no correlation between the ability to displace resident sperm and the ability to resist being displaced by subsequent sperm. This lack of correlation, and the association of Acp alleles with resisting subsequent sperm only, suggests that different mechanisms mediate the two components of sperm displacement. 36 refs., 4 figs., 7 tabs.

  1. Turbulence of swarming sperm

    NASA Astrophysics Data System (ADS)

    Creppy, Adama; Praud, Olivier; Druart, Xavier; Kohnke, Philippa L.; Plouraboué, Franck

    2015-09-01

    Collective motion of self-sustained swarming flows has recently provided examples of small-scale turbulence arising where viscous effects are dominant. We report the first observation of universal enstrophy cascade in concentrated swarming sperm consistent with a body of evidence built from various independent measurements. We found a well-defined k-3 power-law decay of a velocity field power spectrum and relative dispersion of small beads consistent with theoretical predictions in 2D turbulence. Concentrated living sperm displays long-range, correlated whirlpool structures of a size that provides an integral scale of turbulence. We propose a consistent explanation for this quasi-2D turbulence based on self-structured laminated flow forced by steric interactions and alignment, a state of active matter that we call "swarming liquid crystal." We develop scaling arguments consistent with this interpretation.

  2. Turbulence of swarming sperm.

    PubMed

    Creppy, Adama; Praud, Olivier; Druart, Xavier; Kohnke, Philippa L; Plouraboué, Franck

    2015-09-01

    Collective motion of self-sustained swarming flows has recently provided examples of small-scale turbulence arising where viscous effects are dominant. We report the first observation of universal enstrophy cascade in concentrated swarming sperm consistent with a body of evidence built from various independent measurements. We found a well-defined k^{-3} power-law decay of a velocity field power spectrum and relative dispersion of small beads consistent with theoretical predictions in 2D turbulence. Concentrated living sperm displays long-range, correlated whirlpool structures of a size that provides an integral scale of turbulence. We propose a consistent explanation for this quasi-2D turbulence based on self-structured laminated flow forced by steric interactions and alignment, a state of active matter that we call "swarming liquid crystal." We develop scaling arguments consistent with this interpretation. PMID:26465513

  3. Integrated Locomotor Function Tests for Countermeasure Evaluation

    NASA Technical Reports Server (NTRS)

    Bloomberg, J. J.; Mulavara, A. P.; Peters, B. T.; Cohen, H. S.; Landsness, E. C.; Black, F. O.

    2005-01-01

    Following spaceflight crewmembers experience locomotor dysfunction due to inflight adaptive alterations in sensorimotor function. Countermeasures designed to mitigate these postflight gait alterations need to be assessed with a new generation of tests that evaluate the interaction of various sensorimotor sub-systems central to locomotor control. The goal of the present study was to develop new functional tests of locomotor control that could be used to test the efficacy of countermeasures. These tests were designed to simultaneously examine the function of multiple sensorimotor systems underlying the control of locomotion and be operationally relevant to the astronaut population. Traditionally, gaze stabilization has been studied almost exclusively in seated subjects performing target acquisition tasks requiring only the involvement of coordinated eye-head movements. However, activities like walking involve full-body movement and require coordination between lower limbs and the eye-head-trunk complex to achieve stabilized gaze during locomotion. Therefore the first goal of this study was to determine how the multiple, interdependent, full-body sensorimotor gaze stabilization subsystems are functionally coordinated during locomotion. In an earlier study we investigated how alteration in gaze tasking changes full-body locomotor control strategies. Subjects walked on a treadmill and either focused on a central point target or read numeral characters. We measured: temporal parameters of gait, full body sagittal plane segmental kinematics of the head, trunk, thigh, shank and foot, accelerations along the vertical axis at the head and the shank, and the vertical forces acting on the support surface. In comparison to the point target fixation condition, the results of the number reading task showed that compensatory head pitch movements increased, peak head acceleration was reduced and knee flexion at heel-strike was increased. In a more recent study we investigated the adaptive remodeling of the full-body gaze control systems following exposure to visual-vestibular conflict. Subjects walked on a treadmill before and after a 30- minute exposure to 0.5X minifying during which self-generated sinusoidal vertical head rotations were performed while seated. Following exposure to visual-vestibular conflict subjects showed a restriction in compensatory head movements, increased knee and ankle flexion after heel-strike and a decrease in the rate of body loading during the rapid weight transfer phase after the heel strike event. Taken together, results from both studies provide evidence that the full body contributes to gaze stabilization during locomotion, and that different functional elements are responsive to changes in visual task constraints and are subject to adaptive alterations following exposure to visual-vestibular conflict. This information provides the basis for the design of a new generation of integrative tests that incorporate the evaluation of multiple neural control systems relevant to astronaut operational performance.

  4. Male sperm whale acoustic behavior observed from multipaths at a single hydrophone

    NASA Astrophysics Data System (ADS)

    Laplanche, Christophe; Adam, Olivier; Lopatka, Maciej; Motsch, Jean-François

    2005-10-01

    Sperm whales generate transient sounds (clicks) when foraging. These clicks have been described as echolocation sounds, a result of having measured the source level and the directionality of these signals and having extrapolated results from biosonar tests made on some small odontocetes. The authors propose a passive acoustic technique requiring only one hydrophone to investigate the acoustic behavior of free-ranging sperm whales. They estimate whale pitch angles from the multipath distribution of click energy. They emphasize the close bond between the sperm whale's physical and acoustic activity, leading to the hypothesis that sperm whales might, like some small odontocetes, control click level and rhythm. An echolocation model estimating the range of the sperm whale's targets from the interclick interval is computed and tested during different stages of the whale's dive. Such a hypothesis on the echolocation process would indicate that sperm whales echolocate their prey layer when initiating their dives and follow a methodic technique when foraging.

  5. Effect of cryopreservation on fish sperm subpopulations.

    PubMed

    Beirão, J; Cabrita, E; Pérez-Cerezales, S; Martínez-Páramo, S; Herráez, M P

    2011-02-01

    The evaluation of the motility data obtained with a CASA system, applying a Two-Step Cluster analysis, identified in seabream sperm 3 different sperm subpopulations that correlated differently with embryo hatching rates. Hence, we designed an experiment to understand the effect of the application of different cryopreservation protocols in these sperm motility-based subpopulations. We analyzed Sparus aurata frozen/thawed semen motility 15, 30, 45 and 60s after activation, using CASA software. Different protocols were applied for cryopreservation: three different cryoprotectants (Dimethyl Sulfoxide (Me(2)SO), Ethylene Glycol (EG) and Propylene Glycol (PG)) each at two different concentrations and two packaging volumes (0.5ml straws, and 1.8ml cryovials) were tested. Different freezing rates were evaluated corresponding to 1, 2, 3, 4 and 8cm above the liquid nitrogen surface for the straws and 1, 2 and 4cm for the cryovials. Motility parameters rendered by CASA were treated with a Two-Step Cluster analysis. Three different subpopulations were obtained: SP1 - slow non-linear spermatozoa, SP2 - slow linear spermatozoa and SP3 - fast linear spermatozoa. We considered SP3 as the subpopulation of interest and focused further analyses on it. Generally, SP3 was the best represented subpopulation 15s after activation and was also the one showing a greater decrease in time, being the least represented after 60s. According to the applied univariate general linear model, samples frozen in straws with 5% Me(2)SO and in cryovials with 10% Me(2)SO at 2 and 1cm from the LN(2,) respectively, produced the best results (closer to the control). Clustering analysis allowed the detection of fish sperm subpopulations according to their motility pattern and showed that sperm composition in terms of subpopulations was differentially affected by the cryopreservation protocol depending on the cryoprotectant used, freezing rates and packaging systems. PMID:21112321

  6. Isolation and functional characterization for oocyte maturation and sperm motility of the oocyte maturation arresting factor from the Japanese scallop, Patinopecten yessoensis.

    PubMed

    Yuan, Yuan; Tanabe, Toru; Maekawa, Fumihito; Inaba, Kazuo; Maeda, Yusuke; Itoh, Naoki; Takahashi, Keisuke G; Osada, Makoto

    2012-12-01

    In bivalves, serotonin (5-hydroxytriptamine, 5-HT) acts as a major promotional factor in oocyte maturation, sperm motility, and sequential spawning. The previously reported novel neuronal protein, oocyte maturation arresting factor (OMAF) that was found in the central nervous system and hemolymph of the Japanese scallop, Patinopecten yessoensis, has an inhibitory activity in the 5-HT-induced oocyte maturation via a receptor-mediated mechanism, resulting in an arrest of spawning [30]. In this study, OMAF protein was isolated from the supernatant of hemolymph of the scallop using gel and anion-exchange chromatography, and SDS-PAGE. Three digested partial peptides with 4, 11, and 16 amino acid residues were determined through reversed-phase HPLC and amino acid sequencing. The anti-OMAF antibodies generated against the obtained peptides with 11 and 16 amino acid residues were applied to immunohistochemistry and 5-HT-induced spawning and oocyte maturation assays. Fusiform OMAF neurons were localized in the external area of the anterior lobe of the cerebral ganglion, supporting our presumption that OMAF was secreted from the cerebral and pedal ganglia (CPG). Pretreatment with anti-OMAF antibody on three kinds of bivalve species showed a strong in vivo amplification of 5-HT-induced release of egg and sperm, and an in vitro restoration of 5-HT-induced germinal vesicle breakdown (GVBD) from inhibition by the CPG extract, suggesting the release from suppressive activity of OMAF due to the absorption with antibody. These results confirm that the isolated peptides are from OMAF and OMAF acts as an inhibitor of 5-HT-induced oocyte maturation and sperm motility as previously reported. PMID:23000641

  7. Subpopulation distribution of motile sperm relative to activation medium in steelhead (Oncorhynchus mykiss).

    PubMed

    Kanuga, M K; Drew, R E; Wilson-Leedy, J G; Ingermann, R L

    2012-03-15

    In the present study, steelhead sperm were activated in artificial tap water, ovarian fluid, activating saline, or in combinations of these media, and motility characteristics were determined using computer-assisted sperm analysis. Motility characteristics of individual sperm were then assessed to test the hypothesis that motile sperm are distributed among discrete subpopulations and that their distribution is influenced by the activation medium. Analysis with k-means clustering detected three discrete motile sperm subpopulations in steelhead semen, regardless of the activation medium. Based on multivariate analysis of variance, proportions of these subpopulations did not differ between sperm activated with ovarian fluid and activating saline, or any combination of these two media. However, subpopulation distributions for sperm activated with either ovarian fluid or activating saline were influenced by the level of dilution of these media in artificial tap water. There was an increase in the number of sperm in high velocity (curvilinear), high straightness, and high wobble subpopulation with increased levels of ovarian fluid or activating saline. The change in sperm motility characteristics with a change in activation medium may play a role in normal fertilization, as discharged sperm pass from seminal plasma and water through ovarian fluid en route to the egg. PMID:22225678

  8. Quantification of mammalian sperm morphology by slit-scan flow cytometry

    SciTech Connect

    Benaron, D.A.; Gray, J.W.; Gledhill, B.L.; Lake, S.; Wyrobek, A.J.; Young, I.T.

    1982-01-01

    The head shapes of mammalian sperm were measured by slit-scan flow cytometry (SSFCM). Fluorescence profiles were measured for sperm from mice, rabbits, hamsters, and bulls, and for sperm from mice, rabbits, hamsters, and bulls, and for sperm from mice exposed to testicular x-irradiation from 0 to 900 rads. Some of the fluorescence profiles for sperm from the irradiated mice differed significantly from the profiles usually measured for sperm from unexposed mice. An algorithm was developed to determine the frequency of these sperm. The estimated frequencies of atypical profiles correlated well with the frequencies of abnormally shaped sperm determined by microscopic scoring. The maximum SSFCM sensitivity was not as high as that for the visual assay. However, only 100 profiles were measured by SSFCM at each dose while at least 500 sperm were scored visually at each dose. The sensitivity of the SSFCM assay should be increased substantially by measuring more profiles. The objective nature of SSFCM coupled with the high correlation with results from the visually based assay of morphology suggests the use of SSFCM to measure frequencies of misshapen sperm when testing for mutagens or monitoring for effects of environmental contaminants.

  9. Sperm morphology, motility, and velocity in naturally occurring polyploid European weatherfish (Misgurnus fossilis L.).

    PubMed

    Alavi, Sayyed Mohammad Hadi; Drozd, Bo?ek; Hatef, Azadeh; Flajšhans, Martin

    2013-07-15

    Spontaneous polyploidy has been frequently documented in various fish species. This process may lead to disruption in testicular development and function. In the present study, sperm morphology and motility, elements critical to male fertility, were characterized in the naturally occurring triploid (3n) and tetraploid (4n) European weatherfish, Misgurnus fossilis L. (Teleostei, Cobitidae) inhabiting the upper reaches of the Lužnice River in the Czech Republic. Sperm with smaller heads, shorter flagella, and a lower number of mitochondria was observed in 3n specimens compared with 4n, but no differences were observed in size of midpiece or ultrastructure of sperm. Similar to most teleosts, the European weatherfish spermatozoon lacked an acrosome and consisted of a head (containing DNA), a midpiece (containing mitochondria and proximal and distal centrioles), and a flagellum with 9 + 2 microtubular structure. Sperm velocity was significantly lower in individuals with 4n compared with 3n, whereas no difference in sperm motility was observed. The stepwise linear regression reported significant negative correlations between sperm velocity and length of sperm head (r = -0.92, P < 0.01). In conclusion, the data reported no effect of polyploidy on sperm ultrastructure and motility in the European weatherfish, although it can affect sperm velocity, possibly through differences in head size and the number of mitochondria, which provide ATP for sperm movement. PMID:23664795

  10. Hierarchical radial and polar organisation of chromosomes in human sperm.

    PubMed

    Millan, N M; Lau, P; Hann, M; Ioannou, D; Hoffman, D; Barrionuevo, M; Maxson, W; Ory, S; Tempest, H G

    2012-10-01

    It is well established that chromosomes occupy distinct positions within the interphase nuclei, conferring a potential functional implication to the genome. In addition, alterations in the nuclear organisation patterns have been associated with disease phenotypes (e.g. cancer or laminopathies). The human sperm is the smallest cell in the body with specific DNA packaging and the mission of delivering the paternal genome to the oocyte during fertilisation. Studies of nuclear organisation in the sperm have postulated nonrandom chromosome position and have proposed a chromocentre model with the centromeres facing toward the interior and the telomeres toward the periphery of the nucleus. Most studies have assessed the nuclear address in the sperm longitudinally predominantly using centromeric or telomeric probes and to a lesser extent with whole chromosome paints. To date, studies investigating the radial organisation of human sperm have been limited. The purpose of this study was to utilise whole chromosome paints for six clinically important chromosomes (18, 19, 21, 22, X, and Y) to investigate nuclear address by assessing their radial and longitudinal nuclear organisation. A total of 10,800 sperm were analysed in nine normozoospermic individuals. The results have shown nonrandom chromosome position for all chromosomes using both methods of analysis. We present novel radial and polar analysis of chromosome territory localization within the human sperm nucleus. Specifically, a hierarchical organisation was observed radially with chromosomes organised from the interior to the periphery (chromosomes 22, 21, Y, X, 19, and 18 respectively) and polar organisation from the sperm head to tail (chromosomes X, 19, Y, 22, 21, and 18, respectively). We provide evidence of defined nuclear organisation in the human sperm and discuss the function of organisation and potential possible clinical ramifications of these results in regards to male infertility and early human development. PMID:23143649

  11. Beyond Testis Size: Links between Spermatogenesis and Sperm Traits in a Seasonal Breeding Mammal

    PubMed Central

    Pintus, Eliana; Ros-Santaella, José Luis; Garde, José Julián

    2015-01-01

    Spermatogenesis is a costly process that is expected to be under selection to maximise sperm quantity and quality. Testis size is often regarded as a proxy measure of sperm investment, implicitly overlooking the quantitative assessment of spermatogenesis. An enhanced understanding of testicular function, beyond testis size, may reveal further sexual traits involved in sperm quantity and quality. Here, we first estimated the inter-male variation in testicular function and sperm traits in red deer across the breeding and non-breeding seasons. Then, we analysed the relationships between the testis mass, eight parameters of spermatogenic function, and seven parameters of sperm quality. Our findings revealed that the Sertoli cell number and function parameters vary greatly between red deer males, and that spermatogenic activity co-varies with testis mass and sperm quality across the breeding and non-breeding seasons. For the first time in a seasonal breeder, we found that not only is the Sertoli cell number important in determining testis mass (r = 0.619, p = 0.007 and r = 0.248, p = 0.047 for the Sertoli cell number assessed by histology and cytology, respectively), but also sperm function (r = 0.703, p = 0.002 and r = 0.328, p = 0.012 for the Sertoli cell number assessed by histology and cytology, respectively). Testicular histology also revealed that a high Sertoli cell number per tubular cross-section is associated with high sperm production (r = 0.600, p = 0.009). Sperm production and function were also positively correlated (r = 0.384, p = 0.004), suggesting that these traits co-vary to maximise sperm fertilisation ability in red deer. In conclusion, our findings contribute to the understanding of the dynamics of spermatogenesis, and reveal new insights into the role of testicular function and the Sertoli cell number on testis size and sperm quality in red deer. PMID:26430740

  12. Sperm characteristics and heterologous in vitro fertilisation capacity of Iberian ibex (Capra pyrenaica) epididymal sperm, frozen in the presence of the enzymatic antioxidant catalase.

    PubMed

    López-Saucedo, J; Paramio, M T; Fierro, R; Izquierdo, D; Catalá, M G; Coloma, M A; Toledano-Díaz, A; López-Sebastián, A; Santiago-Moreno, J

    2014-06-01

    The aim of this work was to evaluate the protective effect of catalase (CAT) on frozen/thawed ibex epididymal sperm recovered post mortem, and to detect any harmful effect this might have on sperm fertilisation capacity. Epididymal spermatozoa were diluted using a Tris-citric acid-glucose medium (TCG) composed of 3.8% Tris (w/v), 2.2% citric acid (w/v), 0.6% glucose (w/v), 5% glycerol (v/v), and 6% egg yolk (v/v). Sperm masses from the right epididymis were diluted with TCG medium, while those from the left were diluted with TCG medium supplemented with 200IU/mL CAT. Heterologous in vitro fertilisation (IVF) was used to assess the fertilisation capacity of this sperm. The addition of CAT to the extender did not improve frozen/thawed sperm variables. Moreover, a reduced fertilisation capacity was detected: sperm diluted with TCG provided 25.5% 2PN zygotes, while just 13.2% was recorded for that diluted with TCG-CAT (P<0.01). The percentage of cleaved embryos at 48hpi was higher (P<0.01) with the TCG sperm than with the TCG-CAT sperm (16.7% vs. 7.6%). The use of 200IU/mL CAT as an additive cannot, therefore, be recommended for the preservation of ibex epididymal sperm. Other antioxidants should, however, be tested in both this and related wild mountain ungulates. PMID:24699464

  13. Murine Binder of SPerm homolog 2 (BSPH2): the black sheep of the BSP superfamily.

    PubMed

    Plante, Geneviève; Fan, Jinjiang; Manjunath, Puttaswamy

    2014-01-01

    Proteins of the Binder of SPerm superfamily are known to bind choline phospholipids on sperm membrane and promote sperm capacitation. The current study focuses on the biochemical and functional characterization of the murine Binder of SPerm homolog 2 (BSPH2). A recombinant protein (rec-BSPH2) was expressed in Escherichia coli Rosetta-gami B (DE3)pLysS cells using pET32a vector. It was purified by immobilized metal ion affinity chromatography and refolded on column using a decreasing urea gradient. Rec-BSPH2 was found to share some binding characteristics with other BSP proteins, such as binding to gelatin, heparin, and epididymal sperm. Rec-BSPH2 as well as murine recombinant BSPH1 were found to have different immunofluorescence patterns when bound to uncapacitated versus capacitated sperm, indicating a rearrangement of these proteins on sperm surface during or following capacitation. Surprisingly, rec-BSPH2 was unable to bind phosphorylcholine liposomes or promote sperm capacitation. It is the first time that such results are reported for proteins of the BSP family. The results indicate that murine BSPH1 and BSPH2 might not have redundant functions, as is the case with bovine BSPs. This study could lead to a better understanding of the role of BSP proteins in sperm functions and the existence of redundant BSP proteins in the reproductive tract. PMID:24307707

  14. Methods for Cryopreservation of Guinea Fowl Sperm

    PubMed Central

    Váradi, Éva; Végi, Barbara; Liptói, Krisztina; Barna, Judit

    2013-01-01

    Conservation of indigenous poultry species is an important part of the new Hungarian agricultural strategy. Semen cryopreservation is the most practical method for the long term storage of poultry genetic material. The objective was to compare four protocols for cryopreservation of guinea fowl sperm (slow and fast programmable, freezing in nitrogen vapor, and pellet) and three cryoprotectants (10% ethylene glycol, 6% dimethyl-formamide and 6% dimethyl-acetamide). The efficiency of the methods was examined by in vitro tests (subjective motility scoring, sperm concentration, morphological and live/dead sperm analysis with eosin-aniline staining). Thereafter, the two most promising methods were tested by artificial insemination of frozen-thawed semen (3 times a week for 3 weeks using 300 million spermatozoa/hen), followed by candling of incubated eggs, assessment of fertilization, embryonic death, and hatching rate. The survival rate of live, intact spermatozoa was greatest (p?0.05) in pellet method and the slow programmable protocol (with 10% ethylene glycol) (28.6 and 23.5%). The two best protocols (based on in vitro assessment of post-thaw semen quality) were subsequently tested in vivo with artificial insemination. The pellet method yielded a 64% fertility rate compared to slow protocol with only 30% fertility. Regardless, both freezing protocols significantly increased embryonic deaths compared to the control group (16,7; 9,1 and 8,3%, respectively). During the 3-week in vivo trial, fertility increased and early embryonic death decreased over time. According to the results the guinea fowl sperm could tolerate the fast freezing in pellet better than the slower freezing rates and resulted acceptable fertility rate. PMID:23658648

  15. Sperm mitochondria in reproduction: good or bad and where do they go?

    PubMed

    Luo, Shi-Ming; Schatten, Heide; Sun, Qing-Yuan

    2013-11-20

    The mitochondrion is the major energy provider to power sperm motility. In mammals, aside from the nuclear genome, mitochondrial DNA (mtDNA) also contributes to oxidative phosphorylation to impact production of ATP by coding 13 polypeptides. However, the role of sperm mitochondria in fertilization and its final fate after fertilization are still controversial. The viewpoints that sperm bearing more mtDNA will have a better fertilizing capability and that sperm mtDNA is actively eliminated during early embryogenesis are widely accepted. However, this may be not true for several mammalian species, including mice and humans. Here, we review the sperm mitochondria and their mtDNA in sperm functions, and the mechanisms of maternal mitochondrial inheritance in mammals. PMID:24238608

  16. Current knowledge on boar sperm metabolism: Comparison with other mammalian species.

    PubMed

    Rodríguez-Gil, Joan E; Bonet, Sergi

    2016-01-01

    A practical consequence of the specific pig reproductive cycle is that the main functional features that distinguish boar spermatozoa cannot be extrapolated to other species. This prevents an overall picture that explains mammalian sperm function from being assumed. Furthermore, the extraordinary complexity of the molecular mechanisms implied in the control and modulation of mature boar sperm functions makes it impossible to provide a complete description of these mechanisms in the limited space of this chapter. Taking this into account, this chapter centers on the description of three highly important specific aspects of boar sperm function. The first aspect is the mechanisms by which boar sperm cells uptake extracellular energy sources. The second aspect is the necessity of mammalian sperm to use other hexoses than glucose as feasible energy sources. The third aspect would be an analysis of the roles that mitochondria could play in the regulation of the overall boar sperm function. As a whole, this revision intends to be an overall picture of regulatory mechanisms involved in the maintenance of proper energy levels of boar sperm and their relationship with the control of the overall boar sperm function. PMID:26094247

  17. Pattern of sperm transfer in redback spiders: implications for sperm competition and

    E-print Network

    Lovejoy, Nathan

    Pattern of sperm transfer in redback spiders: implications for sperm competition and male sacrifice at Scarborough, 1265 Military Trail, Scarborough, ON, M1C 1A4, Canada Many sperm competition studies have of understanding mechanisms of sperm transfer when attempting to interpret the outcome of sperm competition studies

  18. Effect of male age on sperm traits and sperm competition success in the guppy (Poecilia reticulata)

    E-print Network

    Pilastro, Andrea

    Effect of male age on sperm traits and sperm competition success in the guppy (Poecilia reticulata by Jennions & Petrie, 2000; Simmons, 2005). Polyandry may also benefit females by promoting sperm competition to enhance the probability of fertilizing her eggs with sperm of males that are better-than-average sperm

  19. Involvement of opsins in mammalian sperm thermotaxis

    PubMed Central

    Pérez-Cerezales, Serafín; Boryshpolets, Sergii; Afanzar, Oshri; Brandis, Alexander; Nevo, Reinat; Kiss, Vladimir; Eisenbach, Michael

    2015-01-01

    A unique characteristic of mammalian sperm thermotaxis is extreme temperature sensitivity, manifested by the capacity of spermatozoa to respond to temperature changes of <0.0006?°C as they swim their body-length distance. The identity of the sensing system that confers this exceptional sensitivity on spermatozoa is not known. Here we show that the temperature-sensing system of mammalian spermatozoa involves opsins, known to be G-protein-coupled receptors that act as photosensors in vision. We demonstrate by molecular, immunological, and functional approaches that opsins are present in human and mouse spermatozoa at specific sites, which depend on the species and the opsin type, and that they are involved in sperm thermotaxis via two signalling pathways—the phospholipase C and the cyclic-nucleotide pathways. Our results suggest that, depending on the context and the tissue, mammalian opsins act not only as photosensors but also as thermosensors. PMID:26537127

  20. Delineating the roles of males and females in sperm competition

    PubMed Central

    Evans, Jonathan P.; Rosengrave, Patrice; Gasparini, Clelia; Gemmell, Neil J.

    2013-01-01

    Disentangling the relative roles of males, females and their interactive effects on competitive fertilization success remains a challenge in sperm competition. In this study, we apply a novel experimental framework to an ideally suited externally fertilizing model system in order to delineate these roles. We focus on the chinook salmon, Oncorhynchus tshawytscha, a species in which ovarian fluid (OF) has been implicated as a potential arbiter of cryptic female choice for genetically compatible mates. We evaluated this predicted sexually selected function of OF using a series of factorial competitive fertilization trials. Our design involved a series of 10 factorial crosses, each involving two ‘focal’ rival males whose sperm competed against those from a single ‘standardized’ (non-focal) rival for a genetically uniform set of eggs in the presence of OF from two focal females. This design enabled us to attribute variation in competitive fertilization success among focal males, females (OF) and their interacting effects, while controlling for variation attributable to differences in the sperm competitive ability of rival males, and male-by-female genotypic interactions. Using this experimental framework, we found that variation in sperm competitiveness could be attributed exclusively to differences in the sperm competitive ability of focal males, a conclusion supported by subsequent analyses revealing that variation in sperm swimming velocity predicts paternity success. Together, these findings provide evidence that variation in paternity success can be attributed to intrinsic differences in the sperm competitive ability of rival males, and reveal that sperm swimming velocity is a key target of sexual selection. PMID:24266039

  1. Sperm Proteomics: Road to Male Fertility and Contraception

    PubMed Central

    Rahman, Md Saidur; Lee, June-Sub

    2013-01-01

    Spermatozoa are highly specialized cells that can be easily obtained and purified. Mature spermatozoa are transcriptionally and translationally inactive and incapable of protein synthesis. In addition, spermatozoa contain relatively higher amounts of membrane proteins compared to other cells; therefore, they are very suitable for proteomic studies. Recently, the application of proteomic approaches such as the two-dimensional polyacrylamide gel electrophoresis, mass spectrometry, and differential in-gel electrophoresis has identified several sperm-specific proteins. These findings have provided a further understanding of protein functions involved in different sperm processes as well as of the differentiation of normal state from an abnormal one. In addition, studies on the sperm proteome have demonstrated the importance of spermatozoal posttranslational modifications and their ability to induce physiological changes responsible for fertilization. Large-scale proteomic studies to identify hundreds to thousands of sperm proteins will ultimately result in the development of novel biomarkers that may help to detect fertility, the state of complete contraception, and beyond. Eventually, these protein biomarkers will allow for a better diagnosis of sperm dysfunctions and aid in drug development. This paper reviews the recent scientific publications available from the PubMed database to address sperm proteomics and its potential application to characterize male fertility and contraception. PMID:24363670

  2. Nanomedicine and mammalian sperm: Lessons from the porcine model.

    PubMed

    Barkalina, Natalia; Jones, Celine; Coward, Kevin

    2016-01-01

    Biomedical nanotechnology allows us to engineer versatile nanosized platforms that are comparable in size to biological molecules and intracellular organelles. These platforms can be loaded with large amounts of biological cargo, administered systemically and act at a distance, target specific cell populations, undergo intracellular internalization via endogenous uptake mechanisms, and act as contrast agents or release cargo for therapeutic purposes. Over recent years, nanomaterials have been increasingly viewed as favorable candidates for intragamete delivery. Particularly in the case of sperm, nanomaterial-based approaches have been shown to improve the efficacy of existing techniques such as sperm-mediated gene transfer, loading sperm with exogenous proteins, and tagging sperm for subsequent sex- or function-based sorting. In this short review, we provide an outline of the current state of nanotechnology for biomedical applications in reproductive biology and present highlights from a series of our studies evaluating the use of specialized silica nanoparticles in boar sperm as a potential delivery vehicle into mammalian gametes. The encouraging data obtained already from the porcine model in our laboratory have formed the basis for ethical approval of similar experiments in human sperm, thereby bringing us a step closer toward the potential use of this novel technology in the clinical environment. PMID:26116055

  3. Behavioral mechanisms of mammalian sperm guidance

    PubMed Central

    Pérez-Cerezales, Serafín; Boryshpolets, Sergii; Eisenbach, Michael

    2015-01-01

    In mammals, sperm guidance in the oviduct appears essential for successful sperm arrival at the oocyte. Hitherto, three different potential sperm guidance mechanisms have been recognized: thermotaxis, rheotaxis, and chemotaxis, each of them using specific stimuli – a temperature gradient, fluid flow, and a chemoattractant gradient, respectively. Here, we review sperm behavioral in these mechanisms and indicate commonalities and differences between them. PMID:25999361

  4. SPERM STORAGE IN FEMALE . PLETHODONTIDS WITH ESPECIAL

    E-print Network

    Sever, David M.

    17 SPERM STORAGE IN FEMALE . PLETHODONTIDS WITH ESPECIAL REFERENCE TO THE DESMOGNATHINAE David M, undergo internal fertilization (Duellman and Trueb, 1986). Internal fertilization occurs after a sperm of the female after ritualized courtship (Sever and Houck, 1985). Sperm from the cap migrate into sperm storage

  5. September 2000 DWARF SPERM WHALE (Kogia simus)

    E-print Network

    190 September 2000 DWARF SPERM WHALE (Kogia simus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters. 1991; Southeast Fisheries Science Center, SEFSC, unpublished data). Dwarf sperm whales and pygmy sperm

  6. October 1999 DWARF SPERM WHALE (Kogia simus)

    E-print Network

    176 October 1999 DWARF SPERM WHALE (Kogia simus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters Fisheries Science Center, SEFSC, unpublished data). Dwarf sperm whales and pygmy sperm whales (Kogia

  7. December 2009 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    560 December 2009 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate al. 1991; Mullin and Fulling 2004; Maze- Foley and Mullin 2006). Pygmy sperm whales and dwarf sperm

  8. December 2005 DWARF SPERM WHALE (Kogia sima)

    E-print Network

    256 December 2005 DWARF SPERM WHALE (Kogia sima): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters in oceanic waters (Figure 1; Mullin et al. 1991; Mullin and Fulling 2004). Dwarf sperm whales and pygmy sperm

  9. October 2008 DWARF SPERM WHALE (Kogia sima)

    E-print Network

    260 October 2008 DWARF SPERM WHALE (Kogia sima): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters in oceanic waters (Figure 1; Mullin et al. 1991; Mullin and Fulling 2004). Dwarf sperm whales and pygmy sperm

  10. December 2012 DWARF SPERM WHALE (Kogia sima)

    E-print Network

    275 December 2012 DWARF SPERM WHALE (Kogia sima): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters-Foley and Mullin 2006). Dwarf sperm whales and pygmy sperm whales (Kogia breviceps) are difficult to differentiate

  11. September 2000 DWARF SPERM WHALE (Kogia simus)

    E-print Network

    219 September 2000 DWARF SPERM WHALE (Kogia simus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters. 1991; Southeast Fisheries Science Center, SEFSC, unpublished data). Dwarf sperm whales and pygmy sperm

  12. December 2005 DWARF SPERM WHALE (Kogia sima)

    E-print Network

    354 December 2005 DWARF SPERM WHALE (Kogia sima): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters in oceanic waters (Figure 1; Mullin et al. 1991; Mullin and Fulling 2004). Dwarf sperm whales and pygmy sperm

  13. October 2007 DWARF SPERM WHALE (Kogia sima)

    E-print Network

    305 October 2007 DWARF SPERM WHALE (Kogia sima): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters in oceanic waters (Figure 1; Mullin et al. 1991; Mullin and Fulling 2004). Dwarf sperm whales and pygmy sperm

  14. December 2012 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    281 December 2012 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate al. 1991; Mullin and Fulling 2004; Maze-Foley and Mullin 2006). Pygmy sperm whales and dwarf sperm

  15. September 2000 DWARF SPERM WHALE (Kogia simus)

    E-print Network

    289 September 2000 DWARF SPERM WHALE (Kogia simus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters. 1991; Southeast Fisheries Science Center, SEFSC, unpublished data). Dwarf sperm whales and pygmy sperm

  16. December 2009 DWARF SPERM WHALE (Kogia sima)

    E-print Network

    555 December 2009 DWARF SPERM WHALE (Kogia sima): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters-Foley and Mullin 2006). Dwarf sperm whales and pygmy sperm whales (Kogia breviceps) are difficult to differentiate

  17. December 1998 DWARF SPERM WHALE (Kogia simus)

    E-print Network

    50 December 1998 DWARF SPERM WHALE (Kogia simus): Western North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale (Kogia simus) and the pygmy sperm whale (K. breviceps) appear indicates that in that region K. simus is the nearshore species (Plön et al. 1998). Pygmy sperm whales

  18. December 2009 DWARF SPERM WHALE (Kogia sima)

    E-print Network

    December 2009 DWARF SPERM WHALE (Kogia sima): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale appears to be distributed worldwide in temperate to tropical waters-Foley and Mullin 2006). Dwarf sperm whales and pygmy sperm whales (Kogia breviceps) are difficult to differentiate

  19. December 2009 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    December 2009 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate to tropical waters; Mullin and Fulling 2004; Maze- Foley and Mullin 2006). Pygmy sperm whales and dwarf sperm whales (Kogia

  20. Assessment of different sperm quality parameters to predict in vitro fertility of bulls.

    PubMed

    Tanghe, Sofie; Van Soom, A; Sterckx, V; Maes, D; de Kruif, A

    2002-06-01

    Frozen-thawed semen from six bulls with high (> 60%) and low (20-35%) in vitro fertility was used for studying the predictive value of simple sperm quality tests with respect to in vitro fertilization (IVF) outcome as assessed by pronucleus (PN) formation ability. Sperm quality parameters, such as sperm concentration, motility, progressive motility, live-dead sperm ratio, morphology, membrane integrity, mitochondrial activity and acrosomal status were analysed using both conventional and automatic techniques at three time points during the IVF process, namely after sperm thawing, Percoll differential gradient centrifugation and IVF. Associations between the sperm quality parameters before and after IVF, and PN formation ability were assessed by using linear regression analyses. The percentages of motility, progressive motility and normal morphology determined after sperm thawing, and the percentage of live spermatozoa assessed after Percoll preparation by using nigrosin-eosin (N-E) staining showed a good correlation with PN formation ability, but the regression parameters were borderline not significant. These parameters formed the most reliable basis for predicting IVF outcome. After IVF, the percentage of live spermatozoa determined by using N-E staining was the only sperm quality parameter showing a significant association with the PN formation ability of a given bull. This sperm quality test can be used as a non-invasive method to estimate the PN formation ability of oocytes which are further cultured to assess embryonic development. PMID:12071885

  1. Correlation between the spermatozoal characteristics and sperm penetration distance in polyacrylamide gel and bovine cervical mucus.

    PubMed

    Anilkumar, R; Devanathan, T G; Pattabiraman, S R; Edwin, M J

    2001-01-15

    Correlation between the spermatozoal characteristics and the sperm penetration distance in polyacrylamide gel was assessed, utilizing frozen thawed semen samples obtained from 6 bulls, and it was compared with the correlation between sperm penetration in bovine cervical mucus and spermatozoal characteristics. In vitro sperm penetration tests were performed with mucus and gel. The sperm penetration in gel and mucus was significantly and positively correlated with post-thaw motility (r=0.81; r=0.89:P<0.01) and acrosome integrity (r=0.88; r=0.94:P<0.01). A significant negative correlation with abnormal spermatozoa (r=-0.84;r=0.83:P<0.01) was observed. Both sperm concentration and post-thaw live spermatozoa were not significantly correlated. A significant multiple regression between sperm penetration and the spermatozoal characteristics both in gel (R2=0.87; F=40.27; P<0.01) and mucus (R2=0.91; F=60.48; P<0.01) was observed. The major spermatozoal characteristics determining the capacity of spermatozoa to penetrate gel were post-thaw motility, percentage of abnormal spermatozoa and acrosome integrity. The acrosome integrity has a more significant contribution. The correlation established with sperm penetration in gel was very similar to that of sperm penetration in mucus. The utility of gel as a mucus substitute in in vitro sperm penetration tests was discussed. PMID:11233793

  2. Cryopreservation of sperm of red abalone (Haliotis rufescens)

    USGS Publications Warehouse

    Salinas-Flores, L.; Paniagua-Chavez, C. G.; Jenkins, J.A.; Tiersch, T.R.

    2005-01-01

    Abalone culture, a developing industry in Baja California, Mexico, would benefit from genetic improvement and controlled breeding. The use of cryopreserved sperm would allow germplasm availability, and this study was designed to develop sperm cryopreservation protocols for red abalone Haliotis rufescens. The acute toxic effects of the cryoprotectants dimethyl sulfoxide (DMSO), propylene glycol (PG), and glycerol (GLY) were assessed after suspending sperm in different concentrations, whereby cryoprotectant treatments of 10% DMSO and 10% GLY equilibrated for 10 min yielded the highest range of motile sperm in preliminary freezing trials and were used for cryopreservation studies. To determine effective cooling rates, three freezing chambers were tested. Replicate samples of sperm from 4 males were placed in 0.5-mL French straws and frozen using a commercial freezing chamber (CFC) used for bull sperm, a programmable rate chamber (PRC), and a manually controlled styrofoam chamber (MCC). For the CFC, the cooling rate was 16??C/min, from 4??C to -140??C. For the PRC and MCC, it was 1??C/min, from -20??C to -30??C. The samples were held at -30??C for 5 min before being plunged into liquid nitrogen (-196??C) for storage, and each sample was thawed in a water bath at 45??C for 8 s. The quality of thawed sperm was determined by estimating percent motility, evaluating membrane integrity using a dual-staining technique and flow cytometry, and estimating fertilization rate. Statistical analyses were performed using 2-way ANOVA where chamber and treatment were the independent variables. Sperm quality parameters were independent. For motilities, a significant interaction was noted between the cryoprotective treatment and the chamber type, whereby motilities for DMSO and GLY were higher (P = 0.0055) using MCC. Membrane integrities were significantly lower after using the PRC than the CFC or the MCC (P = 0.0167). The highest post-thaw motility (48 ?? 7%) was found using sperm suspended in 10% glycerol and frozen in the MCC. The highest percent of intact membranes (56 ?? 11%) was for sperm suspended in 10% glycerol and frozen in the CFC. The highest fertilization rate (29 ?? 10%) was with samples frozen with 10% glycerol in the CFC. The use of cryopreserved sperm from red abalone provides an alternative breeding option for culture and the protocols delineated are the first developed for this species.

  3. Analysis of the correlation between sperm DNA integrity and conventional semen parameters in infertile men

    PubMed Central

    Aydos, Oya Sena; Yükselten, Yunus; Kaplan, Fuat; Sunguro?lu, Asuman; Aydos, Kaan

    2015-01-01

    Objective A male factor is responsible in approximately 30–40% of couples receiving infertility treatment. Routinely, such couples undergo semen analysis including parameters such as sperm count, motility and morphology. Generally, the analysis of sperm DNA damage, shown to have a significant clinical importance by many studies, is recognized as an advanced test that is not included in routine infertility tests. Intracytoplasmic sperm injection method, commonly employed in the current infertility treatment protocols, lowers the fertilization rate, however, fertilization can occur even with a damaged DNA which is known to pose a risk in the subsequent pregnancy period. The relation between sperm morphology and the degree of sperm DNA damage has not yet been understood clearly. In this study, we aimed to investigate the association between routine semen analysis and sperm DNA integrity assay, another advanced but costly method. Material and methods The degree of DNA damage was compared with the results of semen analysis, based on the WHO criteria, in 399 male patients who received comet assay for sperm DNA integrity. The statistical correlation analyses were performed with Windows SPPS statistical package program. Results Accordingly, the sperm DNA damage was found to be correlated with all 3 parameters (sperm count, forward motility, and morphology) examined by the semen analysis (p<0.001). Total sperm DNA Damage Count was 226, 216, and 210 arbitrary units in patients with a sperm count <15 mil/mL, forward moving motility <32%, and normal morphology <4%, respectively. The difference with the normal individuals was statistically significant (p<0.001). Conclusion In light of the comet assay results, higher degree of sperm DNA damage is associated with significant impairment of all seminal parameters. PMID:26623148

  4. Test Functions Space in Noncommutative Quantum Field Theory

    E-print Network

    M. Chaichian; M. Mnatsakanova; A. Tureanu; Yu. Vernov

    2008-07-26

    It is proven that the $\\star$-product of field operators implies that the space of test functions in the Wightman approach to noncommutative quantum field theory is one of the Gel'fand-Shilov spaces $S^{\\beta}$ with $\\beta test functions smears the noncommutative Wightman functions, which are in this case generalized distributions, sometimes called hyperfunctions. The existence and determination of the class of the test function spaces in NC QFT is important for any rigorous treatment in the Wightman approach.

  5. Using Dependency Structures for Prioritisation of Functional Test Suites

    E-print Network

    Miller, Tim

    1 Using Dependency Structures for Prioritisation of Functional Test Suites Shifa-e-Zehra Haidry, Australia. 3 Abstract--Test case prioritisation is the process of ordering the exe- cution of test cases software delivery. Many existing test case prioritisation techniques consider that tests can be run in any

  6. Chromosomal abnormalities in human sperm

    SciTech Connect

    Martin, R.H.

    1985-01-01

    The ability to analyze human sperm chromosome complements after penetration of zona pellucida-free hamster eggs provides the first opportunity to study the frequency and type of chromosomal abnormalities in human gametes. Two large-scale studies have provided information on normal men. We have studied 1,426 sperm complements from 45 normal men and found an abnormality rate of 8.9%. Brandriff et al. (5) found 8.1% abnormal complements in 909 sperm from 4 men. The distribution of numerical and structural abnormalities was markedly dissimilar in the 2 studies. The frequency of aneuploidy was 5% in our sample and only 1.6% in Brandriff's, perhaps reflecting individual variability among donors. The frequency of 24,YY sperm was low: 0/1,426 and 1/909. This suggests that the estimates of nondisjunction based on fluorescent Y body data (1% to 5%) are not accurate. We have also studied men at increased risk of sperm chromosomal abnormalities. The frequency of chromosomally unbalanced sperm in 6 men heterozygous for structural abnormalities varied dramatically: 77% for t11;22, 32% for t6;14, 19% for t5;18, 13% for t14;21, and 0% for inv 3 and 7. We have also studied 13 cancer patients before and after radiotherapy and demonstrated a significant dose-dependent increase of sperm chromosome abnormalities (numerical and structural) 36 months after radiation treatment.

  7. Drosophila Sperm Swim Backwards in the Female Reproductive Tract and Are Activated via TRPP2 Ion Channels

    PubMed Central

    Chu, Kristy; Cook, Stacey; Montell, Craig; Watnick, Terry

    2011-01-01

    Background Sperm have but one purpose, to fertilize an egg. In various species including Drosophila melanogaster female sperm storage is a necessary step in the reproductive process. Amo is a homolog of the human transient receptor potential channel TRPP2 (also known as PKD2), which is mutated in autosomal dominant polycystic kidney disease. In flies Amo is required for sperm storage. Drosophila males with Amo mutations produce motile sperm that are transferred to the uterus but they do not reach the female storage organs. Therefore Amo appears to be a mediator of directed sperm motility in the female reproductive tract but the underlying mechanism is unknown. Methodology/Principal Findings Amo exhibits a unique expression pattern during spermatogenesis. In spermatocytes, Amo is restricted to the endoplasmic reticulum (ER) whereas in mature sperm, Amo clusters at the distal tip of the sperm tail. Here we show that flagellar localization of Amo is required for sperm storage. This raised the question of how Amo at the rear end of sperm regulates forward movement into the storage organs. In order to address this question, we used in vivo imaging of dual labelled sperm to demonstrate that Drosophila sperm navigate backwards in the female reproductive tract. In addition, we show that sperm exhibit hyperactivation upon transfer to the uterus. Amo mutant sperm remain capable of reverse motility but fail to display hyperactivation and directed movement, suggesting that these functions are required for sperm storage in flies. Conclusions/Significance Amo is part of a signalling complex at the leading edge of the sperm tail that modulates flagellar beating and that guides a backwards path into the storage organs. Our data support an evolutionarily conserved role for TRPP2 channels in cilia. PMID:21625494

  8. LewisX-containing glycans on the porcine oviductal epithelium contribute to formation of the sperm reservoir.

    PubMed

    Machado, Sergio A; Kadirvel, Govindasamy; Daigneault, Bradford W; Korneli, Claudia; Miller, Paul; Bovin, Nicolai; Miller, David J

    2014-12-01

    In many mammals, after semen deposition, a subpopulation of the sperm is transported to the lower oviduct, or isthmus, to form a functional sperm reservoir that provides sperm to fertilize oocytes. The precise molecular interactions that allow formation of this reservoir are unclear. It is proposed that binding of sperm receptors (lectins) to their oviductal cell ligands is accomplished by glycans. Previous results indicated that Lewis trisaccharides are present in glycosphingolipids and O- and N-linked glycans of the porcine isthmus and that Le(X)-containing molecules bind porcine sperm. Immunohistochemistry indicated that the Lewis structures identified by mass spectrometry were, in fact, Lewis X (Le(X)) trisaccharides. These motifs were localized to the luminal border of the isthmus. Assays using fluoresceinated glycans showed that 3-O-sulfated Le(X) (suLe(X)) bound to receptors localized on the head of nearly 60% of uncapacitated boar sperm but that the positional isomer 3-O-sulfo-Le(A) (suLe(A)) bound to <5% of sperm. Sperm also bound preferentially to suLe(X) made insoluble by coupling to beads. Capacitation reduced the ability of suLe(X) to bind sperm to <10%, perhaps helping to explain why sperm are released at capacitation. Pretreatment of oviduct cell aggregates with the Le(X) antibody blocked 57% of sperm binding to isthmic aggregates. Blocking putative receptors on sperm with soluble Le(X) and suLe(X) glycans specifically reduced sperm binding to oviduct cells up to 61%. These results demonstrate that the oviduct isthmus contains Le(X)-related moieties and that sperm binding to these oviduct glycans is necessary and sufficient for forming the sperm reservoir. PMID:25339106

  9. Screening Test Items for Differential Item Functioning

    ERIC Educational Resources Information Center

    Longford, Nicholas T.

    2014-01-01

    A method for medical screening is adapted to differential item functioning (DIF). Its essential elements are explicit declarations of the level of DIF that is acceptable and of the loss function that quantifies the consequences of the two kinds of inappropriate classification of an item. Instead of a single level and a single function, sets of…

  10. SPERM COUNT, MORPHOLOGY AND FLUORESCENT BODY FREQUENCY IN AUTOPSY SERVICE WORKERS EXPOSED TO FORMALDEHYDE

    EPA Science Inventory

    The ability of a battery of genetic monitoring tests to detect occupational formaldehyde exposure in a population of a hospital autopsy service workers was investigated. Eleven exposed individuals and 11 matched controls were evaluated for sperm count, abnormal sperm morphology a...

  11. Constructing Test Functions for Global Optimization Using Continuous

    E-print Network

    Butenko, Sergiy

    Constructing Test Functions for Global Optimization Using Continuous Formulations of Graph Problems. In particular, global optimization formulations for the max- imum independent set, maximum clique and MAX CUT CUT; Constrained global optimization; Test problems 1 Introduction Good test functions are essential

  12. First State Fitness Test. A Measurement of Functional Health.

    ERIC Educational Resources Information Center

    Brown, Timothy; And Others

    This test is designed to measure the functional health of young people. Functional health refers to those factors relating to personal health that can be improved with regular exercise. This test is unique in comparison to other physical fitness tests because of the absence of motor skill items which have no relationship to an individual's…

  13. Dynamics of the mammalian sperm plasma membrane in the process of fertilization.

    PubMed

    Flesch, F M; Gadella, B M

    2000-11-10

    Sexual reproduction requires the fusion of sperm cell and oocyte during fertilization to produce the diploid zygote. In mammals complex changes in the plasma membrane of the sperm cell are involved in this process. Sperm cells have unusual membranes compared to those of somatic cells. After leaving the testes, sperm cells cease plasma membrane lipid and protein synthesis, and vesicle mediated transport. Biophysical studies reveal that lipids and proteins are organized into lateral regions of the sperm head surface. A delicate reorientation and modification of plasma membrane molecules take place in the female tract when sperm cells are activated by so-called capacitation factors. These surface changes enable the sperm cell to bind to the extra cellular matrix of the egg (zona pellucida, ZP). The ZP primes the sperm cell to initiate the acrosome reaction, which is an exocytotic process that makes available the enzymatic machinery required for sperm penetration through the ZP. After complete penetration the sperm cell meets the plasma membrane of the egg cell (oolemma). A specific set of molecules is involved in a disintegrin-integrin type of anchoring of the two gametes which is completed by fusion of the two gamete plasma membranes. The fertilized egg is activated and zygote formation preludes the development of a new living organism. In this review we focus on the involvement of processes that occur at the sperm plasma membrane in the sequence of events that lead to successful fertilization. For this purpose, dynamics in adhesive and fusion properties, molecular composition and architecture of the sperm plasma membrane, as well as membrane derived signalling are reviewed. PMID:11063883

  14. Sperm-mediated gene transfer: effect on bovine in vitro embryo production.

    PubMed

    Simões, Renata; Nicacio, Alessandra Corallo; Binelli, Mario; de Paula-Lopes, Fabiola Freitas; Milazzotto, Marcella Pecora; Visintin, José Antonio; D'Ávila Assumpção, Mayra Elena Ortiz

    2013-11-01

    The technique of sperm-mediated gene transfer (SMGT) can be used to delivery exogenous DNA into the oocyte. However, it has low repeatability and produces inconsistent results. In order to optimize this technique, it is necessary to study the mechanism by which DNA enters the sperm cell and integrates in the sperm genome. Furthermore, studies must focus in the maintenance of sperm cell viability and function. The aim of this study was to evaluate different SMGT protocols of sperm electroporation or capacitation (CaI) aiming to maintain sperm viability in the production of bovine embryos in vitro. Frozen-thawed semen was divided in two experimental groups (electroporation or CaI) and one control group (non-treated cells). For the electroporation method, five different voltages (100, 500, 750, 1000 or 1500 V) with 25 ?F capacitance were used. For CaI treatment, combinations of two CaI concentrations (250 nM or 500 nM), two incubation periods of sperm cells with CaI (1 or 5 min) and two incubation periods that mimicked time of sperm cell interaction with exogenous DNA molecules (1 or 2 h) were evaluated. According to our data, electroporation and CaI treatments do not prevent sperm penetration and oocyte fertilization and can be an alternative method to achieve satisfactory DNA delivery in SMGT protocols. PMID:22805109

  15. Sperm Competition in Humans: Mate Guarding Behavior Negatively Correlates with Ejaculate Quality

    PubMed Central

    Leivers, Samantha; Rhodes, Gillian; Simmons, Leigh W.

    2014-01-01

    In species where females mate with multiple males, the sperm from these males must compete to fertilise available ova. Sexual selection from sperm competition is expected to favor opposing adaptations in males that function either in the avoidance of sperm competition (by guarding females from rival males) or in the engagement in sperm competition (by increased expenditure on the ejaculate). The extent to which males may adjust the relative use of these opposing tactics has been relatively neglected. Where males can successfully avoid sperm competition from rivals, one might expect a decrease in their expenditure on tactics for the engagement in sperm competition and vice versa. In this study, we examine the relationship between mate guarding and ejaculate quality using humans as an empirical model. We found that men who performed fewer mate guarding behaviors produced higher quality ejaculates, having a greater concentration of sperm, a higher percentage of motile sperm and sperm that swam faster and less erratically. These effects were found independent of lifestyle factors or factors related to male quality. Our findings suggest that male expenditure on mate guarding and on the ejaculate may represent alternative routes to paternity assurance in humans. PMID:25250582

  16. The channel capacity of a diagnostic test as a function of test sensitivity and test specificity.

    PubMed

    Benish, William A

    2015-12-01

    We apply the information theory concept of "channel capacity" to diagnostic test performance and derive an expression for channel capacity in terms of test sensitivity and test specificity. The expected value of the amount of information a diagnostic test will provide is equal to the "mutual information" between the test result and the disease state. For the case in which only two test results and two disease states are considered, mutual information, I(D;R), is a function of sensitivity, specificity, and the pretest probability of disease. The channel capacity of the test is the maximal value of I(D;R) for a given sensitivity and specificity. After deriving an expression for I(D;R) in terms of sensitivity, specificity, and pretest probability, we solve for the value of pretest probability that maximizes I(D;R). Channel capacity is obtained by using this value of pretest probability to calculate I(D;R). Channel capacity provides a convenient and meaningful single parameter measure of diagnostic test performance. It quantifies the upper limit of the amount of information a diagnostic test can be expected to provide about a patient's disease state. PMID:22368178

  17. Acridine Orange and Flow Cytometry: Which Is Better to Measure the Effect of Varicocele on Sperm DNA Integrity?

    PubMed Central

    Mohammed, Essam-Elden M.; Mosad, Eman; Zahran, Asmaa M.; Hameed, Diaa A.; Taha, Emad A.; Mohamed, Mohamed A.

    2015-01-01

    We evaluated the effect of varicocelectomy on semen parameters and levels of sperm DNA damage in infertile men. A total of 75 infertile men with varicocele and 40 fertile men (controls) were included in this study. Semen analysis and sperm DNA damage expressed as the DNA fragmentation index using acridine orange staining and chromatin condensation test by flow cytometry were assessed before and 6 months after varicocelectomy. The patients were also followed up for 1 year for pregnancy outcome. Semen parameters were significantly lower in varicocele patients compared to controls (P < 0.05). Mean percentages of sperm DNA fragmentation and sperm DNA chromatin condensation in patients were significantly higher than those in controls (P < 0.05). After varicocelectomy, sperm DNA fragmentation improved significantly, whereas sperm chromatin condensation was not significantly changed. In 15 out of 75 varicocele patients, clinical pregnancy was diagnosed; those with positive pregnancy outcome had significant improvement in sperm count, progressive sperm motility, and sperm DNA fragmentation, but there was no significant difference in sperm DNA condensation compared to negative pregnancy outcome patients. We concluded from this study that acridine orange stain is more reliable method than flow cytometry in the evaluation of sperm DNA integrity after varicocelectomy. PMID:26681938

  18. Stressful environments induce novel phenotypic variation: hierarchical reaction norms for sperm performance of a pervasive invader

    PubMed Central

    Purchase, Craig F; Moreau, Darek T R

    2012-01-01

    Genetic variation for phenotypic plasticity is ubiquitous and important. However, the scale of such variation including the relative variability present in reaction norms among different hierarchies of biological organization (e.g., individuals, populations, and closely related species) is unknown. Complicating interpretation is a trade-off in environmental scale. As plasticity can only be inferred over the range of environments tested, experiments focusing on fine tuned responses to normal or benign conditions may miss cryptic phenotypic variation expressed under novel or stressful environments. Here, we sought to discern the presence and shape of plasticity in the performance of brown trout sperm as a function of optimal to extremely stressful river pH, and demarcate if the reaction norm varies among genotypes. Our overarching goal was to determine if deteriorating environmental quality increases expressed variation among individuals. A more applied aim was to ascertain whether maintaining sperm performance over a wide pH range could help explain how brown trout are able to invade diverse river systems when transplanted outside of their native range. Individuals differed in their reaction norms of phenotypic expression of an important trait in response to environmental change. Cryptic variation was revealed under stressful conditions, evidenced through increasing among-individual variability. Importantly, data on population averages masked this variability in plasticity. In addition, canalized reaction norms in sperm swimming velocities of many individuals over a very large range in water chemistry may help explain why brown trout are able to colonize a wide variety of habitats. PMID:23145341

  19. Comparison of methods for detecting mitomycin C- and ethyl nitrosourea-induced germ cell damage in mice: sperm enzyme activities, sperm motility, and testis weight

    SciTech Connect

    Ficsor, G.; Oldford, G.M.; Loughlin, K.R.; Panda, B.B.; Dubien, J.L.; Ginsberg, L.C.

    1984-01-01

    Testes weights, sperm motility and enzyme activities in single sperm were compared with respect to their ability to detect either developmental or mutational damage to germ cells. Male mice were injected i.p. with 2.5 mg/kg mitomycin C (MC) or 50 or 100 mg/kg ethylnitrosourea (ENU) or saline and were then killed at times such that sperm derived from treated vas sperm (SZ), spermatids (ST), preleptotene-late-spermatogonial cells (PLSG), spermatogonial cells (SG), or spermatogonial stem cells (SGS) could be evaluated. The authors conclude that testis weight, which is easily obtained, is a sensitive indicator of germ cell damage by these agents. Sperm from each animal were evaluated for sperm motility, acrosin activity, succinic dehydrogenase (SDH) activity with or without the competitive inhibitor malonate or after exposure to 60/sup 8/C for 10 min. The latter two assays were to detect sperm enzymes resistant to the inhibitor or heat. The presence of the acrosin protein was also detected immunologically. Of the sperm assays, acrosin activity proved to be the most sensitive indicator of germ cell damage and was the simplest to measure.

  20. The Molecular Chaperone HSPA2 Plays a Key Role in Regulating the Expression of Sperm Surface Receptors That Mediate Sperm-Egg Recognition

    PubMed Central

    Redgrove, Kate A.; Nixon, Brett; Baker, Mark A.; Hetherington, Louise; Baker, Gordon; Liu, De-Yi; Aitken, R. John

    2012-01-01

    A common defect encountered in the spermatozoa of male infertility patients is an idiopathic failure of sperm–egg recognition. In order to resolve the molecular basis of this condition we have compared the proteomic profiles of spermatozoa exhibiting an impaired capacity for sperm-egg recognition with normal cells using label free mass spectrometry (MS)-based quantification. This analysis indicated that impaired sperm–zona binding was associated with reduced expression of the molecular chaperone, heat shock 70 kDa protein 2 (HSPA2), from the sperm proteome. Western blot analysis confirmed this observation in independent patients and demonstrated that the defect did not extend to other members of the HSP70 family. HSPA2 was present in the acrosomal domain of human spermatozoa as a major component of 5 large molecular mass complexes, the most dominant of which was found to contain HSPA2 in close association with just two other proteins, sperm adhesion molecule 1 (SPAM1) and arylsulfatase A (ARSA), both of which that have previously been implicated in sperm-egg interaction. The interaction between SPAM1, ARSA and HSPA2 in a multimeric complex mediating sperm-egg interaction, coupled with the complete failure of this process when HSPA2 is depleted in infertile patients, provides new insights into the mechanisms by which sperm function is impaired in cases of male infertility. PMID:23209833

  1. Mice lacking FABP9/PERF15 develop sperm head abnormalities but are fertile

    PubMed Central

    Selvaraj, Vimal; Asano, Atsushi; Page, Jennifer L.; Nelson, Jacquelyn L.; Kothapalli, Kumar S. D.; Foster, James A.; Brenna, J. Thomas; Weiss, Robert S.; Travis, Alexander J.

    2010-01-01

    The male germ cell-specific fatty acid binding protein 9 (FABP9/PERF15) is the major component of the murine sperm perforatorium and perinuclear theca. Based on its cytoskeletal association and sequence homology to myelin P2 (FABP8), it has been suggested that FABP9 tethers sperm membranes to the underlying cytoskeleton. Furthermore, its upregulation in apoptotic testicular germ cells and its increased phosphorylation status during capacitation suggested multiple important functions for FABP9. Therefore, we investigated specific functions for FABP9 by means of targeted gene disruption in mice. FABP9?/? mice were viable and fertile. Phenotypic analysis showed that FABP9?/? mice had significant increases in sperm head abnormalities (~8% greater than their WT cohorts); in particular, we observed the reduction or absence of the characteristic structural element known as the “ventral spur” in ~10% of FABP9?/? sperm. However, deficiency of FABP9 neither affected membrane tethering to the perinuclear theca nor the fatty acid composition of sperm. Moreover, epididymal sperm numbers were not affected in FABP9?/? mice. Therefore, we conclude that FABP9 plays only a minor role in providing the murine sperm head its characteristic shape and is not absolutely required for spermatogenesis or sperm function. PMID:20920498

  2. Clinical Background of Patients with Sperm in Their Urinary Sediment

    PubMed Central

    Kikuchi, Eiji; Kabeya, Yusuke; Katsuki, Takeshi; Oikawa, Yoichi; Kato, Kiyoe; Ohashi, Masakazu; Nakamura, So; Oya, Mototsugu; Shimada, Akira

    2015-01-01

    Introduction The detection rate and associated factors of at least one sperm in urinary sediment is not well-known in real clinical practice. Aims The aim of the present study was to evaluate the clinical features associated with the presence of sperm in urinary sediment in a large number of samples. Methods We conducted a cross-sectional study at Tokyo Saiseikai Central Hospital. We identified 5,005 males who were aged ?20 years in whom urinary sedimentation had been performed at least twice between May 2011 and June 2012. The sperm group included patients in whom at least one urinary sediment test performed under a microscope had detected at least one sperm. We evaluated the associations between the presence of at least one sperm in urinary sediment and clinical parameters such as various diseases and the use of particular oral medicines. Main Outcomes In total, 1.6% (339/20,937) of urinary sediment samples contained at least one sperm. The sperm group consisted of 282 subjects (5.6%), and the no-sperm group included 4,723 subjects (94.3%). Results Multivariate analysis demonstrated that younger age (<65) (odds ratio [OR]: 1.71, 95% confidence interval [CI]: 1.32–2.21), the total number of examinations (?4) (OR: 1.46, 95%CI: 1.11–1.92), diabetes (OR: 1.72, 95%CI: 1.31–2.25), a history of pelvic surgery for colon cancer (OR: 4.89, 95%CI: 2.38–10.02), alpha-1 blocker use (OR: 1.55, 95%CI: 1.16–2.08), a history of trans-urethral resection of the prostate (OR: 2.77, 95%CI: 1.46–5.13), and selective serotonin reuptake inhibitor use (OR: 2.12, 95%CI: 1.07–4.19) were independent predictors of the presence of at least one sperm in urinary sediment. Conclusion There is considerable overlap between the factors associated with the presence of at least one sperm in urinary sediment and those that are strongly associated with ejaculatory disorders. PMID:26359862

  3. Relationship between Testicular Volume and Conventional or Nonconventional Sperm Parameters

    PubMed Central

    Condorelli, Rosita; Calogero, Aldo E.; La Vignera, Sandro

    2013-01-01

    Background. Reduced testicular volume (TV) (<12?cm3) is associated with lower testicular function. Several studies explored the conventional sperm parameters (concentration, motility, and morphology) and the endocrine function (gonadotropins and testosterone serum concentrations) in the patients with reduction of TV. No other parameters have been examined. Aim. This study aims at evaluating some biofunctional sperm parameters by flow cytometry in the semen of men with reduced TV compared with that of subjects with normal TV. Methods. 78 patients without primary scrotal disease were submitted to ultrasound evaluation of the testis. They were divided into two groups according to testicular volume: A Group, including 40 patients with normal testicular volume (TV > 15?cm3) and B Group, including 38 patients with reduced testicular volume (TV ? 12?cm3). All patients underwent serum hormone concentration, conventional and biofunctional (flow cytometry) sperm parameters evaluation. Results. With regard to biofunctional sperm parameters, all values (mitochondrial membrane potential, phosphatidylserine externalization, chromatin compactness, and DNA fragmentation) were strongly negatively correlated with testicular volume (P < 0.0001). Conclusions. This study for the first time in the literature states that the biofunctional sperm parameters worsen and with near linear correlation, with decreasing testicular volume. PMID:24089610

  4. Testing quantum nonlocality by generalized quasiprobability functions

    E-print Network

    Lee, Seung-Woo; Jaksch, Dieter; 10.1103/PhysRevA.80.022104

    2009-01-01

    We derive a Bell inequality based on a generalized quasiprobability function which is parameterized by one non-positive real value. Two types of known Bell inequalities formulated in terms of the Wigner and Q functions are included as limiting cases. We investigate violations of our Bell inequalities for single photon entangled states and two-mode squeezed vacuum states when varying the detector efficiency. We show that the Bell inequality for the Q function allows the lowest detection efficiency for violations of local realism.

  5. Identifying X- and Y-chromosome-bearing sperm by DNA content: retrospective perspectives and prospective opinions

    SciTech Connect

    Gledhill, B.L.; Pinkel, D.; Garner, D.L.

    1982-03-05

    Theoretically, since DNA should be the most constant component, quantitatively, of normal sperm, then genotoxic agents arising from energy production and consumption, and chemical and physical mutagens, could be identified by measuring variability in the DNA content of individual sperm from exposed men or test animals. The difference between the DNA content of X and Y sperm seemed a biologically significant benchmark for the measurement technology. Several methods are available for determining the genetic activity of agents in male germ cells, but these tests are generally laborious. Sperm-based methods provide an attractive alternate since they are not invasive, and are directly applicable to the study of human exposure. Slide-based assay of DNA content suggests that human sperm with X, Y, or YY chromosome constitutions can be distinguished by their fluorescence with quinacrine. Subsequent measurement of the dry mass of human sperm heads is performed. Dry mass is proportional to DNA content. While the study showed that human sperm with none and one quinacrine-fluorescent spot are X- and Y-bearing, respectively, the dry mass measurements indicated that many of the sperm with two quinacrine-fluorescent spots are not YY-bearing. While several reports on the initial application of flow cytometry of sperm to the investigation of mammalian infertility have appeared recently, emphasis here has been on the development of an in vivo sperm-based flow cytometric bioassay for mutations, and has not centered on andrological applications. In this review, the ability to differentiate between two equally sized populations of sperm, one bearing X and the other Y chromosomes with mean DNA content differing by about 3 to 4% is described. It has direct application to the preselection of sex of offspring, and could likely have a profound impact on animal improvement. (ERB)

  6. Effect of different extenders on in vitro characteristics of feline epididymal sperm during cryopreservation.

    PubMed

    Jiménez, E; Pérez-Marín, C C; Vizuete, G; Millán, Y; Agüera, E I

    2013-08-01

    To evaluate and compare the efficacy of various extenders for the cryopreservation of epididymal cat spermatozoa, two experiments were planned. Bovine and equine commercial extenders in the experiment 1 and TRIS-egg yolk-based extenders in experiment 2 were separately studied since the number of sperm collected per cat is reduced. Epididymal sperm samples were packaged into 0.25-ml straws and frozen. Vigour, motility, morphology, acrosome status, sperm viability and functional membrane integrity were assessed at collection, after cooling and after thawing, while DNA integrity was evaluated at 0- and 6-h post-thaw. Experiment 1 compared the effect of three non-feline commercial extenders - based on TRIS-egg yolk (Triladyl), egg-yolk-free medium (AndroMed) and skimmed milk-egg yolk (Gent) - on the quality of frozen-thawed epididymal cat sperm. Values for sperm motility and functional membrane integrity in cooled sperm diluted in Triladyl were higher (p < 0.001) than those recorded for Andromed and Gent. Except sperm morphology, the other assessed characteristics showed significant higher values in frozen-thawed sperm diluted in Triladyl than in Andromed and Gent extenders. Experiment 2 analysed the effects of three TRIS-egg yolk-based extenders, one non-feline commercial (Triladyl) and the other two prepared using different monosaccharides (glucose and fructose), on freezing-thawed sperm. Results showed that specifically prepared extenders for cryopreservation of feline spermatozoa performed better than the commercial extender Triladyl, although sperm quality during the freezing-thawing process did not significantly differ associated with the type of monosaccharide (glucose vs fructose) added to the mentioned extenders. Although TRIS-egg yolk-based extenders prepared in experiment 2 improved sperm cryoprotection, Triladyl remains a good option for practitioners who, for ease of use and availability, prefer to work with commercial extenders. PMID:23320450

  7. Sperm environment affects offspring quality in broadcast spawning marine invertebrates

    E-print Network

    Levitan, Don R.

    REPORT Sperm environment affects offspring quality in broadcast spawning marine invertebrates and the abundance of sperm present during fertilization. Larger eggs were fertilized at low sperm concentrations, whilst smaller eggs were successfully fertilized at high sperm concentrations. These differences

  8. Interpreting sperm DNA damage in a diverse range of mammalian sperm by means of the two-tailed comet assay

    PubMed Central

    Cortés-Gutiérrez, Elva I.; López-Fernández, Carmen; Fernández, José Luis; Dávila-Rodríguez, Martha I.; Johnston, Stephen D.; Gosálvez, Jaime

    2014-01-01

    Key Concepts The two-dimensional Two-Tailed Comet assay (TT-comet) protocol is a valuable technique to differentiate between single-stranded (SSBs) and double-stranded DNA breaks (DSBs) on the same sperm cell.Protein lysis inherent with the TT-comet protocol accounts for differences in sperm protamine composition at a species-specific level to produce reliable visualization of sperm DNA damage.Alkaline treatment may break the sugar–phosphate backbone in abasic sites or at sites with deoxyribose damage, transforming these lesions into DNA breaks that are also converted into ssDNA. These lesions are known as Alkali Labile Sites “ALSs.”DBD–FISH permits the in situ visualization of DNA breaks, abasic sites or alkaline-sensitive DNA regions.The alkaline comet single assay reveals that all mammalian species display constitutive ALS related with the requirement of the sperm to undergo transient changes in DNA structure linked with chromatin packing.Sperm DNA damage is associated with fertilization failure, impaired pre-and post- embryo implantation and poor pregnancy outcome.The TT is a valuable tool for identifying SSBs or DSBs in sperm cells with DNA fragmentation and can be therefore used for the purposes of fertility assessment. Sperm DNA damage is associated with fertilization failure, impaired pre-and post- embryo implantation and poor pregnancy outcome. A series of methodologies to assess DNA damage in spermatozoa have been developed but most are unable to differentiate between single-stranded DNA breaks (SSBs) and double-stranded DNA breaks (DSBs) on the same sperm cell. The two-dimensional Two-Tailed Comet assay (TT-comet) protocol highlighted in this review overcomes this limitation and emphasizes the importance in accounting for the difference in sperm protamine composition at a species-specific level for the appropriate preparation of the assay. The TT-comet is a modification of the original comet assay that uses a two dimensional electrophoresis to allow for the simultaneous evaluation of DSBs and SSBs in mammalian spermatozoa. Here we have compiled a retrospective overview of how the TT-comet assay has been used to investigate the structure and function of sperm DNA across a diverse range of mammalian species (eutheria, metatheria, and prototheria). When conducted as part of the TT-comet assay, we illustrate (a) how the alkaline comet single assay has been used to help understand the constitutive and transient changes in DNA structure associated with chromatin packing, (b) the capacity of the TT-comet to differentiate between the presence of SSBs and DSBs (c) and the possible implications of SSBs or DSBs for the assessment of infertility. PMID:25505901

  9. Correlation between sperm ultrastructure in infertile patients with abnormal sperm morphology and DNA damage.

    PubMed

    He, M; Tan, L

    2015-01-01

    This study explored the correlation between sperm ultrastructure in infertile patients with abnormal sperm morphology and DNA damage. Three unusual sperm morphologies were selected for the experimental group namely case 1 (95% headless sperm), case 2 (98% headless sperm), and case 3 (100% headless sperm), and the control group consisted of 2 subjects (20 and 15% headless sperm). For case 1, the patient was negative for sexually transmitted diseases and had normal semen plasma biochemistry, reproductive hormones, peripheral blood chromosomes, and azoospermia factor (AZF). The aneuploid rate of sperm chromosomes was 0.6%, and DNA damage index of sperm nuclei was 84.4%. The partner of this patient did not get pregnant after artificial reproductive technology assistance. For case 2, the aneuploid rate of sperm chromosomes was 0.8% and DNA damage index of sperm nuclei was 95%. This patient and his spouse did not choose assisted reproduction. For case 3, reproductive hormones, peripheral blood chromosomes and AZF were normal and the aneuploid rate of sperm chromosomes was 0.2%. The wife of this patient gave birth to a healthy baby after ova removal, fertilization and transplantation. For the control group, the aneuploid rate of sperm chromosomes and DNA damage index of sperm nuclei were approximately 0.3 and 30%, respectively. To sum up, sperm ultrastructure of infertile patients suffering from unusual sperm morphology is associated with DNA damage to some extent and can cause infertility. However, pregnancy is still possible through intracytoplasmic sperm injection. PMID:26681047

  10. Differences in ATP Generation Via Glycolysis and Oxidative Phosphorylation and Relationships with Sperm Motility in Mouse Species.

    PubMed

    Tourmente, Maximiliano; Villar-Moya, Pilar; Rial, Eduardo; Roldan, Eduardo R S

    2015-08-14

    Mouse sperm produce enough ATP to sustain motility by anaerobic glycolysis and respiration. However, previous studies indicated that an active glycolytic pathway is required to achieve normal sperm function and identified glycolysis as the main source of ATP to fuel the motility of mouse sperm. All the available evidence has been gathered from the studies performed using the laboratory mouse. However, comparative studies of closely related mouse species have revealed a wide range of variation in sperm motility and ATP production and that the laboratory mouse has comparatively low values in these traits. In this study, we compared the relative reliance on the usage of glycolysis or oxidative phosphorylation as ATP sources for sperm motility between mouse species that exhibit significantly different sperm performance parameters. We found that the sperm of species with higher oxygen consumption/lactate excretion rate ratios were able to produce higher amounts of ATP, achieving higher swimming velocities. Additionally, we show that the species with higher respiration/glycolysis ratios have a higher degree of dependence upon active oxidative phosphorylation. Moreover, we characterize for the first time two mouse species in which sperm depend on functional oxidative phosphorylation to achieve normal performance. Finally, we discuss that sexual selection could promote adaptations in sperm energetic metabolism tending to increase the usage of a more efficient pathway for the generation of ATP (and faster sperm). PMID:26048989

  11. Electrophysiological evidence for the presence of cystic fibrosis transmembrane conductance regulator (CFTR) in mouse sperm.

    PubMed

    Figueiras-Fierro, Dulce; Acevedo, Juan José; Martínez-López, Pablo; Escoffier, Jessica; Sepúlveda, Francisco V; Balderas, Enrique; Orta, Gerardo; Visconti, Pablo E; Darszon, Alberto

    2013-03-01

    Mammalian sperm must undergo a maturational process, named capacitation, in the female reproductive tract to fertilize the egg. Sperm capacitation is regulated by a cAMP/protein kinase A (PKA) pathway and involves increases in intracellular Ca(2+), pH, Cl(-), protein tyrosine phosphorylation, and in mouse and some other mammals a membrane potential hyperpolarization. The cystic fibrosis transmembrane conductance regulator (CFTR), a Cl(-) channel modulated by cAMP/PKA and ATP, was detected in mammalian sperm and proposed to modulate capacitation. Our whole-cell patch-clamp recordings from testicular mouse sperm now reveal a Cl(-) selective component to membrane current that is ATP-dependent, stimulated by cAMP, cGMP, and genistein (a CFTR agonist, at low concentrations), and inhibited by DPC and CFTR(inh) -172, two well-known CFTR antagonists. Furthermore, the Cl(-) current component activated by cAMP and inhibited by CFTR(inh) -172 is absent in recordings on testicular sperm from mice possessing the CFTR ?F508 loss-of-function mutation, indicating that CFTR is responsible for this component. A Cl(-) selective like current component displaying CFTR characteristics was also found in wild type epididymal sperm bearing the cytoplasmatic droplet. Capacitated sperm treated with CFTR(inh) -172 undergo a shape change, suggesting that CFTR is involved in cell volume regulation. These findings indicate that functional CFTR channels are present in mouse sperm and their biophysical properties are consistent with their proposed participation in capacitation. PMID:22833409

  12. Electrophysiological Evidence for the Presence of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) in Mouse Sperm

    PubMed Central

    Dulce, Figueiras Fierro; José, Acevedo Juan; Pablo, Martínez; Escoffier, Jessica; Sepúlveda, Francisco V.; Enrique, Balderas; Gerardo, Orta; Pablo, Visconti; Alberto, Darszon

    2014-01-01

    Mammalian sperm must undergo a maturational process, named capacitation, in the female reproductive tract to fertilize the egg. Sperm capacitation is regulated by a cAMP/PKA pathway and involves increases in intracellular Ca2+, pH, Cl?, protein tyrosine phosphorylation, and in mouse and some other mammals a membrane potential hyperpolarization. The cystic fibrosis transmembrane conductance regulator (CFTR), a Cl? channel modulated by cAMP/PKA and ATP, was detected in mammalian sperm and proposed to modulate capacitation. Our whole-cell patch-clamp recordings from testicular mouse sperm now reveal a Cl? selective component to membrane current that is ATP-dependent, stimulated by cAMP, cGMP and genistein (a CFTR agonist, at low concentrations), and inhibited by DPC and CFTRinh-172, two well-known CFTR antagonists. Furthermore, the Cl? current component activated by cAMP and inhibited by CFTRinh-172 is absent in recordings on testicular sperm from mice possessing the CFTR ?F508 loss-of-function mutation, indicating that CFTR is responsible for this component. A Cl? selective like current component displaying CFTR characteristics was also found in wild type epididymal sperm bearing the cytoplasmatic droplet. Capacitated sperm treated with CFTRinh-172 undergo a shape change, suggesting that CFTR is involved in cell volume regulation. These findings indicate that functional CFTR channels are present in mouse sperm and their biophysical properties are consistent with their proposed participation in capacitation. PMID:22833409

  13. Differential proteomics leads to identification of domain-specific epididymal sperm proteins.

    PubMed

    Suryawanshi, Amol R; Khan, Shagufta A; Gajbhiye, Rahul K; Gurav, Mamta Y; Khole, Vrinda V

    2011-01-01

    The alteration in the protein signatures of the testicular sperm during its epididymal sojourn makes it functionally competent for successful fertilization. The present study was undertaken to identify the proteins acquired on its 2 domains, that is, the head and the flagellum, during the epididymal transit using a differential proteomics approach. Testicular sperm proteome was compared with cauda epididymal sperm proteome in rat. The protein spots exclusively present in the cauda epididymal sperm proteome were searched in the cauda sperm head proteome and the cauda sperm flagella proteome, and a total of 335 spots were found by alignment and auto-matching of the gels, of which 140 could be identified by mass spectrometry. Database search revealed that of these 9 proteins were novels. Gene Ontology annotation revealed that the identified proteins were distributed across different cellular components and were primarily involved in metabolic processes. The study also provides information on the localization of these proteins on the sperm domains, which indirectly gives a clue about its putative function. Validation of 3 proteins, namely MMSDH, NDUFS1, and UQCRC2, using antibodies very elegantly demonstrates that the strategy has been very effective. This comprehensive data of domain-specific epididymal sperm proteins will be useful in development of newer targets for posttesticular contraception and diagnostic markers for infertility. PMID:20966424

  14. Characterization and short-term storage of Tasmanian devil sperm collected post-mortem.

    PubMed

    Keeley, T; McGreevy, P D; O'Brien, J K

    2011-09-01

    The Tasmanian devil is suffering from a severe population decline due to the fatal Devil Facial Tumour Disease (DFTD). The development of assisted reproductive technologies such as AI and long-term sperm storage could facilitate genetic management of captive insurance populations. The aim of this study was to characterise semen samples collected post-mortem, and to develop a suitable diluent for short-term preservation of devil sperm. Low numbers of sperm (1.33 ± 0.85 × 10(6) sperm per male) were extracted from the epididymides of 17 males. Devil sperm sample characteristics such as concentration and morphology were similar to other dasyurids. The most commonly observed morphological abnormalities were midpiece separation, tail curling, and tail twisting (on the axial plane). Changes in motility occurred throughout the regions of the epididymis with (mean ± SD) 29.4 ± 16.8, 46.8 ± 13.6 and 29.4 ± 18.1% of sperm exhibiting motility, and 88.9 ± 11.4, 32.0 ± 24.3 and 0.1 ± 0.2% of motile sperm exhibiting forward progressive motility in the cauda, corpus and caput, respectively. Sperm from the cauda and corpus epididymis maintained 31.7 ± 26.6 and 80.6 ± 85.9%, respectively, of initial motility after 12 h at 15 °C in a TEST yolk buffer diluent. These findings provided new information regarding devil sperm biology and short-term sperm storage; such information is necessary for future development of long-term sperm preservation methods in the Tasmanian devil. PMID:21652060

  15. Effect of cryopreservation on the sperm DNA fragmentation dynamics of the bottlenose dolphin (Tursiops truncatus).

    PubMed

    Sánchez-Calabuig, M J; López-Fernández, C; Johnston, S D; Blyde, D; Cooper, J; Harrison, K; de la Fuente, J; Gosálvez, J

    2015-04-01

    Sperm DNA fragmentation is one of the major causes of infertility; the sperm chromatin dispersion test (SCDt) evaluates this parameter and offers the advantage of species-specific validated protocol and ease of use under field conditions. The main purpose of this study was to evaluate sperm DNA fragmentation dynamics in both fresh and post-thaw bottlenose dolphin sperm using the SCDt following different cryopreservation protocols to gain new information about the post-thaw differential sperm DNA longevity in this species. Fresh and cryopreserved semen samples from five bottlenose dolphins were examined for sperm DNA fragmentation dynamics using the SCDt (Halomax(®)). Sperm DNA fragmentation was assessed immediately at collection and following cryopreservation (T0) and then after 0.5, 1, 4, 8, 24, 48 and 72 h incubation at 37°C. Serially collected ejaculates from four dolphins were frozen using different cryopreservation protocols in a TES-TRIS-fructose buffer (TTF), an egg-yolk-free vegetable lipid LP1 buffer (LP1) and human sperm preservation medium (HSPM). Fresh ejaculated spermatozoa initially showed low levels of DNA fragmentation for up to 48 h. Lower Sperm DNA fragmentation (SDF) was found in the second fresh ejaculate compared to the first when more than one sample was collected on the same day (p < 0.05); this difference was not apparent in any other seminal characteristic. While there was no difference observed in SDF between fresh and frozen-thawed sperm using the different cryopreservation protocols immediately after thawing (T0), frozen-thawed spermatozoa incubated at 37°C showed an increase in the rate of SDF after 24 h. Sperm frozen in the LP1(?) buffer had higher levels (p < 0.05) of DNA fragmentation after 24- and 48-h incubation than those frozen in TTF or HSPM. No correlation was found between any seminal characteristic and DNA fragmentation in either fresh and/or frozen-thawed samples. PMID:25604784

  16. Coaching patients during pulmonary function testing: A practical guide

    PubMed Central

    Cheung, Heidi J; Cheung, Lawrence

    2015-01-01

    Pulmonary function tests are an important tool to assist in the diagnosis and management of patients with respiratory disease. Ensuring that the tests are of acceptable quality is vital. Acceptable pulmonary function test quality requires, among others, optimal patient performance. Optimal patient performance, in turn, requires adequate coaching from registered respiratory therapists (RRTs) and other pulmonary function laboratory personnel. The present article provides techniques and tips to help RRTs coach patients during testing. The authors briefly review the components of pulmonary function testing, then describe factors that may hinder a patient’s performance, list common mistakes that patients make during testing, and provide tips that RRTs can use to help patients optimize their performance. PMID:26283871

  17. Sea urchin sperm antigens mediating the acrosome reaction

    SciTech Connect

    Trimmer, J.S.

    1987-01-01

    The study of sea urchin sperm antigens mediating the acrosome reactions (AR) has been undertaken. Monoclonal antibodies (mAbs) have been isolated reacting with a number of sperm surface antigens. These mAbs have been used in functional assays to attempt to infer the roles of these proteins in the induction of the AR. These mAbs have also been used to isolate protein for biochemical characterization and reconstitution studies. mAbs reacting with a 210 kD protein of the sea urchin sperm plasma membrane have been used to identify this protein as playing a role in the regulation of ion fluxes during the induction of the AR. mAbs reacting with certain extracellular regions inhibit the induction of: the AR, the long duration {sup 45}Ca{sup 2+} uptake into the mitochondrion, and H{sup +} efflux. Addition of these same mAbs, however, induces an increase in sperm (Ca{sup 2+}){sub i} to levels much higher than those induced by FSG, as monitored by the fluorescent Ca{sup 2+} indicators fura 2 and indo 1. This (Ca{sup 2+}){sub i} increase occurs without an increase in pH{sub i}, and thus allows for the first time the analysis of the effects of increasing sperm (Ca{sup 2+}){sub i} ion the absence of increased pH{sub i}.

  18. Isolation and Proteomic Characterization of the Mouse Sperm Acrosomal Matrix*

    PubMed Central

    Guyonnet, Benoit; Zabet-Moghaddam, Masoud; SanFrancisco, Susan; Cornwall, Gail A.

    2012-01-01

    A critical step during fertilization is the sperm acrosome reaction in which the acrosome releases its contents allowing the spermatozoa to penetrate the egg investments. The sperm acrosomal contents are composed of both soluble material and an insoluble material called the acrosomal matrix (AM). The AM is thought to provide a stable structure from which associated proteins are differentially released during fertilization. Because of its important role during fertilization, efforts have been put toward isolating the AM for biochemical study and to date AM have been isolated from hamster, guinea pig, and bull spermatozoa. However, attempts to isolate AM from mouse spermatozoa, the species in which fertilization is well-studied, have been unsuccessful possibly because of the small size of the mouse sperm acrosome and/or its fusiform shape. Herein we describe a procedure for the isolation of the AM from caput and cauda mouse epididymal spermatozoa. We further carried out a proteomic analysis of the isolated AM from both sperm populations and identified 501 new proteins previously not detected by proteomics in mouse spermatozoa. A comparison of the AM proteome from caput and cauda spermatozoa showed that the AM undergoes maturational changes during epididymal transit similar to other sperm domains. Together, our studies suggest the AM to be a dynamic and functional structure carrying out a variety of biological processes as implied by the presence of a diverse group of proteins including proteases, chaperones, hydrolases, transporters, enzyme modulators, transferases, cytoskeletal proteins, and others. PMID:22707618

  19. Lithium carbonate inducing disorders in three parameters of rat sperm

    PubMed Central

    Toghyani, Shima; Dashti, Gholam R.; Roudbari, Nasim Hayati; Rouzbehani, Shaila; Monajemi, Ramesh

    2013-01-01

    Background: Lithium has a significant impact in reducing the symptoms of bipolar mania but in long periods of use with therapeutic doses can cause several disorders in various organs including the reproductive system. In this study, the effect of lithium on the sperm concentration and motility and forms of abnormal cells has been examined. Materials and Methods: Male Wistar rats under the 48-day treatment with lithium carbonate at doses of 10, 20, and 30 mg/kg bw/day were kept in standard conditions. At the end of this period, sperm cells isolated from the cauda epididymis were counted, motility was estimated, and stained with smear papanicolaou stain. Results: In lithium-treated groups, the rate of spermatogenesis and sperm quality were reduced and was seen in a dose-dependent manner. Discussion: Lithium alters intracellular signaling pathways such as inositol phosphate metabolic cycle and cyclic adenosine mono phosphate (cAMP) system and adenosine triphosphate (ATP) synthesis. It also interferes in the division of sex cells to produce mature sperm and showed changes in the sperm cell membrane, function, and structure. PMID:24223370

  20. Significance tests for functional data with complex dependence structure

    PubMed Central

    Lahiri, Soumen N.; Carroll, Raymond J.

    2015-01-01

    We propose an L2-norm based global testing procedure for the null hypothesis that multiple group mean functions are equal, for functional data with complex dependence structure. Specifically, we consider the setting of functional data with a multilevel structure of the form groups–clusters or subjects–units, where the unit-level profiles are spatially correlated within the cluster, and the cluster-level data are independent. Orthogonal series expansions are used to approximate the group mean functions and the test statistic is estimated using the basis coefficients. The asymptotic null distribution of the test statistic is developed, under mild regularity conditions. To our knowledge this is the first work that studies hypothesis testing, when data have such complex multilevel functional and spatial structure. Two small-sample alternatives, including a novel block bootstrap for functional data, are proposed, and their performance is examined in simulation studies. The paper concludes with an illustration of a motivating experiment. PMID:26023253

  1. A Generalized DIF Effect Variance Estimator for Measuring Unsigned Differential Test Functioning in Mixed Format Tests

    ERIC Educational Resources Information Center

    Penfield, Randall D.; Algina, James

    2006-01-01

    One approach to measuring unsigned differential test functioning is to estimate the variance of the differential item functioning (DIF) effect across the items of the test. This article proposes two estimators of the DIF effect variance for tests containing dichotomous and polytomous items. The proposed estimators are direct extensions of the…

  2. Human sperm competition: testis size, sperm production and rates of extrapair copulations

    E-print Network

    Clifton, Ken

    Human sperm competition: testis size, sperm production and rates of extrapair copulations LEIGH W the claim that sperm competition is an important selection pressure operating in human populations. We of extrapair paternity are in the region of 2%. These values suggest that the risk of sperm competition

  3. Improvement of Ram Sperm Cryopreservation Protocols Assessed by Sperm Quality Parameters and Heterogeneity Analysis

    E-print Network

    Zaragoza, Universidad de

    Improvement of Ram Sperm Cryopreservation Protocols Assessed by Sperm Quality Parameters, Spain Semen cryopreservation processes generally decrease sperm quality and fertility rate, but this is highly dependent on the specific susceptibility of sperm cells to low temperatures. In order to study

  4. Optimal copula duration in yellow dung flies: ejaculatory duct dimensions and size-dependent sperm displacement.

    PubMed

    Parker, G A; Simmons, L W

    2000-06-01

    We aim to interpret sperm displacement in relation to male size in the yellow dung fly, Scatophaga stercoraria, and to compare the general properties of indirect and direct size-dependent sperm displacement in insects. We examine the hypothesis that male size-dependent sperm displacement in dung flies can be explained by size-dependent increases in the ejaculatory apparatus, allowing greater sperm flow rates in larger males. We expect sperm flow rates to be proportional to the diameter of the aedeagus duct to the power x, where x lies between 2 and 3. We test this hypothesis using a simulation model of indirect sperm displacement that has been developed to accommodate recent observations on sperm transfer, in which sperm flow from the male into the female bursa and are then transferred to the spermathecae by movements of the female tract. The indirect model approximates to the pattern of size-related sperm displacement, with scaling power 3 giving a better fit than power 2. Copula duration shows a male size-dependent decrease in this species. We apply the indirect model of sperm displacement, in conjunction with parameters obtained from field and laboratory data, to predict size-dependent changes in optimal copula duration from the male perspective. This model concurs with the observations by predicting a size-dependent decline in optimal copula duration, as did an earlier model in which displacement was direct (new sperm displace previously stored sperm directly from the sperm stores). Our new approach gives a better fit than the earlier direct model. Thus, both results (displacement rates and copula duration) can be explained by size-dependent changes in the ejaculatory apparatus of the male with the female's exchange rate of sperm (from bursa to spermathecae) remaining constant with respect to male size, although we discuss the possibility that this female process may accelerate with increased male size. In general, where the sperm input rate is around the same magnitude as the exchange rate, indirect displacement will be dependent on the size of the male, as in dung flies, but this dependency is lost if the input rate is very high relative to the exchange rate across the entire range of male size. Size-dependent displacement should always apply for males with direct displacement. PMID:10937265

  5. Effect of Multiple Testing Adjustment in Differential Item Functioning Detection

    ERIC Educational Resources Information Center

    Kim, Jihye; Oshima, T. C.

    2013-01-01

    In a typical differential item functioning (DIF) analysis, a significance test is conducted for each item. As a test consists of multiple items, such multiple testing may increase the possibility of making a Type I error at least once. The goal of this study was to investigate how to control a Type I error rate and power using adjustment…

  6. The Relationship between Seminal Melatonin with Sperm Parameters, DNA Fragmentation and Nuclear Maturity in Intra-Cytoplasmic Sperm Injection Candidates

    PubMed Central

    Sharbatoghli, Mina; Rezazadeh Valojerdi, Mojtaba; Bahadori, Mohammad Hadi; Salman Yazdi, Reza; Ghaleno, Leila Rashki

    2015-01-01

    Objective Melatonin, the chief secretory product of the pineal gland, regulates dynamic physiological adaptations that occur in seasonally breeding mammals as a response to changes in daylight hours. Because of the presence of melatonin in semen and the mem- brane melatonin receptor in spermatozoa, the impact of melatonin on the regulation of male infertility is still questionable. The aim of this study was to determine the effects of endogenous melatonin on human semen parameters (sperm concentration, motility and normal morphology), DNA fragmentation (DF) and nuclear maturity. Materials and Methods In this clinical prospective study, semen samples from 75 infer- tile men were routinely analyzed and assessed for melatonin and total antioxidant capac- ity (TAC) levels using the enzyme-linked immunosorbent assay (ELISA) and colorimetric assay kits, respectively. DF was examined by the sperm chromatin dispersion (SCD) test. Acidic aniline blue staining was used to detect chromatin defects in the sperm nuclei. Results There was no significant correlation between seminal plasma melatonin and TAC with sperm parameters and nuclear maturity. However, we observed a positive significant correlation between DF and melatonin level (r=0.273, P<0.05). Conclusion Melatonin in seminal plasma is positively correlated with damaged sperm DNA of infertile patients. The mechanism of this phenomenon needs further study. PMID:26464827

  7. Extraordinarily long sperm in the socially monogamous cichlid fish Pelvicachromis taeniatus

    NASA Astrophysics Data System (ADS)

    Thünken, Timo; Bakker, Theo C. M.; Kullmann, Harald

    2007-06-01

    The main function of the spermatozoon is the transfer of the male haploid genome during fertilisation. In animals in general and in fishes in particular, there is huge variation in sperm size. In fishes, sperm size ranges from 13 ?m in Mugil cephlus to nearly 100 ?m in the channel catfish Ictalurus punctatus. We examined intra-specific variation in sperm morphometry in the socially monogamous cichlid Pelvicachromis taeniatus using scanning electron microscopy. The mean total sperm length of nearly 70 ?m was extraordinarily large for cichlids. Furthermore, within-male variation was remarkably high. To our knowledge, P. taeniatus produces the longest cichlid sperm ever documented. Several hypotheses concerning the adaptive significance of these results are presented.

  8. Low paternal dietary folate alters the mouse sperm epigenome and is associated with negative pregnancy outcomes

    PubMed Central

    Lambrot, R.; Xu, C.; Saint-Phar, S.; Chountalos, G.; Cohen, T.; Paquet, M.; Suderman, M.; Hallett, M.; Kimmins, S.

    2013-01-01

    Epidemiological studies suggest that a father’s diet can influence offspring health. A proposed mechanism for paternal transmission of environmental information is via the sperm epigenome. The epigenome includes heritable information such as DNA methylation. We hypothesize that the dietary supply of methyl donors will alter epigenetic reprogramming in sperm. Here we feed male mice either a folate-deficient or folate-sufficient diet throughout life. Paternal folate deficiency is associated with increased birth defects in the offspring, which include craniofacial and musculoskeletal malformations. Genome-wide DNA methylation analysis and the subsequent functional analysis identify differential methylation in sperm of genes implicated in development, chronic diseases such as cancer, diabetes, autism and schizophrenia. While >300 genes are differentially expressed in offspring placenta, only two correspond to genes with differential methylation in sperm. This model suggests epigenetic transmission may involve sperm histone H3 methylation or DNA methylation and that adequate paternal dietary folate is essential for offspring health. PMID:24326934

  9. The sperm flagellum counterbend phenomenon

    NASA Astrophysics Data System (ADS)

    Gadelha, Hermes; Gaffney, Eamonn; Goriely, Alain

    2013-11-01

    Recent observations of flagellar counterbend in sperm show that the mechanical induction of curvature in one part of a passive flagellum induces a compensatory countercurvature elsewhere. This apparent paradoxical effect cannot be explained using the standard elastic rod theory of Euler and Bernoulli, or even the more general Cosserat theory of rods. Here, we develop a mechanical model capable of predicting the curvature reversal events observed in eukaryotic flagella. This is achieved by allowing the interaction of deformations in different material directions, by not only accounting for structural bending, but also the elastic forces originating from the cross-linking mechanics. Large amplitude configurations can be described analytically and an excellent match between the model and the observed counterbend deformation was found. This allowed a simultaneous estimation of multiple sperm flagellum material parameters, namely the cross-linking sliding resistance, the bending stiffness and the sperm head junction compliance ratio. Our analysis demonstrates that the counterbend emerges as a fundamental property of sliding resistance, which also suggests that cross-linking proteins may contribute to the regulation of the flagellar waveform in swimming sperm via counterbend mechanics. Finally, we investigate how the counterbend-type dynamics in sperm flagella is affected by viscous dissipation.

  10. Treatment of aromatase (CYP19A1) inhibitor reduces fertility in porcine sperm.

    PubMed

    Oh, Jong-Nam; Hwang, Jae Yeon; Choi, Kwang-Hwan; Lee, Chang-Kyu

    2016-02-01

    To ascertain whether aromatase (CYP19A1) expression is linked to sperm fertility of pigs, the present study determined the expression of the CYP19A1 gene in porcine sperm and its relationship with fertilization in vitro. First, to investigate its role in fertility, the presence of CYP19A1 of mRNA and protein expression in porcine sperm were confirmed by real-time (RT) or quantitative polymerase chain reaction (q-PCR) and by western blots. The expression levels were determined quantitatively using two sperm groups recovered by a Percoll gradient, which revealed that the sperm group with a low density had a higher penetration rate than that of the high-density group (P < 0.05). However, the expression level of CYP19A1 was not significantly different between the two groups. Secondly, to examine the effect of aromatase activity on fertilization, fresh semen was treated with a steroidal inhibitor, exemestane (50 ?M for 0.5 h), followed by the dose- and time-dependent viability test. Our results clearly showed that an exemestane treatment effect (P < 0.05) was found for both the sperm-penetration rate and the oocyte cleavage rate. These results indicated that CYP19A1 could be involved in sperm fertility and its expression in sperm plays an important role in fertilization. PMID:25621494

  11. Meiotic recombination errors, the origin of sperm aneuploidy and clinical recommendations.

    PubMed

    Tempest, Helen G

    2011-02-01

    Since the early 1990s male infertility has successfully been treated by intracytoplasmic sperm injection (ICSI), nevertheless concerns have been raised regarding the genetic risk of ICSI. Chromosome aneuploidy (the presence of extra or missing chromosomes) is the leading cause of pregnancy loss and mental retardation in humans. While the majority of chromosome aneuploidies are maternal in origin, the paternal contribution to aneuploidy is clinically relevant particularly for the sex chromosomes. Given that it is difficult to study female gametes investigations are predominantly conducted in male meiotic recombination and sperm aneuploidy. Research suggests that infertile men have increased levels of sperm aneuploidy and that this is likely due to increased errors in meiotic recombination and chromosome synapsis within these individuals. It is perhaps counterintuitive but there appears to be no selection against chromosomally aneuploid sperm at fertilization. In fact the frequency of aneuploidy in sperm appears to be mirrored in conceptions. Given this information this review will cover our current understanding of errors in meiotic recombination and chromosome synapsis and how these may contribute to increased sperm aneuploidy. Frequencies of sperm aneuploidy in infertile men and individuals with constitutional karyotypic abnormalities are reviewed, and based on these findings, indications for clinical testing of sperm aneuploidy are discussed. In addition, the application of single nucleotide arrays for the analysis of meiotic recombination and identification of parental origin of aneuploidy are considered. PMID:21204593

  12. Effect of proctodeal gland foam on sperm kinetics in Japanese quail (Coturnix japonica).

    PubMed

    Farooq, U; Cho, S; Rybnik-Trzaskowska, P K; Singh, R P; Malecki, I A

    2015-01-15

    The proctodeal gland of the male Japanese quail produces thick foam that accompanies semen when it is transferred to the female. It is thought that this foam enhances fertilization by improving the motility of the sperm, but reports are conflicting because the effect of foam on sperm motility has only been assessed subjectively The velocity of individual sperm was not able to be measured accurately, variations were large, and small changes in motility could not be accurately evaluated. So, we tested the hypothesis that foam affects the motility of spermatozoa of Japanese quail by analyzing motility objectively using computer-assisted semen analysis and determining changes in sperm kinetics in the presence of different concentrations of proctodeal gland foam. The addition of 5% or 10% foam to the sperm suspension increased (P < 0.05) all sperm kinetic parameters (the curvilinear velocity, straight line velocity, the velocity of the average path, linearity, straightness, and beat cross frequency). As a result, the percentage of motile and progressive motile sperm also increased. All these parameters declined (P < 0.05) with a further increase in the concentration of foam to 15% and 20%. Furthermore, this effect was similar in males that were 8, 16, or 26 weeks of age. We conclude that sperm motility is enhanced by proctodeal gland foam, and this enhancement depends on its concentration. PMID:25301249

  13. Maternal effects, but no good or compatible genes for sperm competitiveness in Australian crickets.

    PubMed

    Dowling, Damian K; Nystrand, Magdalena; Simmons, Leigh W

    2010-05-01

    Explanations for the evolution of polyandry often center on the idea that females garner genetic benefits for their offspring by mating multiply. Furthermore, postcopulatory processes are thought to be fundamental to enabling polyandrous females to screen for genetic quality. Much attention has focused on the potential for polyandrous females to accrue such benefits via a sexy- or good-sperm mechanism, whereby additive variation exists among males in sperm competitiveness. Likewise, attention has focused on an alternative model, in which offspring quality (in this context, the sperm competitiveness of sons) hinges on an interaction between parental haplotypes (genetic compatibility). Sperm competitiveness that is contingent on parental compatibility will exhibit nonadditive genetic variation. We tested these models in the Australian cricket, Teleogryllus oceanicus, using a design that allowed us to partition additive, nonadditive genetic, and parental variance for sperm competitiveness. We found an absence of additive and nonadditive genetic variance in this species, challenging the direct relevance of either model to the evolution of sperm competitiveness in particular, and polyandry in general. Instead, we found maternal effects that were possibly sex-linked or cytoplasmically linked. We also found effects of focal male age on sperm competitiveness, with small increments in age conferring more competitive sperm. PMID:20002162

  14. No evidence for a trade-off between sperm length and male premating weaponry.

    PubMed

    Lüpold, S; Simmons, L W; Tomkins, J L; Fitzpatrick, J L

    2015-12-01

    Male ornaments and armaments that mediate success in mate acquisition and ejaculate traits influencing competitive fertilization success are under intense sexual selection. However, relative investment in these pre- and post-copulatory traits depends on the relative importance of either selection episode and on the energetic costs and fitness gains of investing in these traits. Theoretical and empirical work has improved our understanding of how precopulatory sexual traits and investments in sperm production covary in this context. It has recently also been suggested that male weapon size may trade off with sperm length as another post-copulatory sexual trait, but the theoretical framework for this suggestion remains unclear. We evaluated the relationship between precopulatory armaments and sperm length, previously reported in ungulates, in five taxa as well as meta-analytically. Within and between taxa, we found no evidence for a negative or positive relationship between sperm length and male traits that are important in male-male contest competition. It is important to consider pre- and post-copulatory sexual selection together to understand fitness, and to study investments in different reproductive traits jointly rather than separately. A trade-off between pre- and post-copulatory sexual traits may not manifest itself in sperm length but rather in sperm number or function. Particularly in large-bodied taxa such as ungulates, sperm number is more variable interspecifically and likely to be under more intense selection than sperm length. We discuss our and the previous results in this context. PMID:26332435

  15. A role for carbohydrate recognition in mammalian sperm-egg binding

    SciTech Connect

    Clark, Gary F.

    2014-08-01

    Highlights: • Mammalian sperm-egg binding as a carbohydrate dependent species recognition event. • The role of carbohydrate recognition in human, mouse and pig sperm-egg binding. • Historical perspective and future directions for research focused on gamete binding. - Abstract: Mammalian fertilization usually requires three sequential cell–cell interactions: (i) initial binding of sperm to the specialized extracellular matrix coating the egg known as the zona pellucida (ZP); (ii) binding of sperm to the ZP via the inner acrosomal membrane that is exposed following the induction of acrosomal exocytosis; and (iii) adhesion of acrosome-reacted sperm to the plasma membrane of the egg cell, enabling subsequent fusion of these gametes. The focus of this review is on the initial binding of intact sperm to the mammalian ZP. Evidence collected over the past fifty years has confirmed that this interaction relies primarily on the recognition of carbohydrate sequences presented on the ZP by lectin-like egg binding proteins located on the plasma membrane of sperm. There is also evidence that the same carbohydrate sequences that mediate binding also function as ligands for lectins on lymphocytes that can inactivate immune responses, likely protecting the egg and the developing embryo up to the stage of blastocyst hatching. The literature related to initial sperm-ZP binding in the three major mammalian models (human, mouse and pig) is discussed. Historical perspectives and future directions for research related to this aspect of gamete adhesion are also presented.

  16. SPERM COUNT DISTRIBUTIONS IN FERTILE MEN

    EPA Science Inventory

    Sperm concentration and count are often used as indicators of environmental impacts on male reproductive health. Existing clinical databases may be biased towards subfertile men with low sperm counts and less is known about expected sperm count distributions in cohorts of fertil...

  17. October 2008 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    179 October 2008 SPERM WHALE (Physeter macrocephalus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE Sperm whales are found throughout the world's oceans in deep waters to the edge of the ice at both poles (Leatherwood and Reeves 1983; Rice 1989; Whitehead 2002). Sperm whales

  18. December 2012 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    112 December 2012 SPERM WHALE (Physeter macrocephalus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE Sperm whales are found throughout the world's oceans in deep waters to the edge of the ice at both poles (Leatherwood and Reeves 1983; Rice 1989; Whitehead 2002). Sperm whales

  19. October 2007 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    225 October 2007 SPERM WHALE (Physeter macrocephalus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE Sperm whales are found throughout the world's oceans in deep waters surveys confirm that sperm whales are present in the northern Gulf of Mexico in all seasons (Mullin et al

  20. November 2010 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    198 November 2010 SPERM WHALE (Physeter macrocephalus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE Sperm whales are found throughout the world's oceans in deep waters to the edge of the ice at both poles (Leatherwood and Reeves 1983; Rice 1989; Whitehead 2002). Sperm whales

  1. December 2005 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    184 December 2005 SPERM WHALE (Physeter macrocephalus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE Sperm whales are found throughout the world's oceans in deep waters surveys confirm that sperm whales are present in the northern Gulf of Mexico in all seasons (Mullin et al

  2. October 2007 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    47 October 2007 SPERM WHALE (Physeter macrocephalus): North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The distribution of the sperm whale in the U.S. Exclusive Economic Zone (EEZ) occurs and other features. However, the sperm whales that occur in the eastern U.S. Atlantic EEZ likely represent

  3. December 2005 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    357 December 2005 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate whales and dwarf sperm whales (Kogia sima) are difficult to differentiate at sea, and sightings of either

  4. September 2000 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    296 September 2000 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate (Mullin et al. 1991; Southeast Fisheries Science Center, SEFSC, unpublished data). Pygmy sperm whales

  5. SPERM WHALE (Physeter macrocephalus): North Atlantic Stock

    E-print Network

    30 May 2013 SPERM WHALE (Physeter macrocephalus): North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The distribution of the sperm whale in the U.S. Exclusive Economic Zone (EEZ) occurs and other features. However, the sperm whales that occur in the eastern U.S. Atlantic EEZ likely represent

  6. October 2008 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    264 October 2008 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate whales and dwarf sperm whales (Kogia sima) are difficult to differentiate at sea, and sightings of either

  7. October 2007 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    361 October 2007 SPERM WHALE (Physeter macrocephalus): North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The distribution of the sperm whale in the U.S. Exclusive Economic Zone (EEZ) occurs and other features. However, the sperm whales that occur in the eastern U.S. Atlantic EEZ likely represent

  8. September 2000 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    218 September 2000 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate sperm whales have been sighted in the northwestern Gulf of M exico in waters 1000 m deep, on avera ge

  9. October 1999 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    52 October 1999 PYGMY SPERM WHALE (Kogia breviceps): Western North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale (Kogia breviceps) appears to be distributed worldwide in temperate (Hansen et al 1994; Southeast Fisheries Science Center unpublished data). Pygmy sperm whales and dwarf

  10. November 2010 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    249 November 2010 SPERM WHALE (Physeter macrocephalus): Puerto Rico and U.S. Virgin Islands Stock STOCK DEFINITION AND GEOGRAPHIC RANGE Sperm whales are found throughout the world's oceans in deep). Sperm whales throughout the world exhibit a geographic social structure where females and juveniles

  11. SPERM WHALE (Physeter macrocephalus): North Atlantic Stock

    E-print Network

    April 2014 SPERM WHALE (Physeter macrocephalus): North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The distribution of the sperm whale in the U.S. Exclusive Economic Zone (EEZ) occurs and other features. However, the sperm whales that occur in the eastern U.S. Atlantic EEZ likely represent

  12. SPERM WHALE (Physeter macrocephalus): North Atlantic Stock

    E-print Network

    63 May 2015 SPERM WHALE (Physeter macrocephalus): North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The distribution of the sperm whale in the U.S. Exclusive Economic Zone (EEZ) occurs and other features. However, the sperm whales that occur in the eastern U.S. Atlantic EEZ likely represent

  13. December 2005 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    281 December 2005 SPERM WHALE (Physeter macrocephalus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE Sperm whales are found throughout the world's oceans in deep waters surveys confirm that sperm whales are present in the northern Gulf of Mexico in all seasons (Mullin et al

  14. December 2009 SPERM WHALE (Physeter macrocephalus)

    E-print Network

    December 2009 SPERM WHALE (Physeter macrocephalus): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE Sperm whales are found throughout the world's oceans in deep waters to the edge of the ice at both poles (Leatherwood and Reeves 1983; Rice 1989; Whitehead 2002). Sperm whales were commercially

  15. December 2005 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    259 December 2005 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate whales and dwarf sperm whales (Kogia sima) are difficult to differentiate at sea, and sightings of either

  16. September 2000 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    193 September 2000 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate (Mullin et al. 1991; Southeast Fisheries Science Center, SEFSC, unpublished data). Pygmy sperm whales

  17. October 2007 PYGMY SPERM WHALE (Kogia breviceps)

    E-print Network

    309 October 2007 PYGMY SPERM WHALE (Kogia breviceps): Northern Gulf of Mexico Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale appears to be distributed worldwide in temperate whales and dwarf sperm whales (Kogia sima) are difficult to differentiate at sea, and sightings of either

  18. HPV-DNA sperm infection and infertility: from a systematic literature review to a possible clinical management proposal.

    PubMed

    Foresta, C; Noventa, M; De Toni, L; Gizzo, S; Garolla, A

    2015-03-01

    The objectives of this study were to investigate the implications of human papillomavirus (HPV) sperm infection on male fertility, impairment of sperm parameters, and possible alteration of sperm nuclear status and to identify a possible effective management of infertile men with HPV sperm infection. We employed a systematic review and clinical management proposal at the Centers for Reproductive and Health care for treating infertile male patients with HPV infection. Literature search was carried out in electronic databases in the last two decades. We focused our attention on: (i) HPV sperm prevalence (ii) HPV-related alteration of sperm parameters; (iii) molecular mechanisms of HPV semen infection and infertility. The main outcome measures were HPV prevalence in infertile male patients and semen parameters. The prevalence of HPV sperm infection ranges between 2 and 31% in men from general population and between 10 and 35.7% in men affected by unexplained infertility. The presence of HPV in semen is associated with an impairment of sperm motility and the presence of anti-sperm antibodies. The molecular mechanisms underlying impairment of sperm motility apparatus need further evaluations. A greater attention should be applied to assess HPV sperm infection, particularly in men undergoing assisted reproduction techniques cycle for male infertility or sperm banking. It would be useful to perform HPV test and fluorescent in situ hybridization analysis for HPV in semen from these patients both at first admission, to define the possible presence and localization of semen infection, and after 6 months, to assess the possible virus clearance retrieval on normal sperm parameters. PMID:25270519

  19. The Voltage-Gated Sodium Channel Nav1.8 Is Expressed in Human Sperm

    PubMed Central

    Cejudo-Roman, Antonio; Pinto, Francisco M.; Subirán, Nerea; Ravina, Cristina G.; Fernández-Sánchez, Manuel; Pérez-Hernández, Natalia; Pérez, Ricardo; Pacheco, Alberto; Irazusta, Jon; Candenas, Luz

    2013-01-01

    The role of Na+ fluxes through voltage-gated sodium channels in the regulation of sperm cell function remains poorly understood. Previously, we reported that several genes encoding voltage-gated Na+ channels were expressed in human testis and mature spermatozoa. In this study, we analyzed the presence and function of the TTX-resistant VGSC ? subunit Nav1.8 in human capacitated sperm cells. Using an RT-PCR assay, we found that the mRNA of the gene SCN10A, that encode Na v1.8, was abundantly and specifically expressed in human testis and ejaculated spermatozoa. The Na v1.8 protein was detected in capacitated sperm cells using three different specific antibodies against this channel. Positive immunoreactivity was mainly located in the neck and the principal piece of the flagellum. The presence of Na v1.8 in sperm cells was confirmed by Western blot. Functional studies demonstrated that the increases in progressive motility produced by veratridine, a voltage-gated sodium channel activator, were reduced in sperm cells preincubated with TTX (10 ?M), the Na v1.8 antagonist A-803467, or a specific Na v1.8 antibody. Veratridine elicited similar percentage increases in progressive motility in sperm cells maintained in Ca2+-containing or Ca2+-free solution and did not induce hyperactivation or the acrosome reaction. Veratridine caused a rise in sperm intracellular Na+, [Na+]i, and the sustained phase of the response was inhibited in the presence of A-803467. These results verify that the Na+ channel Na v1.8 is present in human sperm cells and demonstrate that this channel participates in the regulation of sperm function. PMID:24086692

  20. Safety and Function Test Report for the SWIFT Wind Turbine

    SciTech Connect

    Mendoza, I.; Hur, J.

    2013-01-01

    This test was conducted as part of the U.S. Department of Energy's (DOE) Independent Testing project. This project was established to help reduce the barriers of wind energy expansion by providing independent testing results for small turbines. Three turbines where selected for testing at the National Wind Technology Center (NWTC) as a part of round two of the Small Wind Turbine Independent Testing project. Safety and Function testing is one of up to 5 tests that may be performed on the turbines. Other tests include power performance, duration, noise, and power quality. The results of the testing will provide the manufacturers with reports that may be used for small wind turbine certification.

  1. A Semi-Automated Functional Test Data Analysis Tool

    SciTech Connect

    Xu, Peng; Haves, Philip; Kim, Moosung

    2005-05-01

    The growing interest in commissioning is creating a demand that will increasingly be met by mechanical contractors and less experienced commissioning agents. They will need tools to help them perform commissioning effectively and efficiently. The widespread availability of standardized procedures, accessible in the field, will allow commissioning to be specified with greater certainty as to what will be delivered, enhancing the acceptance and credibility of commissioning. In response, a functional test data analysis tool is being developed to analyze the data collected during functional tests for air-handling units. The functional test data analysis tool is designed to analyze test data, assess performance of the unit under test and identify the likely causes of the failure. The tool has a convenient user interface to facilitate manual entry of measurements made during a test. A graphical display shows the measured performance versus the expected performance, highlighting significant differences that indicate the unit is not able to pass the test. The tool is described as semiautomated because the measured data need to be entered manually, instead of being passed from the building control system automatically. However, the data analysis and visualization are fully automated. The tool is designed to be used by commissioning providers conducting functional tests as part of either new building commissioning or retro-commissioning, as well as building owners and operators interested in conducting routine tests periodically to check the performance of their HVAC systems.

  2. Thermosensitive ion channel TRPV1 is endogenously expressed in the sperm of a fresh water teleost fish (Labeo rohita) and regulates sperm motility.

    PubMed

    Majhi, Rakesh Kumar; Kumar, Ashutosh; Yadav, Manoj; Swain, Nirlipta; Kumari, Shikha; Saha, Ashish; Pradhan, Avinash; Goswami, Luna; Saha, Somdatta; Samanta, Luna; Maity, Apratim; Nayak, Tapas Kumar; Chattopadhyay, Subhasis; Rajakuberan, Chitra; Kumar, Abhishek; Goswami, Chandan

    2013-01-01

    Sperm cells exhibit extremely high sensitivity in response to slight changes in temperature, osmotic pressure and/or presence of various chemical stimuli. In most cases throughout the evolution, these physico-chemical stimuli trigger Ca (2+)-signaling and subsequently alter structure, cellular function, motility and survival of the sperm cells. Few reports have recently demonstrated the presence of Transient Receptor Potential (TRP) channels in the sperm cells from higher eukaryotes, mainly from higher mammals. In this work, we have explored if the sperm cells from lower vertebrates can also have thermo-sensitive TRP channels. In this paper, we demonstrate the endogenous presence of one specific thermo-sensitive ion channel, namely Transient Receptor Potential Vanilloid family member sub type 1 (TRPV1) in the sperm cells collected from fresh water teleost fish, Labeo rohita. By using western blot analysis, fluorescence assisted cell sorting (FACS) and confocal microscopy; we confirm the presence of this non-selective cation channel. Activation of TRPV1 by an endogenous activator NADA significantly increases the quality as well as the duration of fish sperm movement. The sperm cell specific expression of TRPV1 matches well with our in silico sequence analysis. The results demonstrate that TRPV1 gene is conserved in various fishes, ranging from 1-3 in copy number, and it originated by fish-specific duplication events within the last 320 million years (MY). To the best of our knowledge, this is the first report demonstrating the presence of any thermo-sensitive TRP channels in the sperm cells of early vertebrates as well as of aquatic animals, which undergo external fertilization in fresh water. This observation may have implications in the aquaculture, breeding of several fresh water and marine fish species and cryopreservation of fish sperms. PMID:23912940

  3. Identification of multiple HPV types on spermatozoa from human sperm donors.

    PubMed

    Kaspersen, Maja D; Larsen, Peter B; Ingerslev, Hans Jakob; Fedder, Jens; Petersen, Gert Bruun; Bonde, Jesper; Höllsberg, Per

    2011-01-01

    Human papillomaviruses (HPV) may cause sexually transmitted disease. High-risk types of HPV are involved in the development of cervical cell dysplasia, whereas low-risk types may cause genital condyloma. Despite the association between HPV and cancer, donor sperm need not be tested for HPV according to European regulations. Consequently, the potential health risk of HPV transmission by donor bank sperm has not been elucidated, nor is it known how HPV is associated with sperm. The presence of 35 types of HPV was examined on DNA from semen samples of 188 Danish sperm donors using a sensitive HPV array. To examine whether HPV was associated with the sperm, in situ hybridization were performed with HPV-6, HPV-16 and -18, and HPV-31-specific probes. The prevalence of HPV-positive sperm donors was 16.0% and in 66.7% of these individuals high-risk types of HPV were detected. In 5.3% of sperm donors, two or more HPV types were detected. Among all identified HPV types, 61.9% were high-risk types. In situ hybridization experiments identified HPV genomes particularly protruding from the equatorial segment and the tail of the sperm. Semen samples from more than one in seven healthy Danish donors contain HPV, most of them of high-risk types binding to the equatorial segment of the sperm cell. Most HPV-positive sperm showed decreased staining with DAPI, indicative of reduced content of DNA. Our data demonstrate that oncogenic HPV types are frequent in men. PMID:21479232

  4. Rescue of germline transmission from chimeras by IVF after sperm analysis.

    PubMed

    Li, Ming-Wen; Willis, Brandon J; Evans, Kristin D; Araiza, Renee S; Lee, Angus Yiu-Fai; Lloyd, K C Kent

    2015-02-01

    Successful production of genetically modified mouse lines is dependent on germline transmission (GLT) of mutant alleles from chimeras. When natural mating fails to achieve GLT due to male infertility, sickness, or other problems, sperm can be harvested from chimeras and used for assisted reproductive technologies such as in vitro fertilization (IVF) to attempt to "rescue" GLT. However, a rational, evidence-based approach to determine if such extraordinary efforts should be attempted on a chimera has not been established. Therefore, in the present study we assessed the production, quality and genotype of epididymal sperm harvested from male chimeras generated by blastocyst or morula microinjection of gene targeted embryonic stem (ES) cell clones containing a LacZ expression cassette and that failed to achieve GLT. Results of this analysis enabled us to determine the cause of GLT failure, correlate coat color chimerism with the proportion of LacZ-positive sperm, and test the likelihood of achieving GLT by IVF. In 415 chimeras, 332 (80%) produced no offspring by natural mating ("infertile"), while 83 (20%) produced only wildtype offspring ("fertile"). Of the 332 infertile chimeras, 209 (63%) failed to produce any sperm whatsoever, 48 (15%) had extremely poor quality sperm, and 75 (23%) had good quality sperm. These results indicate that most chimeras that do not achieve GLT by natural mating are infertile, and the primary cause of infertility is failed spermatogenesis. Genotyping of sperm from 519 chimeras revealed a significant positive linear correlation between coat color chimerism and mean percentage of LacZ-positive sperm (R(2) = 0.95). Finally, IVF using good quality, LacZ-positive sperm from fertile and infertile chimeras "rescued" GLT for 19 out of 56 genes. We conclude that an assessment of coat color chimerism together with sperm quality and genotype can better inform the selection of chimeras for IVF to rescue GLT than coat color chimerism alone. PMID:25080098

  5. Nuclear microscopy of sperm cell elemental structure

    SciTech Connect

    Bench, G.S.

    1994-12-31

    Theories have suggested that there is a link between protamine concentrations in individual sperm and sperm fertility. At present, biochemical analyses have only been performed on bulk populations and existing methods have not been able to determine what percentage of morphologically normal sperm are biochemically defective and potentially infertile. As part of an investigation into male sperm fertility, nuclear microscopy has been utilized to measure elemental profiles at the single sperm level. By measuring the ratio of Phosphorus to Sulfur the authors have been able to determine the amount of protamine 1 and protamine 2 in individual cells from bulk fertile samples of bull and mouse sperm. Preliminary results show that, for each species, the relative amounts of protamine 1 and protamine 2 in morphologically normal sperm agree well with expected values.

  6. IDENTIFICATION AND CHARACTERIZATION OF DISEASE USING PULMONARY FUNCTION TESTS

    EPA Science Inventory

    Abstract
    Pulmonary function testing is used routinely in human medicine to objectively define functional deficits in individuals with respiratory disease. Despite the fact that respiratory disease is a common problem in veterinary medicine, evaluation of the small animal pa...

  7. Effect of transient scrotal hyperthermia on sperm parameters, seminal plasma biochemical markers, and oxidative stress in men.

    PubMed

    Rao, Meng; Zhao, Xiao-Ling; Yang, Jing; Hu, Shi-Fu; Lei, Hui; Xia, Wei; Zhu, Chang-Hong

    2015-01-01

    In this experimental prospective study, we aimed to analyze the effect of transient scrotal hyperthermia on the male reproductive organs, from the perspective of sperm parameters, semen plasma biochemical markers, and oxidative stress, to evaluate whether different frequencies of heat exposure cause different degrees of damage to spermatogenesis. Two groups of volunteers (10 per group) received testicular warming in a 43°C water bath 10 times, for 30 min each time: group 1: 10 consecutive days; group 2: once every 3 days. Sperm parameters, epididymis and accessory sex gland function, semen plasma oxidative stress and serum sex hormones were tested before treatment and in the 16-week recovery period after treatment. At last, we found an obvious reversible decrease in sperm concentration (P = 0.005 for Group 1 and P= 0.008 for Group 2 when the minimums were compared with baseline levels, the same below), motility (P = 0.009 and 0.021, respectively), the hypoosmotic swelling test score (P = 0.007 and 0.008, respectively), total acrosin activity (P = 0.018 and 0.009, respectively), and an increase in the seminal plasma malondialdehyde concentration (P = 0.005 and 0.017, respectively). The decrease of sperm concentration was greater for Group 2 than for Group 1 (P = 0.031). We concluded that transient scrotal hyperthermia seriously, but reversibly, negatively affected the spermatogenesis, oxidative stress may be involved in this process. In addition, intermittent heat exposure more seriously suppresses the spermatogenesis compared to consecutive heat exposure. This may be indicative for clinical infertility etiology analysis and the design of contraceptive methods based on heat stress. PMID:25652627

  8. Comparison among different cryoprotectants for cryopreservation of epididymal sperm from agouti (Dasyprocta leporina).

    PubMed

    Castelo, T S; Silva, A M; Bezerra, L G P; Costa, C Y M; Lago, A E A; Bezerra, J A B; Campos, L B; Praxedes, E C G; Silva, A R

    2015-12-01

    We verify the effects of different cryoprotectants on the cryopreservation of agouti (Dasyprocta leporina) epididymal sperm. We used 16 pairs of testes-epididymis complexes of sexually mature animals. We immediately evaluated epididymal sperm obtained by retrograde flushing for concentration, motility, vigor, viability, osmotic response, and morphology. Samples were extended in a coconut water extender plus 20% egg yolk, containing glycerol, ethylene glycol, dimethylsulfoxide - DMSO, or dimethylformamide. Finally, samples were stored in 0.25 mL straws, frozen in liquid nitrogen, and thawed after one week, being reevaluated and assessed for membrane integrity using fluorescent probes. The higher values for postthawing sperm motility, vigor, and membrane integrity were achieved by the usage of glycerol, when compared to ethylene glycol and dimethylformamide (P < 0.05); however, no differences were found between glycerol and DMSO (P > 0.05). All cryoprotectants provided a similar effect on the preservation of sperm morphology, osmotic response, and viability (P > 0.05). Therefore, here onwards, there was testing of glycerol and DMSO at 3 and 6% concentrations using the same freezing-thawing protocol reported previously. As the main result, DMSO at 6% concentration provided a decrease in sperm parameters, as well as in the chromatin integrity and in the binding capability of sperm. In conclusion, glycerol 3 or 6% and DMSO 3% can be used as alternative cryoprotectants for agouti epididymal sperm cryopreservation. PMID:26408846

  9. Sperm DNA Fragmentation and Standard Semen Parameters in Algerian Infertile Male Partners

    PubMed Central

    Boushaba, Sarah

    2015-01-01

    Purpose To date, standard semen parameters have been the only parameters investigated in sperm samples of infertile men in Algeria. We investigated, for the first time, semen parameters according to sperm DNA fragmentation (SDF) in these subjects. Materials and Methods SDF was determined by a validated sperm chromatin dispersion test in 26 infertile men. Patients were split into two groups according to the SDF level estimated by the DNA fragmentation index (DFI): the low fragmentation group (LFG; LFG with DFI ?18%) and high fragmentation group (HFG; HFG with DFI >18%). The standard semen parameters were measured in both groups. Results We found that semen concentration and motility were negatively correlated with DFI (r=-0.65, r=-0.45, respectively; p<0.05), while morphology and semen volume were not correlated with it (r=0.24, r=-0.18, respectively; p>0.05). Comparison of the sperm concentration revealed that it was significantly higher in LFG than in HFG (37.57%±13.16% vs. 7.32%±3.59%, respectively; p<0.05), whereas no significant difference was observed regarding sperm motility and morphology. Conclusions Our findings suggest that SDF correlates well with both sperm motility and concentration but not with morphology. Thus, we conclude that SDF evaluation provides additional information regarding sperm quality, and should be used as a complementary test for assessing semen characteristics in infertile males. PMID:25927056

  10. Studies on Brahma rasayana in male swiss albino mice: Chromosomal aberrations and sperm abnormalities

    PubMed Central

    Guruprasad, K. P.; Mascarenhas, Roshan; Gopinath, P. M.; Satyamoorthy, K.

    2010-01-01

    Ayurveda, the Indian holistic healthcare system encompasses traditional medicines with a principle of creating harmony and maintaining balance within the natural rhythms of the body. Rasayana is one of the branches of Ayurveda frequently used as rejuvenant therapy to overcome many discomforts and prevent diseases. It has been reported that rasayanas have immunomodulatory, antioxidant and antitumor functions. However, the genotoxic potential of many rasayanas remains to be evaluated. The present study was undertaken to assess the role of Brahma rasayana(BR) on genotoxicity in vivo in a mouse test system. The older mice (9 months) were orally fed with rasayana for 8 weeks. The treated groups showed no signs of dose-dependent toxicity at the dosage levels tested. The body weight loss/gain and feed consumption were unaffected at tested doses. Furthermore, sperm abnormalities and chromosomal aberrations were insignificant in the treatment group when compared to controls. However, there was a marginal increase in sperm count in the BR treated animals. These findings clearly indicate that there are no observed adverse genotoxic effects elicited by BR in experimental animals such as mice. PMID:21829300

  11. Studies on Brahma rasayana in male swiss albino mice: Chromosomal aberrations and sperm abnormalities.

    PubMed

    Guruprasad, K P; Mascarenhas, Roshan; Gopinath, P M; Satyamoorthy, K

    2010-01-01

    Ayurveda, the Indian holistic healthcare system encompasses traditional medicines with a principle of creating harmony and maintaining balance within the natural rhythms of the body. Rasayana is one of the branches of Ayurveda frequently used as rejuvenant therapy to overcome many discomforts and prevent diseases. It has been reported that rasayanas have immunomodulatory, antioxidant and antitumor functions. However, the genotoxic potential of many rasayanas remains to be evaluated. The present study was undertaken to assess the role of Brahma rasayana(BR) on genotoxicity in vivo in a mouse test system. The older mice (9 months) were orally fed with rasayana for 8 weeks. The treated groups showed no signs of dose-dependent toxicity at the dosage levels tested. The body weight loss/gain and feed consumption were unaffected at tested doses. Furthermore, sperm abnormalities and chromosomal aberrations were insignificant in the treatment group when compared to controls. However, there was a marginal increase in sperm count in the BR treated animals. These findings clearly indicate that there are no observed adverse genotoxic effects elicited by BR in experimental animals such as mice. PMID:21829300

  12. Sperm in "parhenogenetic" freshwater gastrotrichs.

    PubMed

    Weiss, M J; Levy, D P

    1979-07-20

    Freshwater members of the phylum Gastrotricha have been considered obligate parthenogens. In Lepidodermelia squammata, the species for which there is most evidence for parthenogenesis, sperm have been discovered. This finding will necessitate reexamination of the nature of sexuality and life cycles and of the concept of "species" in freshwater gastrotrichs. PMID:17747043

  13. Branchial Cilia and Sperm Flagella Recruit Distinct Axonemal Components

    PubMed Central

    Konno, Alu; Shiba, Kogiku; Cai, Chunhua; Inaba, Kazuo

    2015-01-01

    Eukaryotic cilia and flagella have highly conserved 9 + 2 structures. They are functionally diverged to play cell-type-specific roles even in a multicellular organism. Although their structural components are therefore believed to be common, few studies have investigated the molecular diversity of the protein components of the cilia and flagella in a single organism. Here we carried out a proteomic analysis and compared protein components between branchial cilia and sperm flagella in a marine invertebrate chordate, Ciona intestinalis. Distinct feature of protein recruitment in branchial cilia and sperm flagella has been clarified; (1) Isoforms of ?- and ?-tubulins as well as those of actins are distinctly used in branchial cilia or sperm flagella. (2) Structural components, such as dynein docking complex, tektins and an outer dense fiber protein, are used differently by the cilia and flagella. (3) Sperm flagella are specialized for the cAMP- and Ca2+-dependent regulation of outer arm dynein and for energy metabolism by glycolytic enzymes. Our present study clearly demonstrates that flagellar or ciliary proteins are properly recruited according to their function and stability, despite their apparent structural resemblance and conservation. PMID:25962172

  14. Flow cytometric evaluation of antibiotic effects on viability and mitochondrial function of refrigerated spermatozoa of Nile tilapia

    USGS Publications Warehouse

    Segovia, M.; Jenkins, J.A.; Paniagua-Chavez, C.; Tiersch, T.R.

    2000-01-01

    Improved techniques for storage and evaluation of fish sperm would enhance breeding programs around the world. The goal of this study was to test the effect of antibiotics on refrigerated sperm from Nile tilapia (Oreochromis niloticus) by use of flow cytometry with 2 dual-staining protocols for objective assessment of sperm quality. Concentrations of 1 x 109 sperm/mL were suspended in Ringer's buffer at 318 mOsmol/kg (pH 8.0). The fluorescent stains Sybr 14 (10 ??M), propidium iodide (2.4 mM), and rhodamine 123 (0.13 ??M) were used to assess cell viability and mitochondrial function. Three concentrations of ampicillin, gentamicin, and an antibiotic/antimycotic solution were added to fresh spermatozoa. Motility estimates and flow cytometry measurements were made daily during 7 d of refrigerated storage (4 ??C). The highest concentrations of gentamicin and antibiotic/antimycotic and all 3 concentrations of ampicillin significantly reduced sperm viability. The highest of each of the 3 antibiotic concentrations significantly reduced mitochondrial function. This study demonstrates that objective sperm quality assessments can be made using flow cytometry and that addition of antibiotics at appropriate concentrations can lengthen refrigerated storage time for tilapia spermatozoa. With minor modifications, these protocols can be adapted for use with sperm from other species and with other tissue types.

  15. Sperm release pathway

    MedlinePLUS

    The key male reproductive organs include the testes, epididymis, urethra, vas deferens, prostate gland, seminal vesicle, and penis. The testes are composed of coiled structures called seminiferous tubules, which are the ...

  16. An update on contraindications for lung function testing.

    PubMed

    Cooper, Brendan G

    2011-08-01

    Guidelines on contraindications for lung function tests have been based on expert opinion from >30 years ago. High-risk contraindications to lung function testing are associated with cardiovascular complications such as myocardial infarct, pulmonary embolism or ascending aortic aneurysm. Slightly less risky but still serious contraindications are predominantly centred on recovery from major thoracic, abdominal or head surgery. Less serious surgical procedures will present a possible risk, but the RR depends upon whether the lung function is essential or can wait until the patient's condition improves. In recent decades there have been moves towards less invasive surgical techniques, keyhole surgery and new technology such as laser surgery which minimise the amount of collateral damage to surrounding tissues. In thoracic surgery there is a shift in emphasis to quicker postsurgical mobility. Furthermore there has been little analysis of the scientific facts behind the current recommendations and contraindications. The principle absolute and relative contraindications are in need of revision, and recommended times of abstaining from lung function tests needs to be reviewed. This review aims to outline the key issues and suggests newer recommendations for contraindications for performing lung function using a risk matrix, as well as offering alternative approaches to testing patients who may be at risk of complication from testing. In general, the previous recommendation of waiting for 6 weeks after surgical procedures or medical complications before performing lung function can often now be reduced to <3 weeks with modern less invasive surgical techniques. PMID:20671309

  17. Republished review: An update on contraindications for lung function testing.

    PubMed

    Cooper, Brendan G

    2011-10-01

    Guidelines on contraindications for lung function tests have been based on expert opinion from >30 years ago. High-risk contraindications to lung function testing are associated with cardiovascular complications such as myocardial infarct, pulmonary embolism or ascending aortic aneurysm. Slightly less risky but still serious contraindications are predominantly centred on recovery from major thoracic, abdominal or head surgery. Less serious surgical procedures will present a possible risk, but the RR depends upon whether the lung function is essential or can wait until the patient's condition improves. In recent decades there have been moves towards less invasive surgical techniques, keyhole surgery and new technology such as laser surgery which minimise the amount of collateral damage to surrounding tissues. In thoracic surgery there is a shift in emphasis to quicker postsurgical mobility. Furthermore there has been little analysis of the scientific facts behind the current recommendations and contraindications. The principle absolute and relative contraindications are in need of revision, and recommended times of abstaining from lung function tests needs to be reviewed. This review aims to outline the key issues and suggests newer recommendations for contraindications for performing lung function using a risk matrix, as well as offering alternative approaches to testing patients who may be at risk of complication from testing. In general, the previous recommendation of waiting for 6 weeks after surgical procedures or medical complications before performing lung function can often now be reduced to <3 weeks with modern less invasive surgical techniques. PMID:21954034

  18. Lifestyle factors and sperm aneuploidy.

    PubMed

    Jurewicz, Joanna; Radwan, Micha?; Sobala, Wojciech; Radwan, Pawe?; Jakubowski, Lucjusz; Hawu?a, Wanda; Ula?ska, Anna; Hanke, Wojciech

    2014-09-01

    Different environmental and lifestyle factors may interfere with the normal disjunction of sister chromatids/chromosomes during meiosis and may cause aneuploidy. The aim of the study was to examine the association between lifestyle factors and sperm aneuploidy. The study population consisted of 212 healthy men under 45 years of age attending an infertility clinic for diagnostic purposes and who had a normal semen concentration of 20-300×10?mL or slight oligozoospermia (semen concentration of 15-20×10?/mL). All participants were interviewed and provided a semen sample. Sperm aneuploidy was assessed using multicolor FISH (DNA probes specific for chromosomes X, Y, 18, 13, 21). Results from the study suggest that lifestyle factors are related to sperm aneuploidy. A positive relationship was found between coffee drinking everyday and the lack of chromosome X or Y, as well as coffee drinking 1-6 times per week and additional chromosome 18. Wearing boxer shorts decrease the copy number changes in the whole chromosome 18, the number of additional chromosome 18 and the lack of chromosome 13. Additionally, obesity (BMI 30-40 kg/m²) was positively associated with additional chromosome 21 after being adjusted for potential confounders. These findings demonstrate that changing the men's lifestyle habits may contribute to reduction of the incidence of sperm aneuploidy. It is necessary that men continue to follow sensible health advice concerning excess weight, coffee drinking and wearing tight fitting underwear. As this is the first such study to examine different lifestyle factors and sperm aneuploidy, the results need to be confirmed on larger population. PMID:25152516

  19. In Vitro Effect of Cell Phone Radiation on Motility, DNA Fragmentation and Clusterin Gene Expression in Human Sperm

    PubMed Central

    Zalata, Adel; El-Samanoudy, Ayman Z; Shaalan, Dalia; El-Baiomy, Youssef; Mostafa, Taymour

    2015-01-01

    Background Use of cellular phones emitting radiofrequency electromagnetic field (RF-EMF) has been increased exponentially and become a part of everyday life. This study aimed to investigate the effects of in vitro RF-EMF exposure emitted from cellular phones on sperm motility index, sperm DNA fragmentation and seminal clusterin (CLU) gene expression. Materials and Methods In this prospective study, a total of 124 semen samples were grouped into the following main categories: i. normozoospermia (N, n=26), ii. asthenozoospermia (A, n=32), iii. asthenoteratozoospermia (AT, n=31) and iv. oligoasthenoteratozoospermia (OAT, n=35). The same semen samples were then divided into two portions non-exposed and exposed samples to cell phone radiation for 1 hour. Before and immediately after exposure, both aliquots were subjected to different assessments for sperm motility, acrosin activity, sperm DNA fragmentation and CLU gene expression. Statistical differences were analyzed using paired t student test for comparisons between two sub-groups where p<0.05 was set as significant. Results There was a significant decrease in sperm motility, sperm linear velocity, sperm linearity index, and sperm acrosin activity, whereas there was a significant increase in sperm DNA fragmentation percent, CLU gene expression and CLU protein levels in the exposed semen samples to RF-EMF compared with non-exposed samples in OAT>AT>A>N groups, respectively (p<0.05). Conclusion Cell phone emissions have a negative impact on exposed sperm motility index, sperm acrosin activity, sperm DNA fragmentation and seminal CLU gene expression, especially in OAT cases. PMID:25918601

  20. Regulation of salmonid fish sperm motility by osmotic shock-induced water influx across the plasma membrane.

    PubMed

    Takei, Gen Leon; Mukai, Chinatsu; Okuno, Makoto

    2015-04-01

    The motility of salmonid fish sperm is initiated by a decrease in the extracellular K(+) concentration. However, our previous studies revealed that salmonid fish sperm motility could be initiated in the presence of an inhibitory concentration of K(+) by drastic osmotic shock induced by suspension in a hypertonic glycerol solution and subsequent dilution in a hypotonic solution (glycerol-treatment). In the present study, we examined if an osmotic shock-induced water influx is involved in the regulation of salmonid fish sperm motility. HgCl2, a common inhibitor of aquaporins (AQPs), decreased the duration of salmonid fish sperm motility. Dilution of sperm cells in a hypotonic solution increased the cellular volume, whereas HgCl2 inhibited such an increase in cellular volume. Furthermore, the expression of AQP 1a and 10 in rainbow trout testes was confirmed. In contrast, HgCl2 did not affect glycerol-treated sperm motility, indicating that AQPs are not involved in glycerol-treated sperm motility. We also explored the possibility of aquaporin-independent water influx in glycerol-treated sperm by assessing the sperm membrane permeability using propidium iodide. The plasma membrane of glycerol-treated sperm was considerably permeabilized. The cellular volume was decreased in a hypertonic glycerol solution and increased upon subsequent hypoosmotic shock, indicating an AQP-independent water flux across the plasma membrane upon glycerol-treatment. Taken together, these results showed that water influx across the plasma membrane via AQP is crucial for the maintenance of salmonid fish sperm motility under normal conditions, whereas water influx by osmotic shock-induced membrane permeation is critical for the initiation of glycerol-treated sperm motility. PMID:25522712

  1. Seasonal changes of sperm storage and correlative structures in male and female soft-shelled turtles, Trionyx sinensis.

    PubMed

    Xiangkun, Han; Li, Zhang; Meiying, Li; Huijun, Bao; Nainan, Hei; Qiusheng, Chen

    2008-11-01

    Reproductive ducts of male and female soft-shelled turtles, Trionyx sinensis were examined throughout the year (March, May, September, December) using brightfield and electron microscopes (TEM and SEM), to determine the location and histomorphological characteristics of sperm storage structures as well as their changes at different phases of the seasonal reproductive cycle. Sperm stored in the epididymis were also examined. In the male, spermatogenesis is initiated in spring (May), and then the mature sperm are released in autumn as an episodic event. Spermatogenesis is inactive in winter. However, in this species, the epididymis contains sperm throughout the entire year. Sperm observed in the epididymis are intact and some structures are uniquely different from other reptiles, and is characterized by 35-40 concentric mitochondria with a dense core in the centre. Many glycogen granules are observed in the cytoplasm of the midpiece. However, the epithelial cell type of epididymal duct change in different seasons. The cells are fully developed with a highly secretory activity in September. The materials secreted from the epithelium might have the function as nourishment for the stored sperm. Sperm storage structures in the form of tubules are observed in the wall of the isthmus of the oviduct in hibernating females but are absent in the groups of May and September. These tubules develop either by folding or fusion of the oviductal mucosal folds and are lined by both ciliated and secretory cells. These tubules might provide a microenvironment for the sperm to enable its long-term storage. After being separated 4 months (December-March) from the male, sperm are observed in the tubules of the isthmus of the oviduct. The unique character of the sperm combined with the special sperm storage structures enable the sperm to maintain fertility and activity during their storage. PMID:17997057

  2. Sperm competition and the evolution of precopulatory weapons: Increasing male density promotes sperm competition and reduces selection on arm strength in a chorusing frog.

    PubMed

    Buzatto, Bruno A; Roberts, J Dale; Simmons, Leigh W

    2015-10-01

    Sperm competition theory assumes a trade-off between precopulatory traits that increase mating success and postcopulatory traits that increase fertilization success. Predictions for how sperm competition might affect male expenditure on these traits depend on the number of competing males, the advantage gained from expenditure on weapons, and the level of sperm competition. However, empirical tests of sperm competition theory rarely examine precopulatory male expenditure. We investigated how variation in male density affects precopulatory sexual selection on male weaponry and the level of sperm competition in the chorusing frog Crinia georgiana, where males use their arms as weapons in male-male combat. We measured body size and arm girth of 439 males, and recorded their mating success in the field. We found density-dependent selection acting on arm girth. Arm girth was positively associated with mating success, but only at low population densities. Increased male density was associated with higher risk and intensity of sperm competition arising from multimale amplexus, and a reversal in the direction of selection on arm girth. Opposing patterns of pre- and postcopulatory selection may account for the negative covariation between arm girth and testes across populations of this species. PMID:26375605

  3. Potential Changes in Rat Spermatogenesis and Sperm Parameters after Inhalation of Boswellia papyrifera and Boswellia carterii Incense

    PubMed Central

    Ahmed, Mukhtar; Al-Daghri, Nasser; Alokail, Majed S.; Hussain, Tajamul

    2013-01-01

    In this study the effect of Boswellia papyrifera (B. papyrifera) and Boswellia carterii (B. carterii) smoke exposure on spermatogenesis and sperm parameters in male albino rats was investigated. Rats (n = 11) were exposed daily in smoking chambers to smoke emanated by burning 4 g each of either B. papyrifera or B. carterii for 48 days. At the end of exposure duration rats were killed, and the testes were excised and analysed for histopathological and ultrastructural changes. Sperm analysis including total sperm count, motility, velocity and relative percentage of abnormal sperms were recorded. Rats exposed to B. papyrifera and B. carterii showed significant disturbances in spermatogenetic patterns and changes in sperm kinetics compared to unexposed rats. Atrophied seminiferous tubules with dynamic changes were also noticed. The boundaries of intercellular and intracellular vacuoles were seen in the Sertoli cells. Furthermore, in spermatids acrosomal vesicles were not fully formed. Degenerating spermatids were devoid of their nuclear membrane with electron dense matrix and vacuolization. Structural changes in Leydig cells were observed. Sperm analysis in exposed rats exhibited significant decrease in the sperm count, motility, speed and an increase in sperm anomalies when compare to controls. These findings demonstrate that the B. papyrifera and B. carterii smoke affects the process of spermatogenesis and sperm parameters and indicate the detrimental effects of these incense materials on human reproductive system. PMID:23449005

  4. Potential changes in rat spermatogenesis and sperm parameters after inhalation of Boswellia papyrifera and Boswellia carterii incense.

    PubMed

    Ahmed, Mukhtar; Al-Daghri, Nasser; Alokail, Majed S; Hussain, Tajamul

    2013-03-01

    In this study the effect of Boswellia papyrifera (B. papyrifera) and Boswellia carterii (B. carterii) smoke exposure on spermatogenesis and sperm parameters in male albino rats was investigated. Rats (n = 11) were exposed daily in smoking chambers to smoke emanated by burning 4 g each of either B. papyrifera or B. carterii for 48 days. At the end of exposure duration rats were killed, and the testes were excised and analysed for histopathological and ultrastructural changes. Sperm analysis including total sperm count, motility, velocity and relative percentage of abnormal sperms were recorded. Rats exposed to B. papyrifera and B. carterii showed significant disturbances in spermatogenetic patterns and changes in sperm kinetics compared to unexposed rats. Atrophied seminiferous tubules with dynamic changes were also noticed. The boundaries of intercellular and intracellular vacuoles were seen in the Sertoli cells. Furthermore, in spermatids acrosomal vesicles were not fully formed. Degenerating spermatids were devoid of their nuclear membrane with electron dense matrix and vacuolization. Structural changes in Leydig cells were observed. Sperm analysis in exposed rats exhibited significant decrease in the sperm count, motility, speed and an increase in sperm anomalies when compare to controls. These findings demonstrate that the B. papyrifera and B. carterii smoke affects the process of spermatogenesis and sperm parameters and indicate the detrimental effects of these incense materials on human reproductive system. PMID:23449005

  5. Laser radiation and motility patterns of human sperm

    SciTech Connect

    Lenzi, A.; Claroni, F.; Gandini, L.; Lombardo, F.; Barbieri, C.; Lino, A.; Dondero, F. )

    1989-01-01

    Human sperm were exposed in vitro to laser radiation. An increase in progressive sperm motility was associated with a faster rate of sperm ATP consumption. Computer-assisted analysis of sperm motility confirmed the positive effect of laser irradiation on velocity and linearity of sperm.

  6. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... conditions: (a) The sperm oil is derived from rendering the fatty tissue of the sperm whale or is prepared by synthesis of fatty acids and fatty alcohols derived from the sperm whale. The sperm oil obtained by... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Hydrogenated sperm oil. 173.275 Section...

  7. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... conditions: (a) The sperm oil is derived from rendering the fatty tissue of the sperm whale or is prepared by synthesis of fatty acids and fatty alcohols derived from the sperm whale. The sperm oil obtained by... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Hydrogenated sperm oil. 173.275 Section...

  8. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... conditions: (a) The sperm oil is derived from rendering the fatty tissue of the sperm whale or is prepared by synthesis of fatty acids and fatty alcohols derived from the sperm whale. The sperm oil obtained by... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Hydrogenated sperm oil. 173.275 Section...

  9. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... conditions: (a) The sperm oil is derived from rendering the fatty tissue of the sperm whale or is prepared by synthesis of fatty acids and fatty alcohols derived from the sperm whale. The sperm oil obtained by... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Hydrogenated sperm oil. 173.275 Section...

  10. Copulatory behavior in a pholcid spider: males use specialized genitalic movements for sperm removal and copulatory courtship

    NASA Astrophysics Data System (ADS)

    Calbacho-Rosa, Lucía; Galicia-Mendoza, Ivette; Dutto, María Sofía; Córdoba-Aguilar, Alex; Peretti, Alfredo V.

    2013-05-01

    Sexual selection may operate on pre-copulatory, copulatory, and post-copulatory traits. An example of a copulatory target of sexual selection is the genitalic movements a male performs during copulation. These movements may function either to prevent sperm competition or to influence a female's fertilization decision. Here we investigated how copulation duration, pedipalp movements, and abdominal movements that males of the pholcid spider Holocnemus pluchei produce during copulation influence sperm removal and/or patterns of successful sperm transfer. We compared mating events with virgin and mated females for differences in copulatory and post-copulatory behavior. We expected longer copulation duration, longer pedipalp movement duration, and more complex and frequent pedipalp and abdominal movements when males mated with mated females compared to virgin females. Except for abdominal movements, our results corroborated these predictions. Furthermore, when we investigated mating events with mated females, we observed sperm mass ejection from the female gonopore and physical removal of sperm by males' procursi. Females with interrupted second mating events showed a significant reduction of stored sperm masses compared to females with completed mating events. We suggest that males use alternating pedipalp movements to remove most of the rival sperm stored by mated females prior to sperm transfer. Copulation duration and pedipalp movements can be further used to transfer sperm and/or as a form of genitalic copulatory courtship.

  11. Integrated morphophysiological assessment of two methods for sperm selection in bovine embryo production in vitro.

    PubMed

    Cesari, A; Kaiser, G G; Mucci, N; Mutto, A; Vincenti, A; Fornés, M W; Alberio, R H

    2006-09-15

    Extensive work was done regarding the ability of Swim up and Percoll gradient to select functional sperm for in vitro embryo production (IVP) systems. The aim of this work was to compare Swim up and Percoll as methods of sperm selection by ultrastructural, biochemical and functional studies. Frozen-thawed semen from two bulls (Experiments 1 and 2, respectively) were treated using Swim up or Percoll discontinuous gradients. Motility, sperm membrane ultrastructure, sperm proteins, in vitro embryo production (insemination doses, cleavage, embryo yield and quality) and embryo sex ratio were scored and compared. Electron transmission microscopy of outer sperm membranes showed higher (P<0.05) percentage of sperm with lost acrosomes in Percoll treated samples compared to Swim up. A differential protein pattern was also detected. When in vitro embryo production was performed, Percoll gradient produced higher (P<0.05) number of fertilizing doses (7.6 versus 5.9, Bull 1; 13.5 versus 7.8, Bull 2) and higher sperm motility (90% versus 76.6%, Bull 1; 81.7% versus 68.3%, Bull 2) than Swim up. The percentage of cleavage (Day 3) was similar in both treatment groups, whereas embryo production rate (Day 7) was higher (39.4% versus 30.2%, Bull 1; 38% versus 32.4%, Bull 2; P<0.05) when Percoll gradient was used. The percentage of hatched embryos (Day 11) and sex ratio did not differ. Total cell counting and embryo differential staining (inner cell mass and trophoblast cells) of Day 7 embryos showed that Percoll treated sperm produced better quality embryos compared to Swim up. We concluded that Percoll had a better performance selecting sperm and an enhanced capacity for embryo production when compared with the Swim up procedure; this could be attributed to a better acrosome exocytosis, associated to the absence of certain membrane proteins. PMID:16647751

  12. Insights into female sperm storage from the spermathecal fluid proteome of the honeybee Apis mellifera

    PubMed Central

    Baer, Boris; Eubel, Holger; Taylor, Nicolas L; O'Toole, Nicholas; Millar, A Harvey

    2009-01-01

    Background Female animals are often able to store sperm inside their body - in some species even for several decades. The molecular basis of how females keep non-own cells alive is largely unknown, but since sperm cells are reported to be transcriptionally silenced and, therefore, limited in their ability to maintain their own function, it is likely that females actively participate in sperm maintenance. Because female contributions are likely to be of central importance for sperm survival, molecular insights into the process offer opportunities to observe mechanisms through which females manipulate sperm. Results We used the honeybee, Apis mellifera, in which queens are highly polyandrous and able to maintain sperm viable for several years. We identified over a hundred proteins representing the major constituents of the spermathecal fluid, which females contribute to sperm in storage. We found that the gel profile of proteins from spermathecal fluid is very similar to the secretions of the spermathecal gland and concluded that the spermathecal glands are the main contributors to the spermathecal fluid proteome. A detailed analysis of the spermathecal fluid proteins indicate that they fall into a range of different functional groups, most notably enzymes of energy metabolism and antioxidant defense. A metabolic network analysis comparing the proteins detected in seminal fluid and spermathecal fluid showed a more integrated network is present in the spermathecal fluid that could facilitate long-term storage of sperm. Conclusions We present a large-scale identification of proteins in the spermathecal fluid of honeybee queens and provide insights into the molecular regulation of female sperm storage. PMID:19538722

  13. Ion Permeabilities in Mouse Sperm Reveal an External Trigger for SLO3-Dependent Hyperpolarization

    PubMed Central

    Chávez, Julio C.; de la Vega-Beltrán, José L.; Escoffier, Jessica; Visconti, Pablo E.; Treviño, Claudia L.; Darszon, Alberto; Salkoff, Lawrence; Santi, Celia M.

    2013-01-01

    Unlike most cells of the body which function in an ionic environment controlled within narrow limits, spermatozoa must function in a less controlled external environment. In order to better understand how sperm control their membrane potential in different ionic conditions, we measured mouse sperm membrane potentials under a variety of conditions and at different external K+ concentrations, both before and after capacitation. Experiments were undertaken using both wild-type, and mutant mouse sperm from the knock-out strain of the sperm-specific, pH-sensitive, SLO3 K+ channel. Membrane voltage data were fit to the Goldman-Hodgkin-Katz equation. Our study revealed a significant membrane permeability to both K+ and Cl? before capacitation, as well as Na+. The permeability to both K+ and Cl? has the effect of preventing large changes in membrane potential when the extracellular concentration of either ion is changed. Such a mechanism may protect against undesired shifts in membrane potential in changing ionic environments. We found that a significant portion of resting membrane potassium permeability in wild-type sperm was contributed by SLO3 K+ channels. We also found that further activation of SLO3 channels was the essential mechanism producing membrane hyperpolarization under two separate conditions, 1) elevation of external pH prior to capacitation and 2) capacitating conditions. Both conditions produced a significant membrane hyperpolarization in wild-type which was absent in SLO3 mutant sperm. Hyperpolarization in both conditions may result from activation of SLO3 channels by raising intracellular pH; however, demonstrating that SLO3-dependent hyperpolarization is achieved by an alkaline environment alone shows that SLO3 channel activation might occur independently of other events associated with capacitation. For example sperm may undergo stages of membrane hyperpolarization when reaching alkaline regions of the female genital tract. Significantly, other events associated with sperm capacitation, occur in SLO3 mutant sperm and thus proceed independently of hyperpolarization. PMID:23577126

  14. Influence of osmolality and ions on the activation and characteristics of zebrafish sperm motility.

    PubMed

    Wilson-Leedy, J G; Kanuga, M K; Ingermann, R L

    2009-04-15

    Despite the prevalence of zebrafish as a model scientific organism, understanding sperm function in this species is essentially limited to observations that osmotic shock initiates motility. During natural spawning, sperm encounter a range of environmental salinities as well as freshwater mixed with egg-associated ovarian fluid (OF), thus sperm are likely to be exposed to saline prior to egg contact. Effects of saline on sperm function in this model species are unknown, but likely to be important. Using computer assisted sperm analysis, this study addressed the effects of osmolality of spawning media and ionic composition and pH on the proportion of sperm becoming motile at activation (motility), as well as sperm velocity and path. When activated with tap water, motility was maximal (80%) at 10s (earliest time measured), declining to 5% by 87 s postactivation. With activation at moderate osmolalities ( approximately 160-200 mmol/kg) initial motility was decreased relative to low osmolality, increased from 10 to 30s, and subsequently declined less rapidly (motility in 80 mM NaCl was 35%, 80%, and 60% at 10, 30 and 147 s, respectively). Thus, moderate osmolality increased duration, but introduced a temporal lag in motility onset. With moderate osmolalities, the rate of velocity decay was less than that with tap water activation. Sodium chloride and sucrose similarly impacted both motility and velocity. Replacement of NaCl with KCl, pH values ranging from 6.8 to 8.4, or the presence of gadolinium were without effect. Motility, but not velocity, was slightly supressed by Ca(2+). Therefore, whereas pH and concentrations of Ca(2+) or K(+) of OF are unlikely to impact fertility via sperm motility, the OF contribution to spawning media osmolality may have pronounced effects on motility and velocity of sperm, factors previously correlated with fertility in other species. PMID:19185341

  15. An automated miniaturized Haploscope for testing binocular visual function

    NASA Technical Reports Server (NTRS)

    Decker, T. A.; Williams, R. E.; Kuether, C. L.; Wyman-Cornsweet, D.

    1976-01-01

    A computer-controlled binocular vision testing device has been developed as one part of a system designed for NASA to test the vision of astronauts during spaceflight. The device, called the Mark III Haploscope, utilizes semi-automated psychophysical test procedures to measure visual acuity, stereopsis, phorias, fixation disparity and accommodation/convergence relationships. All tests are self-administered, yield quantitative data and may be used repeatedly without subject memorization. Future applications of this programmable, compact device include its use as a clinical instrument to perform routine eye examinations or vision screening, and as a research tool to examine the effects of environment or work-cycle upon visual function.

  16. Psychometric functions for efficient and effective psychophysical testing of sensory function.

    PubMed

    Vriens, J P M; Strijbos, S A; van der Glas, H W

    2011-08-01

    An efficient test for sensory function has been reported using random order pairs of real and sham stimuli. The constant magnitude of the real stimulus is chosen such that the first error in a patient's forced-choice report on the order of real and sham stimulation, immediately indicates abnormal sensory function. This magnitude just exceeds a critical large percentile of the psychometric function (i.e. the relationship between percentage of detection and stimulus magnitude in healthy subjects). The aim was to determine psychometric functions for one tongue site and six facial sites for adjusting three variants of the real/sham stimulus method (i.e. for light touch, cold sensation and for two-point discrimination). All 150 healthy subjects participated in testing for light touch sensation, 100 subjects additionally participated in testing two-point discrimination and 50 subjects participated additionally in testing cold sensation. The stimulus magnitude was varied using a staircase-limits procedure. Following curve fitting with a Boltzmann function, 90th, 95th or 99th percentiles of the psychometric functions were determined. A set of at least two real/sham tests, one testing the function of large nerve fibres and one for small fibres, allows quick assessment of a patient's disturbed sensory function including its fibre pathology. PMID:21600735

  17. Chromosomal aberrations induced by in vitro irradiation: comparisons between human sperm and lymphocytes

    SciTech Connect

    Brandriff, B.F.; Gordon, L.A.; Ashworth, L.K.; Carrano, A.V.

    1988-01-01

    Types and frequencies of structural aberrations in human sperm and lymphocyte chromosomes from one donor were compared after in vitro irradiation with 100, 200, and 400 rad in order to determine if cells with dramatically different chromatin configurations are similarly affected and to investigate the feasibility of using lymphocytes as surrogates for germ cells in risk estimation. Sperm chromosomes were analyzed after fusion with eggs from the golden hamster. Total frequencies of induced aberrations were similar in the two cell types. However, the relative frequencies of rejoined lesions (dicentrics), compared with unrejoined lesions (chromosome breaks and acentric fragments), were different. At the three doses tested, a constant ratio of 5 dicentrics in lymphocytes for every dicentric in sperm was induced. Conversely, for every chromosome break or acentric fragment induced in lymphocytes, 1.7 such events were induced in sperm at the three doses tested.

  18. Nucleotide sequence of the D-loop region of the sperm whale (Physeter macrocephalus) mitochondrial genome.

    PubMed

    Dillon, M C; Wright, J M

    1993-03-01

    We have amplified, by the polymerase chain reaction, and have sequenced the D-loop region of the mitochondrial DNA from the sperm whale (Physeter macrocephalus). The sperm whale D-loop was aligned with D-loop sequences from four other cetaceans (Commerson's dolphin, orca, fin whale, and minke whale) and an out-group (cow). This alignment showed the sperm whale sequence to be larger than that of other cetaceans. In addition, some sequence blocks were highly conserved among all six species, suggesting roles in the functioning of mitochondrial DNA. Other blocks that were previously reported to be well conserved among cetaceans showed little sequence conservation with the sperm whale D-loop, which argues against the functional importance of these sequence blocks in cetaceans. PMID:8487632

  19. Sperm competition games: Sperm size (mass) and number under raffle and displacement, and the evolution of P2

    E-print Network

    Pitnick, Scott

    Sperm competition games: Sperm size (mass) and number under raffle and displacement 2010 Keywords: Ejaculate expenditure Giant sperm Size­number trade off Sperm precedence a b s t r a c t We examine models for evolution of sperm size (i.e. mass m) and number (s) under three mechanisms

  20. A Study of the Antioxidant Effect of Alpha Lipoic Acids on Sperm Quality

    PubMed Central

    Ibrahim, Siti Fatimah; Osman, Khairul; Das, Srijit; Othman, Abas Mazni; Majid, Norzaiti Abdul; Rahman, Mohd Padzil Abdul

    2008-01-01

    OBJECTIVE Assisted reproductive techniques are useful in helping infertile couples achieve successful conception. Initial studies have shown that sperm cryopreservation, one step in assisted reproduction, causes a dramatic reduction in sperm quality. This has been attributed to, among other things, free radical activities. The aim of the present study was to minimize this oxidative attack by adding an antioxidant into the sperm microenvironment. Alpha lipoic acids were selected for this purpose for their efficient free radical scavenging properties and solubility in lipid and aqueous phases. METHODS For this investigation, semen from six Boer bucks was pooled. Seminal analysis of the baseline prior to incubation of samples with different concentrations of Alpha lipoic acids (0.00625, 0.0125, 0.025, 0.05, 0.1 mmol/ml) was performed, and post-seminal analysis was conducted after a one-hour incubation. The comet assay was used to observe the effect of Alpha lipoic acids on sperm DNA integrity. Statistical analysis using an unpaired t-test with a significance level of p<0.05 was then performed. RESULTS Our results indicate that the sperm motility rate was improved after incubation with Alpha lipoic acids at a concentration of 0.02 mmol/ml. This concentration was also capable of reducing DNA damage. CONCLUSION In conclusion, Alpha lipoic acids renders cryoprotection to sperm, thereby improving sperm quality. PMID:18719769

  1. Seminal CD38 Enhances Human Sperm Capacitation through Its Interaction with CD31

    PubMed Central

    Kim, Byung-Ju; Park, Dae-Ryoung; Nam, Tae-Sik; Lee, Seo Ho; Kim, Uh-Hyun

    2015-01-01

    Human sperm have to undergo a maturational process called capacitation in the female reproductive tract. Capacitation confers upon the sperm an ability to gain hypermotility and undergo acrosome reaction. Previous studies have suggested that seminal plasma proteins induce the capacitation of sperm in the female reproductive tract for the successful fertilization of the oocyte. However, the function of seminal plasma proteins in capacitation remains largely unclear. To the end, we found that soluble CD38 (sCD38) in seminal plasma increases the capacitation of sperm via specific interactions between sCD38 and the CD31 on the sperm. Upon the association of sCD38 with CD31, tyrosine kinase Src phosphorylates CD31, a process blocked by Src inhibitors. Shc, SHP-2, Grb2, and SOS, as well as Src kinase were found to associate with the phosphorylated CD31. The sCD38-induced phosphorylation of CD31 initiates a cascade reaction through the phosphorylation of Erk1/2, which results in the acrosome reaction, and sperm hypermotility. These processes were prevented by Src, Ras and MEK inhibitors. Taken together, these data indicate that the sCD38 present in seminal plasma plays a critical role in the capacitation of sperm. PMID:26407101

  2. [Ovum fertilization using surgically obtained sperm: additional risk to offspring unlikely with meticulous procedure].

    PubMed

    Meuleman, E J; van Moorselaar, R J

    1998-01-17

    In the Netherlands a presumed higher risk of transmission of genetic abnormalities to the offspring in programmes of intracytoplasmatic sperm injection (ICSI) using surgically retrieved sperm (microscopic epididymal sperm aspiration (MESA) or testicular sperm extraction (TESE)) has triggered a moratorium. However, the ICSI-MESA/TESE-programme should be resumed under conditions for the following reasons: the source of the sperm is not a accurate standard of its genetic contents: the cause of the male infertility and the morphology and function of the injected spermatozoon appear to be better criteria; animal experiments and clinical results are reassuring as to the proposed risk; it is uncertain whether the complex genetical mechanisms involved in spermiogenesis and sperm maturation can be unraveled in the laboratory in due course; the exodus of patients to neighbouring countries will be stopped and biological material necessary for human research will become available. The conditions are that (a) only morphologically normal and motile sperm should be used, (b) ICSI should be preceded by genetical screening and succeeded by long term follow up of the offspring. PMID:9557005

  3. Executive function on the Psychology Experiment Building Language tests

    PubMed Central

    Li, Victoria; Eiwaz, Massarra A.; Kobel, Yuliyana V.; Benice, Ted S.; Chu, Alex M.; Olsen, Reid H. J.; Rice, Douglas Z.; Gray, Hilary M.; Mueller, Shane T.

    2013-01-01

    The measurement of executive function has a long history in clinical and experimental neuropsychology. The goal of the present report was to determine the profile of behavior across the lifespan on four computerized measures of executive function contained in the recently developed Psychology Experiment Building Language (PEBL) test battery http://pebl.sourceforge.net/ and evaluate whether this pattern is comparable to data previously obtained with the non-PEBL versions of these tests. Participants (N = 1,223; ages, 5–89 years) completed the PEBL Trail Making Test (pTMT), the Wisconsin Card Sort Test (pWCST; Berg, Journal of General Psychology, 39, 15–22, 1948; Grant & Berg, Journal of Experimental Psychology, 38, 404–411, 1948), the Tower of London (pToL), or a time estimation task (Time-Wall). Age-related effects were found over all four tests, especially as age increased from young childhood through adulthood. For several tests and measures (including pToL and pTMT), age-related slowing was found as age increased in adulthood. Together, these findings indicate that the PEBL tests provide valid and versatile new research tools for measuring executive functions. PMID:21534005

  4. Executive function on the Psychology Experiment Building Language tests.

    PubMed

    Piper, Brian J; Li, Victoria; Eiwaz, Massarra A; Kobel, Yuliyana V; Benice, Ted S; Chu, Alex M; Olsen, Reid H J; Rice, Douglas Z; Gray, Hilary M; Mueller, Shane T; Raber, Jacob

    2012-03-01

    The measurement of executive function has a long history in clinical and experimental neuropsychology. The goal of the present report was to determine the profile of behavior across the lifespan on four computerized measures of executive function contained in the recently developed Psychology Experiment Building Language (PEBL) test battery http://pebl.sourceforge.net/ and evaluate whether this pattern is comparable to data previously obtained with the non-PEBL versions of these tests. Participants (N = 1,223; ages, 5-89 years) completed the PEBL Trail Making Test (pTMT), the Wisconsin Card Sort Test (pWCST; Berg, Journal of General Psychology, 39, 15-22, 1948; Grant & Berg, Journal of Experimental Psychology, 38, 404-411, 1948), the Tower of London (pToL), or a time estimation task (Time-Wall). Age-related effects were found over all four tests, especially as age increased from young childhood through adulthood. For several tests and measures (including pToL and pTMT), age-related slowing was found as age increased in adulthood. Together, these findings indicate that the PEBL tests provide valid and versatile new research tools for measuring executive functions. PMID:21534005

  5. Chd5 orchestrates chromatin remodelling during sperm development.

    PubMed

    Li, Wangzhi; Wu, Jie; Kim, Sang-Yong; Zhao, Ming; Hearn, Stephen A; Zhang, Michael Q; Meistrich, Marvin L; Mills, Alea A

    2014-01-01

    One of the most remarkable chromatin remodelling processes occurs during spermiogenesis, the post-meiotic phase of sperm development during which histones are replaced with sperm-specific protamines to repackage the genome into the highly compact chromatin structure of mature sperm. Here we identify Chromodomain helicase DNA binding protein 5 (Chd5) as a master regulator of the histone-to-protamine chromatin remodelling process. Chd5 deficiency leads to defective sperm chromatin compaction and male infertility in mice, mirroring the observation of low CHD5 expression in testes of infertile men. Chd5 orchestrates a cascade of molecular events required for histone removal and replacement, including histone 4 (H4) hyperacetylation, histone variant expression, nucleosome eviction and DNA damage repair. Chd5 deficiency also perturbs expression of transition proteins (Tnp1/Tnp2) and protamines (Prm1/2). These findings define Chd5 as a multi-faceted mediator of histone-to-protamine replacement and depict the cascade of molecular events underlying this process of extensive chromatin remodelling. PMID:24818823

  6. Chd5 orchestrates chromatin remodeling during sperm development

    PubMed Central

    Li, Wangzhi; Wu, Jie; Kim, Sang-Yong; Zhao, Ming; Hearn, Stephen A.; Zhang, Michael Q.; Meistrich, Marvin L.

    2014-01-01

    One of the most remarkable chromatin remodeling processes occurs during spermiogenesis, the post-meiotic phase of sperm development during which histones are replaced with sperm-specific protamines to repackage the genome into the highly compact chromatin structure of mature sperm. Here we identify Chromodomain helicase DNA binding protein 5 (Chd5) as a master regulator of the histone-to-protamine chromatin remodeling process. Chd5 deficiency leads to defective sperm chromatin compaction and male infertility in mice, mirroring the observation of low CHD5 expression in testes of infertile men. Chd5 orchestrates a cascade of molecular events required for histone removal and replacement, including histone 4 (H4) hyperacetylation, histone variant expression, nucleosome eviction, and DNA damage repair. Chd5 deficiency also perturbs expression of transition proteins (Tnp1/Tnp2) and protamines (Prm1/2). These findings define Chd5 as a multi-faceted mediator of histone-to-protamine replacement and depict the cascade of molecular events underlying chromatin remodeling during this process of extensive chromatin remodeling. PMID:24818823

  7. The Importance of Functional Tests in Personalized Medicine

    PubMed Central

    Widmer, R. Jay; Lerman, Amir

    2013-01-01

    Cardiovascular disease is the most prevalent disease mainly in the Western society and becoming the leading cause of death worldwide. Standard methods by which healthcare providers screen for cardiovascular disease have only minimally reduced the burden of disease while exponentially increasing costs. As such, more specific and individualized methods for functionally assessing cardiovascular threats are needed to identify properly those at greatest risk, and appropriately treat these patients so as to avoid a fate such as heart attack, stroke, or death. Currently, endothelial function testing—in both the coronary and peripheral circulation—is well established as being associated with the disease process and future cardiovascular events. Improving such testing can lead to a reduction in the risk of future events. Combining this functional assessment of vascular fitness with other, more personalized, testing methods should serve to identify those at the greatest risk of cardiovascular disease earlier and subsequently reduce the affliction of such diseases worldwide. PMID:23908864

  8. The effects of radiation on sperm swimming behavior depend on plasma oxidative status in the barn swallow (Hirundo rustica).

    PubMed

    Bonisoli-Alquati, Andrea; Møller, Anders Pape; Rudolfsen, Geir; Saino, Nicola; Caprioli, Manuela; Ostermiller, Shanna; Mousseau, Timothy A

    2011-06-01

    Sperm are highly susceptible to reactive oxygen species (ROS) that can damage sperm DNA and structure, resulting in reduced fertilizing capacity. Exposure to radioactive contamination can also impair sperm swimming behavior and fertilizing ability, both through a reduction of sperm DNA integrity and via an increased generation of reactive oxygen species (ROS). However, the relationship between individual oxidative status and sperm swimming behavior has never been investigated in any wild population of animals exposed to radioactive contamination. We studied the motility of sperm collected from barn swallows, Hirundo rustica, breeding under different levels of radioactive contamination following the Chernobyl accident in 1986, in relation to individual oxidative status. We tested the hypothesis that the degree of impairment of sperm swimming behavior by radioactive contamination depended on plasma antioxidant capacity, the level of reactive oxygen metabolites (ROMs) and oxidative stress (sensu Costantini et al. 2006), a better oxidative status being associated with higher sperm motility. Sperm behavior parameters were subjected to principal component (PC) analysis, which extracted four PCs explaining 86% of the variance in sperm motility. PC2, representing sperm with high track velocity and ample lateral head displacement, was significantly predicted by the interaction between radiation level and either oxidative damage or oxidative stress. Contrary to our predictions, the highest values of PC2 were associated with relatively high radiation levels, particularly for high levels of either ROMs or oxidative stress. In addition, there was a tendency for values of PC3 (representing the percent of motile sperm) and PC4 (representing slow sperm with high beat cross frequency) to depend on the interaction between radiation level and total plasma antioxidant protection. Our results confirm the importance of oxidative status in determining the genetic and physiological outcome of exposure to radioactive contamination, complementing previous studies relating sperm abnormality to circulating levels of specific antioxidants. Our results also complement previous evidence that oxidative damage of sperm was negatively related to sperm motility, thus indicating a possible trade-off in quenching pro-oxidant compounds in the plasma and the seminal fluid. PMID:21295152

  9. Sperm selection based on electrostatic charge.

    PubMed

    Simon, Luke; Ge, Shao-Qin; Carrell, Douglas T

    2013-01-01

    Charge is a fundamental property of all forms of matter that exhibit attraction or repulsion in the presence of another charged particle. This electrokinetic property occurs when the particles exhibiting a net negative or positive charge are subjected to an external electric field that exerts an electrostatic force between them. Sperm surface membranes exhibit varying levels of electrostatic potential that are proportional to the levels of sialic acid residue acquired on the cell surface during maturation. Electrostatic charge-based sperm separation is a recently developed technique that uses an electric field to isolate mature sperm with reduced levels of DNA fragmentation. Two methods for the separation of sperm based on electrostatic charge, the Zeta method and a commercially available electrophoretic method using the SpermSep Cell Sorter 10, are discussed in this chapter including a detailed protocol for sperm separation based on the Zeta method. PMID:22992922

  10. Ultrastructural organization of the sperm nuclear matrix.

    PubMed

    Santi, S; Rubbini, S; Cinti, C; Squarzoni, S; Matteucci, A; Caramelli, E; Guidotti, L; Maraldi, N M

    1995-01-01

    The organization of DNA-protamine complexes and their association with the nuclear matrix have been analyzed in sperm nuclei by in situ Nick Translation at the electron microscope. The data obtained indicate that the chromatin organization in sperm nuclei is maintained during the sperm condensation by means of interaction with the nuclear matrix at fixed sites. The fine structure of the sperm nucleus and sperm nuclear matrix, investigated by sectioning and replica of freeze-fractured specimens, suggests that the lamellar array observed by freeze-fracturing in the sperm nuclei, could depend on the inner matrix which presents a regular organization of globular structures involved in the maintenance of chromatin domains. PMID:11322315

  11. Nuclear microscopy of sperm cell elemental structure

    SciTech Connect

    Bench, G.S.; Balhorn, R.; Friz, A.M.; Freeman, S.P.H.T.

    1994-09-28

    Theories suggest there is a link between protamine concentrations in individual sperm and male fertility. Previously, biochemical analyses have used pooled samples containing millions of sperm to determine protamine concentrations. These methods have not been able to determine what percentage of morphologically normal sperm are biochemically defective and potentially infertile. Nuclear microscopy has been utilized to measure elemental profiles at the single sperm level. By measuring the amount of phosphorus and sulfur, the total DNA and protamine content in individual sperm from fertile bull and mouse semen have been determined. These values agree with results obtained from other biochemical analyses. Nuclear microscopy shows promise for measuring elemental profiles in the chromatin of individual sperm. The technique may be able to resolve theories regarding the importance of protamines to male fertility and identify biochemical defects responsible for certain types of male infertility.

  12. GROUND-WATER MODEL TESTING: SYSTEMATIC EVALUATION AND TESTING OF CODE FUNCTIONALITY AND PERFORMANCE

    EPA Science Inventory

    Effective use of ground-water simulation codes as management decision tools requires the establishment of their functionality, performance characteristics, and applicability to the problem at hand. This is accomplished through application of a systematic code-testing protocol and...

  13. Development of a specific method to evaluate 8-hydroxy, 2-deoxyguanosine in sperm nuclei: relationship with semen quality in a cohort of 94 subjects.

    PubMed

    Cambi, M; Tamburrino, L; Marchiani, S; Olivito, B; Azzari, C; Forti, G; Baldi, E; Muratori, M

    2013-03-01

    Oxidative stress (OS) is involved in many disoders including male infertility. Human spermatozoa are very sensitive targets of reactive oxygen species (ROS) and most sperm functions are impaired in the case of OS. In addition unbalanced production of ROS is considered one of the most important causes of sperm DNA fragmentation, a semen trait of infertile men. The relationship between oxidative damage and semen quality is partially controversial, probably due to the different methods and/or targets used to reveal the OS. In this study, by fluorescence microscopy and flow cytometry, we compared two methods to reveal 8-hydroxy,2-deoxyguanosine (8-OHdG), the hallmark of oxidative DNA damage: an immunofluorescence method and the commercial OxyDNA kit. We found that although both methods localized the labelling in sperm nuclei they yielded different measures, and only with the immunofluorescence method was the labelling specific for sperm 8-OHdG. The immunofluorescence method, coupled to flow cytometry, was thus selected to analyse the 8-OHdG content in semen samples from 94 subfertile patients and to investigate the relationship with semen quality. We found that the percentages of spermatozoa with 8-OHdG (mean±s.d., 11.4±6.9%) were related to sperm count (Pearson's correlation coefficient (r)=-0.27, P=0.04 (ANOVA and student's t-test)), motility (progressive: r=-0.22, P=0.04; non-progressive: r=0.25, P=0.01), and normal morphology (r=-0.27, P=0.01). In conclusion, we demonstrate that immunofluorescence/flow cytometry is a reliable and specific method to detect 8-OHdG at single-cell level and show that oxidative damage only partially overlaps poor semen quality, suggesting that it could provide additional information on male fertility with respect to routine semen analysis. PMID:23457372

  14. Decreased activity of superoxide dismutase in the seminal plasma of infertile men correlates with increased sperm deoxyribonucleic acid fragmentation during the first hours after sperm donation.

    PubMed

    Wdowiak, Artur; Bakalczuk, Szymon; Bakalczuk, Grzegorz

    2015-07-01

    Sperm DNA fragmentation varies between individuals and is more pronounced with increased patient age and time after sperm donation. The intensification of DNA fragmentation depends on the balance of the oxidoreductive system, which is regulated mainly by two enzymes - superoxide dismutase (SOD) and catalase. The objective of this study was to determine the relationship between sperm DNA fragmentation dynamics, fertility and seminal SOD and catalase activity. The study was conducted in 2013 and 2014 at the Non-Public Health Care Unit 'Ovum Reproduction and Andrology' in Lublin, Lublin, Poland, and covered 218 men aged 25-35 (85 fertile and 133 patients treated for infertility). Percentage of fragmented DNA was measured in a modified chromatin dispersion test at four time points after sperm donation (t = 0, 3, 6, 12 h). SOD and catalase activities were determined spectrophotometrically. We confirmed that the activity of SOD in the seminal plasma of men with reproductive disorders was lower compared with fertile men. Conversely, no significant correlations were found between fertility and catalase activity. Sperm DNA of infertile males was initially more fragmented than fertile male sperm DNA. SOD and catalase activity did not correlate with the degree of DNA fragmentation in fertile men. In men with reproductive disorders, the rate of DNA fragmentation was slow within first 3 h after sperm donation and then increased between 6 and 12 h. In this group of infertile men, those with higher SOD activity had a lower DNA fragmentation index (DFI) after 12 h, and a reduced rate of intensity of fragmentation from 6 to 12 h. Alternatively, higher catalase activity among men treated for infertility was accompanied by higher initial DFI and higher rate of DNA fragmentation from 6 to 12 h. These results highlight the importance of determining a proper time window between sperm donation and procedures of assisted reproductive technology. PMID:26198800

  15. An Acute-phase Protein as a Regulator of Sperm Survival in the Bovine Oviduct: Alpha 1-acid-glycoprotein Impairs Neutrophil Phagocytosis of Sperm In Vitro

    PubMed Central

    LIU, Jinghui; MAREY, Mohamed A.; KOWSAR, Rasoul; HAMBRUCH, Nina; SHIMIZU, Takashi; HANEDA, Shingo; MATSUI, Motozumi; SASAKI, Motoki; HAYAKAWA, Hiroyuki; PFARRER, Christiane; MIYAMOTO, Akio

    2014-01-01

    We have previously shown that polymorphonuclear neutrophils (PMNs) are present in bovine oviduct fluid under physiological conditions, and that the oviduct provides a microenvironment that protects sperm from phagocytosis by PMNs. Alpha 1-acid glycoprotein (AGP) is a major acute-phase protein produced mainly in the liver that has immunomodulatory functions. AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary. Therefore, in this study, we investigated, 1) the local production of AGP in the bovine oviduct, 2) the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production and 3) the impact of AGP desialylation on the PMN phagocytosis of sperm. The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to produce prostaglandin E2 (PGE2) which has been shown to partially contribute to the regulation of sperm phagocytosis in the bovine oviduct. AGP and PGE2 at concentrations detected in the oviducts additively suppressed sperm phagocytosis by PMNs. These results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct. PMID:24931131

  16. Sperm volumetric dynamics during in vitro capacitation process in bovine spermatozoa.

    PubMed

    García-Herreros, M; Leal, C L V

    2015-06-01

    Previous studies have demonstrated that sperm head morphometry can be used as a potential diagnostic tool for detecting biophysical changes associated with sperm viability in bovine spermatozoa. In this study, sperm head morphometry was used to investigate its value as a biophysical marker for detecting volumetric changes in bovine spermatozoa under in vitro capacitating and non-capacitating incubation conditions. To further test this hypotesis, aliquots of pooled, washed bovine sperm were incubated in either Tyrode's complete medium with heparin (TCMH; a capacitating medium containing Ca2+, NaHCO3 and heparin), Tyrode's complete medium heparin-free (TCM; a medium containing just Ca2+ and NaHCO3) or Tyrode's basal medium (TBM; a non-capacitating medium free of Ca2+, NaHCO3 and heparin, used as control). Aliquots of sperm were processed for morphometric analysis at different incubation-time intervals (0, 3 and 6 h at 38°C), and the chlortetracycline assay was used simultaneously to confirm the ability of the sperm to undergo capacitation (B pattern) and the acrosome reaction (AR pattern) status in each medium. After 3 h of incubation under TCMH conditions, a significant increase was observed in the percentage of B and AR patterns and a significant decrease was found in all sperm morphometric parameters (P<0.01). Interestingly, after 6 h of incubation in TCMH, the percentage of B and AR patterns increased drastically over time and marked differences were found in the dimensional and shape parameters, which were significantly smaller compared with TBM or TCM media (P<0.001). Significant correlations were observed between sperm size and AR pattern (r=-0.875, P<0.01). In conclusion, sperm head morphometry can be used as a potential biophysical marker for detecting volumetric changes during capacitation process in bovine spermatozoa. PMID:25684453

  17. Microscopic Varicocelectomy Significantly Decreases the Sperm DNA Fragmentation Index in Patients with Infertility

    PubMed Central

    Kadioglu, Teoman Cem; Aliyev, Emin; Celtik, Murad

    2014-01-01

    Background. Varicocele is associated with high levels of DNA damage in spermatozoa due to oxidative stress and elevated levels of sperm DNA fragmentation, which has been currently proposed to be an essential additional diagnostic test to be recommended for patients with clinical varicocele. The aim of this study was to evaluate the parameters of semen and the DNA fragmentation index (DFI) in patients with varicocele before and after varicocelectomy. Methods. The details of 92 consecutive patients were retrospectively analyzed from January 2010 to December 2012. The sperm samples were evaluated according to the World Health Organization Guidelines. Sperm DNA damage, characterized as DFI, was evaluated by sperm chromatin structure assay using flow cytometry. Results. There was a statistically significant improvement in the semen concentration, the total motile count, the total normal sperm count, and the sperm DNA fragmentation index (DFI; the percentage of sperm with denatured DNA) after varicocelectomy. There was a large decrease in DFI from a preoperative mean of 42.6% to a postoperative mean of 20.5% (P < 0.001). A higher preoperative DFI was associated with a larger decrease in postoperative DFI, and significant negative correlations were observed between the DFI and sperm motility (r = ?0.42, P < 0.01). Conclusion. Our data suggest that varicocelectomy can improve multiple semen parameters and sperm DNA damage in infertile men with varicocele. The patients with preoperative defects in those parameters showed greater improvement postoperatively. Further research in this area is needed to understand the exact mechanisms of DNA damage in infertile men with varicocele. PMID:24712000

  18. Sperm competition in a fish with external fertilization: the contribution of sperm number, speed and length

    E-print Network

    Neff, Bryan D.

    was investigated using in vitro fertilization experiments with bluegill (Lepomis macrochirus). Bluegill males useSperm competition in a fish with external fertilization: the contribution of sperm number, speed for the fertilization of a given set of eggs (Hildemann & Wagner, 1954; Parker, 1970). Sperm competition is widespread

  19. UBIQUITINATION OF BOAR SPERM: WHAT DOES THIS REVEAL ABOUT SPERM PHYSIOLOGY?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Limited data on sperm ubiquitination have shown negative correlations (bovine and human) and positive correlations (human) with sperm quality, but its effects on boar sperm are largely unknown. This research determined the influence of ubiquitination on motility, mitochondrial activity, membrane fl...

  20. Autoradiographic visualization of the mouse egg's sperm receptor bound to sperm

    SciTech Connect

    Bleil, J.D.; Wassarman, P.M.

    1986-04-01

    The extracellular coat, or zona pellucida, of mammalian eggs contains species-specific receptors to which sperm bind as a prelude to fertilization. In mice, ZP3, one of only three zona pellucida glycoproteins, serves as sperm receptor. Acrosome-intact, but not acrosome-reacted, mouse sperm recognize and interact with specific O-linked oligosaccharides of ZP3 resulting in sperm-egg binding. Binding, in turn, causes sperm to undergo the acrosome reaction; a membrane fusion event that results in loss of plasma membrane at the anterior region of the head and exposure of inner acrosomal membrane with its associated acrosomal contents. Bound, acrosome-reacted sperm are able to penetrate the zona pellucida and fuse with the egg's plasma membrane (fertilization). In the present report, we examined binding of radioiodinated, purified, egg ZP3 to both acrosome intact and acrosome reacted sperm by whole-mount autoradiography. Silver grains due to bound 125I-ZP3 were found localized to the acrosomal cap region of heads of acrosome-reacted sperm. Under the same conditions, 125I-fetuin bound at only background levels to heads of both acrosome-intact and -reacted sperm, and 125I-ZP2, another zona pellucida glycoprotein, bound preferentially to acrosome-reacted sperm. These results provide visual evidence that ZP3 binds preferentially and specifically to heads of acrosome intact sperm; properties expected of the mouse egg's sperm receptor.

  1. Protective effects of vitamin E and Cornus mas fruit extract on methotrexate-induced cytotoxicity in sperms of adult mice

    PubMed Central

    Zarei, Leila; Sadrkhanlou, Rajabali; Shahrooz, Rasoul; Malekinejad, Hassan; Eilkhanizadeh, Behroz; Ahmadi, Abbas

    2014-01-01

    This study was aimed to assess the protective effects of Cornus mas fruit extract (CMFE) and vitamin E (Vit E) on sperm quality parameters in the methotrexate (MTX)-treated mice. Forty-eight young adult male mice (8-12 weeks) were randomly divided into six groups including control and test groups. The control group received normal saline orally , and the test groups were treated MTX (20 mg kg-1, ip, once weekly), MTX + CMFE (250 mg kg-1), MTX + CMFE (500 mg kg-1), MTX + CMFE (1000 mg kg-1), and MTX + Vit E (100 IU kg-1, po) for 35 consecutive days. On day 35, after euthanasia the epididymal sperms were isolated. Then the total mean sperm count, sperm viability and motility were determined. The total antioxidant capacity (TAOC) of all experimental groups were also evaluated. The MTX-treated animals showed a significant changes in all parameters of sperm quality assessment compared to the control group. Both Vit E and CMFE were able to protect from MTX-induced effects on sperm maturity and DNA damage. Co-administration of MTX and CMFE and/or Vit E resulted in protection from MTX-reduced TAOC. In conclusion, these data suggested that MTX administration could adversely affect the sperm quality. Moreover, the protective effect of Vit E and CMFE on MTX-induced sperm toxicity was also documented. PMID:25568688

  2. [Pulmonary function testing: history up to the present].

    PubMed

    Tojo, Naoko

    2014-12-01

    The main roles of pulmonary function testing are: aiding in the diagnosis, assessing disease severity, and monitoring the progress and effects of treatment. The spirometric measurements of highest clinical value are the vital capacity(VC) and forced expiratory volume in one second(FEV1). Measurements of VC were reported in both healthy subjects and patients with respiratory disease during the mid-19th century. The VC, however, was little used in clinical medicine until the second half of the 20th century. The FEV1, FEV1/VC ratio, and/or FEV1/forced vital capacity (FVC) ratio all came into existence after 1950. Pulmonary function testing evolved over time into a more complex set of lung function measurements using various techniques and devices. If the variability of the results can be minimized and the measurement accuracy can be improved, abnormalities will be more easily detected. In 2005, the American Thoracic Society (ATS) and European Respiratory Society (ERS) together published an updated set of standards for pulmonary function testing that could be applied more widely. These standards are structured to cover definitions, equipment, and patient-related procedures commonly applied for many methods of lung function testing. In 2004, the Japanese Respiratory Society issued an initial set of recommendations for the standardized measurement of VC, FVC, and the single-breath carbon monoxide-diffusing capacity. These recommendations have not been updated, and the prediction equations for pulmonary function testing have not been fully established. The statements need to be periodically updated in accordance with new developments. (Review). PMID:25823236

  3. High total antioxidant capacity of the porcine seminal plasma (SP-TAC) relates to sperm survival and fertility.

    PubMed

    Barranco, Isabel; Tvarijonaviciute, Asta; Perez-Patiño, Cristina; Parrilla, Inmaculada; Ceron, Jose J; Martinez, Emilio A; Rodriguez-Martinez, Heriberto; Roca, Jordi

    2015-01-01

    The study attempted to clarify the role of total antioxidant capacity of seminal plasma (SP-TAC) on boar sperm survival and fertility after artificial insemination (AI). SP-TAC differed (P?sperm-peak portion of the sperm rich ejaculate fraction (SRF) had the highest SP-TAC. SP-TAC was not correlated with sperm quality (motility and viability) or functionality (intracellular ROS generation and lipid peroxidation) of liquid AI-semen samples stored at 17?°C for 72?h (90 AI-samples), but the decline in sperm quality was larger (P?sperm cryosurvival rates (14 ejaculates from 7 boars), showing sperm from sperm-peak portion better (P?functionality than those from the entire ejaculate (mainly post-SRF). Boars (n°?=?18) with high SP-TAC (hierarchically grouped) had higher (P?

  4. High total antioxidant capacity of the porcine seminal plasma (SP-TAC) relates to sperm survival and fertility

    PubMed Central

    Barranco, Isabel; Tvarijonaviciute, Asta; Perez-Patiño, Cristina; Parrilla, Inmaculada; Ceron, Jose J.; Martinez, Emilio A.; Rodriguez-Martinez, Heriberto; Roca, Jordi

    2015-01-01

    The study attempted to clarify the role of total antioxidant capacity of seminal plasma (SP-TAC) on boar sperm survival and fertility after artificial insemination (AI). SP-TAC differed (P?sperm-peak portion of the sperm rich ejaculate fraction (SRF) had the highest SP-TAC. SP-TAC was not correlated with sperm quality (motility and viability) or functionality (intracellular ROS generation and lipid peroxidation) of liquid AI-semen samples stored at 17?°C for 72?h (90 AI-samples), but the decline in sperm quality was larger (P?sperm cryosurvival rates (14 ejaculates from 7 boars), showing sperm from sperm-peak portion better (P?functionality than those from the entire ejaculate (mainly post-SRF). Boars (n°?=?18) with high SP-TAC (hierarchically grouped) had higher (P?

  5. The Functional Task Test (FTT): An Interdisciplinary Testing Protocol to Investigate the Factors Underlying Changes in Astronaut Functional Performance

    NASA Technical Reports Server (NTRS)

    Bloomberg, J. J.; Lawrence, E. L.; Arzeno, N. M.; Buxton, R. E.; Feiveson, A. H.; Kofman, I. S.; Lee, S. M. C.; Mulavara, A. P.; Peters, B. T.; Platts. S. H.; Ploutz-Snyder, L. L.; Reschke, M. F.; Ryder, J. W.; Spiering, B. A.; Stenger, M. B.; Taylor, L. C.; Wood, S. J.

    2011-01-01

    Exposure to space flight causes adaptations in multiple physiological systems including changes in sensorimotor, cardiovascular, and neuromuscular systems. These changes may affect a crewmember s ability to perform critical mission tasks immediately after landing on a planetary surface. The overall goal of this project is to determine the effects of space flight on functional tests that are representative of high priority exploration mission tasks and to identify the key underlying physiological factors that contribute to decrements in performance. To achieve this goal we developed an interdisciplinary testing protocol (Functional Task Test, FTT) that evaluates both astronaut functional performance and related physiological changes. Functional tests include ladder climbing, hatch opening, jump down, manual manipulation of objects and tool use, seat egress and obstacle avoidance, recovery from a fall and object translation tasks. Physiological measures include assessments of postural and gait control, dynamic visual acuity, fine motor control, plasma volume, orthostatic intolerance, upper- and lower-body muscle strength, power, endurance, control, and neuromuscular drive. Crewmembers perform this integrated test protocol before and after short (Shuttle) and long-duration (ISS) space flight. Data are collected on two sessions before flight, on landing day (Shuttle only) and 1, 6 and 30 days after landing. Preliminary results from both Shuttle and ISS crewmembers indicate decrement in performance of the functional tasks after both short and long-duration space flight. On-going data collection continues to improve the statistical power required to map changes in functional task performance to alterations in physiological systems. The information obtained from this study will be used to design and implement countermeasures that specifically target the physiological systems most responsible for the altered functional performance associated with space flight.

  6. Melatonin-enhanced hyperactivation of hamster sperm.

    PubMed

    Fujinoki, Masakatsu

    2008-11-01

    The effects of melatonin on reproductive function were examined using hamster spermatozoa. When 1 pM to 1 microM melatonin was added to the mTALP medium, hyperactivation was significantly enhanced. Antagonists and agonists of the melatonin receptor (i.e., MT1 and MT2) were added to the medium. Luzindole, an MT1 and MT2 competitive antagonist, significantly inhibited melatonin-induced hyperactivation, whereas the MT2-specific antagonists, 4-phenyl-2-propionamidotetralin and N-pentanoyl-2-benzyltryptamine, had no effect. Moreover, hyperactivation was significantly enhanced when non-specific agonists, such as 6-chloromelatonin and 2-iodomelatonin, were added to the medium. 8-Methoxy-2-propionamidotetralin, which is a strong MT2 agonist and a weak MT1 agonist, significantly increased hyperactivation, although the effect was weak. Therefore, it is likely that melatonin enhances sperm hyperactivation via the MT1 receptor. PMID:18715981

  7. A Role for the Chemokine Receptor CCR6 in Mammalian Sperm Motility and Chemotaxis

    PubMed Central

    Caballero-Campo, Pedro; Buffone, Mariano G.; Benencia, Fabian; Conejo-García, José R.; Rinaudo, Paolo F.; Gerton, George L.

    2013-01-01

    Although recent evidence indicates that several chemokines and defensins, well-known as inflammatory mediators, are expressed in the male and female reproductive tracts, the location and functional significance of chemokine networks in sperm physiology and sperm reproductive tract interactions are poorly understood. To address this deficiency in our knowledge, we examined the expression and function in sperm of CCR6, a receptor common to several chemoattractant peptides, and screened several reproductive tract fluids for the presence of specific ligands. CCR6 protein is present in mouse and human sperm and mainly localized in the sperm tail with other minor patterns in sperm from mice (neck and acrosomal region) and men (neck and midpiece regions). As expected from the protein immunoblotting and immunofluorescence results, mouse Ccr6 mRNA is expressed in the testis. Furthermore, the Defb29 mRNA encoding the CCR6 ligand, ?-defensin DEFB29, is expressed at high levels in the epididymis. As determined by protein chip analysis, several chemokines (including some that act through CCR6, such as CCL20/MIP-3? (formerly Macrophage Inflammatory Protein 3?) and protein hormones were present in human follicular fluid, endometrial secretions, and seminal plasma. In functional chemotaxis assays, capacitated human sperm exhibited a directional movement towards CCL20, and displayed modifications in motility parameters. Our data indicate that chemokine ligand/receptor interactions in the male and female genital tracts promote sperm motility and chemotaxis under non-inflammatory conditions. Therefore, some of the physiological reactions mediated by CCR6 ligands in male reproduction extend beyond a pro-inflammatory response and might find application in clinical reproduction and/or contraception. PMID:23765988

  8. Study of Sperm Reproductive Parameters in Mature Zanjani Viper

    PubMed Central

    Moshiri, Malihe; Todehdehghan, Fatemeh; Shiravi, Abdolhossein

    2014-01-01

    Objective Zanjani viper (Vipera albicornuta) is an endemic venomous snake in East Azerbai- jan Province, Iran which is medically important due to its application for antivenin production in the laboratory. We need to produce this snake in captivity. This study was conducted to charac- terize mature male Zanjani viper and to evaluate its sperm reproductive parameters. Materials and Methods This applied- descriptive study was conducted on twenty Zan- jani viper samples collected from Ag Dag Mountain in East Azarbaijan Province, Iran, between September and October 2010. After the snakes were anesthetized and sacrificed humanly, their morphometric specifications and sperm reproductive parameters, including concentration, motility, vitality, morphology, and survival time, were measured. Results Morphometric specifications and evaluation of sperms of the snake showed the following information: Zanjani male viper, body length of 73.65 ± 4.35 cm, tail length of 5.465 ± 0.48 cm, and mature snakes with testicular volumes of 0.61 ± 0.81 ml (right) and of 0.46 ± 0.17 ml (left). Our findings revealed average sperm concen- tration of 0.47 ± 0.1 ×106ml-1, motility of 49 -55 %, vitality of 46.11 ± 9.63 %, normal morphology of 61.71 ± 5.3%, and survival time of 6 ± 2 hours at the laboratory tem- perature. Statistical analyses were performed using Student’s t test for comparison of two values, and one-way ANOVA was applied where three values were compared. Conclusion Results suggest that mature Zanjani male viper with mature sperms in its vas deferens is present in late summer and early autumn seasons in Bostanabad County, Iran. PMID:24567940

  9. MUSCULOSKELETAL MODELING OF SMILODON FATALIS FOR VIRTUAL FUNCTIONAL PERFORMANCE TESTING

    E-print Network

    Krovi, Venkat

    MUSCULOSKELETAL MODELING OF SMILODON FATALIS FOR VIRTUAL FUNCTIONAL PERFORMANCE TESTING by KIRAN in studying the behavioral patterns of extinct animals. In the absence of living specimens, typically two approaches can be adopted. One approach is to study the behavior of a living animal from a closely related

  10. Testing for Differential Item Functioning with Measures of Partial Association

    ERIC Educational Resources Information Center

    Woods, Carol M.

    2009-01-01

    Differential item functioning (DIF) occurs when an item on a test or questionnaire has different measurement properties for one group of people versus another, irrespective of mean differences on the construct. There are many methods available for DIF assessment. The present article is focused on indices of partial association. A family of average…

  11. Changes in the motility patterns of spermatozoa from the rabbit epididymis as assessed by computer-aided sperm motion analysis.

    PubMed

    Pérez-Sánchez, F; Tablado, L; Yeung, C H; Cooper, T G; Soler, C

    1996-11-01

    Sperm maturation in the epididymis includes changes in their potential for motility that enables spermatozoa to reach the egg and penetrate its investments. The motility characteristics of spermatozoa from the testis, the epididymis, and vas deferens of the rabbit were investigated by computer-assisted sperm analysis (CASA). Various forms of motility were displayed by sperm from different regions of the epididymis released into incubation medium. Testicular sperm were motile, although nonprogressive. The maximum percentage motility was expressed by sperm in the proximal cauda epididymidis, and forward progression was developed by spermatozoa from the distal caput. Once forward progression was established, the curvilinear velocity was about the same for sperm from all regions of the tract, whereas straight-line velocity increased between the mid-corpus and cauda and paralleled the decline in lateral displacement of the head. The maintenance of motility in vitro was best maintained by sperm from the distal regions of the tract although sperm from the distal caput maintained motility better than sperm from the proximal and midcorpus regions. Analysis of the motile sperm cells revealed several types of trajectories ("irregular," "small circular," "large circular and arcs," "jagged" and "straight-line") that were analyzed by discriminant analysis using the variables generated by CASA. Accuracy of classification varied from 70% to 96%, depending on the type of track. The classification function was then applied to the changes that occurred during incubation and showed that irregular trajectories gave way to small and then large circular tracks and progressive forms as sperm matured. PMID:8916048

  12. Fertilization of sea urchin eggs and sperm motility are negatively impacted under low hypergravitational forces significant to space flight

    NASA Technical Reports Server (NTRS)

    Tash, J. S.; Kim, S.; Schuber, M.; Seibt, D.; Kinsey, W. H.

    2001-01-01

    Sperm and other flagellates swim faster in microgravity (microG) than in 1 G, raising the question of whether fertilization is altered under conditions of space travel. Such alterations have implications for reproduction of plant and animal food and for long-term space habitation by man. We previously demonstrated that microG accelerates protein phosphorylation during initiation of sperm motility but delays the sperm response to the egg chemotactic factor, speract. Thus sperm are sensitive to changes in gravitational force. New experiments using the NiZeMi centrifugal microscope examined whether low hypergravity (hyperG) causes effects opposite to microG on sperm motility, signal transduction, and fertilization. Sperm % motility and straight-line velocity were significantly inhibited by as little as 1.3 G. The phosphorylation states of FP130, an axonemal phosphoprotein, and FP160, a cAMP-dependent salt-extractable flagellar protein, both coupled to motility activation, showed a more rapid decline in hyperG. Most critically, hyperG caused an approximately 50% reduction in both the rate of sperm-egg binding and fertilization. The similar extent of inhibition of both fertilization parameters in hyperG suggests that the primary effect is on sperm rather than eggs. These results not only support our earlier microG data demonstrating that sperm are sensitive to small changes in gravitational forces but more importantly now show that this sensitivity affects the ability of sperm to fertilize eggs. Thus, more detailed studies on the impact of space flight on development should include studies of sperm function and fertilization.

  13. Fluorescent sperm in a transparent worm: validation of a GFP marker to study sexual selection

    PubMed Central

    2014-01-01

    Background Sexual selection has initially been thought to occur exclusively at the precopulatory stage in terms of contests among males and female mate choice, but research over the last four decades revealed that it often continues after copulation through sperm competition and cryptic female choice. However, studying these postcopulatory processes remains challenging because they occur internally and therefore are often difficult to observe. In the transparent free-living flatworm Macrostomum lignano, a recently established transgenic line that expresses green fluorescent protein (GFP) in all cell types, including sperm, offers a unique opportunity to non-invasively visualise and quantify the sperm of a GFP-expressing donor inside the reproductive tract of wild-type recipients in vivo. We here test several aspects of the reproductive performance of the transgenic individuals and the accuracy of the techniques involved in assessing the GFP-expressing worms and their sperm. We then show the usefulness of these methods in a study on sperm displacement. Results GFP-expressing worms do not differ from wild-type worms in terms of morphology, mating rate and reproductive success. In addition, we show that the GFP signal is reliably and unequivocally expressed by all GFP-expressing individuals observed under epifluorescence illumination. However, the intensity of the GFP signal emitted by sperm of GFP expressing donors can vary (which we show to be at least in part due to sperm ageing) and the GFP marker is inherited according to Mendel’s laws in most, but not all, of the individuals. Nevertheless, we argue these two issues can be addressed with an appropriate experimental design. Finally, we demonstrate the value of the GFP-techniques by comparing the number of GFP-expressing sperm in a wild-type recipient before and after mating with a competing sperm donor, providing clear experimental evidence for sperm displacement in M. lignano. This result suggests that sperm donors can displace previously stored sperm and replace it with their own. Conclusion The availability of the GFP-techniques in a transparent organism provide unique opportunities to visualise and quantify internal processes in the female reproductive tract after mating, which opens new avenues in the study of sexual selection. PMID:24980980

  14. The sperm ultrastructure and spermiogenesis of Tribolium castaneum (Coleoptera: Tenebrionidae) with evidence of cyst degeneration.

    PubMed

    Dias, Glenda; Lino-Neto, José; Mercati, David; Dallai, Romano

    2015-06-01

    Previous studies on the spermatogenesis of tenebrionid beetles showed the unusual formation of two antiparallel sperm bundles per cyst. In this work we reported this feature also in Tribolium castaneum using light and transmission electron microscopy. The sperm structure of T. castaneum, similar to other tenebrionids, consists of a three-layered acrosome, an elongated nucleus and a flagellum with a 9+9+2 axoneme, two accessory bodies and two asymmetric mitochondrial derivatives. The presence of two antiparallel sperm bundles per cyst also in Meloidae and Rhipiphoridae suggests that it is a strong trait synapomorphic for Tenebrionoidea. The huge degeneration of whole sperm cells in several cysts of testes during spermiogenesis is also described. PMID:25867758

  15. Ability of Catalonian donkey sperm to penetrate zona pellucida-free bovine oocytes matured in vitro.

    PubMed

    Taberner, E; Morató, R; Mogas, T; Miró, J

    2010-04-01

    An experiment was designed to study the interaction between fresh/frozen-thawed donkey spermatozoa and zona pellucida (ZP)-free bovine oocytes in an attempt to develop a model for assessing cryopreserved Catalonian donkey sperm function. Semen from five donkeys was collected using an artificial vagina. Sperm motility and viability were immediately assessed and the semen sample cryopreserved. Sperm viability and motility were then reassessed immediately after thawing. The motion characteristics of the fresh and frozen-thawed spermatozoa were determined using a computer-assisted sperm analysis system. In vitro-matured cow oocytes were inseminated with different percent live donkey sperm (high (>60%) or low (<40%) viability donkey sperm). After 18h of co-incubation, the oocytes were fixed, stained with 4',6-diamidino-2-phenylindole (DAPI) and examined for sperm penetration, the number of penetrated spermatozoa per oocyte, and male pronucleus formation. Frozen-thawed spermatozoa from high viability semen showed significantly lower VCL, VAP and mean ALH values than did high viability fresh spermatozoa. In contrast, frozen-thawed spermatozoa of low viability had significantly higher velocity values than fresh spermatozoa of low viability. A significant positive correlation (P<0.01) was detected between percentage fertilization and viability (r=0.84), and between percentage fertilization and certain CASA parameters (VAP, r=0.56; VCL, r=0.61 and mean ALH, r=0.68). Fresh or frozen-thawed high viability spermatozoa penetrated 90.1% and 85.4% of bovine oocytes respectively. Lower rates of penetration were observed for fresh and frozen-thawed low viability spermatozoa (34% and 22.5% respectively). The donkey spermatozoa were able to fuse with the oolema and even to decondense and form the male pronucleus (85-94%). Larger numbers of penetrated spermatozoa per oocyte were recorded when high viability sperm samples were used, whether fresh (3.02 vs. 1.12 for low viability sperm) or frozen-thawed (3.41 vs. 1.47). Consequently, low viability sperm samples showed higher percentages of monospermic penetration (91.17% and 61.97% for fresh and frozen-thawed sperm samples respectively). These findings suggest that bovine oocytes provide a useful model for assessing the penetration potential of frozen-thawed donkey sperm. PMID:19748750

  16. Glucose Regulated Protein 78 Phosphorylation in Sperm Undergoes Dynamic Changes during Maturation

    PubMed Central

    Lobo, Vivian; Rao, Parimala; Gajbhiye, Rahul; Kulkarni, Vijay; Parte, Priyanka

    2015-01-01

    GRP78, a resident endoplasmic reticulum (ER) chaperone involved in protein transport, folding and assembly, has been reported in sperm. It is shown to be localized in the neck region of human sperm. We have previously reported GRP78 to be less phosphorylated in asthenozoosperm.The present study aimed to determine whether sperm GRP78 undergoes phosphorylation changes during epididymal maturation and whether there are any differences in GRP78 phosphoforms in asthenozoosperm vis-à-vis normozoosperm. Testicular- and cauda epididymal- sperm from adult male Holtzman rats, and semen ejaculates collected from normal and asthenozoospermic individuals were investigated. DIGE carried out to determine phosphorylation of GRP78 in asthenozoosperm and normal sperm reveals a shift in the location of GRP78 of asthenozoosperm towards the alkaline pH, indicative of reduced GRP78 phosphorylation. Immunoprecipitation studies using antibodies specific to GRP78, serine-, threonine-, and tyrosine phosphorylation and Pan phospho antibody demonstrates GRP78 to be phosphorylated at all three residues in rat spermatozoa. Phosphatase assays using Calf intestinal alkaline phosphatase and Lambda protein phosphatase followed by nanofluidic proteomic immunoassay (NIA) show that in rat, GP4.96, GP4.94 and GP4.85 are the three phosphoforms in mature (caudal) sperm as against two phosphoforms GP4.96and GP4.94in immature (testicular) sperm. In mature human sperm GP5.04, GP4.96, and GP4.94were the 3 phosphoforms observed. GP4.94[P = 0.014]andGP5.04 [P = 0.02] are significantly reduced in asthenozoosperm. Ours is the first report indicating GRP78 in sperm to be phosphorylated at serine, threonine and tyrosine residues contrary to published literature reporting GRP78 not to be tyrosine phosphorylated. We report the presence of GRP78 phosphoforms in rat- and human- sperm and our data suggest that GRP78 phosphorylation in sperm undergoes spatial reorganization during epididymal maturation. Significant differences observed in 2 out of 3 phosphoforms in asthenozoosperm suggest that GRP78 phosphorylation may have functional relevance in sperm with consequent clinical implications. PMID:26618558

  17. Disruption of histone methylation in developing sperm impairs offspring health transgenerationally.

    PubMed

    Siklenka, Keith; Erkek, Serap; Godmann, Maren; Lambrot, Romain; McGraw, Serge; Lafleur, Christine; Cohen, Tamara; Xia, Jianguo; Suderman, Matthew; Hallett, Michael; Trasler, Jacquetta; Peters, Antoine H F M; Kimmins, Sarah

    2015-11-01

    A father's lifetime experiences can be transmitted to his offspring to affect health and development. However, the mechanisms underlying paternal epigenetic transmission are unclear. Unlike in somatic cells, there are few nucleosomes in sperm, and their function in epigenetic inheritance is unknown. We generated transgenic mice in which overexpression of the histone H3 lysine 4 (H3K4) demethylase KDM1A (also known as LSD1) during spermatogenesis reduced H3K4 dimethylation in sperm. KDM1A overexpression in one generation severely impaired development and survivability of offspring. These defects persisted transgenerationally in the absence of KDM1A germline expression and were associated with altered RNA profiles in sperm and offspring. We show that epigenetic inheritance of aberrant development can be initiated by histone demethylase activity in developing sperm, without changes to DNA methylation at CpG-rich regions. PMID:26449473

  18. Sperm Dynamics in Spiders (Araneae): Ultrastructural Analysis of the Sperm Activation Process in the Garden Spider Argiope bruennichi (Scopoli, 1772)

    PubMed Central

    Vöcking, Oliver; Uhl, Gabriele; Michalik, Peter

    2013-01-01

    Storage of sperm inside the female genital tract is an integral phase of reproduction in many animal species. The sperm storage site constitutes the arena for sperm activation, sperm competition and female sperm choice. Consequently, to understand animal mating systems information on the processes that occur from sperm transfer to fertilization is required. Here, we focus on sperm activation in spiders. Male spiders produce sperm whose cell components are coiled within the sperm cell and that are surrounded by a proteinaceous sheath. These inactive and encapsulated sperm are transferred to the female spermathecae where they are stored for later fertilization. We analyzed the ultrastructural changes of sperm cells during residency time in the female genital system of the orb-web spider Argiope bruennichi. We found three clearly distinguishable sperm conditions: encapsulated sperm (secretion sheath present), decapsulated (secretion sheath absent) and uncoiled sperm (cell components uncoiled, presumably activated). After insemination, sperm remain in the encapsulated condition for several days and become decapsulated after variable periods of time. A variable portion of the decapsulated sperm transforms rapidly to the uncoiled condition resulting in a simultaneous occurrence of decapsulated and uncoiled sperm. After oviposition, only decapsulated and uncoiled sperm are left in the spermathecae, strongly suggesting that the activation process is not reversible. Furthermore, we found four different types of secretion in the spermathecae which might play a role in the decapsulation and activation process. PMID:24039790

  19. Off-axis rotation as a test of otolith function.

    PubMed

    Blakely, B W

    1995-12-01

    This experiment was performed to determine whether linear acceleration causes a change in the vestibulo-linear reflex (VOR) that could be exploited as a research and/or clinical test of otolith function. The effect of otolith stimulation on the VOR was studied in the horizontal and vertical planes in 10 cats. The VOR was induced by pulses of angular acceleration (velocity ramps). During some of the test runs, a centripetal linear acceleration stimulus was also applied to stimulate the otolith organs. Using the stimuli combination as tested in this project, the authors found that otolith stimulation affected the vertical VOR but not the horizontal VOR. The vertical VOR was asymmetric. While the methods used in this study are useful and the equipment requirements make these techniques generally impractical for clinical testing. PMID:8523989

  20. Sperm production responds to perceived sperm competition risk in male Drosophila melanogaster.

    PubMed

    Moatt, Joshua P; Dytham, Calvin; Thom, Michael D F

    2014-05-28

    Postcopulatory sexual selection arising from female multiple mating leads to the evolution of ejaculates that maximize a male's reproductive success under sperm competition. Where the risk of sperm competition is variable, optimal fitness may be achieved by plastically altering ejaculate characteristics in response to the prevailing sperm competition environment. In the model species Drosophila melanogaster, males expecting to encounter sperm competition mate for longer and transfer more accessory proteins and sperm. Here we show that after being housed with a single rival for one week, the seminal vesicles of male D. melanogaster contain a significantly greater proportion of live sperm than those of males maintained alone, indicating adaptive adjustment of sperm quality in response to the perceived risk of sperm competition. This effect is due to an increase in the number of live sperm produced, indicating that males upregulate sperm production in response to the presence of rivals. Our data suggest that males show plasticity in the rate of spermatogenesis that is adaptive in the context of a fluctuating sperm competition environment. PMID:24769021

  1. Abstract Most comparative analyses of relative testes mass find that testes are larger in species in which more

    E-print Network

    Pyron, Mark

    in which more sperm competition is predicted (multiple males mate with individual females). I tested sperm competition in min- now species. Possible explanations for the lack of the ex- pected relationship between testes mass and mating sys- tems in minnows are presented. Key words Minnows · Testes · Sperm

  2. Morphology of Mammalian Sperm Membranes During Differentiation, Maturation, and Capacitation

    PubMed Central

    BEARER, ELAINE L.; FRIEND, DANIEL S.

    2015-01-01

    The mammalian spermatozoon is a highly polarized cell whose surface membrane can be divided into five functionally, structurally, and biochemically distinct domains. These domains are formed during spermatogenesis, continue to be modified during passage through the epididymis, and are further refined in the female reproductive tract. The integrity of these domains appears to be necessary for the sperm to perform its function—fusion with the egg and subsequent fertilization. The domains can be identified morphologically by their surface contours and texture, the content, distribution, and organization of intramembranous particles after freeze-fracture, and by the density of surface and cytoplasmic electron-dense coatings in thin sections. By using a variety of labels that stain carbohydrates (lectins), lipids (filipin and polymyxin B), and monoclonal antibodies to specific membrane constituents, the biochemical composition of these contiguous membrane regions has also been partly elucidated. We review here what is known about the structure, composition, and behavior of each membrane domain in the mature sperm and include some information regarding domain formation during spermatogenesis. The sperm is an excellent model system to study the creation and maintenance of cell polarity, granule exocytosis, and fertilization. Hopefully this review will provide impetus for future studies aimed more directly at addressing the relationship of its morphology to its functions. PMID:2250184

  3. Male Investments in High Quality Sperm Improve Fertilization Success, but May Have Negative Impact on Offspring Fitness in Whitefish

    PubMed Central

    Kekäläinen, Jukka; Soler, Carles; Veentaus, Sami; Huuskonen, Hannu

    2015-01-01

    Many ejaculate traits show remarkable variation in relation to male social status. Males in disfavoured (subordinate) mating positions often invest heavily on sperm motility but may have less available resources on traits (e.g., secondary sexual ornaments) that improve the probability of gaining matings. Although higher investments in sperm motility can increase the relative fertilization success of subordinate males, it is unclear whether status-dependent differences in sperm traits could have any consequences for offspring fitness. We tested this possibility in whitefish (Coregonus lavaretus L.) by experimentally fertilizing the eggs of 24 females with the sperm of either highly-ornamented (large breeding tubercles, dominant) or less-ornamented (small tubercles, subordinate) males (split-clutch breeding design). In comparison to highly-ornamented individuals, less-ornamented males had higher sperm motility, which fertilized the eggs more efficiently, but produced embryos with impaired hatching success. Also offspring size and body condition were lower among less-ornamented males. Furthermore, sperm motility was positively associated with the fertilization success and offspring size, but only in highly-ornamented males. Together our results indicate that male investments on highly motile (fertile) sperm is not necessarily advantageous during later offspring ontogeny and that male status-dependent differences in sperm phenotype may have important effects on offspring fitness in different life-history stages. PMID:26389594

  4. Validation of a field based chromatin dispersion assay to assess sperm DNA fragmentation in the bottlenose dolphin (Tursiops truncatus).

    PubMed

    Sánchez-Calabuig, M-J; López-Fernández, C; Martínez-Nevado, E; Pérez-Gutiérrez, J F; de la Fuente, J; Johnston, S D; Blyde, D; Harrison, K; Gosálvez, J

    2014-10-01

    Over the last two decades, there have been significant advances in the use of assisted reproductive technology for genetic and reproductive management of captive dolphin populations, including evaluation of sperm DNA quality. This study validated a customized sperm chromatin dispersion test (SCDt) for the bottlenose dolphin (Tursiops truncatus) as a means of assessing sperm DNA damage both in the field and in the laboratory. After performing the SCDt, two different sperm morphotypes were identified: (i) sperm with fragmented DNA showed large haloes of dispersed DNA fragments emerging from a compact sperm nucleoid core and (ii) sperm containing non-fragmented DNA displayed small compact haloes surrounded by a dense core of non-dispersed DNA and protein complex. Estimates of sperm DNA fragmentation by means of SCDt were directly comparable to results obtained following a two-tailed comet assay and showed a significant degree of correlation (r = 0.961; p < 0.001). This investigation also revealed that the SCDt, with minor modifications to the standard protocol, can be successfully conducted in the field using a LED florescence microscopy obtaining a high correlation (r = 0.993; p = 0.01) between the data obtained in the laboratory and in the field. PMID:25130370

  5. Mammalian sperm interactions with the female reproductive tract.

    PubMed

    Suarez, Susan S

    2016-01-01

    The mammalian female reproductive tract interacts with sperm in various ways in order to facilitate sperm migration to the egg while impeding migrations of pathogens into the tract, to keep sperm alive during the time between mating and ovulation, and to select the fittest sperm for fertilization. The two main types of interactions are physical and molecular. Physical interactions include the swimming responses of sperm to the microarchitecture of walls, to fluid flows, and to fluid viscoelasticity. When sperm encounter walls, they have a strong tendency to remain swimming along them. Sperm will also orient their swimming into gentle fluid flows. The female tract seems to use these tendencies of sperm to guide them to the site of fertilization. When sperm hyperactivate, they are better able to penetrate highly viscoelastic media, such as the cumulus matrix surrounding eggs. Molecular interactions include communications of sperm surface molecules with receptors on the epithelial lining of the tract. There is evidence that specific sperm surface molecules are required to enable sperm to pass through the uterotubal junction into the oviduct. When sperm reach the oviduct, most bind to the oviductal epithelium. This interaction holds sperm in a storage reservoir until ovulation and serves to maintain the fertilization competence of stored sperm. When sperm are released from the reservoir, they detach from and re-attach to the epithelium repeatedly while ascending to the site of fertilization. We are only beginning to understand the communications that may pass between sperm and epithelium during these interactions. PMID:26183721

  6. Validation of Cardiovascular Parameters During NASA's Functional Task Test

    NASA Technical Reports Server (NTRS)

    Arzeno, N. M.; Stenger, M. B.; Bloomberg, J. J.; Platts, Steven H.

    2008-01-01

    Microgravity-induced physiological changes, including cardiovascular deconditioning may impair crewmembers f capabilities during exploration missions on the Moon and Mars. The Functional Task Test (FTT), which will be used to assess task performance in short and long duration astronauts, consists of 7 functional tests to evaluate crewmembers f ability to perform activities to be conducted in a partial-gravity environment or following an emergency landing on Earth. The Recovery from Fall/Stand Test (RFST) tests both the subject fs ability to get up from a prone position and orthostatic intolerance. PURPOSE: Crewmembers have never become presyncopal in the first 3 min of quiet stand, yet it is unknown whether 3 min is long enough to cause similar heart rate fluctuations to a 5-min stand. The purpose of this study was to validate and test the reliability of heart rate variability (HRV) analysis of a 3-min quiet stand. METHODS: To determine the validity of using 3 vs. 5-min of standing to assess HRV, 7 healthy subjects remained in a prone position for 2 min, stood up quickly and stood quietly for 6 min. ECG and continuous blood pressure data were recorded. Mean R-R interval and spectral HRV were measured in minutes 0-3 and 0-5 following the heart rate transient due to standing. Significant differences between the segments were determined by a paired t-test. To determine the reliability of the 3-min stand test, 13 healthy subjects completed 3 trials of the complete FTT on separate days, including the RFST with a 3-min stand test. Analysis of variance (ANOVA) was performed on the HRV measures. RESULTS: Spectral HRV measures reflecting autonomic activity were not different (p>0.05) during the 0-3 and 0-5 min segment (mean R-R interval: 738+/-74 ms, 728+/-69 ms; low frequency to high frequency ratio: 6.5+/-2.2, 7.7+/-2.7; normalized high frequency: 0.19+/-0.03, 0.18+/-0.04). The average coefficient of variation for mean R-R interval, systolic and diastolic blood pressures in the prone position and stand test were less than 8% for the test sessions. ANOVA results yielded a greater inter-subject variability (p.0.006) than inter-session variability (p>0.05) for HRV in the stand test. CONCLUSION: These studies show that a 3 minute stand delivers repeatable cardiovascular heart rate and BP data in the context of this larger series of tests such as the FTT.

  7. August 1997 DWARF SPERM WHALE (Kogia simus)

    E-print Network

    in the northern Gulf of Mexico occur primarily along the continental shelf edge and over the deeper waters off the continental shelf (Mullin et al. 1991; NMFS unpublished data). Pygmy sperm whales and dwarf sperm whales observed by NMFS Sea Samplers in the pelagic drift gillnet fishery, but no mortalities or serious injuries

  8. October 1999 DWARF SPERM WHALE (Kogia simus)

    E-print Network

    49 October 1999 DWARF SPERM WHALE (Kogia simus): Western North Atlantic Stock STOCK DEFINITION population. In a recent study using hematological and stable-isotope data, Barroset al.(1998)speculatedthatdwarfspermwhalesmayhaveamorepelagic distribution than pygmy sperm whales, and/or dive deeper during feeding bouts. POPULATION SIZE

  9. Supporting Information: Chemotaxis of sperm cells

    E-print Network

    Jülicher, Frank

    Supporting Information: Chemotaxis of sperm cells Benjamin M. Friedrich and Frank J¨ulicher Max) A sperm moving along r(t) samples according to equation (2) a time-dependent stimulus s(t) = c0 + r0 c1

  10. REGULATION OF SPERM NUCLEAR REACTIVATION DURING FERTILIZATION

    EPA Science Inventory

    Upon fusion of sperm and oocyte at fertilization, a series of events is initiated whereby the highly compacted sperm nucleus expands and is transformed into a male pronucleus capable of DNA synthesis. The regulation of these early post-fusion fertilization events has been studied...

  11. Six-Minute Walk Test Vs. Three-Minute Step Test for Measuring Functional Endurance.

    PubMed

    Bohannon, Richard W; Bubela, Deborah J; Wang, Ying-Chih; Magasi, Susan S; Gershon, Richard C

    2015-11-01

    Bohannon, RW, Bubela, DJ, Wang, Y-C, Magasi, SS, and Gershon, RC. Six-minute walk test vs. three-minute step test for measuring functional endurance. J Strength Cond Res 29(11): 3240-3244, 2015-The purpose of this study was to compare 2 practical measures of functional endurance. Specifically, the six-minute walk test (SMWT) and three-minute step test (TMST) were compared to determine their appropriateness for use as field tests and inclusion in the NIH Toolbox for the assessment of neurological and behavioral function. Individuals between 14 and 85 years performed both the SMWT and TMST in random order. We documented completion rates, criterion performance, heart rate responses, and subjective exertion associated with the 2 tests. All 189 participants completed the SMWT, but only 73.0% completed the TMST. Those completing the TMST were more likely to be male, report better health, and have a younger age and lower body mass index. The SMWT distance was greater for those who did vs. those who did not complete the TMST. For those completing both tests, the average distance walked in 6 minutes was 595.9 m; the average cumulative heart beats during the minute after the TMST was 107.4. Distance walked and cumulative heart beats correlated weakly. Average heart rate and perceived exertion were significantly higher after the TMST than the SMWT. Posttest heart rate and perceived exertion for the 2 tests correlated significantly but not strongly. We conclude that the SMWT is more likely to be completed and is usually less stressful physiologically than the TMST and therefore may be a better option for field testing functional endurance and inclusion in the NIH Toolbox. PMID:24077375

  12. Towards a Semen Proteome of the Dengue Vector Mosquito: Protein Identification and Potential Functions

    PubMed Central

    Sirot, Laura K.; Ribeiro, José M. C.; Kimura, Mari; Deewatthanawong, Prasit; Wolfner, Mariana F.; Harrington, Laura C.

    2011-01-01

    Background No commercially licensed vaccine or treatment is available for dengue fever, a potentially lethal infection that impacts millions of lives annually. New tools that target mosquito control may reduce vector populations and break the cycle of dengue transmission. Male mosquito seminal fluid proteins (Sfps) are one such target since these proteins, in aggregate, modulate the reproduction and feeding patterns of the dengue vector, Aedes aegypti. As an initial step in identifying new targets for dengue vector control, we sought to identify the suite of proteins that comprise the Ae. aegypti ejaculate and determine which are transferred to females during mating. Methodology and Principal Findings Using a stable-isotope labeling method coupled with proteomics to distinguish male- and female-derived proteins, we identified Sfps and sperm proteins transferred from males to females. Sfps were distinguished from sperm proteins by comparing the transferred proteins to sperm-enriched samples derived from testes and seminal vesicles. We identified 93 male-derived Sfps and 52 predicted sperm proteins that are transferred to females during mating. The Sfp protein classes we detected suggest roles in protein activation/inactivation, sperm utilization, and ecdysteroidogenesis. We also discovered that several predicted membrane-bound and intracellular proteins are transferred to females in the seminal fluids, supporting the hypothesis that Ae. aegypti Sfps are released from the accessory gland cells through apocrine secretion, as occurs in mammals. Many of the Ae. aegypti predicted sperm proteins were homologous to Drosophila melanogaster sperm proteins, suggesting conservation of their sperm-related function across Diptera. Conclusion and Significance This is the first study to directly identify Sfps transferred from male Ae. aegypti to females. Our data lay the groundwork for future functional analyses to identify individual seminal proteins that may trigger female post-mating changes (e.g., in feeding patterns and egg production). Therefore, identification of these proteins may lead to new approaches for manipulating the reproductive output and vectorial capacity of Ae. aegypti. PMID:21423647

  13. Why so many sperm cells?

    E-print Network

    Schuss, K Reynaud Z; Holcman, D

    2014-01-01

    A key limiting step in fertility is the search for the oocyte by spermatozoa. Initially, there are tens of millions of sperm cells, but a single one will make it to the oocyte. This may be one of the most severe selection processes designed by evolution, whose role is yet to be understood. Why is it that such a huge redundancy is required and what does that mean for the search process? we propose to discuss here these questions and consequently a new line of interdisciplinary research needed to find possible answers.

  14. NICMOS Filter Wheel/Mechanisms Mini-Functional Test

    NASA Astrophysics Data System (ADS)

    Schneider, Glenn

    2001-07-01

    This is an early engineering test to verify the aliveness, functionality, operability, and electro-mechanical calibration of the NICMOS filter wheel motors and assembly. This is the FIRST use of the NICMOS filter wheel mechanisms since they were disabled by ground command in January, 1999. This test has been designed to obviate concerns over possible deformation or breakage of the fitter wheel "soda-straw" shafts due to excess rotational drag torque and/or bending moments which may be imparted due to changes in the dewar metrology from warm-up/cool-down. This test should be executed after the VCS {and filter wheel housing} has reached and approximately equilibrated to its nominal Cycle 11 operating temperature.

  15. [Respiratory function testing in infants: recommendations on normal values].

    PubMed

    Amsallem, F; Gauthier, R; Ramonatxo, M; Counil, F; Voisin, M; Denjean, A; Matecki, S

    2008-04-01

    The present document is being produced on behalf of the French Society of the Physiology Task Force on standards for Infant Respiratory Function Testing whose aim is to provide guidelines for good laboratory practices according to the latest international recommendations. Application of such recommendations could be of particular value when attempting to develop standardized protocols in the scope of multi-centre trials. The first part resume these recommendations about apparatus, acquisition system and software for Infant Respiratory Function Testing. The second part focuses on physiological principles and practical considerations: calibration procedure, infant conditioning, tidal breathing measurements, and occlusion techniques for assessing passive respiratory mechanics, plethysmographic measurements of lung volume and airway resistance and forced expiratory flows measurements. The major problem when collecting lung function data is that of predicted values. Valid reference data, set up according to these recommendations, are, to date, still to be established. The last part of the document provides a review of the literature concerning infant respiratory function reference data and a resume of the most used of them. This document will clearly need to be updated regularly in response to advances in knowledge in this field. PMID:18536627

  16. Adenine nucleotide metabolism and a role for AMP in modulating flagellar waveforms in mouse sperm.

    PubMed

    Vadnais, Melissa L; Cao, Wenlei; Aghajanian, Haig K; Haig-Ladewig, Lisa; Lin, Angel M; Al-Alao, Osama; Gerton, George L

    2014-06-01

    While most ATP, the main energy source driving sperm motility, is derived from glycolysis and oxidative phosphorylation, the metabolic demands of the cell require the efficient use of power stored in high-energy phosphate bonds. In times of high energy consumption, adenylate kinase (AK) scavenges one ATP molecule by transphosphorylation of two molecules of ADP, simultaneously yielding one molecule of AMP as a by-product. Either ATP or ADP supported motility of detergent-modeled cauda epididymal mouse sperm, indicating that flagellar AKs are functional. However, the ensuing flagellar waveforms fueled by ATP or ADP were qualitatively different. Motility driven by ATP was rapid but restricted to the distal region of the sperm tail, whereas ADP produced slower and more fluid waves that propagated down the full flagellum. Characterization of wave patterns by tracing and superimposing the images of the flagella, quantifying the differences using digital image analysis, and computer-assisted sperm analysis revealed differences in the amplitude, periodicity, and propagation of the waves between detergent-modeled sperm treated with either ATP or ADP. Surprisingly, addition of AMP to the incubation medium containing ATP recapitulated the pattern of sperm motility seen with ADP alone. In addition to AK1 and AK2, which we previously demonstrated are present in outer dense fibers and mitochondrial sheath of the mouse sperm tail, we show that another AK, AK8, is present in a third flagellar compartment, the axoneme. These results extend the known regulators of sperm motility to include AMP, which may be operating through an AMP-activated protein kinase. PMID:24740601

  17. Effect of curcumin on rat sperm morphology after the freeze-thawing process

    PubMed Central

    Soleimanzadeh, Ali; Saberivand, Adel

    2013-01-01

    Reactive oxygen species (ROS) generation, induced by the cryopreservation process, can be responsible for mammalian sperm damage. Curcumin is known as an effective antioxidant against oxidative stress. The aim of this study was to evaluate the effects of curcumin on sperm count, motility and viability, semen total antioxidant capacity and DNA integrity of rat spermatozoa during semen freeze-thawing process. Sperm collected from 10 adult rats were divided into two groups (n=10 for each group): control and a test group supplemented with 2.5 mM curcumin. After freezing-thawing, the number of spermatozoa, motility, viability, total antioxidant capacity (TAC), and DNA integrity of the sperm were analyzed. Motility, viability and DNA integrity of sperm were significantly preserved in treatment groups compared to the control (p < 0.05) after freezing-thawing. Following cryopreservation, TAC was significantly preserved in thawing semen supplemented with curcumin compared to the control group (p<0.05). Based on our results, it is concluded that curcumin addition during freezing resulted in positive effects on sperm parameters after thawing in adult rats. PMID:25653795

  18. ZP-binding peptides identified via phage display stimulate production of sperm antibodies in dogs.

    PubMed

    Samoylova, Tatiana I; Cox, Nancy R; Cochran, Anna M; Samoylov, Alexandre M; Griffin, Brenda; Baker, Henry J

    2010-07-01

    Zona pellucida (ZP) glycoproteins play a central role in sperm-oocyte binding and fertilization. Sperm protein sequences that are involved in sperm-ZP recognition and have an important role in fertilization represent attractive targets for development of contraceptive vaccines, yet are currently unknown. To identify peptide sequences that recognize and bind to ZP proteins, we developed a novel selection procedure from phage display libraries that utilizes intact oocytes surrounded by ZP proteins. The major advantage of this procedure is that ZP proteins remain in their native conformation unlike a selection protocol previously published that utilized solubilized ZP on artificial solid support. Several peptides of 7 and 12 amino acids with binding specificity to canine ZP proteins were identified. Four of them (LNSFLRS, SSWYRGA, YLPIYTIPSMVY, and NNQSPILKLSIH) plus a control ZP-binding peptide (YLPVGGLRRIGG) from the literature were synthesized and tested for antigenic properties in dogs. NNQSPILKLSIH peptide stimulated production of anti-peptide antibodies. These antibodies bind to the acrosomal region of the canine sperm cell, demonstrating ability to act as sperm antibodies. The identified ZP-binding peptides (mimicking sperm cell surface antigens) may be useful in the design of immunocontraceptive agents for dogs. PMID:20434854

  19. Sneaker "jack" males outcompete dominant "hooknose" males under sperm competition in Chinook salmon (Oncorhynchus tshawytscha).

    PubMed

    Young, Brent; Conti, David V; Dean, Matthew D

    2013-12-01

    In a variety of taxa, males deploy alternative reproductive tactics to secure fertilizations. In many species, small "sneaker" males attempt to steal fertilizations while avoiding encounters with larger, more aggressive, dominant males. Sneaker males usually face a number of disadvantages, including reduced access to females and the higher likelihood that upon ejaculation, their sperm face competition from other males. Nevertheless, sneaker males represent an evolutionarily stable strategy under a wide range of conditions. Game theory suggests that sneaker males compensate for these disadvantages by investing disproportionately in spermatogenesis, by producing more sperm per unit body mass (the "fair raffle") and/or by producing higher quality sperm (the "loaded raffle"). Here, we test these models by competing sperm from sneaker "jack" males against sperm from dominant "hooknose" males in Chinook salmon. Using two complementary approaches, we reject the fair raffle in favor of the loaded raffle and estimate that jack males were ?1.35 times as likely as hooknose males to fertilize eggs under controlled competitive conditions. Interestingly, the direction and magnitude of this skew in paternity shifted according to individual female egg donors, suggesting cryptic female choice could moderate the outcomes of sperm competition in this externally fertilizing species. PMID:24455130

  20. Sneaker “jack” males outcompete dominant “hooknose” males under sperm competition in Chinook salmon (Oncorhynchus tshawytscha)

    PubMed Central

    Young, Brent; Conti, David V; Dean, Matthew D

    2013-01-01

    In a variety of taxa, males deploy alternative reproductive tactics to secure fertilizations. In many species, small “sneaker” males attempt to steal fertilizations while avoiding encounters with larger, more aggressive, dominant males. Sneaker males usually face a number of disadvantages, including reduced access to females and the higher likelihood that upon ejaculation, their sperm face competition from other males. Nevertheless, sneaker males represent an evolutionarily stable strategy under a wide range of conditions. Game theory suggests that sneaker males compensate for these disadvantages by investing disproportionately in spermatogenesis, by producing more sperm per unit body mass (the “fair raffle”) and/or by producing higher quality sperm (the “loaded raffle”). Here, we test these models by competing sperm from sneaker “jack” males against sperm from dominant “hooknose” males in Chinook salmon. Using two complementary approaches, we reject the fair raffle in favor of the loaded raffle and estimate that jack males were ?1.35 times as likely as hooknose males to fertilize eggs under controlled competitive conditions. Interestingly, the direction and magnitude of this skew in paternity shifted according to individual female egg donors, suggesting cryptic female choice could moderate the outcomes of sperm competition in this externally fertilizing species. PMID:24455130

  1. Sperm motility inversely correlates with seminal leptin levels in idiopathic asthenozoospermia

    PubMed Central

    Guo, Jianhua; Zhao, Yang; Huang, Weiying; Hu, Wei; Gu, Jianjun; Chen, Chuhong; Zhou, Juan; Peng, Yubing; Gong, Min; Wang, Zhong

    2014-01-01

    Background: Asthenozoospermia is one kind cause of male infertility. Nevertheless, no specific etiology can be identified by routine tests in some cases. Recently, it has been shown that leptin plays a critical role in male fertility. However, the link between leptin and sperm motility is yet to be determined. The aim of this study was to explore association between seminal and serum leptin levels and sperm motility in idiopathic asthenozoospermia. Methods: Our study included 79 asthenozoospermic men and 77 normozoospermic men. Semen was assessed by volume, sperm concentration, motility and morphology. Serum gonadotropic and sex hormones were determined by a chemiluminescent assay. The leptin levels in serum and seminal plasma were detected with ELISA. Results: The mean seminal leptin level in asthenozoospermic group was significantly higher than that in control group, but there was no significant difference in the serum leptin levels between these two groups. The serum leptin had no significant correlation with sperm motility. The seminal leptin had significantly negative correlation with sperm progressive motility and serum total testosterone. Conclusions: The findings indicate a pathophysiological relevance of seminal leptin in sperm motility. PMID:25419396

  2. Sperm-Storage Defects and Live Birth in Drosophila Females Lacking Spermathecal Secretory Cells

    PubMed Central

    Schnakenberg, Sandra L.; Matias, Wilfredo R.; Siegal, Mark L.

    2011-01-01

    Male Drosophila flies secrete seminal-fluid proteins that mediate proper sperm storage and fertilization, and that induce changes in female behavior. Females also produce reproductive-tract secretions, yet their contributions to postmating physiology are poorly understood. Large secretory cells line the female's spermathecae, a pair of sperm-storage organs. We identified the regulatory regions controlling transcription of two genes exclusively expressed in these spermathecal secretory cells (SSC): Spermathecal endopeptidase 1 (Send1), which is expressed in both unmated and mated females, and Spermathecal endopeptidase 2 (Send2), which is induced by mating. We used these regulatory sequences to perform precise genetic ablations of the SSC at distinct time points relative to mating. We show that the SSC are required for recruiting sperm to the spermathecae, but not for retaining sperm there. The SSC also act at a distance in the reproductive tract, in that their ablation: (1) reduces sperm motility in the female's other sperm-storage organ, the seminal receptacle; and (2) causes ovoviviparity—the retention and internal development of fertilized eggs. These results establish the reproductive functions of the SSC, shed light on the evolution of live birth, and open new avenues for studying and manipulating female fertility in insects. PMID:22087073

  3. Protein and carbohydrate intake influence sperm number and fertility in male cockroaches, but not sperm viability.

    PubMed

    Bunning, Harriet; Rapkin, James; Belcher, Laurence; Archer, C Ruth; Jensen, Kim; Hunt, John

    2015-03-01

    It is commonly assumed that because males produce many, tiny sperm, they are cheap to produce. Recent work, however, suggests that sperm production is not cost-free. If sperm are costly to produce, sperm number and/or viability should be influenced by diet, and this has been documented in numerous species. Yet few studies have examined the exact nutrients responsible for mediating these effects. Here, we quantify the effects of protein (P) and carbohydrate (C) intake on sperm number and viability in the cockroach Nauphoeta cinerea, as well as the consequences for male fertility. We found the intake of P and C influenced sperm number, being maximized at a high intake of diets with a P : C ratio of 1 : 2, but not sperm viability. The nutritional landscapes for male fertility and sperm number were closely aligned, suggesting that sperm number is the major determinant of male fertility in N. cinerea. Under dietary choice, males regulate nutrient intake at a P : C ratio of 1 : 4.95, which is midway between the ratios needed to maximize sperm production and pre-copulatory attractiveness in this species. This raises the possibility that males regulate nutrient intake to balance the trade-off between pre- and post-copulatory traits in this species. PMID:25608881

  4. INTRODUCTION The fertilizing sperm activates all vertebrate and invertebrate

    E-print Network

    Newcastle upon Tyne, University of

    INTRODUCTION The fertilizing sperm activates all vertebrate and invertebrate eggs by generating [Ca: sperm may bind to a receptor, generating inositol 1,4,5-triphosphate (InsP3) via a G-protein pathway on the idea that an acti- vating diffusible messenger is introduced into the egg by the sperm when the sperm

  5. Effect of different extenders and storage temperatures on sperm viability

    E-print Network

    Zaragoza, Universidad de

    Effect of different extenders and storage temperatures on sperm viability of liquid ram semen Heiko at 5 and 20 8C, respectively. We evaluated sperm viability after 0, 6, 12, 24 and 30 h of storage. We assessed sperm motility subjectively, and we determined sperm membrane integrity using both the hypo

  6. SNAREs in Mammalian Sperm: Possible Implications for Fertilization

    E-print Network

    Wessel, Gary M.

    SNAREs in Mammalian Sperm: Possible Implications for Fertilization Joa~o Ramalho-Santos,*, Ricardo attachment protein receptor (SNARE) proteins are present in mammalian sperm and could be involved in critical sperm (human, rhesus monkey, bull, hamster, mouse). Sperm SNAREs are sloughed off during the acrosome

  7. VALIDATION OF THE SPERM MOBILITY ASSAY IN BOARS AND STALLIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The sperm mobility assay measures the rate of sperm penetration in a biologically inert cell separation solution (Accudenz). When a sample of sperm is overlaid in a cuvette containing Accudenz, the sperm penetrates the solution and the absorbance of the sample can be measured using a spectrophotome...

  8. Sperm Aggregations in Female Agkistrodon piscivorus (Reptilia: Squamata): A Histological

    E-print Network

    Sever, David M.

    Sperm Aggregations in Female Agkistrodon piscivorus (Reptilia: Squamata): A Histological, sperm migrate up the oviduct to sperm stor- age tubules (SSTs) in the posterior infundibulum that of the epithelium lining the lumen of the posterior infundibu- lum. Sperm pass through an area composed primarily

  9. Females become infertile as the stored sperm's oxygen radicals increase

    E-print Network

    Richner, Heinz

    Females become infertile as the stored sperm's oxygen radicals increase Klaus Reinhardt1,2,3 & Anne-vitro studies suggest that oxidative sperm damage causes infertility. Oxidative sperm damage can be reduced via should evolve frequently, especially during female sperm storage. However, in-vivo evidence linking

  10. DWARF SPERM WHALE (Kogia sima): Western North Atlantic Stock

    E-print Network

    April 2014 DWARF SPERM WHALE (Kogia sima): Western North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale (Kogia sima) is distributed worldwide in temperate to tropical waters). Dwarf sperm whales and pygmy sperm whales (K. breviceps) are difficult to differentiate at sea (Caldwell

  11. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... from rendering the fatty tissue of the sperm whale or is prepared by synthesis of fatty acids and fatty alcohols derived from the sperm whale. The sperm oil obtained by rendering is refined. The oil is... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Hydrogenated sperm oil. 173.275 Section...

  12. 61(1), 1999 59 The Sperm Whale

    E-print Network

    61(1), 1999 59 The Sperm Whale Introduction The sperm whale, Physeter macro- cephalus Linnaeus at birth have an average length of 4 m. The most distinctive physical feature of the sperm whale as ambergris, is produced in the sperm whale's intestines. Near the an- terior of the head and to the left

  13. PYGMY SPERM WHALE (Kogia breviceps): Western North Atlantic Stock

    E-print Network

    43 July 1995 PYGMY SPERM WHALE (Kogia breviceps): Western North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The dwarf sperm whale (Kogia breviceps) and the pygmy sperm whale (K. simus) appear whales and dwarf sperm whales are difficult to distinguish and sightings of either species are often

  14. PYGMY SPERM WHALE (Kogia breviceps): Western North Atlantic Stock

    E-print Network

    April 2014 PYGMY SPERM WHALE (Kogia breviceps): Western North Atlantic Stock STOCK DEFINITION AND GEOGRAPHIC RANGE The pygmy sperm whale (Kogia breviceps) is distributed worldwide in temperate to tropical (Willis and Baird 1998). Pygmy sperm whales and dwarf sperm whales (K. sima) are difficult

  15. Effect of cholesterol-loaded cyclodextrin on cryosurvival and fertility of cryopreserved carp (Cyprinus carpio) sperm.

    PubMed

    Yildiz, Cengiz; Yavas, Ilker; Bozkurt, Yusuf; Aksoy, Melih

    2015-04-01

    Addition of cholesterol-loaded cyclodextrin (CLC) to the diluents of mammalian semen increased stability and rigidity of phospholipid hydrocarbon chains of plasma membrane during sperm cryopreservation process. CLC has been tested successfully as cryoprotectant in various livestock sperm cryopreservation protocols but its efficacy for cryopreserving of fish sperm has not previously been tested. In the present study, different cholesterol loaded cyclodextrin concentrations were evaluated for the cryopreservation of carp (Cyprinus carpio) sperm. Sexually mature fish were induced to spermiation and ovulation with Ovopel. The extenders were prepared by using 300 mM glucose and 10% DMSO supplemented with different concentrations of CLC (0.5, 1.0, 1.5, 2.0, 2.5, and 3.0mg per 120×10(6) spermatozoa) and without CLC (control). The pooled semen was diluted separately at a ratio of 1:3 (v/v) by using CLC extenders. Diluted semen placed into 0.25 ml straws were equilibrated at 4°C for 15 min and frozen in liquid nitrogen vapor. Fertilization was conducted using a ratio of 1×10(5) spermatozoa/egg. Fresh sperm with no treatment showed the greatest sperm motility, duration of motility, viability, and fertilization results compared to the other tested cryopreserved and control groups (p<0.05). Supplementation of 1.5 mg CLC to the extender showed the best cryoprotective effect for sperm motility, duration of motility, and viability against freezing damage in comparison to extenders containing 2.5 mg, 3.0 mg CLC, and control group (p<0.05). Cryopreserved sperm containing 1.5 mg CLC provided greater result in term of fertilization success when compared to other extenders containing 0.5, 2.5, and 3.0 mg CLC or control (p<0.05). The amount of CLC effected post-thaw sperm quality and fertility as a dose-dependent manner. It is concluded that treatment of cholesterol-loaded cyclodextrin for carp sperm cryopreservation significantly improves cell cryosurvival and fertilization. PMID:25661710

  16. Fishery induces sperm depletion and reduction in male reproductive potential for crab species under male-biased harvest strategy.

    PubMed

    Pardo, Luis Miguel; Rosas, Yenifer; Fuentes, Juan Pablo; Riveros, Marcela Paz; Chaparro, Oscar Roberto

    2015-01-01

    Sperm depletion in males can occur when polygynous species are intensively exploited under a male-biased management strategy. In fisheries involving crabs species, the effects of this type of management on the reproductive potential is far from being understood. This study tests whether male-biased management of the principal Chilean crab fishery is able to affect the potential capacity of Metacarcinus edwardsii males to transfer sperm to females. Five localities in southern Chile, recording contrasting crab fishery landing, were selected to assess the potential of sperm depletion triggered by fishery. Seasonally, male crabs from each locality were obtained. Dry weight and histological condition of vasa deferentia and the Vaso-Somatic Index (VSI) were determined in order to use them as proxies for sperm depletion and male reproductive condition. A manipulative experiment was performed in the laboratory to estimate vasa deferentia weight and VSI from just-mated males in order to obtain a reference point for the potential effects of the fishery on sperm reserves. Sperm storage capacity is significantly affected by fisheries; during the mating season vasa deferentia from localities with low fishery intensity were heavier than those from high intensity fisheries, and these differences were even more evident in large males. Histological section showed that this disparity in vasa deferentia weight was explained principally by differences in the quantity of spermatophores rather than other seminal material. VSI was always higher in males from localities with low fishery intensity. Males from localities with high fishery intensity showed little capacity to recover sperm reserves and the VSI of these males remained below the values of the just-mated males. Detriment in the capacity of males to transfer sperm is the first step to sperm limitation in an exploited population, thus detection of sperm depletion can be an alert to introduce changes in the current management of crabs. PMID:25768728

  17. Use of penicillin and streptomycin to reduce spread of bacterial coldwater disease I: antibiotics in sperm extenders.

    PubMed

    Oplinger, Randall W; Wagner, Eric J

    2015-03-01

    Bacterial coldwater disease caused by Flavobacterium psychrophilum has led to the loss of significant numbers of hatchery-reared salmonids. The bacteria can be spread from parent to progeny within contaminated sperm and ovarian fluid. Methods for disinfecting ovarian fluid and unfertilized eggs are available, but methods for disinfecting sperm have not been described. In this study we determined whether sperm extenders containing a mixture of penicillin and streptomycin can be used to eliminate F. psychrophilum. In vitro trials demonstrated that when Rainbow Trout Oncorhynchus mykiss sperm is mixed with an extender, a 15-min exposure to 0.197 mg penicillin plus 0.313 mg/mL streptomycin is effective at killing the bacteria and has no effect on sperm motility. Small-scale trials showed that egg fertilization rates were not reduced when sperm held in an extender solution containing the same antibiotic mixture for 15 min was used to fertilize eggs. Production-scale trials, however, showed a roughly 18% decrease in egg fertilization rate when sperm stored in an antibiotic containing extender was used. To determine why a reduction in fertilization capacity was observed, a small-scale experiment testing the fertilization of eggs with larger quantities of sperm was performed and showed that increasing the volume of sperm used did not increase fertilization rates. Our results demonstrate that extenders containing penicillin and streptomycin can be used to disinfect sperm, especially when small quantities of eggs are fertilized, but factors negatively affecting egg fertilization and survival on a production scale still need further investigation. PMID:25581335

  18. SLC6 family transporter SNF-10 is required for protease-mediated activation of sperm motility in C. elegans

    PubMed Central

    Fenker, Kristin E.; Hansen, Angela A.; Chong, Conrad A.; Jud, Molly C.; Duffy, Brittany A.; Norton, J. Paul; Hansen, Jody M.; Stanfield, Gillian M.

    2014-01-01

    Summary Motility of sperm is crucial for their directed migration to the egg. The acquisition and modulation of motility are regulated to ensure that sperm move when and where needed, thereby promoting reproductive success. One specific example of this phenomenon occurs during differentiation of the amoeboid sperm of C. elegans as they activate from a round spermatid to a mature, crawling spermatozoon. Sperm activation is regulated by redundant pathways to occur at a specific time and place for each sex. Here, we report the identification of the solute carrier 6 (SLC6) transporter protein SNF-10 as a key regulator of C. elegans sperm activation in response to male protease activation signals. We find that SNF-10 is present in sperm and is required for activation by the male but not by the hermaphrodite. Loss of both snf-10 and a hermaphrodite activation factor render sperm completely insensitive to activation. Using in vitro assays, we find that snf-10 mutant sperm show a specific deficit in response to protease treatment but not to other activators. Prior to activation, SNF-10 is present in the plasma membrane, where it represents a strong candidate to receive signals that lead to subcellular morphogenesis. After activation, it shows polarized localization to the cell body region that is dependent on membrane fusions mediated by the dysferlin FER-1. Our discovery of snf-10 offers insight into the mechanisms differentially employed by the two sexes to accomplish the common goal of producing functional sperm, as well as how the physiology of nematode sperm may be regulated to control motility as it is in mammals. PMID:24929237

  19. Reliability of the Function in Sitting Test (FIST)

    PubMed Central

    Gorman, Sharon L.; Rivera, Monica; McCarthy, Lise

    2014-01-01

    The function in sitting test (FIST) is a newly developed, performance-based measure examining deficits in seated postural control. The FIST has been shown to be internally consistent and valid in persons with neurological dysfunction but intra- and interrater reliability and test-retest reliability have not been previously described. Seven patients with chronic neurologic dysfunction were tested and videotaped performing the FIST on two consecutive days. Seventeen acute care and inpatient rehabilitation physical therapist raters scored six of the videotaped performance of the FIST on two occasions at least 2 weeks apart. Intraclass correlation coefficients were used to calculate the test-retest and intra- and interrater reliability of the FIST. ICC of 0.97 (95% CI 0.847–0.995) indicated excellent test-retest reliability of the FIST. Intra- and interrater reliability was also excellent with ICCs of 0.99 (95% CI 0.994–0.997) and 0.99 (95% CI 0.988–0.994), respectively. Physical therapists and other rehabilitation professionals can confidently use the FIST in a variety of clinical practice and research settings due to its favorable reliability characteristics. More studies are needed to describe the responsiveness and minimal clinically important level of change in FIST scores to further enhance clinical usefulness of this measure. PMID:24757566

  20. Estradiol blood test

    MedlinePLUS

    E2 test ... temporarily stop taking certain medicines that may affect test results. Be sure to tell your provider about ... helps prevent sperm from dying too early. This test may be ordered to check: How well your ...