Sample records for functional sperm tests

  1. Sperm functional tests.

    PubMed

    Oehninger, Sergio; Franken, Daniel R; Ombelet, Willem

    2014-12-01

    Several semen parameters are used to discriminate the fertile male from the subfertile male. The most widely used parameters are sperm concentration, motility, progressive motility, and sperm morphology. Semen analysis is usually applied as described in the World Health Organization manual for semen analysis. In addition to a routine semen analysis, sperm functional tests have been described for many years, which in most cases are regarded as research tools and not part of the routine semen testing in an infertility clinic. In this review we report on the value of four sperm function tests: the sperm penetration assay, the sperm-zona pellucida binding tests, the acrosome reaction, and the hyaluronan binding assay. For each test we describe the current value, the indication for performing the test, how to interpret the results, and its therapeutic implications. Our data show that sperm functional assays are highly predictive of IVF outcome results and have the potential to assist in clinical decision making, especially to avoid the current long-standing treatment with IUI and to direct the patients to intracytoplasmic sperm injection without delay when sperm functional testing fails. We believe that advances in molecular biology techniques will allow us to develop simpler sperm function assays in the near future. This will undoubtedly help clinicians in optimizing male factor infertility diagnosis and treatment. PMID:25450304

  2. 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.

  3. 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

  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. Mitochondria functionality and sperm quality.

    PubMed

    Amaral, Alexandra; Lourenço, Bárbara; Marques, Mónica; Ramalho-Santos, João

    2013-01-01

    Although mitochondria are best known for being the eukaryotic cell powerhouses, these organelles participate in various cellular functions besides ATP production, such as calcium homoeostasis, generation of reactive oxygen species (ROS), the intrinsic apoptotic pathway and steroid hormone biosynthesis. The aim of this review was to discuss the putative roles of mitochondria in mammalian sperm function and how they may relate to sperm quality and fertilisation ability, particularly in humans. Although paternal mitochondria are degraded inside the zygote, sperm mitochondrial functionality seems to be critical for fertilisation. Indeed, changes in mitochondrial integrity/functionality, namely defects in mitochondrial ultrastructure or in the mitochondrial genome, transcriptome or proteome, as well as low mitochondrial membrane potential or altered oxygen consumption, have been correlated with loss of sperm function (particularly with decreased motility). Results from genetically engineered mouse models also confirmed this trend. On the other hand, increasing evidence suggests that mitochondria derived ATP is not crucial for sperm motility and that glycolysis may be the main ATP supplier for this particular aspect of sperm function. However, there are contradictory data in the literature regarding sperm bioenergetics. The relevance of sperm mitochondria may thus be associated with their role in other physiological features, particularly with the production of ROS, which in controlled levels are needed for proper sperm function. Sperm mitochondria may also serve as intracellular Ca²? stores, although their role in signalling is still unclear. PMID:23901129

  6. How accurate is sperm morphology as an indicator of sperm function?

    PubMed

    Franken, D R

    2015-08-01

    Sperm morphology has been consistently correlated with fertilisation success or failure. The clinical relevance of the percentage normal spermatozoa has been a widely discussed topic amongst infertility specialists and scientists. This study aimed to evaluate the role of sperm morphology as an indicator of additional sperm functions among 114 andrology referrals. The sperm functions that were investigated included chromatin packaging quality (CMA3 test (n = 109), zona-induced acrosome reaction (ZIAR test; n = 36), hemizona assay (HZI; n = 36) and progressive motility (n = 47). Chromatin packaging quality had a negative and significant (P = 0.0001, r = -0.74) correlation with the percentage normal spermatozoa, while progressive motility had a significant and positive correlation (P = 0.0001, 0.59). Accurate sperm morphology scoring as described by the WHO 2010 manual can therefore be used as an indicator of specific sperm functions. PMID:25130990

  7. Relationship between in vitro sperm functional tests and in vivo fertility of rams following cervical artificial insemination of ewes with frozen-thawed semen

    Microsoft Academic Search

    C. M. O’ Meara; J. P. Hanrahan; N. S. Prathalingam; J. S. Owen; A. Donovan; S. Fair; F. Ward; M. Wade; A. C. O. Evans; P. Lonergan

    2008-01-01

    Several procedures have been proposed to assess structural and functional characteristics of cryopreserved ram semen but none so far have yielded consistent relationships with in vivo fertility. The objectives of this study were to evaluate several sperm function tests as potential markers of in vivo ram fertility (determined by pregnancy rate in ewes) using frozen-thawed semen. In experiment 1, frozen-thawed

  8. Toona sinensis Leaf Aqueous Extract Improves the Functions of Sperm and Testes via Regulating Testicular Proteins in Rats under Oxidative Stress

    PubMed Central

    Yu, Bu-Chin; Yu, Wen-Jen; Huang, Chun-Yung; Chen, Ying-Hsin; Tsai, Yun-Ching; Chang, Chen-Chin; Chang, Sue-Joan

    2012-01-01

    Toona sinensis leaf (TSL) is commonly used as a vegetable and in spice in Asia. In this study, feeding with aqueous extract of TSL (TSL-A) alleviated oxidative stress and recovered the motility and functions of sperm in rats under oxidative stress. Protein expressions in testes identified by proteomic analysis and verified by Western blot demonstrated that TSL-A not only downregulated the level of glutathione transferase mu6 (antioxidant system), heat shock protein 90?kDa-? (protein misfolding repairing system), cofilin 2 (spermatogenesis), and cyclophilin A (apoptosis) but also upregulated crease3-hydroxy-3-methylglutaryl-coenzyme A synthase 2 (steroidogenesis), heat shock glycoprotein 96, and pancreatic trypsin 1 (sperm-oocyte interaction). These results indicate that TSL-A promotes the functions of sperm and testes via regulating multiple testicular proteins in rats under oxidative stress, suggesting that TSL-A is a valuable functional food supplement to improve functions of sperm and testes for males under oxidative stress. PMID:23093985

  9. Human sperm chromosomes obtained from hamster eggs after sperm capacitation in TEST-yolk buffer

    SciTech Connect

    Brandriff, B.; Gordon, L.; Watchmaker, G.

    1985-01-01

    Human sperm chromosomes were obtained after capacitation with TES-Tris (TEST) yolk buffer and fusion with Syrian hamster eggs. Semen samples could be stored at 4/sup 0/C for 3 days and remain functional in the assay system. The efficiency of TEST yolk buffer for obtaining karyotypes was as good as, or greater than, the efficiency of standard BWW medium containing human serum albumin. 16 references, 3 tables.

  10. Human sperm chromosomes obtained from hamster eggs after sperm capacitation in TEST-yolk buffer

    Microsoft Academic Search

    B. Brandriff; L. Gordon; G. Watchmaker

    1985-01-01

    Human sperm chromosomes were obtained after capacitation with TES-Tris (TEST) yolk buffer and fusion with Syrian hamster eggs. Semen samples could be stored at 4°C for 3 days and remain functional in the assay system. The efficiency of TEST yolk buffer for obtaining karyotypes was as good as, or greater than, the efficiency of standard BWW medium containing human serum

  11. Emodin inhibits human sperm functions by reducing sperm [Ca(2+)]i and tyrosine phosphorylation.

    PubMed

    Luo, Tao; Li, Na; He, Yuan-Qiao; Weng, Shi-Qi; Wang, Tao; Zou, Qian-Xing; Zeng, Xu-Hui

    2015-01-01

    Emodin, a bioactive anthraquinone widely used in Chinese traditional medicine, disrupts mouse testicular gene expression in vivo. In this study, we investigated the toxicity of emodin to human sperm in vitro. Different doses of emodin (25, 50, 100, 200 and 400?M) were applied to ejaculated human sperm. The results indicated that 100, 200 and 400?M emodin significantly inhibited the total motility, progressive motility and linear velocity of human sperm. In addition, sperm's ability to penetrate viscous medium together with progesterone induced capacitation and acrosome reaction was also adversely affected by emodin. In contrast, emodin did not affect sperm viability. Furthermore, intracellular Ca(2+) concentration ([Ca(2+)]i) and tyrosine phosphorylation, which serve as key regulators of sperm function, were dose-dependently reduced by emodin (50-400?M). These results suggest that emodin inhibits human sperm functions by reducing sperm [Ca(2+)]i and suppressing tyrosine phosphorylation in vitro. PMID:25463531

  12. 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

  13. 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

  14. Relationship between in vitro sperm functional tests and in vivo fertility of rams following cervical artificial insemination of ewes with frozen-thawed semen.

    PubMed

    O' Meara, C M; Hanrahan, J P; Prathalingam, N S; Owen, J S; Donovan, A; Fair, S; Ward, F; Wade, M; Evans, A C O; Lonergan, P

    2008-03-01

    Several procedures have been proposed to assess structural and functional characteristics of cryopreserved ram semen but none so far have yielded consistent relationships with in vivo fertility. The objectives of this study were to evaluate several sperm function tests as potential markers of in vivo ram fertility (determined by pregnancy rate in ewes) using frozen-thawed semen. In experiment 1, frozen-thawed straws (n=3 per ram) of semen from three high and three low fertility rams were assessed using fluorescent microscopy for (1) progressive motility, (2) viability and, (3) acrosomal status. In experiment 2, frozen-thawed straws (n=3 per ram) of semen from 18 rams of known fertility were analysed using either computer-assisted sperm analysis (CASA) for eight motion characteristics or flow cytometric staining for: (1) viability and acrosomal status, (2) plasma membrane status and capacitation-like changes, and (3) live cells following an osmotic resistance test (ORT). In experiment 3, platelet-activating factor (PAF) was isolated from straws (n=2 per ram) of semen using high-pressure liquid chromatography (HPLC) and quantified using HPLC-tandem mass spectrometry for 18 rams. In experiment 1, no association was found between motility, viability (% live) or acrosomal status (% damaged, % intact and % reacted) and in vivo fertility. In experiment 2, no correlation was found between motility (CASA), viability (% live), acrosomal status (% live, % live intact and % reacted), capacitation status (% capacitated, % non-capacitated), plasma membrane stability (% dead) and % live cells following ORT and ram in vivo fertility. In experiment 3, there was no relationship between PAF content in spermatozoa and ram fertility. In conclusion, we were unable to relate the in vivo fertility of rams with in vitro functional tests of their frozen-thawed semen and suggest that the fertility of a given semen sample cannot easily be quantified using available in vitro tests. PMID:18248736

  15. Do Candidate Genes Mediating Conspecific Sperm Precedence Affect Sperm Competitive Ability Within Species? A Test Case in Drosophila

    PubMed Central

    Civetta, Alberto; Finn, Scott

    2014-01-01

    When females mate to multiple males, the last male to mate fathers the majority of progeny. When males of different species inseminate a female, the sperm of the male conspecific to the female is favored in fertilization in a process known as conspecific sperm precedence (CSP). A large number of studies in Drosophila have assayed the genetic basis of sperm competition, with a main focus on D. melanogaster and accessory gland protein genes. Only a few studies have attempted to disentangle the genetic basis of CSP between related species of Drosophila. Although there is no a priori reason to believe that genes influencing intraspecific sperm competitive ability might also mediate conspecific sperm precedence, no study has addressed the question. Here, we test a group of candidate CSP genes between D. simulans and D. mauritiana for their effect on sperm competition in D. melanogaster. The use of P-element insertion lines identified CG14891 gene disruption as the only one causing a significant decrease in second male paternity success relative to wild-type and ebony tester males. The gene disruption affected both sperm displacement and the sperm fertilizing ability. Out of five genes tested using RNA interference, only gene knockdown of CG6864 (Mst89B) significantly reduced the male’s ability to father progeny when second to mate. Our results suggest that CG14891 and CG6864 might have been co-opted from an intraspecies gene function (i.e., sperm competition) into an interspecies avoidance phenotype (i.e., CSP). Alternatively, the dual role of these genes could be a consequence of their pleiotropic roles. PMID:25031180

  16. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

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

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

    Code of Federal Regulations, 2013 CFR

    2013-04-01

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

  18. Sperm competition: linking form to function

    PubMed Central

    2008-01-01

    Background Using information from physics, biomechanics and evolutionary biology, we explore the implications of physical constraints on sperm performance, and review empirical evidence for links between sperm length and sperm competition (where two or more males compete to fertilise a female's eggs). A common theme in the literature on sperm competition is that selection for increased sperm performance in polyandrous species will favour the evolution of longer, and therefore faster swimming, sperm. This argument is based on the common assumption that sperm swimming velocity is directly related to sperm length, due to the increased thrust produced by longer flagella. Results We critically evaluate the evidence for links between sperm morphology and swimming speed, and draw on cross-disciplinary studies to show that the assumption that velocity is directly related to sperm length will rarely be satisfied in the microscopic world in which sperm operate. Conclusion We show that increased sperm length is unlikely to be driven by selection for increased swimming speed, and that the relative lengths of a sperm's constituent parts, rather than their absolute lengths, are likely to be the target of selection. All else being equal, we suggest that a simple measure of the ratio of head to tail length should be used to assess the possible link between morphology and speed. However, this is most likely to be the case for external fertilizers in which females have relatively limited opportunity to influence a sperm's motility. PMID:19032741

  19. The regulation of spermatogenesis and sperm function in nematodes.

    PubMed

    Ellis, Ronald E; Stanfield, Gillian M

    2014-05-01

    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

  20. 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

  1. Morphology and function of the reproductive tract of the spider crab Libinia spinosa (Crustacea, Brachyura, Majoidea): pattern of sperm storage

    Microsoft Academic Search

    M. P. Sal Moyano; M. A. Gavio; E. I. Cuartas

    2010-01-01

    Morphology and function of the male reproductive tract, female spermatheca and patterns of sperm storage were assessed in the crab Libinia spinosa using histological methods. Testes are characterized by the presence of peripheral spermatogonia and different sequences of sperm maturity. Spermatophores begin to be packed in the last portion. The vas deferens consists of three sections: anterior, with undeveloped spermatophores

  2. Characterisation of a subpopulation of sperm with massive nuclear damage, as recognised with the sperm chromatin dispersion test.

    PubMed

    Gosálvez, J; Rodríguez-Predreira, M; Mosquera, A; López-Fernández, C; Esteves, S C; Agarwal, A; Fernández, J L

    2014-08-01

    Assessment of human sperm DNA fragmentation by the sperm chromatin dispersion (SCD) test is based on the detection of haloes of spreading DNA loops after sequential DNA denaturing and protamine removal. After the SCD test, sperm without DNA fragmentation show chromatin haloes emerging from the central nuclear core, while sperm containing fragmented DNA present small or no haloes. The nuclear degraded sperm are recognised as a differentiated category within the sperm with fragmented DNA, whose cores appear irregularly and/or faintly stained. This subpopulation is more prevalent in patients with varicocele. Protein staining with 2.7-dibrom-4-hydroxy-mercury-fluorescein demonstrated that degraded sperm intensely lose nuclear core proteins after the SCD processing. Moreover, degraded sperm are 65% more faintly labelled for DNA breaks after in situ nick translation (ISNT) on average, due to extensive DNA loss. A two-dimensional comet assay under sequential neutral and alkaline conditions demonstrated that degraded sperm contain both massive double- and single-strand DNA breaks. The degraded sperm appear as a subpopulation with stronger nuclear damage, affecting both DNA and protein fractions, possibly due to intense intratesticular oxidative stress, what could explain its higher proportion in patients with varicocele. PMID:23710631

  3. Sperm competition and the evolution of testes size in birds

    Microsoft Academic Search

    T. E. PITCHER; P. O. DUNN; L. A. WHITTINGHAM

    2005-01-01

    Comparative analyses suggest that a variety of ecological and behavioural factors contribute to the tremendous variability in extrapair mating among birds. In an analysis of 1010 species of birds, we examined several ecological and behavioural factors in relation to testes size; an index of sperm competition and the extent of extrapair mating. In univariate and multivariate analyses, testes size was

  4. 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

  5. Regulation of Sperm Function by Oviduct Fluid and the Epithelium: Insight into the Role of Glycans.

    PubMed

    Miller, D J

    2015-07-01

    In many vertebrates, females store sperm received at mating in specialized reservoirs until fertilization. In some species, sperm are routinely stored for up to a decade. But the structures used to store sperm vary considerably across taxa, suggesting the underlying mechanisms might be equally variable. In mammals, after mating, sperm pass through the utero-tubal junction and bind to epithelial cells of the oviduct isthmus to form a reservoir. This reservoir regulates sperm function, including viability and capacitation, ultimately affecting sperm lifespan. In addition, sperm binding to oviduct cells influences oviduct cell gene transcription and translation, perhaps to aid sperm storage and fertility. The sperm reservoir allows successful reproduction in species in which semen deposition and ovulation are not always synchronized. The focus of this review is on recent studies of the functions of oviduct fluid and of the adhesion molecules that allow sperm to adhere to the oviduct epithelium. The important of glycans on the oviduct epithelium is highlighted. PMID:26174917

  6. Sperm competition selects for increased testes mass in Australian frogs

    Microsoft Academic Search

    P. G. B YRNE; J. D. R OBERTS; L. W. S IMMONS

    2002-01-01

    Game theory predicts that investment in spermatogenesis will increase with the risk and intensity of sperm competition. Widespread support for this prediction has come from comparative studies of internal fertilizing species reporting positive associations between testes mass and the probability that females mate with more than one male. Data for external fertilizers have generated conflicting results. We investigated how risk

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

    PubMed

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

    2014-09-19

    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

  8. Stallion Sperm Transcriptome Comprises Functionally Coherent Coding and Regulatory RNAs as Revealed by Microarray Analysis and RNA-seq

    PubMed Central

    Das, Pranab J.; McCarthy, Fiona; Vishnoi, Monika; Paria, Nandina; Gresham, Cathy; Li, Gang; Kachroo, Priyanka; Sudderth, A. Kendrick; Teague, Sheila; Love, Charles C.; Varner, Dickson D.; Chowdhary, Bhanu P.; Raudsepp, Terje

    2013-01-01

    Mature mammalian sperm contain a complex population of RNAs some of which might regulate spermatogenesis while others probably play a role in fertilization and early development. Due to this limited knowledge, the biological functions of sperm RNAs remain enigmatic. Here we report the first characterization of the global transcriptome of the sperm of fertile stallions. The findings improved understanding of the biological significance of sperm RNAs which in turn will allow the discovery of sperm-based biomarkers for stallion fertility. The stallion sperm transcriptome was interrogated by analyzing sperm and testes RNA on a 21,000-element equine whole-genome oligoarray and by RNA-seq. Microarray analysis revealed 6,761 transcripts in the sperm, of which 165 were sperm-enriched, and 155 were differentially expressed between the sperm and testes. Next, 70 million raw reads were generated by RNA-seq of which 50% could be aligned with the horse reference genome. A total of 19,257 sequence tags were mapped to all horse chromosomes and the mitochondrial genome. The highest density of mapped transcripts was in gene-rich ECA11, 12 and 13, and the lowest in gene-poor ECA9 and X; 7 gene transcripts originated from ECAY. Structural annotation aligned sperm transcripts with 4,504 known horse and/or human genes, rRNAs and 82 miRNAs, whereas 13,354 sequence tags remained anonymous. The data were aligned with selected equine gene models to identify additional exons and splice variants. Gene Ontology annotations showed that sperm transcripts were associated with molecular processes (chemoattractant-activated signal transduction, ion transport) and cellular components (membranes and vesicles) related to known sperm functions at fertilization, while some messenger and micro RNAs might be critical for early development. The findings suggest that the rich repertoire of coding and non-coding RNAs in stallion sperm is not a random remnant from spermatogenesis in testes but a selectively retained and functionally coherent collection of RNAs. PMID:23409192

  9. Stallion sperm transcriptome comprises functionally coherent coding and regulatory RNAs as revealed by microarray analysis and RNA-seq.

    PubMed

    Das, Pranab J; McCarthy, Fiona; Vishnoi, Monika; Paria, Nandina; Gresham, Cathy; Li, Gang; Kachroo, Priyanka; Sudderth, A Kendrick; Teague, Sheila; Love, Charles C; Varner, Dickson D; Chowdhary, Bhanu P; Raudsepp, Terje

    2013-01-01

    Mature mammalian sperm contain a complex population of RNAs some of which might regulate spermatogenesis while others probably play a role in fertilization and early development. Due to this limited knowledge, the biological functions of sperm RNAs remain enigmatic. Here we report the first characterization of the global transcriptome of the sperm of fertile stallions. The findings improved understanding of the biological significance of sperm RNAs which in turn will allow the discovery of sperm-based biomarkers for stallion fertility. The stallion sperm transcriptome was interrogated by analyzing sperm and testes RNA on a 21,000-element equine whole-genome oligoarray and by RNA-seq. Microarray analysis revealed 6,761 transcripts in the sperm, of which 165 were sperm-enriched, and 155 were differentially expressed between the sperm and testes. Next, 70 million raw reads were generated by RNA-seq of which 50% could be aligned with the horse reference genome. A total of 19,257 sequence tags were mapped to all horse chromosomes and the mitochondrial genome. The highest density of mapped transcripts was in gene-rich ECA11, 12 and 13, and the lowest in gene-poor ECA9 and X; 7 gene transcripts originated from ECAY. Structural annotation aligned sperm transcripts with 4,504 known horse and/or human genes, rRNAs and 82 miRNAs, whereas 13,354 sequence tags remained anonymous. The data were aligned with selected equine gene models to identify additional exons and splice variants. Gene Ontology annotations showed that sperm transcripts were associated with molecular processes (chemoattractant-activated signal transduction, ion transport) and cellular components (membranes and vesicles) related to known sperm functions at fertilization, while some messenger and micro RNAs might be critical for early development. The findings suggest that the rich repertoire of coding and non-coding RNAs in stallion sperm is not a random remnant from spermatogenesis in testes but a selectively retained and functionally coherent collection of RNAs. PMID:23409192

  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. Acetylproteomic analysis reveals functional implications of lysine acetylation in human spermatozoa (sperm).

    PubMed

    Yu, Heguo; Diao, Hua; Wang, Chunmei; Lin, Yan; Yu, Fudong; Lu, Hui; Xu, Wei; Li, Zheng; Shi, Huijuan; Zhao, Shimin; Zhou, Yuchuan; Zhang, Yonglian

    2015-04-01

    Male infertility is a medical condition that has been on the rise globally. Lysine acetylation of human sperm, an essential posttranslational modification involved in the etiology of sperm abnormality, is not fully understood. Therefore, we first generated a qualified pan-anti-acetyllysine monoclonal antibody to characterize the global lysine acetylation of uncapacitated normal human sperm with a proteomics approach. With high enrichment ratios that were up to 31%, 973 lysine-acetylated sites that matched to 456 human sperm proteins, including 671 novel lysine acetylation sites and 205 novel lysine-acetylated proteins, were identified. These proteins exhibited conserved motifs XXXKYXXX, XXXKFXXX, and XXXKHXXX, were annotated to function in multiple metabolic processes, and were localized predominantly in the mitochondrion and cytoplasmic fractions. Between the uncapacitated and capacitated sperm, different acetylation profiles in regard to functional proteins involved in sperm capacitation, sperm-egg recognition, sperm-egg plasma fusion, and fertilization were observed, indicating that acetylation of functional proteins may be required during sperm capacitation. Bioinformatics analysis revealed association of acetylated proteins with diseases and drugs. Novel acetylation of voltage-dependent anion channel proteins was also found. With clinical sperm samples, we observed differed lysine acetyltransferases and lysine deacetylases expression between normal sperm and abnormal sperm of asthenospermia or necrospermia. Furthermore, with sperm samples impaired by epigallocatechin gallate to mimic asthenospermia, we observed that inhibition of sperm motility was partly through the blockade of voltage-dependent anion channel 2 Lys-74 acetylation combined with reduced ATP levels and mitochondrial membrane potential. Taken together, we obtained a qualified pan-anti-acetyllysine monoclonal antibody, analyzed the acetylproteome of uncapacitated human sperm, and revealed associations between functional protein acetylation and sperm functions. PMID:25680958

  12. Morphology and function of the reproductive tract of the spider crab Libinia spinosa (Crustacea, Brachyura, Majoidea): pattern of sperm storage

    Microsoft Academic Search

    M. P. Sal Moyano; M. A. Gavio; E. I. Cuartas

    2010-01-01

    Morphology and function of the male reproductive tract, female spermatheca and patterns of sperm storage were assessed in\\u000a the crab Libinia spinosa using histological methods. Testes are characterized by the presence of peripheral spermatogonia and different sequences\\u000a of sperm maturity. Spermatophores begin to be packed in the last portion. The vas deferens consists of three sections: anterior,\\u000a with undeveloped spermatophores

  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. Environ Toxicol Chem 2015;34:1617-1622. © 2015 SETAC. PMID:25728985

  14. Functional features and protein network of human sperm-egg interaction.

    PubMed

    Sabetian, Soudabeh; Shamsir, Mohd Shahir; Abu Naser, Mohammed

    2014-12-01

    Elucidation of the sperm-egg interaction at the molecular level is one of the unresolved problems in sexual reproduction, and understanding the molecular mechanism is crucial in solving problems in infertility and failed in vitro fertilization (IVF). Many molecular interactions in the form of protein-protein interactions (PPIs) mediate the sperm-egg membrane interaction. Due to the complexity of the problem such as difficulties in analyzing in vivo membrane PPIs, many efforts have failed to comprehensively elucidate the fusion mechanism and the molecular interactions that mediate sperm-egg membrane fusion. The main purpose of this study was to reveal possible protein interactions and associated molecular function during sperm-egg interaction using a protein interaction network approach. Different databases have been used to construct the human sperm-egg interaction network. The constructed network revealed new interactions. These included CD151 and CD9 in human oocyte that interact with CD49 in sperm, and CD49 and ITGA4 in sperm that interact with CD63 and CD81, respectively, in the oocyte. These results showed that the different integrins in sperm may be involved in human sperm-egg interaction. It was also suggested that sperm ADAM2 plays a role as a protein candidate involved in sperm-egg membrane interaction by interacting with CD9 in the oocyte. Interleukin-4 receptor activity, receptor signaling protein tyrosine kinase activity, and manganese ion transmembrane transport activity are the major molecular functions in sperm-egg interaction protein network. The disease association analysis indicated that sperm-egg interaction defects are also reflected in other disease networks such as cardiovascular, hematological, and breast cancer diseases. By analyzing the network, we identified the major molecular functions and disease association genes in sperm-egg interaction protein. Further experimental studies will be required to confirm the significance of these new computationally resolved interactions and the genetic links between sperm-egg interaction abnormalities and the associated disease. PMID:25222562

  15. Picomolar gradients of progesterone select functional human sperm even in subfertile samples.

    PubMed

    Gatica, L V; Guidobaldi, H A; Montesinos, M M; Teves, M E; Moreno, A I; Uñates, D R; Molina, R I; Giojalas, L C

    2013-09-01

    More than 1 million infertility treatments are practiced around the world per year, but only 30% of the couples succeed in taking a baby home. Reproductive technology depends in part on sperm quality, which influences not only fertilization but also embryo development and implantation. In order to provide a better quality sperm subpopulation, innovative sperm selection techniques based on physiological sperm features are needed. Spermatozoa at an optimum state may be selected by following an increasing concentration gradient of picomolar progesterone, a steroid secreted by the cumulus cells at the time of ovulation. In this study we developed a method to recruit spermatozoa at the best functional state, based on sperm guidance toward progesterone. The sperm selection assay (SSA) consists of a device with two wells connected by a tube. One well was filled with the sperm suspension and the other with picomolar progesterone, which diffused inside the connecting tube as a gradient. The sperm quality after the SSA was analyzed in normal and subfertile semen samples. Several sperm parameters indicative of sperm physiological state were determined before and after the SSA: capacitation, DNA integrity and oxidative stress. After the SSA, the mean level of capacitated spermatozoa increased three times in normal and in subfertile samples. The level of sperm with intact DNA was significantly increased, while sperm oxidative stress was decreased after sperm selection. Interestingly, the exposure to a progesterone gradient stimulated the completion of capacitation in some spermatozoa that could not do it by themselves. Thus, the SSA supplies a sperm population enriched with spermatozoa at an optimum physiological state that may improve the assisted reproductive technology outcome. PMID:23729411

  16. Effect of immunization against prostate- and testis-expressed (PATE) proteins on sperm function and fecundity in the rat.

    PubMed

    Rajesh, Angireddy; Yenugu, Suresh

    2015-08-01

    Evaluating the immunocontraceptive potential of sperm-bound proteins is an active area of investigation. In this study, we analyzed the role of prostate- and testes-expressed (PATE) and PATE-F proteins in sperm function. Capacitation was measured as a function of tyrosine phosphorylation of sperm membrane proteins. Ionophore-induced acrosome reaction was assessed by measuring the fluorescence intensity of calcium-bound Fluo 3-AM and sperm-bound PNA-FITC in a flow cytometer. Rat spermatozoa subjected to capacitation and acrosome reaction in vitro displayed changes in the PATE and PATE-F protein localization on their surface, indicating the role of these proteins in sperm function. Capacitation and ionophore-induced acrosome reaction in vitro were inhibited in spermatozoa pre-incubated with antiserum raised in rabbit against PATE or PATE-F. Male rats were immunized with PATE proteins to assess their role in sperm function and fecundity. Antibody titer in the serum, testicular, and epididymal fluid was measured by ELISA. The motility parameters were recorded using CASA. High antibody titer was observed in serum, epididymal, and testicular fluid in rats immunized with PATE or PATE-F protein. Immunization did not cause any structural damage and inflammation in the testis and epididymis. PATE and PATE-F antisera obtained from the immunized rats inhibited acrosome reaction. Motility parameters, capacitation, acrosome reaction, and fecundity were compromised in PATE-F-immunized rats, whereas the same were not affected in rats immunized with PATE. These results suggest that PATE-F might play an important role in sperm function and fecundity and can be explored further to determine its immunocontraceptive potential. PMID:25899154

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

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test Systems § 866.5800 Seminal fluid (sperm) immunological test system. (a)...

  18. Human Lactate Dehydrogenase A (LDHA) Rescues Mouse Ldhc-Null Sperm Function1

    PubMed Central

    Tang, Huanghui; Duan, Chongwen; Bleher, Reiner; Goldberg, Erwin

    2013-01-01

    ABSTRACT By targeted disruption of the lactate dehydrogenase c (Ldhc) gene, we demonstrated that spermatozoa require Ldhc for capacitation, motility, and fertilizing capacity. Ldhc expression is restricted to the developing germ cells that, however, are apparently not compromised by the lack of the LDHC isozyme. Because LDHC is abundant in spermatozoa that utilize aerobic glycolysis for energy requirements, its main function was presumed to be the interconversion of pyruvate to lactate with the concomitant oxidation/reduction of NADH to NAD+. We found that sperm without LDHC were still able to convert lactate to pyruvate as mediated by LDHA that is tightly bound to the fibrous sheath. It was assumed that the level of glycolysis was insufficient to power motility and the subsequent fertilizing capacity of the mutated sperm. To investigate whether LDHC possesses certain unique characteristics essential for fertility, human LDHA was introduced as a transgene to Ldhc-null mice. We report here that the exogenous LDHA rescued the phenotype of the Ldhc-null males. Sperm from the LDHA transgenic males with the Ldhc deletion (LDHA+/Ldhc?/?) are motile, capable of protein tyrosine phosphorylation, and able to fertilize, thus restoring these properties to LDHC-null sperm. However, the lactate and ATP levels in the rescued sperm did not differ significantly from sperm lacking LDHC. We suggest that it is the localization of the transgene to the sperm cytosol that is mainly responsible for restoration of sperm function and fertility. PMID:23467744

  19. Extreme—Pressure lubricant tests on jojoba and sperm whale oils

    Microsoft Academic Search

    T. K. Miwa; J. A. Rothfus; E. Dimitroff

    1979-01-01

    Laboratory and simulated in-use lubricant tests were performed on sulfurized jojoba oil and on reference sulfurized sperm\\u000a whale oil. Data from these comprehensive tests indicated sulfurized jojoba oil prepared from heat-treated filtered oil to\\u000a be comparable or superior to sulfurized sperm whale oil as an extreme-pressure additive for motor oils, gear lubricants, and\\u000a automotive transmission fluids.

  20. 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

  1. The effect of cryoprotectant on kangaroo sperm ultrastructure and mitochondrial function.

    PubMed

    McClean, Rhett; Holt, William V; Zee, Yeng Peng; Lisle, Allan; Johnston, Stephen D

    2008-12-01

    This study examined the effect of cryoprotectants (20% DMSO, a 10% DMSO/10% glycerol mixture, 20% glycerol and 1M sucrose solution) on kangaroo sperm structure and function, along with the effect of varying concentrations of glycerol on sperm mitochondrial function. Eastern grey kangaroo cauda epididymidal spermatozoa were incubated for 10 min at 35 degrees C in each cryoprotectant and the plasma membrane integrity (PMI) and motility assessed using light microscopy. The same samples were fixed for TEM and the ultrastructural integrity of the spermatozoa examined. To investigate the effect of glycerol on the kangaroo sperm mitochondrial function, epididymidal spermatozoa were incubated with JC-1 in Tris-citrate media at 35 degrees C for 20 min in a range of glycerol concentrations (0%, 5%, 10%, 15% and 20%) and the mitochondrial membrane potential (MMP) and plasma membrane integrity determined. As expected, incubation of spermatozoa in 20% glycerol for 10 min resulted in a significant reduction in motility, PMI and ultrastructural integrity. Interestingly, incubation in 20% DMSO resulted in no significant reduction in motility or PMI but a significant loss of structural integrity when compared to the control spermatozoa (0% cryoprotectant). However, 20% DMSO was overall less damaging to sperm ultrastructure than glycerol, a combination of 10% glycerol and 10% DMSO, and sucrose. While all glycerol concentrations had an adverse effect on mitochondrial function, the statistical models presented for the relationship between MMP and glycerol predicted that spermatozoa, when added to 20% glycerol, would lose half of their initial MMP immediately at 35 degrees C and MMP would halve after 19.4 min at 4 degrees C. Models for the relationship between PMI and glycerol predicted that spermatozoa would lose half of their initial PMI after 1.8 min at 35 degrees C and PMI would halve after 21.1 min at 4 degrees C. These results suggest that if glycerol is to be used as a cryoprotectant for kangaroo spermatozoa then it is best administered at 4 degrees C and that mitochondrial function is more sensitive to glycerol than PMI. Future research should be directed at investigating strategies that reduce exposure of spermatozoa to glycerol during processing and that test the cryoprotective properties of 20% DMSO for kangaroo spermatozoa. PMID:18805409

  2. 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.

  3. Sperm DNA integrity testing: big halo is a good predictor of embryo quality and pregnancy after conventional IVF.

    PubMed

    Tandara, M; Baji?, A; Tandara, L; Bili?-Zulle, L; Šunj, M; Kozina, V; Goluža, T; Juki?, M

    2014-09-01

    Sperm DNA integrity is a sperm functional parameter of male fertility evaluation. Two parameters of sperm DNA integrity were observed: DNA damage expressed as DNA fragmentation index (DFI) and percentage of the DNA undamaged spermatozoa expressed as big halo. Halosperm test was used for sperm DNA integrity determination. The aim of this study was to evaluate which DNA integrity parameter is better as an embryo quality and pregnancy prognostic parameter after the conventional IVF. We evaluated two embryo groups (positive and negative group) according to the 3rd day cumulative embryo score. Big halo and DFI, as we expected, showed good correlation (r = -0.69; p < 0.001). Receiver operating characteristic (ROC) analyses show that DFI and big halo are significant (p < 0.001) as prognostic parameters of embryo quality. ROC curves comparison of DFI and big halo revealed the AUC value for big halo to be significantly higher (DFI AUC = 0.71 vs. big halo AUC = 0.83; p = 0.025) than for DFI. Big halo was found to be the only independent predictor of embryo quality. Sperm DNA integrity both parameters are good prognostic parameters of embryo quality after the conventional IVF where big halo seems to be better. ROC analyses show DFI and big halo as significant prognostic parameters for achieved pregnancy (AUC ± SE for DFI was 0.67 ± 0.06 and 0.75 ± 0.06 for big halo). To our knowledge, this is the first study demonstrating the correlation between sperm DNA undamaged rate expressed as big halo parameter and semen characteristics as well as the influence on fertilization rate, embryo quality and pregnancy in conventional IVF. PMID:24947544

  4. Identification, characterization, and functional correlation of calmodulin-dependent protein phosphatase in sperm

    PubMed Central

    1988-01-01

    Preliminary data demonstrated that the inhibition of reactivated sperm motility by calcium was correlated with inhibited protein phosphorylation. The inhibition of phosphorylation by Ca2+ was found to be catalyzed by the calmodulin-dependent protein phosphatase (calcineurin). Sperm from dog, pig, and sea urchin contain both the Ca2+-binding B subunit of the enzyme (Mr 15,000) and the calmodulin- binding A subunit with an Mr of 63,000. The sperm A subunit is slightly higher in Mr than reported for other tissues. Inhibition of endogenous calmodulin-dependent protein phosphatase activity with a monospecific antibody revealed the presence of 14 phosphoprotein substrates in sperm for this enzyme. The enzyme was localized to both the flagellum and the postacrosomal region of the sperm head. The flagellar phosphatase activity was quantitatively extracted with 0.6 M KCl from isolated flagella from dog, pig, and sea urchin sperm. All salt-extractable phosphatase activity was inhibited with antibodies against the authentic enzyme. Preincubation of sperm models with the purified phosphatase stimulated curvolinear velocity and lateral head amplitude (important components of hyperactivated swimming patterns) and inhibited beat cross frequency suggesting a role for this enzyme in axonemal function. Our results suggest that calmodulin-dependent protein phosphatase plays a major role in the calcium-dependent regulation of flagellar motility. PMID:2836436

  5. The micronutrient supplements, zinc sulphate and folic acid, did not ameliorate sperm functional parameters in oligoasthenoteratozoospermic men.

    PubMed

    Raigani, M; Yaghmaei, B; Amirjannti, N; Lakpour, N; Akhondi, M M; Zeraati, H; Hajihosseinal, M; Sadeghi, M R

    2014-11-01

    We investigated the effects of folic acid and zinc sulphate supplementation on the improvement of sperm function in subfertile oligoasthenoteratozoospermic (OAT) men. Eighty-three OAT men participated in a 16-week intervention randomised, double-blind clinical trial with daily treatment of folic acid (5 mg day(-1) ) and zinc sulphate (220 mg day(-1) ), or placebo. Before and after treatment, semen and blood samples were obtained for determining sperm concentration, motility, and morphology, sperm viability, sperm mitochondrial function, sperm chromatin status using toluidine blue, aniline blue, acridine orange and chromomycin A3 staining; and semen and blood folate, zinc, B12 , total antioxidant capacity (TAC) and malondialdehyde (MDA) concentrations. Sperm concentration (×10(6)  ml(-1) ) increased in subfertile men receiving the combined treatment of folic acid and zinc sulphate and also in the group receiving only folic acid treatment; however, it was not statistically significant (P = 0.056 and P = 0.05, respectively). Sperm chromatin integrity (%) increased significantly in subfertile men receiving only zinc sulphate treatment (P = 0.048). However, this improvement in sperm quality was not significant after adjusting placebo effect. This study showed that zinc sulphate and folic acid supplementation did not ameliorate sperm quality in infertile men with severely compromised sperm parameters, OAT. Male infertility is a multifactorial disorder, and also nutritional factors play an important role in results of administration of supplementation on sperm parameters. However, these results should be confirmed by multiple studies in larger populations of OAT men. PMID:24147895

  6. Sperm competition enhances functional capacity of mammalian spermatozoa

    E-print Network

    Museo Nacional de Ciencias Naturales

    coevolution between the sexes and may contribute to the explanation of the rapid divergence observed the rules of the competition and thus determine which ejaculate features will improve their competitiveness and enhanced sperm production in taxa as diverse as mammals (reviewed in ref. 3), birds (4), butterflies (5

  7. A sperm viability test using SYBR-14/propidium iodide flow cytometry as a tool for rapid screening of primary ciliary dyskinesia patients and for choosing sperm sources for intracytoplasmic sperm injection.

    PubMed

    Niu, Zhi-hong; Huang, Xue-feng; Jia, Xiao-feng; Zheng, Jufen; Yuan, Yao; Shi, Ting-yan; Diao, Hua; Yu, He-guo; Sun, Fei; Zhang, Hui-qing; Shi, Hui-juan; Feng, Yun

    2011-01-01

    Spermatozoa viability tests based on dual-color flow cytometry after staining with Sybr-14/propidium iodide were performed on 44 men with complete asthenospermia for primary ciliary dyskinesia (PCD) screening, and seven were identified with PCD by electron microscopy of ultrastructural ciliary defects. Six PCD patients underwent eight intracytoplasmic sperm injection therapy cycles using ejaculated sperm or testicular sperm, obtaining a mean fertilization rate of 46.6%, with three healthy babies born and one in utero at the time of writing. PMID:20797703

  8. Sperm competition and the evolution of gamete morphology in frogs

    Microsoft Academic Search

    Phillip G. Byrne; Leigh W. Simmons; J. Dale Roberts

    2003-01-01

    Despite detailed knowledge of the ultrastructure of spermatozoa, there is a paucity of information on the selective pressures that influence sperm form and function. Theoretical models for both internal and external fertilizers predict that sperm competition could favour the evolution of longer sperm. Empirical tests of the external-fertilization model have been restricted to just one group, the fishes, and these

  9. 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

  10. In vitro exposure to the organochlorine p,p'-DDE affects functional human sperm parameters.

    PubMed

    Tavares, Renata S; Amaral, Sandra; Paiva, Carla; Baptista, Marta; Ramalho-Santos, João

    2015-02-01

    Although no information exists regarding the levels of p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) on reproductive fluids of heavily exposed populations, they are possibly quite high given the serum levels reported so far. In these populations altered semen quality has been reported, although the direct effects of this DDT metabolite on crucial sperm parameters remain largely unexplored. With this in mind, a long-term in vitro incubation that better mimics the putative continuous exposure of spermatozoa to p,p'-DDE in the female reproductive tract in vivo was used. Before compromising sperm viability, continuous p,p'-DDE exposure remarkably decreased sperm motility, possibly due to the combined reduction in the proportion of sperm with high mitochondrial membrane potential and cellular ATP levels, all of which were clearly more affected at 50 and 100 ?M p,p'-DDE. Moreover, 25 ?M p,p'-DDE was also able to promote a decline in sperm with high MMP, however without significantly affecting motility. On the other hand, p,p'-DDE at the highest concentration strongly inhibited the process of capacitation following 24h of incubation. In conclusion, human sperm function is affected by continuous high p,p'-DDE exposure which may ultimately compromise male fertility. Given our previously findings that showed a non-regulated Ca(2+) entry in the presence of p,p'-DDE, we suggest that this organochlorine may promote mitochondrial Ca(2+) overload which may culminate in a general mitochondrial dysfunction and cellular ATP depletion, thus affecting sperm fertilizing potential. Our findings suggest a broader understanding of the non-genomic mechanism of p,p'-DDE action in human sperm. PMID:25240159

  11. Cytotoxic Effects of Benzene Metabolites on Human Sperm Function: An In Vitro Study

    PubMed Central

    Mandani, Priyanka; Desai, Ketki; Highland, Hyacinth

    2013-01-01

    In recent years, individuals are rampantly exposed to vapours of benzene, through paint, plastic, petroleum industries, fuel exhaust, and tobacco smoke. Hence the present investigation was directed towards determining the effect of benzene metabolites, namely, phenol-hydroquinone and catechol, on the motility, viability, and nuclear integrity of the human spermatozoa. From the results obtained it was clear that exposure to phenol-hydroquinone caused a significant decline in both, sperm motility and viability. Exposure to a phenol-hydroquinone (Phase I) microenvironment may therefore inhibit metabolically active enzymes, thus impeding ATP production, and in turn lowers sperm motility and viability. In addition, the present study also revealed that both metabolites of benzene caused significant denaturation of sperm nuclear DNA. Hence, exposure to phenol-hydroquinone in vitro could have resulted in generation of free radicals and altered membrane function, which is reflected by a decline in the motility, viability, and loss of sperm nuclear DNA integrity. In Phase II, the exposure of human sperm in vitro to varied concentrations of catechol caused only insignificant changes in sperm motility and viability as compared to those observed on exposure to phenol-hydroquinone. Hence, exposure to catechol appeared to have less toxic effects than those of phenol-hydroquinone. PMID:24416599

  12. Central role of 5'-AMP-activated protein kinase in chicken sperm functions.

    PubMed

    Nguyen, Thi Mong Diep; Alves, Sabine; Grasseau, Isabelle; Métayer-Coustard, Sonia; Praud, Christophe; Froment, Pascal; Blesbois, Elisabeth

    2014-11-01

    Avian gametes present specific features related to their internal long-term mode of fertilization. Among other central actors of energetic metabolism control, it has been suspected that 5'-AMP-activated protein kinase (AMPK) influences sperm functions and thus plays a key role in fertilization success. In the present work, we studied AMPK localization and function in chicken sperm incubated in vitro. Effects of the pharmacological AMPK activators (AICAR, metformin) and the AMPK inhibitor compound C were assessed by evaluating AMPKalpha (Thr(172)) phosphorylation (by Western blotting), semen quality (by viability, motility, and ability to perform acrosome reaction), and energetic metabolism indicators (lactate, ATP). Localization of AMPK in subcellular sperm compartments was evaluated by immunocytochemistry. Total AMPK was found in all compartments except for the nucleus, but the phosphorylated form phospho-Thr(172)-AMPK was essentially localized in the flagellum and acrosome. AMPK activators significantly improved AMPK phosphorylation, sperm motility (increased by 40% motile, 90% progressive, and 60% rapid sperm), acrosome reaction and lactate production (increased by 40%) and viability. The AMPK inhibitor significantly reduced AMPK phosphorylation and percentages of motility (decrease by 25%), progressive energy (decrease by 35%), and rapid sperm (decreased by 30%), acrosome reaction, lactate production, and ATP release. The two activators differed in their effect on ATP concentration: AICAR stimulated ATP formation, whereas metformin did not. Our results indicate that AMPK plays a key role in the regulation of chicken sperm functions and metabolism. This action differs from that suggested in mammals, mainly by its crucial involvement in the acrosome reaction process. PMID:25297543

  13. 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

  14. Is the Function of the Porcine Sperm Reservoir Restricted to the Ovulatory Period?

    PubMed Central

    BRÜSSOW, Klaus-Peter; EGERSZEGI, Istvan; RÁTKY, Jozsef

    2014-01-01

    The uterotubal junction (UTJ) and caudal isthmus are recognized as a functional pre-ovulatory sperm reservoir (SR). Spermatozoa are released from the SR in a complex and concerted action. However, whether this functionality is restricted only to the ovulatory period is still open to debate. Our study was aimed to analyze the presence of spermatozoa within the UTJ (SR), isthmus (ISTH) and ampulla (AMP) after laparoscopic intrauterine insemination (LIUI) either in the peri- (PERI) or post-ovulatory (POST) period or at mid cycle (MID). Each uterine horn of estrus synchronized gilts (n=12) was inseminated with 20 ml sperm (29.5×106 cells/ml). Oviducts were recovered 7 h after LIUI and separated into the UTJ, ISTH and AMP, and sections were flushed with 10 ml PBS+EDTA solution. After centrifugation, the sperm pellet was evaluated by ?e?ovský staining. The median sperm numbers in the PERI, POST and MID groups were 578, 171 and 789 in the UTJ; 545, 233 and 713 in the ISTH; and 496, 280 and 926 in the AMP, respectively, and there were differences between the POST and MID groups (P<0.05) but not between the oviductal sections of each group (P>0.05). Compared with the MID group, the percent of intact sperm cells was higher (P<0.01) in the PERI and POST groups (32.8 vs. 66.4 and 76.8%). Also, the percentages of aberrations in the acrosome and tail were higher (P<0.05) in the MID group. Based on this, it can be assumed that the sperm reservoir is active during different phases of the estrus cycle. However, the mid-cycle oviduct environment considerably impairs sperm cell quality. PMID:24964752

  15. 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

  16. 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

  17. The use of the hypo-osmotic swelling test, water test, and supravital staining in the evaluation of drone sperm

    E-print Network

    of drone sperm Zekariya NUR 1 , Selvinar SEVEN-CAKMAK 2 , Burcu USTUNER 1 , Ibrahim CAKMAK 3 , Melih ERTURK was obtained from mature drones (16 days or older) collected from four colonies. The means of the per-drone evaluation in mammals. bee semen / HOS test / water test / viability 1. INTRODUCTION The quality of drone

  18. Rapid Evolution of a Primate Sperm Protein: Relaxation of Functional Constraint or Positive Darwinian Selection?

    Microsoft Academic Search

    Alejandro P. Rooney; Jianzhi Zhang

    Protamines are arginine-rich proteins that replace histones and bind sperm DNA during spermatogenesis in verte- brates. Previous studies have shown that protamine exons evolve faster than does the protamine intron. It has been suggested that this is a result of a relaxation of functional constraint. However, a more likely explanation is that the evolutionary rate of exons has been accelerated

  19. Enhanced fertility prediction of cryopreserved boar spermatozoa using novel sperm function assessment.

    PubMed

    Daigneault, B W; McNamara, K A; Purdy, P H; Krisher, R L; Knox, R V; Rodriguez-Zas, S L; Miller, D J

    2015-05-01

    Due to reduced fertility, cryopreserved semen is seldom used for commercial porcine artificial insemination (AI). Predicting the fertility of individual frozen ejaculates for selection of higher quality semen prior to AI would increase overall success. Our objective was to test novel and traditional laboratory analyses to identify characteristics of cryopreserved spermatozoa that are related to boar fertility. Traditional post-thaw analyses of motility, viability, and acrosome integrity were performed on each ejaculate. In vitro fertilization, cleavage, and blastocyst development were also determined. Finally, spermatozoa-oviduct binding and competitive zona-binding assays were applied to assess sperm adhesion to these two matrices. Fertility of the same ejaculates subjected to laboratory assays was determined for each boar by multi-sire AI and defined as (i) the mean percentage of the litter sired and (ii) the mean number of piglets sired in each litter. Means of each laboratory evaluation were calculated for each boar and those values were applied to multiple linear regression analyses to determine which sperm traits could collectively estimate fertility in the simplest model. The regression model to predict the percent of litter sired by each boar was highly effective (p < 0.001, r(2)  = 0.87) and included five traits; acrosome-compromised spermatozoa, percent live spermatozoa (0 and 60 min post-thaw), percent total motility, and the number of zona-bound spermatozoa. A second model to predict the number of piglets sired by boar was also effective (p < 0.05, r(2)  = 0.57). These models indicate that the fertility of cryopreserved boar spermatozoa can be predicted effectively by including traditional and novel laboratory assays that consider functions of spermatozoa. PMID:25914302

  20. High glucose concentrations per se do not adversely affect human sperm function in vitro.

    PubMed

    Portela, J M D; Tavares, R S; Mota, P C; Ramalho-Santos, J; Amaral, S

    2015-07-01

    Diabetes mellitus (DM) represents one of the greatest concerns to global health and it is associated with diverse clinical complications, including reproductive dysfunction. Given the multifactorial nature of DM, the mechanisms that underlie reproductive dysfunction remain unclear. Considering that hyperglycemia has been described as a major effector of the disease pathophysiology, we used an in vitro approach to address the isolated effect of high glucose conditions on human sperm function, thus avoiding other in vivo confounding players. We performed a complete and integrated analysis by measuring a variety of important indicators of spermatozoa functionality (such as motility, viability, capacitation status, acrosomal integrity, mitochondrial superoxide production and membrane potential) in human sperm samples after incubation with d- and l-glucose (5, 25, or 50?mM) for 24 and 48?h. No direct effects promoted by 25 or 50?mM d-glucose were found for any of the parameters assessed (P>0.05), except for the acrosome reaction, which was potentiated after 48?h of exposure to 50?mM d-glucose (P<0.05). Interestingly, non-metabolizable l-glucose drastically increased superoxide production (P<0.05) and suppressed sperm motility (P<0.05) and capacitation (P<0.05) after 24?h of treatment, whereas mitochondrial membrane potential (P<0.05), acrosomal integrity (P<0.01) and viability (P<0.05) were later decreased. The overall results suggest that high glucose levels per se do not influence human sperm function in vitro, which stresses the importance of other factors involved in DM pathology. Nevertheless, the absence of metabolizable glucose contributes to a severe impairment of sperm function and thus compromises male fertility.Free Portuguese abstract: A Portuguese translation of this abstract is freely available at http://www.reproduction-online.org/content/150/1/77/suppl/DC1. PMID:25926692

  1. Human sperm competition: ejaculate adjustment by males and the function of masturbation (1993)

    Microsoft Academic Search

    R. Robin Baker; Mark A. Bellis

    Sperm competition theory argues that the number of sperm inseminated into a female by a male is a trade-off between two opposing pressures. On the one hand, the risk that sperm may find themselves in competition with the sperm from another male favours the male inseminating more sperm. On the other hand, ejaculates are costly to produce and males are

  2. DNA integrity of canine spermatozoa during chill storage assessed by the sperm chromatin dispersion test using bright-field or fluorescence microscopy.

    PubMed

    Hidalgo, M; Urbano, M; Ortiz, I; Demyda-Peyras, S; Murabito, M R; Gálvez, M J; Dorado, J

    2015-08-01

    The objective of this study was to evaluate the effect of chill storage on canine sperm DNA fragmentation assessed by the sperm chromatin dispersion test using bright-field microscopy with Wright solution (sDF-B) or fluorescence microscopy with propidium iodide (sDF-F). The relationship and agreement between the results obtained with both staining methods were analyzed. The values of DNA fragmentation indexes (sDF-F and sDF-B) were compared at each time of chill storage (0, 24, 48, 72, and 96 hours). Additionally, the sperm DNA fragmentation rate (slope) was compared between the methods during chill storage. Good agreement and no significant differences between values obtained with both staining procedures were observed. Finally, the effect of chill storage for up to 96 hours was assessed on sperm motility parameters and DNA fragmentation indexes. Significant differences were found after 48 hours of chill storage, obtaining greater values of fragmented DNA. Progressive sperm motility was lower just after 96 hours of chill storage, and no effect was found in total sperm motility. In conclusion, the Sperm-Halomax kit, developed for canine semen and based on the sperm chromatin dispersion test, can be used accurately under bright-field or fluorescence microscopy to assess the sperm DNA integrity of canine semen during chill storage. The sperm DNA fragmentation index increased after 48 hours of chill storage, thereby detecting sperm damage earlier than other routine sperm parameters, such as sperm motility. PMID:25963130

  3. Nigerian Bonny Light Crude Oil Disrupts Antioxidant Systems in Testes and Sperm of Rats

    Microsoft Academic Search

    Ebenezer O. FarombiIsaac; Isaac A. Adedara; Azubike P. Ebokaiwe; Roy Teberen; Theresa Ehwerhemuepha

    2010-01-01

    Nigerian Bonny light crude oil (BLCO) is commonly used by the local population in folklore medicine for the management of\\u000a various forms of gastrointestinal problems and male reproductive capacity. The study investigated the effects of BLCO on the\\u000a antioxidant systems of the testes and epidydimal sperm in rats by oral exposure to 0, 200, 400 and 800 mg\\/kg BLCO for 7 days.

  4. COMPARATIVE EVALUATION OF THREE RAPID MARINE TOXICITY TESTS: SEA URCHIN EARLY EMBRYO GROWTH TEST, SEA URCHIN SPERM CELL TOXICITY TEST AND MICROTOX

    EPA Science Inventory

    Three rapid marine toxicity tests were evaluated to determine their potential usefulness in a toxicity testing program: early embryo growth test and sperm cell toxicity test, both using the sea urchin Arbacia punctulata, and Microtox. Toxicity values (EC50s) were derived for eigh...

  5. Fenitrothion induced oxidative stress and morphological alterations of sperm and testes in male sprague-dawley rats

    PubMed Central

    Taib, Izatus Shima; Budin, Siti Balkis; Ghazali, Ahmad Rohi; Jayusman, Putri Ayu; Louis, Santhana Raj; Mohamed, Jamaludin

    2013-01-01

    OBJECTIVE: Fenitrothion residue is found primarily in soil, water and food products and can lead to a variety of toxic effects on the immune, hepatobiliary and hematological systems. However, the effects of fenitrothion on the male reproductive system remain unclear. This study aimed to evaluate the effects of fenitrothion on the sperm and testes of male Sprague-Dawley rats. METHODS: A 20 mg/kg dose of fenitrothion was administered orally by gavages for 28 consecutive days. Blood sample was obtained by cardiac puncture and dissection of the testes and cauda epididymis was performed to obtain sperm. The effects of fenitrothion on the body and organ weight, biochemical and oxidative stress, sperm characteristics, histology and ultrastructural changes in the testes were evaluated. RESULTS: Fenitrothion significantly decreased the body weight gain and weight of the epididymis compared with the control group. Fenitrothion also decreased plasma cholinesterase activity compared with the control group. Fenitrothion altered the sperm characteristics, such as sperm concentration, sperm viability and normal sperm morphology, compared with the control group. Oxidative stress markers, such as malondialdehyde, protein carbonyl, total glutathione and glutathione S-transferase, were significantly increased and superoxide dismutase activity was significantly decreased in the fenitrothion-treated group compared with the control group. The histopathological and ultrastructural examination of the testes of the fenitrothion-treated group revealed alterations corresponding with the biochemical changes compared with the control group. CONCLUSION: A 20 mg/kg dose of fenitrothion caused deleterious effects on the sperm and testes of Sprague-Dawley rats. PMID:23420164

  6. Detection of oxidative DNA damage in human sperm and its association with sperm function and male infertility

    Microsoft Academic Search

    Han-Ming Shen; Choon-Nam Ong

    2000-01-01

    The expanding research interest in the last two decades on reactive oxygen species (ROS), oxidative stress, and male infertility has led to the development of various techniques for evaluating oxidative DNA damage in human spermatozoa. Measurement of 8-hydroxydeoxyguanosine (8-OHdG) offers a specific and quantitative biomarker on the extent of oxidative DNA damage caused by ROS in human sperm. The close

  7. Reproductive function in male endurance athletes: sperm analysis and hormonal profile.

    PubMed

    Lucía, A; Chicharro, J L; Pérez, M; Serratosa, L; Bandrés, F; Legido, J C

    1996-12-01

    The purpose of this investigation was to study the effects of endurance exercise on male reproductive function (sex hormones and seminograms). Professional cyclists [n = 12; mean age 24 +/- 2 (SD) yr], elite triathletes (n = 9; 26 +/- 3 yr), recreational marathon runners (n = 10; 32 +/- 6 yr), and sedentary subjects (control group; n = 9; 30 +/- 4 yr) were selected as subjects. for each group, the following parameters were measured three times during the sports season (training period: winter; competition period: spring; resting period: fall): percentage of body fat, hormonal profile (resting levels of follicle-stimulating hormone, luteinizing hormone, total and free testosterone, and cortisol), and seminograms (quantitative parameters sperm volume and sperm count; qualitative parameters: sperm motality and morphology). The following comparisons were made in the measured parameters: 1) within groups (longitudinal design) and 2) between groups in each of the three periods (cross-sectional design) and over time (mixed design). In addition, both the volume and the intensity of training of each subject during the season (except for the control group) were quantified. Despite significant differences in training characteristics and in body fat percent, in general no significant differences (P > 0.05) were found in hormonal profiles or in semen characteristics between or within groups. A lower sperm motility (46.2 +/- 19.5%), however, was observed in the cyclists during the competition period when compared either with the other groups during this same period (P < 0.05) or with themselves during the other two periods of study (P < 0.01). In any case, the later phenomenon was attributed to physical factors associated with cycling, such as mechanical trauma to the testis and/or increased gonadal temperature. In conclusion, our findings suggest that endurance exercise does not adversely affect the hypothalamic-pituitary-testis axis. PMID:9018515

  8. 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

  9. 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.

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

    E-print Network

    Pina-Guzman, Belem

    2010-01-01

    acid fragmentation in human sperm: correlation withSperm chromatin alteration and DNA damage by methyl-parathion, chlorpyrifos and diazinon and their oxon metabolites in humanhuman reproduction has increased because of its genotoxic effect in somatic (Rupa et al. , 1990) and sperm

  11. Thyroid function tests

    MedlinePLUS

    ... common thyroid function tests are: Total, or free T4 (the main thyroid hormone in your blood) TSH ( ... pituitary gland that stimulates the thyroid to produce T4) T3 (also included sometimes) Other thyroid tests include: ...

  12. SPERM WHALE TRUMPET SOUNDS

    Microsoft Academic Search

    V. TELONI; W. M. X. ZIMMER; P. L. TYACK

    2005-01-01

    Sperm whale Physeter macrocephalus L. clicks have been studied for nearly fifty years, during which time great efforts have been made to understand the functions and production mechanisms of this sound. Other than clicks, sperm whales may also produce low intensity sounds arranged in short sequences, named trumpets, which have been recorded occasionally in the past by few groups of

  13. Validation of a spectrophotometer-based method for estimating daily sperm production and deferent duct transit.

    PubMed

    Froman, D P; Rhoads, D D

    2012-10-01

    The objectives of the present work were 3-fold. First, a new method for estimating daily sperm production was validated. This method, in turn, was used to evaluate testis output as well as deferent duct throughput. Next, this analytical approach was evaluated in 2 experiments. The first experiment compared left and right reproductive tracts within roosters. The second experiment compared reproductive tract throughput in roosters from low and high sperm mobility lines. Standard curves were constructed from which unknown concentrations of sperm cells and sperm nuclei could be predicted from observed absorbance. In each case, the independent variable was based upon hemacytometer counts, and absorbance was a linear function of concentration. Reproductive tracts were excised, semen recovered from each duct, and the extragonadal sperm reserve determined by multiplying volume by sperm cell concentration. Testicular sperm nuclei were procured by homogenization of a whole testis, overlaying a 20-mL volume of homogenate upon 15% (wt/vol) Accudenz (Accurate Chemical and Scientific Corporation, Westbury, NY), and then washing nuclei by centrifugation through the Accudenz layer. Daily sperm production was determined by dividing the predicted number of sperm nuclei within the homogenate by 4.5 d (i.e., the time sperm with elongated nuclei spend within the testis). Sperm transit through the deferent duct was estimated by dividing the extragonadal reserve by daily sperm production. Neither the efficiency of sperm production (sperm per gram of testicular parenchyma per day) nor deferent duct transit differed between left and right reproductive tracts (P > 0.05). Whereas efficiency of sperm production did not differ (P > 0.05) between low and high sperm mobility lines, deferent duct transit differed between lines (P < 0.001). On average, this process required 2.2 and 1.0 d for low and high lines, respectively. In summary, we developed and then tested a method for quantifying male reproductive tract throughput. This method makes the study of semen production amenable to systems biology. PMID:22991549

  14. Morphology and function of male genitalia (spermatophores) in Euscorpius italicus (Euscorpiidae, Scorpiones): complex spermatophore structures enable safe sperm transfer.

    PubMed

    Jacob, Alain; Gantenbein, Iris; Braunwalder, Matt E; Nentwig, Wolfgang; Kropf, Christian

    2004-04-01

    The structure and function of the spermatophore of Euscorpius italicus are analyzed. We show how the spermatophore gets shaped from two hemispermatophores and for the first time the sperm transfer mechanism is shown in detail, illustrating function and importance of all complex lobe structures of an euscorpiid spermatophore. A detailed description of the interaction of spermatophore and female genitalia is given. The capsular region of the spermatophore bears different lobes: The distal and basal lobes hook into two cavities on the inner side of the female's genital operculum. A so-called "crown-like structure" hooks into a membranous area in the genital atrium. During sperm transfer, these crown-like structures move backwards, in this way widening the female's genital opening. The sperm duct of the spermatophore is coated with numerous spicules on its outer side, which could serve as a sealing mechanism and/or may stimulate the female. Furthermore, we conclude that "safeguarding of sperm transfer" is one driving force for evolution of male genital complexity in scorpions, but also sexual selection by cryptic female choice could partly play a role. PMID:15052598

  15. 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

  16. Sperm morphometric subpopulations are differentially distributed in rams with different maturity age in cryopreserved ejaculates.

    PubMed

    Martí, J I; Aparicio, I M; García-Herreros, M

    2011-07-01

    It is widely accepted that sperm morphology is a good indicator of fertility and it has been proposed that sperm quality may be related to subtle changes in sperm head morphology. However, a precise estimation of the morphology of ram sperm would be very useful to improve reproductive success in ovine. Computer-assisted morphometric analysis and clustering analysis have been important tools to study sperm subpopulations in domestic animals. However, to the best of our knowledge, no data exist studing morphometric differences regarding to sperm subpopulations within the ovine ejaculate. The aim of this study was to test the presence and distribution of sperm morphometric subpopulations in cryopreserved ejaculates from yearling and mature rams using an objective method by computer analysis system and to establish the relationship between the distribution of the subpopulations found and sperm quality in each individual ram. Principal component analysis revealed that three principal components for yearlings and four components for mature rams that represented more than 84% of the cumulative variance in both cases. After cluster analysis, three sperm morphometric subpopulations for yearlings (CLY) and four for mature (CLM) rams were identified with defined sperm dimensions and shapes. CLY1 included big, round and short sperm (37%), CLY2 included average size and slightly elliptical and elongated sperm (48%), CLY3 included small, long, elliptical and elongated sperm cells (15%). CLM1 consisted of average size and moderate elliptical and elongated (26%), CLM2 consisted of small, long, elliptical and elongated (31%), CLM3 consisted of small and round (32%) and CLM4 included big, short and round (8%) spermatozoa respectively. There were significant differences in the distribution of the three subpopulations (P < 0.001) as well as in the sperm concentration, total motility (%), sperm viability (%) and the overall (P < 0.05) in the ejaculates among the four yearling rams tested. Same results were found for the four subpopulations and the different sperm quality parameters in the ejaculates among the four mature rams tested. In conclusion, cryopreserved ram semen showed a specific structure with regard to sperm morphometric subpopulations. In addition, the distribution of these subpopulations seems to be related to stud maturity age and the ejaculate quality which would be a very important indicator of sperm function. Thus, analysis of sperm morphometric subpopulation structure together with functional tests could provide valuable information to assess the cryoresistence of ram spermatozoa. PMID:21396697

  17. Gamete evolution and sperm numbers: sperm competition versus sperm limitation.

    PubMed

    Parker, Geoff A; Lehtonen, Jussi

    2014-09-22

    Both gamete competition and gamete limitation can generate anisogamy from ancestral isogamy, and both sperm competition (SC) and sperm limitation (SL) can increase sperm numbers. Here, we compare the marginal benefits due to these two components at any given population level of sperm production using the risk and intensity models in sperm economics. We show quite generally for the intensity model (where N males compete for each set of eggs) that however severe the degree of SL, if there is at least one competitor for fertilization (N - 1 ? 1), the marginal gains through SC exceed those for SL, provided that the relationship between the probability of fertilization (F) and increasing sperm numbers (x) is a concave function. In the risk model, as fertility F increases from 0 to 1.0, the threshold SC risk (the probability q that two males compete for fertilization) for SC to be the dominant force drops from 1.0 to 0. The gamete competition and gamete limitation theories for the evolution of anisogamy rely on very similar considerations: our results imply that gamete limitation could dominate only if ancestral reproduction took place in highly isolated, small spawning groups. PMID:25100694

  18. 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 ( ...

  19. Assessment of sperm quality: a flow cytometric approach.

    PubMed

    Graham, J K

    2001-12-01

    For many years, scientists have sought to develop laboratory assays that accurately predict the fertilizing capacity of a semen sample. This goal, however, has proven elusive and will most likely be very difficult to achieve, due to the complex nature of the problem. Part of the problem results from the many attributes that a spermatozoon must possess to fertilize an egg, and how laboratory assays can evaluate all of these attributes simultaneously. The percentage of motile sperm in a sample is most commonly used to evaluate semen quality. This assay, however, is not highly correlated with the fertilizing capacity of semen samples. One reason motion assays do not correlate well with fertility is that we are evaluating only one of many attributes that a sperm must possess to fertilize an oocyte. One of the problems of measuring multiple sperm attributes is the time and cost required. Using flow cytometric assays, multiple sperm attributes, including cell viability, acrosomal integrity, and mitochondrial function, can be measured simultaneously in sperm cells. In addition, the ability of sperm to undergo capacitation and the acrosome reaction, as well as the chromosomal integrity of sperm can be measured using flow cytometry. Flow cytometry permits us to evaluate 50,000 sperm in less then 1 min and at reasonable cost. Although flow cytometry is a powerful tool for evaluating many sperm attributes, it cannot evaluate all of the attributes a sperm cell requires to fertilize an oocyte. Therefore, laboratory assays are also being developed to evaluate the ability of sperm: (1) to bind to the oocyte, by evaluating the ability of sperm to bind to the perivitelline membrane of the hen egg in vitro; (2) to undergo an acrosome reaction in vitro, after treatment with membrane destabilizing compounds; and (3) to penetrate oocytes in vitro. When data from multiple sperm assays are used, higher correlations with the fertilizing potential of a semen sample is achieved. For example, in a study conducted utilizing five stallions, the percentage of motile sperm in semen samples correlated poorly with fertility (r(2)=0.22), however, when data for sperm motility, viability and penetration rates into zona-free hamster oocytes were utilized together, these data explained 72% of the differences in the fertility of the stallions (r=0.849; [Theriogenology 46 (1996) 559]). Armed with a battery of tests, which evaluate many different sperm attributes, researchers should be able to more accurately estimate the fertilizing potential of semen samples. PMID:11744268

  20. 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.

  1. Evolution of larger sperm in response to experimentally increased sperm competition in Caenorhabditis elegans.

    PubMed Central

    LaMunyon, Craig W; Ward, Samuel

    2002-01-01

    Sperm morphology evolves rapidly, resulting in an exceptional diversity of sperm size and shape across animal phyla. This swift evolution has been thought to prevent fertilizations between closely related species. Alternatively, recent correlative analyses suggest that competition among sperm from more than one male may cause sperm diversity, but these hypotheses have not been tested. Here, we test experimentally the effect of sperm competition on sperm-size evolution using the nematode Caenorhabditis elegans. This worm has a three day generation time, which allowed the study to cover many generations. Sperm volume increased nearly 20% over 60 generations in lines genetically induced to have high levels of sperm competition compared with those of control lines. These results show that sperm competition can and does cause morphological evolution of sperm and, therefore, can explain much of the diversity in sperm morphology. PMID:12061955

  2. 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

  3. A new method for sperm characterization for infertility treatment: hypothesis testing by using combination of watershed segmentation and graph theory.

    PubMed

    Shojaedini, Seyed Vahab; Heydari, Masoud

    2014-10-01

    Shape and movement features of sperms are important parameters for infertility study and treatment. In this article, a new method is introduced for characterization sperms in microscopic videos. In this method, first a hypothesis framework is defined to distinguish sperms from other particles in captured video. Then decision about each hypothesis is done in following steps: Selecting some primary regions as candidates for sperms by watershed-based segmentation, pruning of some false candidates during successive frames using graph theory concept and finally confirming correct sperms by using their movement trajectories. Performance of the proposed method is evaluated on real captured images belongs to semen with high density of sperms. The obtained results show the proposed method may detect 97% of sperms in presence of 5% false detections and track 91% of moving sperms. Furthermore, it can be shown that better characterization of sperms in proposed algorithm doesn't lead to extracting more false sperms compared to some present approaches. PMID:25426431

  4. Testing Security Policies: Going Beyond Functional Testing

    Microsoft Academic Search

    Yves Le Traon; Tejeddine Mouelhi; Benoit Baudry

    2007-01-01

    While important efforts are dedicated to system functional testing, very few works study how to test specifically security mechanisms, implementing a security policy. This paper introduces security policy testing as a specific target for testing. We propose two strategies for producing security policy test cases, depending if they are built in complement of existing functional test cases or independently from

  5. Germ cell-specific localization of immunoreactive riboflavin carrier protein in the male golden hamster: appearance during spermatogenesis and role in sperm function.

    PubMed

    Sreekumar, A; Acharya, K K; Lalitha, H S; Indi, S S; Bali, P; Seshagiri, P B

    2005-05-01

    Riboflavin carrier protein (RCP) is a phosphoglycoprotein (37 kDa) that is well studied in chicken. An immunologically cross-reacting protein was identified in mammals and active immunization of male rats and bonnet monkeys with chicken RCP lead to an approximately 80% reduction in fertility. However, the physiological mechanism responsible for inhibition of male fertility has not been investigated. Moreover, information on the cell type-specific localization and the origin of immunoreactive RCP during spermatogenesis is extremely limited. Hence, studies were carried out to determine the pattern of expression of immunoreactive RCP during spermatogenesis and its role in sperm function in the golden hamster. Immunoreactive RCP was germ cell-specific, found to be associated with the acrosome-organizing region of early spermatids and showed interesting patterns of immunolocalization during late stages of spermiogenesis. Mature spermatozoa exhibited acrosome-specific localization, mainly in the peri-acrosomal membrane. The immunoreactive protein was undetectable in (non)gonadal somatic cells tested. The protein had a molecular mass of 45-55 kDa and was biosynthesized by round spermatids. The acrosome-specific localization of immunoreactive RCP was unchanged during capacitation, but it was substantially lost during acrosome reaction. Functional studies indicated that treatment of spermatozoa with anti-RCP antibodies did not have any effect on either capacitation or acrosome reaction, but markedly reduced the rate of sperm penetration into zona-free hamster oocytes. These results show the existence of male germ cell-specific immunoreactive RCP, having a potential role in sperm-egg interaction in hamsters. Also the pattern of immunoreactive-RCP localization makes it an ideal marker to monitor development of acrosome in mammalian spermatozoa. PMID:15855621

  6. Infant Pulmonary Function Testing

    PubMed Central

    Davis, Stephanie D.

    2011-01-01

    Infant pulmonary function testing has evolved from a research technique into a diagnostic tool. As such, new current procedural terminology (CPT) codes have been created and are available for use as of January 1, 2010. The technology now available has a range of applications. Through a series of vignettes, this article illustrates the methodology of the tests, some of their applications, and how to code and bill for the procedures. PMID:21540218

  7. 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

  8. 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 involved in sperm and seminal fluid production (Wyckoff et al. 2000). Humans have moderately sized testes

  9. Sperm swimming speed and energetics vary with sperm competition risk in bluegill ( Lepomis macrochirus )

    Microsoft Academic Search

    Gary Burness; Stephen J. Casselman; Albrecht I. Schulte-Hostedde; Christopher D. Moyes; Robert Montgomerie

    2004-01-01

    Under sperm competition, a male’s fertilization success depends largely on the ejaculate characteristics of competing males. Theoretical models predict that, in external fertilizers, increased risk of sperm competition should result in selection for increased sperm swimming speed. To test this prediction, we studied the behavior of sperm from parental and sneaker male bluegill ( Lepomis macrochirus), a fish species characterized

  10. 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.

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

    SciTech Connect

    Spriggs, E.L.; Martin, R.H. [Univ. of Calgary, Alberta (Canada)] [and others

    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.

  12. 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

  13. Mammalian Sperm Proteins Are Rapidly Evolving: Evidence of Positive Selection in Functionally Diverse Genes

    Microsoft Academic Search

    Dara G. Torgerson; Rob J. Kulathinal; Rama S. Singh

    A growing number of genes involved in sex and reproduction have been demonstrated to be rapidly evolving. Here, we show that genes expressed solely in spermatozoa represent a highly diverged subset among mouse and human tissue-specific orthologs. The average rate of nonsynonymous substitutions per site (Ka) is significantly higher in sperm proteins (mean Ka 5 0.18; N 5 35) than

  14. Human sperm competition: ejaculate manipulation by females and a function for the female orgasm (1993)

    Microsoft Academic Search

    R. Robin Baker; Mark A. Bellis

    Behavioural ecologists view monogamy as a subtle mixture of conflict and cooperation between the sexes. In part, conflict and cooperation is cryptic, taking place within the female’s reproductive tract. In this paper the cryptic interaction for humans was analysed using data from both a nationwide survey and counts of sperm inseminated into, and ejected by, females. On average, 35% of

  15. 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

  16. Sperm cleanup and centrifugation processing for cryopreservation.

    PubMed

    Sieme, Harald; Oldenhof, Harriëtte

    2015-01-01

    Fertility rates with artificial insemination are highest with good-quality sperm samples. Therefore, nonviable sperm, cellular debris, and seminal plasma are preferably removed from semen samples prior to use or for preservation. Such compounds are sources where reactive oxygen species are generated during storage or upon cryopreservation, impairing sperm function. In this chapter we describe methods to remove seminal plasma and cellular debris from sperm samples, and for selecting morphologically normal motile sperm. The methods that are described here include: ordinary centrifugation, sperm swim-up, glass wool and Sephadex filtration/adherence, and single-layer as well as discontinuous two-layer iodixanol density gradient centrifugation. PMID:25428016

  17. 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

  18. Maintenance of steelhead trout (Oncorhynchus mykiss) sperm at different in vitro oxygen tensions alters ATP levels and cell functional characteristics

    Microsoft Academic Search

    D. C. Bencic; M. Krisfalusi; J. G. Cloud; R. L. Ingermann

    1999-01-01

    Adenosine triphosphate (ATP) levels in sperm from steelhead trout (Oncorhynchus mykiss) were found to be 12.0 ± 1.4 pmol ATP per 106 sperm cells. Sperm were stored at 0–2 °C for up to 72 h under 100 and 21% O2, and 100% N2. The sperm ATP content of samples maintained under 100 and 21% O2 both decreased to about 70 and 50% of

  19. Cryopreservation of epididymal stallion sperm.

    PubMed

    Olaciregui, M; Gil, L; Montón, A; Luño, V; Jerez, R A; Martí, J I

    2014-02-01

    Any event that makes semen collection or mating impossible, such as death, castration, or injury, may terminate a stallion's breeding career. Fortunately, stallion sperm which are capable of fertilization can be harvested from the epididymis, and frozen for future use. However, the fertility of frozen-thawed epididymal sperm has been found to be lower than that of ejaculated sperm. Therefore, this study aimed to optimize the fertility of frozen epididymal stallion sperm by investigating the effects of different cryoprotectants and freezing protocols on sperm quality. Dimethylformamide was tested alone or combination with pasteurized egg yolk as substitute of fresh egg yolk. In addition, the effect of the pre-freeze stabilization on sperm quality was analyzed. Heterospermic samples obtained from stallion epididymis were collected and cryopreserved in lactose-egg-yolk extender or in the same extender with varying content of cryoprotectant and content of egg yolk, stabilized and no-stabilized. Sperm motility, viability, hypoosmotic swelling test (HOST) and acrosome integrity were evaluated post-thawing. No improvement was observed on the replacement of fresh yolk by pasteurized egg yolk, whereas the results suggest that dimethylformamide is a cryoprotectant suitable for cryopreservation of equine epididymal semen, even better than glycerol. In addition, we found that the stabilization before freezing on epididymal stallion sperm, can improve sperm quality parameters. PMID:24412395

  20. Sperm release evoked by electrical stimulation of the fish brain: a functional-anatomical study.

    PubMed

    Demski, L S; Bauer, D H; Gerald, J W

    1975-02-01

    Acute brain stimulation experiments were carried out in anesthetized male green sunfish, Lepomis cyanellus. Semen discharge was evoked consistently by low level electrical stimulation (100 muA or less) in the following areas: the preoptic region, dorsal hypothalamus, thalamus, midbrain tegmentum and the basolateral midbrain and medulla. Areas which were stimulated repeatedly at 100 muA and were always negative for sperm release included: the telencephalon with the exception of the preoptic region, the optic tectum, the cerebellum, the inferior lobe of the hypothalamus, the nucleus rotundus and the dorsal medulla. Removal of most of the optic tectum and cerebellum failed to block reponses evoked from the preoptic area; however, they were usually eliminated by transecting the rostral spinal cord. Electrical stimulation of an isolated 4 mm segment of spinal cord located at the third vertebral level resulted in sperm release, indicating that adequate mechanisms for semen discharge are present within the upper spinal cord. The results of this study suggest that a sperm release system in the green sunfish extends from the preoptic area to the spinal cord passing through the hypothalamus, midbrain tegmentum and basal midbrain and medulla. PMID:1113069

  1. Leukocytospermia and sperm preparation - a flow cytometric study

    PubMed Central

    2009-01-01

    Background Leukocytes represent the predominant source of reactive oxygen species both in seminal plasma and in sperm suspensions and have been demonstrated to negatively influence sperm function and fertilization rate in assisted reproduction procedures. Peroxidase test is the standard method recommended by WHO to detect semen leukocytes but it may be inaccurate. The aims of this study were (i) to compare the efficiency of swim-up and density-gradient centrifugation techniques in removing seminal leukocytes, (ii) to examine the effect of leukocytes on sperm preparation, and (iii) to compare flow cytometry and peroxidase test in determining leukocyte concentration in semen using a multiparameter flow cytometric method. Methods Semen samples from 126 male partners of couples undergoing infertility investigations were analyzed for leukocytospermia using standard optical microscopy and flow cytometry. Sixty-nine out of 126 samples were also processed using simultaneously the swim-up and density-gradient centrifugation techniques. A multiparameter flow cytometric analysis to assess simultaneously sperm concentration, sperm viability, sperm apoptosis, and leukocyte concentration was carried out on neat and prepared sperm. Results Both sperm preparation methods removed most seminal leukocytes. However, the concentration of leukocytes was significantly lower after swim-up compared to that after density-gradient centrifugation preparation. Leukocytes concentration, either initial or in prepared fractions, was not correlated with sperm parameters (optical microscopy and flow cytometry parameters) after semen processing. There was no correlation between leukocyte concentration in the ejaculate and sperm recovery rate, whereas a significant correlation was found between the concentration of the residual leukocytes in prepared fractions and viable sperm recovery rate. Although the overall concordance between the flow cytometry and the optical microscopy was satisfactory, the sensitivity of peroxidase test for the detection of leukocytospermia resulted low. Conclusion Seminal leukocytes do not seem to influence sperm preparation results. However, for assisted conception, semen samples containing leukocytes should be processed using swim-up method. Although peroxidase-test is recommended by WHO as the standard method for determining semen leukocytes, it should not be used in clinical research study. PMID:19925640

  2. Linking sperm length and velocity: the importance of intramale variation

    PubMed Central

    Fitzpatrick, John L.; Garcia-Gonzalez, Francisco; Evans, Jonathan P.

    2010-01-01

    Selection imposed through sperm competition is commonly thought to promote the evolution of longer sperm, since sperm length is assumed to be positively associated with sperm swimming velocity. Yet, the basis for this assumption remains controversial, and there is surprisingly little intraspecific evidence demonstrating such a link between sperm form and function. Here, we show that sperm length and velocity are highly correlated in the sea urchin Heliocidaris erythrogramma, but importantly we report that failure to account for within-male variation in these sperm traits can obscure this relationship. These findings, in conjunction with the mounting evidence for extremely high levels of intra-specific variance in sperm traits, suggest that a functional link between sperm morphology and velocity may be more prevalent than what current evidence suggests. Our findings also suggest that selection for faster swimming sperm may promote the evolution of longer sperm, thereby supporting recent findings from macroevolutionary studies. PMID:20484233

  3. 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.

  4. Sperm storage in caecilian amphibians

    PubMed Central

    2012-01-01

    Background Female sperm storage has evolved independently multiple times among vertebrates to control reproduction in response to the environment. In internally fertilising amphibians, female salamanders store sperm in cloacal spermathecae, whereas among anurans sperm storage in oviducts is known only in tailed frogs. Facilitated through extensive field sampling following historical observations we tested for sperm storing structures in the female urogenital tract of fossorial, tropical caecilian amphibians. Findings In the oviparous Ichthyophis cf. kohtaoensis, aggregated sperm were present in a distinct region of the posterior oviduct but not in the cloaca in six out of seven vitellogenic females prior to oviposition. Spermatozoa were found most abundantly between the mucosal folds. In relation to the reproductive status decreased amounts of sperm were present in gravid females compared to pre-ovulatory females. Sperm were absent in females past oviposition. Conclusions Our findings indicate short-term oviductal sperm storage in the oviparous Ichthyophis cf. kohtaoensis. We assume that in female caecilians exhibiting high levels of parental investment sperm storage has evolved in order to optimally coordinate reproductive events and to increase fitness. PMID:22672478

  5. Structural test with functional characteristics

    Microsoft Academic Search

    B. Nadeau-Dosti; Muhiuzzaman Mahmud; Jean-François Côté; F. Maamaril

    2005-01-01

    A novel structural test clocking architecture is proposed to reproduce characteristics of the functional mode of operation that are essential to a high-quality test. Bursts of functional mode clocks are controlled to provide accurate timing tests taking into account functional timing constraints such as multi-cycle paths and false paths. Any mix of asynchronous and synchronous clocks is supported. Long term

  6. Sperm-Associated Antigen 6 (SPAG6) Deficiency and Defects in Ciliogenesis and Cilia Function: Polarity, Density, and Beat

    PubMed Central

    Teves, Maria E.; Sears, Patrick R.; Li, Wei; Zhang, Zhengang; Tang, Waixing; van Reesema, Lauren; Costanzo, Richard M.; Davis, C. William; Knowles, Michael R.; Strauss, Jerome F.; Zhang, Zhibing

    2014-01-01

    SPAG6, an axoneme central apparatus protein, is essential for function of ependymal cell cilia and sperm flagella. A significant number of Spag6-deficient mice die with hydrocephalus, and surviving males are sterile because of sperm motility defects. In further exploring the ciliary dysfunction in Spag6-null mice, we discovered that cilia beat frequency was significantly reduced in tracheal epithelial cells, and that the beat was not synchronized. There was also a significant reduction in cilia density in both brain ependymal and trachea epithelial cells, and cilia arrays were disorganized. The orientation of basal feet, which determines the direction of axoneme orientation, was apparently random in Spag6-deficient mice, and there were reduced numbers of basal feet, consistent with reduced cilia density. The polarized epithelial cell morphology and distribution of intracellular mucin, ?-tubulin, and the planar cell polarity protein, Vangl2, were lost in Spag6-deficient tracheal epithelial cells. Polarized epithelial cell morphology and polarized distribution of ?-tubulin in tracheal epithelial cells was observed in one-week old wild-type mice, but not in the Spag6-deficient mice of the same age. Thus, the cilia and polarity defects appear prior to 7 days post-partum. These findings suggest that SPAG6 not only regulates cilia/flagellar motility, but that in its absence, ciliogenesis, axoneme orientation, and tracheal epithelial cell polarity are altered. PMID:25333478

  7. 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.

  8. Phenotypic engineering of sperm-production rate confirms evolutionary predictions of sperm competition theory

    PubMed Central

    Sekii, Kiyono; Vizoso, Dita B.; Kuales, Georg; De Mulder, Katrien; Ladurner, Peter; Schärer, Lukas

    2013-01-01

    Sperm production is a key male reproductive trait and an important parameter in sperm competition theory. Under sperm competition, paternity success is predicted to depend strongly on male allocation to sperm production. Furthermore, because sperm production is inherently costly, individuals should economize in sperm expenditure, and conditional adjustment of the copulation frequency according to their sperm availability may be expected. However, experimental studies showing effects of sperm production on mating behaviour and paternity success have so far been scarce, mainly because sperm production is difficult to manipulate directly in animals. Here, we used phenotypic engineering to manipulate sperm-production rate, by employing dose-dependent RNA interference (RNAi) of a spermatogenesis-specific gene, macbol1, in the free-living flatworm Macrostomum lignano. We demonstrate (i) that our novel dose-dependent RNAi approach allows us to induce high variability in sperm-production rate; (ii) that a reduced sperm-production rate is associated with a decreased copulation frequency, suggesting conditional adjustment of mating behaviour; and (iii) that both sperm production and copulation frequency are important determinants of paternity success in a competitive situation, as predicted by sperm competition theory. Our study clearly documents the potential of phenotypic engineering via dose-dependent RNAi to test quantitative predictions of evolutionary theory. PMID:23446521

  9. 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.

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

    PubMed

    Blanco, Juan M; Long, Julie A; Gee, George; Donoghue, Ann M; Wildt, David E

    2008-02-01

    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 degrees C versus 21 degrees C; (2) pre-equilibrating with dimethylacetamide (DMA) or dimethylsulfoxide (Me2SO) at either 4 degrees C or 21 degrees 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 degrees 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. PMID:18005955

  11. Functions and Vertical Line Test

    NSDL National Science Digital Library

    2010-01-01

    This lesson is designed to introduce students to the vertical line test for functions as well as practice plotting points and drawing simple functions. The lesson provides links to discussions and activities related to the vertical line test and functions as well as suggested ways to integrate them into the lesson.

  12. Evaluating the function of calcium antagonist on the Cd-induced stress in sperm of Russian sturgeon, Acipenser gueldenstaedtii.

    PubMed

    Li, Zhi-Hua; Li, Ping; Rodina, Marek; Randak, Tomas

    2010-11-15

    In the current study, the sperm of Russian sturgeon (Acipenser gueldenstaedtii) was used to evaluate the roles of Verapamil (VRP), a calcium channel blocker, against cadmium (Cd)-induced stress. Sturgeon sperm were exposed for 2h at 50?g/L VRP, 5.0?g/L Cd, the mixture of 50?g/L VRP+5.0?g/L Cd, 50?g/L Cd and the mixture of 50?g/L VRP+50?g/L Cd. After exposure, the sperm motility parameters (motility and velocity), oxidative stress levels (lipid peroxidation [LPO] and carbonyl protein [CP]) and antioxidant enzyme activities (superoxide dismutase [SOD], glutathione reductase [GR], glutathione peroxidase [GPx]) were measured in sturgeon sperm. Compared to the control, Cd-induced stress was apparent as reflected by depressed motility parameters, induced oxidative stress and inhibited antioxidant enzyme activities at both Cd concentrations. In the presence of VRP, Cd-induced stress was reduced in sturgeon sperm, especially all the measured parameters in the sperm exposed at 5.0?g/L Cd returned to control levels, expect for the sperm motility. The present results indicate that VRP can reduce the Cd-induced stress in sturgeon sperm and suggest that using of sperm in vitro assays may provide a novel and efficient means for evaluating the effects of residual metals in the aquatic environment of sturgeon. PMID:20832874

  13. Why small males have big sperm: dimorphic squid sperm linked to alternative mating behaviours

    PubMed Central

    2011-01-01

    Background Sperm cells are the target of strong sexual selection that may drive changes in sperm structure and function to maximize fertilisation success. Sperm evolution is regarded to be one of the major consequences of sperm competition in polyandrous species, however it can also be driven by adaptation to the environmental conditions at the site of fertilization. Strong stabilizing selection limits intra-specific variation, and therefore polymorphism, among fertile sperm (eusperm). Here we analyzed reproductive morphology differences among males employing characteristic alternative mating behaviours, and so potentially different conditions of sperm competition and fertilization environment, in the squid Loligo bleekeri. Results Large consort males transfer smaller (average total length = 73 ?m) sperm to a female's internal sperm storage location, inside the oviduct; whereas small sneaker males transfer larger (99 ?m) sperm to an external location around the seminal receptacle near the mouth. No significant difference in swimming speed was observed between consort and sneaker sperm. Furthermore, sperm precedence in the seminal receptacle was not biased toward longer sperm, suggesting no evidence for large sperm being favoured in competition for space in the sperm storage organ among sneaker males. Conclusions Here we report the first case, in the squid Loligo bleekeri, where distinctly dimorphic eusperm are produced by different sized males that employ alternative mating behaviours. Our results found no evidence that the distinct sperm dimorphism was driven by between- and within-tactic sperm competition. We propose that presence of alternative fertilization environments with distinct characteristics (i.e. internal or external), whether or not in combination with the effects of sperm competition, can drive the disruptive evolution of sperm size. PMID:21831296

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

    PubMed

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

    2014-09-15

    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

  15. Liver function tests

    MedlinePLUS

    ... laboratory results. In: McPherson RA, Pincus MR, eds. Henry's Clinical Diagnosis and Management by Laboratory Methods . 22nd ... liver function. In: McPherson RA, Pincus MR, eds. Henry's Clinical Diagnosis and Management by Laboratory Methods . 22nd ...

  16. 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.

  17. Pulmonary Function Tests

    MedlinePLUS

    ... tests to find out how well you move air in and out of your lungs and how well oxygen enters your body. The most common PFT’s are spirometry (spy-RAH-me-tree), diffusion studies and body plethysmography (ple-thiz-MA- ...

  18. Mating behavior and the evolution of sperm design

    PubMed Central

    Schärer, Lukas; Littlewood, D. Timothy J.; Waeschenbach, Andrea; Yoshida, Wataru; Vizoso, Dita B.

    2011-01-01

    Sperm are the most diverse of all animal cell types, and much of the diversity in sperm design is thought to reflect adaptations to the highly variable conditions under which sperm function and compete to achieve fertilization. Recent work has shown that these conditions often evolve rapidly as a consequence of multiple mating, suggesting a role for sexual selection and sexual conflict in the evolution of sperm design. However, very little of the striking diversity in sperm design is understood functionally, particularly in internally fertilizing organisms. We use phylogenetic comparative analyses covering 16 species of the hermaphroditic flatworm genus Macrostomum to show that a complex sperm design is associated with reciprocal mating and that this complexity is lost secondarily when hypodermic insemination—sperm injection through the epidermis—evolves. Specifically, the complex sperm design, which includes stiff lateral bristles, is likely a male persistence trait associated with sexual conflicts over the fate of received ejaculates and linked to female resistance traits, namely an intriguing postcopulatory sucking behavior and a thickened epithelium of the sperm-receiving organ. Our results suggest that the interactions between sperm donor, sperm, and sperm recipient can change drastically when hypodermic insemination evolves, involving convergent evolution of a needle-like copulatory organ, a simpler sperm design, and a simpler female genital morphology. Our study documents that a shift in the mating behavior may alter fundamentally the conditions under which sperm compete and thereby lead to a drastic change in sperm design. PMID:21220334

  19. Markers of renal function tests

    PubMed Central

    Gowda, Shivaraj; Desai, Prakash B.; Kulkarni, Shruthi S.; Hull, Vinayak V.; Math, Avinash A. K.; Vernekar, Sonal N.

    2010-01-01

    Background: The markers of renal function test assess the normal functioning of kidneys. These markers may be radioactive and non radioactive. They indicate the glomerular filtration rate, concentrating and diluting capacity of kidneys (tubular function). If there is an increase or decrease in the valves of these markers it indicates dysfunction of kidney. Aim: The aim of this review is to compare and analyze the present and newer markers of renal function tests which help in diagnosis of clinical disorders. Material & Methods: An extensive literature survey was done aiming to compare and compile renal function tests makers required in diagnosis of diseases. Results: Creatinine, urea, uric acid and electrolytes are makers for routine analysis whereas several studies have confirmed and consolidated the usefulness of markers such as cystatin C and ?-Trace Protein. Conclusion: We conclude that further investigation is necessary to define these biomarkers in terms of usefulness in assessing renal function. PMID:22624135

  20. Synergistic effect of gramicidin and EDTA in inhibiting sperm motility and cervical mucus penetration in vitro

    Microsoft Academic Search

    Aldar S. Bourinbaiar; Chi-Hyun Lee

    1996-01-01

    Gramicidin, a linear polypeptide with antiviral and antimicrobial properties, was compared in vitro with a commonly used spermicidal detergent—nonoxynol-9 (N9). The inhibition of sperm functions was evaluated by computer-assisted semen analysis (CASA) for sperm motility, in cervical mucus penetration assay, and by colorimetric tetrazolium salt and lactate dehydrogenase release assays routinely employed for testing the toxicity of drugs. The effective

  1. Sperm competitiveness in frogs: slow and steady wins the race

    PubMed Central

    Dziminski, Martin A.; Roberts, J. Dale; Beveridge, Maxine; Simmons, Leigh W.

    2009-01-01

    When sperm compete to fertilize available ova, selection is expected to favour ejaculate traits that contribute to a male's fertilization success. While there is much evidence to show that selection favours increased numbers of sperm, only a handful of empirical studies have examined how variation in sperm form and function contributes to competitive fertilization success. Here, we examine selection acting on sperm form and function in the externally fertilizing myobatrachid frog, Crinia georgiana. Using in vitro fertilization techniques and controlling for variation in the number of sperm contributed by males in competitive situations, we show that males with a greater proportion of motile sperm, and motile sperm with slower swimming velocities, have an advantage when competing for fertilizations. Sperm morphology and the degree of genetic similarity between putative sires and the female had no influence on competitive fertilization success. These unusual patterns of selection might explain why frog sperm typically exhibit relatively slow swimming speeds and sustained longevity. PMID:19710059

  2. Ejaculated mouse sperm enter cumulus-oocyte complexes more efficiently in vitro than epididymal sperm.

    PubMed

    Li, Honggang; Hung, Pei-Hsuan; 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

  3. 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

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

    Microsoft Academic Search

    Juan M. Blanco; Julie A. Long; George Gee; Ann M. Donoghue; David E. Wildt

    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 (300–3000mOsm\\/kg) was evaluated after: (1) equilibration times of 2, 10, 45 and 60min 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

  5. Investigation of the association between the outcomes of sperm chromatin condensation and decondensation tests, and assisted reproduction techniques.

    PubMed

    Irez, T; Sahmay, S; Ocal, P; Goymen, A; Senol, H; Erol, N; Kaleli, S; Guralp, O

    2015-05-01

    The main purpose of this prospective study is to examine possible influences of abnormalities of sperm nuclear condensation and chromatin decondensation with sodium dodecyl sulphate (SDS)-EDTA on outcomes of intrauterine insemination (IUI) or intracytoplasmic sperm injection (ICSI) cycles. Semen samples from 122 IUI and 236 ICSI cycles were evaluated. Before semen preparation for IUI or ICSI, basic semen analysis was performed and a small portion from each sample was spared for fixation. The condensation of sperm nuclear chromatin was evaluated with acidic aniline blue, followed by sperm chromatin decondensation by SDS-EDTA and evaluation under light microscope. Ongoing pregnancy rate was 24% and 26.2% in the IUI and ICSI groups respectively. The chromatin condensation rate was significantly higher in the ongoing pregnancy-positive group compared to the negative group, both in IUI (P = 0.042) and ICSI groups (P = 0.027), and it was positively correlated with ongoing pregnancy rate in both IUI and ICSI groups (P = 0.015, r = 0.214 and P = 0.014, r = 0.312 respectively). Chromatin decondensation rates were not significantly different in neither of the groups. These results indicate that IUI and ICSI outcome is influenced by the rate of spermatozoa with abnormal chromatin condensation. Sperm chromatin condensation with aniline blue is useful for selecting assisted reproduction techniques (ART) patients. PMID:24766543

  6. Female presence influences sperm velocity in the guppy

    PubMed Central

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

    2009-01-01

    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

  7. Alterations to the Bull Sperm Surface Proteins That Bind Sperm to Oviductal Epithelium1

    PubMed Central

    Hung, Pei-hsuan; Suarez, Susan S.

    2012-01-01

    ABSTRACT Three Binder of SPerm proteins (BSP1, BSP3, BSP5) are secreted by bovine seminal vesicles into seminal plasma and adsorbed onto sperm. When sperm inseminated into the female reach the oviduct, the BSP proteins bind them to its epithelial lining, forming a sperm storage reservoir. Previously, we reported that binding of capacitated sperm to oviductal epithelium in vitro is lower than that of uncapacitated sperm and we proposed that reduced binding was due to loss of BSP proteins during capacitation. Because of differences in amino acid sequences, we predicted that each BSP would respond differently to capacitating conditions. To test whether all three BSP proteins were lost from sperm during capacitation and whether the kinetics of loss differed among the three BSP proteins, ejaculated bull sperm were incubated under various capacitating conditions, and then the amounts of BSP proteins remaining on the sperm were assayed by Western blotting. Capacitation was assayed by analysis of protein tyrosine phosphorylation. While loss of BSP1 was not detected, most of the BSP5 was lost from sperm during incubation in TALP medium, even without addition of the capacitation enhancers heparin and dbcAMP-IBMX. Surprisingly, a smaller molecular mass was detected by anti-BSP3 antibodies in extracts of incubated sperm. Its identity was confirmed as BSP3 by mass spectrometry, indicating that BSP3 undergoes modification on the sperm surface. These changes in the composition of BSP proteins on sperm could play a role in releasing sperm from the storage reservoir by modifying sperm interactions with the oviductal epithelium. PMID:22837481

  8. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...purposes to identify and differentiate animal and human semen. The test results may be used as court evidence in alleged instances of rape and other sex-related crimes. (b) Classification. Class I (general controls). The device is exempt from the...

  9. 21 CFR 866.5800 - Seminal fluid (sperm) immunological test system.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...purposes to identify and differentiate animal and human semen. The test results may be used as court evidence in alleged instances of rape and other sex-related crimes. (b) Classification. Class I (general controls). The device is exempt from the...

  10. Functional role of feline zona pellucida protein 4 trefoil domain: a sperm receptor or structural component of the domestic cat zona pellucida?

    PubMed

    Braun, B C; Ringleb, J; Waurich, R; Viertel, D; Jewgenow, K

    2009-07-01

    The trefoil domain (TFD) is a protein structure characterized by six cysteines, which form a typical three-loop structure by three disulphide bridges. It is assumed that two of these loops generate a hydrophobic groove, which could be a binding site for carbohydrate residues or proteins. The zona pellucida (ZP) protein, ZP4, contains such a TFD. The carbohydrate-/protein-binding property of TFD allows us to assume a potential sperm receptor function of this domain. Additionally, gastrointestinal trefoil peptides are stable against proteases; therefore, a structural role of TFD within the ZP might also be possible. We were able to show that the synthesized and natural folded feline TFD (fTFD) expresses the typical protease resistance that vanished under reducing conditions and after substitution of cysteine residues within the peptide. Furthermore, an antibody directed against the first loop of fTFD was almost unable to bind to intact in vitro mature cat oocytes. Pre-incubation of oocytes in the reducing agent (DDT), however, improved antibody binding substantially. Therefore, we suggest structural masking of the fTFD domain within the intact ZP. An interaction between fTFD and feline sperm cells was examined using several methods, including immunocytochemistry, immunoelectron microscopy, co-immunoprecipitation and far western blot, but we found no indication for an involvement of TFD in the primary sperm binding to the ZP. To summarize, there is increasing evidence that the TFD of fZP4 has a structural rather than a sperm-binding function. PMID:19754576

  11. How to select the spermatozoon for intracytoplasmic sperm injection in 2015?

    PubMed

    Herbemont, C; Sifer, C

    2015-04-01

    The selection of the individual spermatozoon in intracytoplasmic sperm injection (ICSI) is routinely performed by the observation of its motility and morphology. However, in case of severe oligoasthenozoospermia or non-obstructive azoospermia requiring the use of testicular sperm, other methods are necessary to help the embryologist making this choice. According to some authors, sperm processing before ICSI seems to limit the DNA fragmentation index, and in this way improves ICSI outcomes. Moreover, intracytoplasmic morphologically selected sperm injection is potentially a good option in some specific indications such as severe teratozoospermia, or repeated ICSI failures. Other methods based on sperm structure, as sperm head birefringence observation, or based on its function, like the hyaluronic acid or zona pellucida binding capacity, could be of interest, but still need to be evaluated. Finally, in case of akinetozoospermia, the use of functional tests, such as pentoxifylline test, HOS-test, or to a lesser extent laser touch, makes the selection of viable spermatozoa easier. Nevertheless, studies on larger series have to be conducted to evaluate and precise the interest of each of these methods and their indications, before considering an application on larger scale. PMID:25581325

  12. Sperm DNA fragmentation, recurrent implantation failure and recurrent miscarriage.

    PubMed

    Coughlan, Carol; Clarke, Helen; Cutting, Rachel; Saxton, Jane; Waite, Sarah; Ledger, William; Li, Tinchiu; Pacey, Allan A

    2015-01-01

    Evidence is increasing that the integrity of sperm DNA may also be related to implantation failure and recurrent miscarriage (RM). To investigate this, the sperm DNA fragmentation in partners of 35 women with recurrent implantation failure (RIF) following in vitro fertilization, 16 women diagnosed with RM and seven recent fathers (control) were examined. Sperm were examined pre- and post-density centrifugation by the sperm chromatin dispersion (SCD) test and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. There were no significant differences in the age of either partner or sperm concentration, motility or morphology between three groups. Moreover, there were no obvious differences in sperm DNA fragmentation measured by either test. However, whilst on average sperm DNA fragmentation in all groups was statistically lower in prepared sperm when measured by the SCD test, this was not seen with the results from the TUNEL assay. These results do not support the hypothesis that sperm DNA fragmentation is an important cause of RIF or RM, or that sperm DNA integrity testing has value in such patients. It also highlights significant differences between test methodologies and sperm preparation methods in interpreting the data from sperm DNA fragmentation tests. PMID:25814156

  13. How nematode sperm crawl

    Microsoft Academic Search

    Dean Bottino; Alexander Mogilner; Tom Roberts; Murray Stewart; George Oster

    Sperm of the nematode, Ascaris suum, crawl using lamellipodial protrusion, adhesion and retraction, a process analogous to the amoeboid motility of other eukaryotic cells. However, rather than employing an actin cytoskeleton to generate locomotion, nematode sperm use the major sperm protein (MSP). Moreover, nematode sperm lack detectable molecular motors or the battery of actin-binding proteins that characterize actin-based motility. The

  14. Competition drives cooperation among closely related sperm of deer mice

    E-print Network

    Hoekstra, Hopi E.

    with unrelated conspecific sperm. In contrast, in the highly promiscuous deer mouse, Peromyscus maniculatus of deer mice (genus Peromyscus) form motile aggregations, then we use this system to test predictions. In a monogamous species lacking sperm competition, Peromyscus polionotus, sperm indiscrimi- nately group

  15. Sperm maturation in the domestic cat

    Microsoft Academic Search

    Eva Axnér

    2006-01-01

    The epididymis is essential for sperm development and maturation, and, subsequently, the ability of spermatozoa to penetrate and fertilize the female gamete. Functional differences in segments of the long tubule are reflected by histological differences among epididymal regions. The feline epididymis can be divided into six different regions according to their histological differences. A marked increase in sperm concentration occurs

  16. Sperm competition and ejaculate investment in red squirrels ( Tamiasciurus hudsonicus )

    Microsoft Academic Search

    Vanessa L. Bonanno; Albrecht I. Schulte-Hostedde

    2009-01-01

    Sperm competition is widespread in mammals and occurs when ejaculates from two or more males compete within the female’s reproductive\\u000a tract to fertilize the ova. Enlarged testes are associated with sperm competition because they produce sperm, but the accessory\\u000a glands produce fluids and proteins that are also important for fertilization success. Sperm morphology can also have consequences\\u000a for fertilization success

  17. Characterisation of the Manduca sexta sperm proteome: Genetic novelty underlying sperm composition in Lepidoptera.

    PubMed

    Whittington, Emma; Zhao, Qian; Borziak, Kirill; Walters, James R; Dorus, Steve

    2015-07-01

    The application of mass spectrometry based proteomics to sperm biology has greatly accelerated progress in understanding the molecular composition and function of spermatozoa. To date, these approaches have been largely restricted to model organisms, all of which produce a single sperm morph capable of oocyte fertilisation. Here we apply high-throughput mass spectrometry proteomic analysis to characterise sperm composition in Manduca sexta, the tobacco hornworm moth, which produce heteromorphic sperm, including one fertilisation competent (eupyrene) and one incompetent (apyrene) sperm type. This resulted in the high confidence identification of 896 proteins from a co-mixed sample of both sperm types, of which 167 are encoded by genes with strict one-to-one orthology in Drosophila melanogaster. Importantly, over half (55.1%) of these orthologous proteins have previously been identified in the D. melanogaster sperm proteome and exhibit significant conservation in quantitative protein abundance in sperm between the two species. Despite the complex nature of gene expression across spermatogenic stages, a significant correlation was also observed between sperm protein abundance and testis gene expression. Lepidopteran-specific sperm proteins (e.g., proteins with no homology to proteins in non-Lepidopteran taxa) were present in significantly greater abundance on average than those with homology outside the Lepidoptera. Given the disproportionate production of apyrene sperm (96% of all mature sperm in Manduca) relative to eupyrene sperm, these evolutionarily novel and highly abundant proteins are candidates for possessing apyrene-specific functions. Lastly, comparative genomic analyses of testis-expressed, ovary-expressed and sperm genes identified a concentration of novel sperm proteins shared amongst Lepidoptera of potential relevance to the evolutionary origin of heteromorphic spermatogenesis. As the first published Lepidopteran sperm proteome, this whole-cell proteomic characterisation will facilitate future evolutionary genetic and developmental studies of heteromorphic sperm production and parasperm function. Furthermore, the analyses presented here provide useful annotation information regarding sex-biased gene expression, novel Lepidopteran genes and gene function in the male gamete to complement the newly sequenced and annotated Manduca genome. PMID:25731083

  18. DNA fragmentation and sperm head morphometry in cat epididymal spermatozoa.

    PubMed

    Vernocchi, Valentina; Morselli, Maria Giorgia; Lange Consiglio, Anna; Faustini, Massimo; Luvoni, Gaia Cecilia

    2014-10-15

    Sperm DNA fragmentation is an important parameter to assess sperm quality and can be a putative fertility predictor. Because the sperm head consists almost entirely of DNA, subtle differences in sperm head morphometry might be related to DNA status. Several techniques are available to analyze sperm DNA fragmentation, but they are labor-intensive and require expensive instrumentations. Recently, a kit (Sperm-Halomax) based on the sperm chromatin dispersion test and developed for spermatozoa of different species, but not for cat spermatozoa, became commercially available. The first aim of the present study was to verify the suitability of Sperm-Halomax assay, specifically developed for canine semen, for the evaluation of DNA fragmentation of epididymal cat spermatozoa. For this purpose, DNA fragmentation indexes (DFIs) obtained with Sperm-Halomax and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) were compared. The second aim was to investigate whether a correlation between DNA status, sperm head morphology, and morphometry assessed by computer-assisted semen analysis exists in cat epididymal spermatozoa. No differences were observed in DFIs obtained with Sperm-Halomax and TUNEL. This result indicates that Sperm-Halomax assay provides a reliable evaluation of DNA fragmentation of epididymal feline spermatozoa. The DFI seems to be independent from all the measured variables of sperm head morphology and morphometry. Thus, the evaluation of the DNA status of spermatozoa could effectively contribute to the completion of the standard analysis of fresh or frozen semen used in assisted reproductive technologies. PMID:25129872

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

    Microsoft Academic Search

    Andrea Pilastro; Marta Scaggiante; Maria B. Rasotto

    2002-01-01

    Sperm competition theory predicts that males should strategically allocate their sperm reserves according to the level of sperm competition, defined as the probability that the sperm of two males compete for fertilizing a given set of ova. Substantial evidence from numerous animal taxa suggests that, at the individual level, sperm expenditure increases when the risk of sperm competition is greater.

  20. 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

  1. Characterization of sperm motility in sea bass: the effect of heavy metals and physicochemical

    E-print Network

    Villefranche sur mer

    Characterization of sperm motility in sea bass: the effect of heavy metals and physicochemical of several physicochemical variables and heavy metals on sperm swimming performance. Duration of sperm. Two of the heavy metals tested, Cu2þ and Pb2þ , did not affect sperm motility when the activating

  2. Sperm from tw32/+ mice: capacitation is normal, but hyperactivation is premature and nonhyperactivated sperm are slow.

    PubMed

    Olds-Clarke, P

    1989-02-01

    The t complex in the mouse is a large group of linked genes that affect sperm function in fertilization. In t/+ males, sperm carrying the t complex (t sperm) have normal fertilizing ability, while sperm carrying the normal homolog (+t sperm) are dysfunctional (P. Olds-Clarke and B. Peitz, 1985, Genet. Res. 47, 49). The specific step in fertilization which is dysfunctional, however, is not known. Two characteristics of fertilizing sperm, capacitation (the process by which sperm become capable of undergoing the acrosome reaction) and hyperactivation (the change in swimming behavior concomitant with capacitation), were assayed by objective methods in epididymal sperm from tw32/+ males of two strains, and compared to sperm from +/+ males of the same strains. Capacitated and acrosome-reacted sperm were identified by a chlortetracycline assay (C.R. Ward and B.T. Storey, 1984, Dev. Biol. 104, 287). Hyperactivated sperm were identified by their path shape and swimming speed, using a computer-assisted motion-analysis system (J.M. Neill and P. Olds-Clarke, 1987, Gamete Res. 18, 121). Hyperactivation occurred significantly sooner among sperm from tw32/+ mice than among sperm from +/+ mice of the same strain, while the rates and maximal levels of capacitation and spontaneous acrosome reactions were normal. Of the nonhyperactivated motile sperm from tw32/+ mice, almost all were slower than sperm from +/+ mice of the same strain. While the effect of premature hyperactivation on fertilization is not clear, slow movements are likely to impair fertilizing ability. These results raise the possibility that the slow sperm are the dysfunctional +t sperm. PMID:2912803

  3. A Systematic Analysis of a Deep Mouse Epididymal Sperm Proteome1

    PubMed Central

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

    2012-01-01

    ABSTRACT 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 therefore is theoretically possible to understand the protein networks contained in a sperm cell and to deduce its cellular function capabilities. To this end, we performed a proteomic analysis of mouse sperm isolated from the cauda epididymis and confidently identified 2850 proteins, which to our knowledge 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, beta-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. PMID:23115268

  4. A novel acrosomal protein, IQCF1, involved in sperm capacitation and the acrosome reaction.

    PubMed

    Fang, P; Xu, W; Li, D; Zhao, X; Dai, J; Wang, Z; Yan, X; Qin, M; Zhang, Y; Xu, C; Wang, L; Qiao, Z

    2015-03-01

    On the basis of the unknown tags in the mature human sperm serial analysis of gene expression library constructed by our laboratory, some transcripts were cloned, including Iqcf1 (IQ motif containing F1). To investigate the function of sperm-retained Iqcf1 in spermatogenesis and fertilization of mice, we investigated the spatial and temporal expression of IQCF1. By using the (transcription activator-like effector nuclease) strategy, Iqcf1-knockout mice were produced, and the phenotypes of the Iqcf1(-/-) mice were analyzed. The results showed that IQCF1 was localized in the acrosome of spermatozoa and spermatids; the expression of IQCF1 in testes was associated with spermatogenic capacity. The Iqcf1(-/-) mice were significantly less fertile than the wild-type mice (p = 0.0057) because of reduced sperm motility (p = 0.0094) and the acrosome reaction (AR) (p = 0.0093). In spermatozoa, IQCF1 interacted with calmodulin (CaM) and possibly participated in the tyrosine phosphorylation of sperm proteins during capacitation. In conclusion, a newly identified acrosomal protein, IQCF1, is closely related to sperm capacitation and AR; in particular, it is involved in tyrosine phosphorylation of sperm proteins through interaction with CaM. Research into the function of IQCF1 during fertilization could facilitate the investigation of the molecular mechanism of capacitation, which is unclear. PMID:25380116

  5. Evaluation of maize grain and polyunsaturated fatty acid (PUFA) as energy sources for breeding rams based on hormonal, sperm functional parameters and fertility.

    PubMed

    Selvaraju, Sellappan; Raju, Priyadarshini; Rao, Somu Bala Nageswara; Raghavendra, Subbarao; Nandi, Sumantha; Dineshkumar, Dhanasekaran; Thayakumar, Allen; Parthipan, Shivashanmugam; Ravindra, Janivara Parameswaraiah

    2012-01-01

    The objective of the present study was to elucidate the effect of different sources of dietary energy (maize vs polyunsaturated fatty acid (PUFA) on semen functional parameters and fertility of adult rams. Eighteen adult rams were divided into two groups (maize and PUFA, n=9). The main energy source for the rams in the maize group was coarsely ground maize grain, whereas in the PUFA group it was sunflower oil (rich in 18:2 linoleic acid, an omega-6 acid). The ration was fed for a minimum period of 60 days and thereafter semen was collected for evaluation. The proportion of progressive forward motility was significantly (P<0.05) higher in the PUFA group compared with the maize group. Sperm lipid peroxidation as measured by malondialdehyde formation (µM per 1×10(9) spermatozoa) was significantly (P<0.05) higher in the PUFA group compared with the maize group. When the semen was diluted with Tris-egg yolk-citrate buffer and incubated for 24h at 4°C, the proportions of plasmalemma integrity, the sperm subpopulation positive for functional membrane and acrosomal integrities, and mitochondrial membrane potential were significantly (P<0.05) higher in PUFA-fed than in maize-fed animals. The different sources of energy did not influence the serum and seminal plasma IGF-I levels. The cleavage rate (percentage) did not differ significantly between PUFA- (45.4±4.91) and maize- (44.63±6.8) fed animals. In conclusion, PUFA feeding influenced sperm quality by altering or stabilising membrane integrity. The present study indicates that PUFA may improve semen quality but did not improve in vitro fertilisation. PMID:22697117

  6. EVIDENCE FAVORING SPERM SELECTION OVER SPERM COMPETITION IN THE INTERACTION BETWEEN HUMAN SEMINAL PLASMA AND SPERM MOTILITY IN VITRO

    Microsoft Academic Search

    K. Jaffe; M. I. Camejo; T. E. Carrillo; M. Weffer; M. G. Muñoz

    2006-01-01

    & The effect on sperm motility of sperm-sperm and sperm-seminal plasma interactions was stud- ied among homologous and heterologous sperm. There were no significant interactions between sperm in vitro, but it was found that seminal plasmas of different donors have different effects on sperm motility, and different sperm react differently to the same seminal plasma. Sperm showed higher motility in

  7. Sperm chromatin structure assay (SCSA®).

    PubMed

    Evenson, Donald P

    2013-01-01

    The SCSA(®) is the pioneering assay for the detection of damaged sperm DNA and altered proteins in sperm nuclei via flow cytometry of acridine orange (AO) stained sperm. The SCSA(®) is considered to be the most precise and repeatable test providing very unique, dual parameter data (red vs. green fluorescence) on a 1,024 × 1,024 channel scale, not only on DNA fragmentation but also on abnormal sperm characterized by lack of normal exchange of histones to protamines. Raw semen/sperm aliquots or purified sperm can be flash frozen, placed in a box with dry ice and shipped by overnight courier to an experienced SCSA(®) lab. The samples are individually thawed, prepared, and analyzed in ?10 min. Of significance, data on 5,000 individual sperm are recorded on a 1,024 × 1,024 dot plot of green (native DNA) and red (broken DNA) fluorescence. Repeat measurements have virtually identical dot plot patterns demonstrating that the low pH treatment that opens up the DNA strands at the sites of breaks and staining by acridine orange (AO) are highly precise and repeatable (CVs of 1-3%) and the same between fresh and frozen samples. SCSAsoft(®) software transforms the X-Y data to total DNA stainability versus red/red + green fluoresence (DFI) providing a more accurate determination of % DFI as well as the more sensitive value of standard deviation of DFI (SD DFI) as demonstrated by animal fertility and dose-response toxicology studies. The current established clinical threshold is 25% DFI for placing a man into a statistical probability of the following: (a) longer time to natural pregnancy, (b) low odds of IUI pregnancy, (c) more miscarriages, or (d) no pregnancy. Changes in lifestyle as well as medical intervention can lower the %DFI to increase the probability of natural pregnancy. Couples of men with >25% DFI are counseled to try ICSI and when in the >50% range may consider TESE/ICSI. The SCSA(®) simultaneously determines the % of sperm with high DNA stainability (%HDS) related to retained nuclear histones consistent with immature sperm; high HDS values are predictive of pregnancy failure.The SCSA(®) is considered to be the most technician friendly, time- and cost-efficient, precise and repeatable DNA fragmentation assay, with the most data and the only fragmentation assay with an accepted clinical threshold for placing a man at risk for infertility. SCSA(®) data are more predictive of male factor infertility than classical semen analyses. PMID:22992911

  8. Methods for human and murine sperm assays

    SciTech Connect

    Wyrobek, A.J.

    1980-01-01

    Human sperm assays in the context of an epidemiological study provide an effective means to identify agents that affect testicular function. Four assays for human sperm anomaly (counts, motility, morphology, and double-fluorescence bodies) have been successfully used for a variety of human occupational and drug-related exposures. However, more research is needed to validate the genetic implications of sperm defects induced by exposure to chemicals in vivo. The F/sub 0/ and F/sub 1/ mouse-sperm assay, especially for induced morphological defects, provides a method of screening agents that may be potentially harmful to man and provides a means to assess the antifertility effects and consequences of induced sperm defect.

  9. Functional significance of the outer dense fibers of mammalian sperm examined by computer simulations with the geometric clutch model.

    PubMed

    Lindemann, C B

    1996-01-01

    The flagella of mammalian sperm possess certain structural characteristics that distinguish them from simple flagella. Most notable of these features are the sheath (surrounding the axoneme), the outer dense fibers of ODFs (that are attached to the outer doublets), and the connecting piece (which anchors the ODFs at the base of the flagellum). In this study, the significance of these specialized axonemal elements is explored. Their impact on microtubule sliding and force production within the axoneme is specifically analyzed. A working hypothesis is developed based on the premise that forces produced by interdoublet sliding are transferred to the ODFs. In this way, the torque required to bend the flagellum is developed between the ODFs, which are anchored in the connecting piece. This working hypothesis was incorporated into the pre-existing "geometric clutch" model that earlier simulated only cilia and simple flagella. The characteristic length and stiffness of bovine sperm flagella were specified as modelling parameters. Additionally, the inter-ODF spacing of bull sperm was incorporated to calculate doublet sliding and bending torque. The resultant computer-simulated pattern of flagellar beating possesses many of the attributes of the beat of a live bull sperm flagellum. Notably, this life-like simulation can be produced using parameters for the central axonemal "motor" that are comparable to those effective in modelling a simple flagellum. In the proposed scheme, the accessory structures of the mammalian sperm axoneme provide increased stiffness while at the same time providing a means to proportionately raise the bending torque to overcome that additional flexural rigidity. This capacity is due to the inter-ODF distances being larger than the corresponding interdoublet spacings. If force is transmitted to the flagellar base by way of the ODFs, then the larger effective diameter generates both a greater bending torque and increased interdoublet sliding. This has the interesting effect of consolidating the energy from more dynein cross-bridges into the production of a single bend. Consequently. greater bending torque development is permitted than would be possible in a simple flagellum. In This way, the same 9 + 2 organization of a simple flagellum can power a much larger (and stiffer) version than would otherwise be possible. PMID:8871813

  10. Sperm output of older men

    Microsoft Academic Search

    K. K. Ng; R. Donat; L. Chan; A. Lalak; I. Di Pierro; D. J. Handelsman

    2004-01-01

    BACKGROUND: Declining fertility of couples from the fourth decade of life is largely attributable to the drop in female fertility. However, increasing numbers of men, whose fertility theoretically lasts until death, are seeking fer- tility treatment at older ages, yet there is little information on sperm production and function past the age of 50 years. The few studies of such

  11. Discriminating males alter sperm production between species.

    PubMed

    Aspbury, Andrea S; Gabor, Caitlin R

    2004-11-01

    Prezygotic reproductive isolation and its importance in speciation is traditionally approached from the viewpoint of those events that occur before mating. However, recent interest in sperm competition theory has shown that prezygotic isolation can be affected by mechanisms that occur after mating but before fertilization. One neglected aspect of these studies is how the cost of sperm production might play a role in species isolation. We examined differential sperm production in a species whose males are sexually parasitized by a unisexual gynogenetic species. Gynogens are clonal females that require sperm from males of closely related bisexual species to initiate embryogenesis. We tested for differential sperm production by male sailfin mollies (Poecilia latipinna) when they were in the presence of either a heterospecific, gynogenetic female (Poecilia formosa, Amazon molly) or a conspecific female. We found that previously demonstrated male mate choice for conspecific over heterospecific females also is revealed in sperm production. Males from both an allopatric and a sympatric population produce more sperm when in the presence of a conspecific female than when in the presence of a heterospecific female. We suggest that differential sperm production also could play a role in prezygotic reproductive isolation in bisexual species complexes that occur in sympatry. PMID:15520395

  12. Discriminating males alter sperm production between species

    PubMed Central

    Aspbury, Andrea S.; Gabor, Caitlin R.

    2004-01-01

    Prezygotic reproductive isolation and its importance in speciation is traditionally approached from the viewpoint of those events that occur before mating. However, recent interest in sperm competition theory has shown that prezygotic isolation can be affected by mechanisms that occur after mating but before fertilization. One neglected aspect of these studies is how the cost of sperm production might play a role in species isolation. We examined differential sperm production in a species whose males are sexually parasitized by a unisexual gynogenetic species. Gynogens are clonal females that require sperm from males of closely related bisexual species to initiate embryogenesis. We tested for differential sperm production by male sailfin mollies (Poecilia latipinna) when they were in the presence of either a heterospecific, gynogenetic female (Poecilia formosa, Amazon molly) or a conspecific female. We found that previously demonstrated male mate choice for conspecific over heterospecific females also is revealed in sperm production. Males from both an allopatric and a sympatric population produce more sperm when in the presence of a conspecific female than when in the presence of a heterospecific female. We suggest that differential sperm production also could play a role in prezygotic reproductive isolation in bisexual species complexes that occur in sympatry. PMID:15520395

  13. 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.

  14. Direct action of endocrine disrupting chemicals on human sperm

    PubMed Central

    Schiffer, Christian; Müller, Astrid; Egeberg, Dorte L; Alvarez, Luis; Brenker, Christoph; Rehfeld, Anders; Frederiksen, Hanne; Wäschle, Benjamin; Kaupp, U Benjamin; Balbach, Melanie; Wachten, Dagmar; Skakkebaek, Niels E; Almstrup, Kristian; Strünker, Timo

    2014-01-01

    Synthetic endocrine disrupting chemicals (EDCs), omnipresent in food, household, and personal care products, have been implicated in adverse trends in human reproduction, including infertility and increasing demand for assisted reproduction. Here, we study the action of 96 ubiquitous EDCs on human sperm. We show that structurally diverse EDCs activate the sperm-specific CatSper channel and, thereby, evoke an intracellular Ca2+ increase, a motility response, and acrosomal exocytosis. Moreover, EDCs desensitize sperm for physiological CatSper ligands and cooperate in low-dose mixtures to elevate Ca2+ levels in sperm. We conclude that EDCs interfere with various sperm functions and, thereby, might impair human fertilization. PMID:24820036

  15. Human sperm DNA integrity in normal and abnormal semen samples and its correlation with sperm characteristics

    Microsoft Academic Search

    A. C. Varghese; F. M. Bragais; D. Mukhopadhyay; S. Kundu; M. Pal; A. K. Bhattacharyya; A. Agarwal

    2009-01-01

    Summary Reports indicate an increase in the incidence of DNA fragmentation in male factor infertility and its role in the outcome of assisted reproductive techniques (ART). However, reports are conflicting between the relationships of sperm DNA integrity with conventional semen parameters. We examined the relation- ship between conventional sperm parameters and DNA integrity using acridine orange (AO) test. The study

  16. 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?

  17. The future of computer-aided sperm analysis.

    PubMed

    Mortimer, Sharon T; Horst, Gerhard van der; 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

  18. 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

  19. Nickel deficiency diminishes sperm quantity and movement in rats.

    PubMed

    Yokoi, Katsuhiko; Uthus, Eric O; Nielsen, Forrest H

    2003-01-01

    Early studies on nickel essentiality with rats and goats indicated that nickel deprivation impaired reproductive performance. Nickel also has been found to influence cyclic nucleotide gated channels (CNG); these types of channels are important in sperm physiology. Thus, two experiments were conducted to test the hypothesis that nickel deficiency affects sperm physiology in a manner consistent with nickel having an essential function related to CNG channel functions. The experiments were factorially arranged with four treatment groups of eight weanling rats in each. In experiment 1, the treatments were supplemental dietary nickel of 0 and 1 mg/kg and N(omega)-nitro-L-arginine methyl ester (L-NAME, a nitric oxide synthase inhibitor) added to the drinking water (50 mg/100 mL) the last 3 wk of an 8-wk experiment. In experiment 2, the treatments were supplemental dietary nickel at 0 and 1 mg/kg and supplemental dietary sodium chloride (NaCl) at 0 and 80 g/kg. The NaCl and L-NAME variables were included to act as stressors affecting CNG channel activity. The basal diet contained per kilogram about 27 microg of nickel and 1 g of sodium. After 8 wk in experiment 1 and 16 wk in experiment 2, urine while fasting and testes and epididymis in both experiments, and seminal vesicles and prostates in experiment 2 were harvested for analysis. Nickel deprivation significantly decreased spermatozoa motility and density in the epididymides, epididymal transit time of spermatozoa, and testes sperm production rate. Nickel deficiency also significantly decreased the weights of the seminal vesicles and prostate glands. Excessive NaCl had no effect on sperm physiology; however, it decreased prostate gland weights. The findings support the hypothesis that nickel has an essential function that possibly could affect reproductive performance in higher animals, perhaps through affecting a CNG channel function. PMID:12835498

  20. Sperm competition, male prudence, and sperm-limited females (2002)

    Microsoft Academic Search

    Nina Wedell; Matthew Gage; Geoffrey Parker

    Sperm are produced in astronomical numbers compared with eggs, and there is good evidence that sperm competition is the force behind the evolution of many tiny sperm. However, sperm production inevitably has costs. Recent research shows that male ejaculate expenditure is dynamic in both time and space, and that males are sensitive to risks of sperm competition and can vary

  1. 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.

  2. Intra-specific variation of sperm length in the malaria vector Anopheles gambiae: males with shorter sperm have higher reproductive success

    PubMed Central

    Voordouw, Maarten J; Koella, Jacob C; Hurd, Hilary

    2008-01-01

    Background Intra-specific variation in sperm length influences male reproductive success in several species of insects. In males of the malaria vector Anopheles gambiae, sperm length is highly variable but the significance of this variation is unknown. Understanding what determines the reproductive success of male mosquitoes is critical for controlling malaria, and in particular for replacing natural populations with transgenic, malaria-resistant mosquitoes. Methods A laboratory population of A. gambiae males was tested for intra-specific variation in sperm length. A full-sib quantitative genetic design was used to test for a genetic component of sperm length in A. gambiae males and estimate its heritability. This study also tested for a relationship between sperm length and male reproductive success in A. gambiae. Male reproductive success was measured as the proportions of inseminated and ovipositing females. Results There was intra-specific variation of sperm length in A. gambiae. There was no significant genetic variation in sperm length and its heritability was low (h2 = 0.18) compared to other insects. Sperm length was correlated with male body size (measured as wing length). Males with short sperm had significantly higher reproductive success than males with long sperm and this was independent of body size. Conclusion This is the first study to demonstrate intra-specific variation in sperm length in A. gambiae and that males with short sperm have higher reproductive success. That sperm length influences female oviposition is important for any strategy considering the release of transgenic males. PMID:18939985

  3. 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

  4. 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

  5. Sexual rest and post-meiotic sperm ageing in house mice.

    PubMed

    Firman, R C; Young, F J; Rowe, D C; Duong, H T; Gasparini, C

    2015-07-01

    Fertilization by aged sperm can result in adverse fitness consequences for both males and females. Sperm storage during male sexual rest could provide an environment for post-meiotic sperm senescence causing a deterioration in the quality of stored sperm, possibly impacting on both sperm performance (e.g. swimming ability) and DNA quality. Here, we compared the proportion of sperm with fragmented DNA, an indicator of structural damage of DNA within the sperm cell, among males that had been sexually rested for approximately 2 months, to that of males that had mated recently. We found no evidence of intra-epididymal sperm DNA damage or any impairment in sperm performance, and consequently no evidence of post-meiotic sperm senescence. Our results suggest that male house mice are likely to possess mechanisms that function to ensure that their sperm reserves remain stocked with 'young', viable sperm during periods of sexual inactivity. We also discuss the possibility that our experimental design leads to no difference in the age of sperm among males from the two mating treatments. Post-meiotic sperm senescence is especially relevant under sperm competition. Thus, we sourced mice from populations that differed in their levels of post-copulatory sexual selection, enabling us to gain insight into how selection for higher sperm production influences the rate of sperm ageing and levels of DNA fragmentation. We found that males from the population that produced the highest number of sperm also had the smallest proportion of DNA-fragmented sperm and discuss this outcome in relation to selection acting upon males to ensure that they produce ejaculates with high-quality sperm that are successful in achieving fertilizations under competitive conditions. PMID:26012513

  6. The Sperm Epigenome

    Microsoft Academic Search

    Donovan Chan; Jacquetta Trasler

    \\u000a The development of male germ cells from the primordial germ cell stage to that of the mature spermatozoon is a key time of\\u000a epigenetic reprogramming. Orchestrated by specialized enzymes, DNA methylation and histone modifications undergo dynamic changes\\u000a throughout gametogenesis. Alterations to any level of the sperm epigenetic coding may affect fertility and the sperm’s contribution\\u000a to normal embryo development. In

  7. CAENORHABDITIS ELEGANS Fertilization-Defective Mutants with Abnormal Sperm

    PubMed Central

    Argon, Yair; Ward, Samuel

    1980-01-01

    Seven new fertilization-defective mutants of C. elegans have been isolated and characterized; six are temperature sensitive, one is absolute and all are autosomal recessive. One mutation is in a previously described gene, while the other six define six new fer genes that appear to code for sperm-specific functions necessary for normal fertilization. In all fer mutants, both males and hermaphrodites accumulate sperm in near normal numbers. In hermaphrodites, mutant sperm contact the oocytes, but fail to fertilize them. Instead, the sperm are swept into the uterus by the passing oocytes and are expelled when oocytes are laid. Males of two fer mutants do not transfer sperm during copulation, but the other mutant males transfer sperm that fail to move to the spermatheca. Spermatozoa from fer-1 and fer-4 mutants are motility-defective in vitro as well as in vivo, and their pseudopods have an altered morphology. The period of development during which mutant hermaphrodites are temperature sensitive for fertility overlaps the time of sperm development. Some mutants are temperature sensitive throughout the entire period, and others are temperature sensitive during or just prior to spermiogenesis. In fer-4/+ and fer-7/+ males, the fertility of the mutation-bearing sperm is diminished, reducing the transmission ratio. This implies some post-meiotic expression of these genes.—This set of mutants provides a variety of functional and structural alterations in nematode sperm that should help identify and analyze gene products involved in sperm morphogenesis and motility. PMID:7196361

  8. Testing Boolean Function Isomorphism Noga Alon1,

    E-print Network

    Alon, Noga

    is to characterize the query complexity for every graph or boolean function. 2. Testing isomorphism to the hardestTesting Boolean Function Isomorphism Noga Alon1, and Eric Blais2 1 Schools of Mathematics: eblais@cs.cmu.edu. Abstract. Two boolean functions f, g : {0, 1}n {0, 1} are isomorphic

  9. Testing Boolean Function Isomorphism Noga Alon1,

    E-print Network

    Shamir, Ron

    the query complexity for every graph or boolean function. 2. Testing isomorphism to the hardest known objectTesting Boolean Function Isomorphism Noga Alon1, and Eric Blais2 1 Schools of Mathematics: eblais@cs.cmu.edu. Abstract. Two boolean functions f, g : {0, 1}n {0, 1} are isomorphic

  10. 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

  11. Male Genital Tract Antioxidant Enzymes: Their Source, Function in the Female, and Ability to Preserve Sperm DNA Integrity in the Golden Hamster

    Microsoft Academic Search

    HONG CHEN; PAK HAM CHOW; SO KWAN CHENG; ANNIE L. M. CHEUNG; LYDIA Y. L. CHENG; WAI-SUM O

    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

  12. Using neural networks for functional testing

    Microsoft Academic Search

    Larry V. Kirkland; R. G. Wright

    1995-01-01

    This paper discusses using Neural Networks for diagnosing circuit faults. As a circuit is tested, the output signals from a Unit Under Test can vary as different functions are invoked by the test. When plotted against time, these signals create a characteristic trace for the test performed. Sensors in the ATS can be used to monitor the output signals during

  13. Correlation between serum lipids profile with sperm parameters of infertile men with abnormal semen analysis

    Microsoft Academic Search

    Mohammad Ali Khalili; Najmeh Zare-Zadeh; Hamideh Hashemi

    2009-01-01

    Background: One of the main laboratory tests for evaluation of infertility is semen analysis (SA). However, additional tests may be suggested for further diagnosis of male fertility potentials. The seminal fluid contains sperms, non-sperm cells, and various types of lipids and glucose. Objective: The objective of this cross-sectional study was to correlate the sperm parameters with concentrations of cholesterol, triglyceride,

  14. Functional interaction of phospholipid hydroperoxide glutathione peroxidase with sperm mitochondrion-associated cysteine-rich protein discloses the adjacent cysteine motif as a new substrate of the selenoperoxidase.

    PubMed

    Maiorino, Matilde; Roveri, Antonella; Benazzi, Louise; Bosello, Valentina; Mauri, Pierluigi; Toppo, Stefano; Tosatto, Silvio C E; Ursini, Fulvio

    2005-11-18

    The mitochondrial capsule is a selenium- and disulfide-rich structure enchasing the outer mitochondrial membrane of mammalian spermatozoa. Among the proteins solubilized from the sperm mitochondrial capsule, we confirmed, by using a proteomic approach, the presence of phospholipid hydroperoxide glutathione peroxidase (PHGPx) as a major component, and we also identified the sperm mitochondrion-associated cysteine-rich protein (SMCP) and fragments/aggregates of specific keratins that previously escaped detection (Ursini, F., Heim, S., Kiess, M., Maiorino, M., Roveri, A., Wissing, J., and Flohé, L. (1999) Science 285, 1393-1396). The evidence for a functional association between PHGPx, SMCP, and keratins is further supported by the identification of a sequence motif of regularly spaced Cys-Cys doublets common to SMCP and high sulfur keratin-associated proteins, involved in bundling hair shaft keratin by disulfide cross-linking. Following the oxidative polymerization of mitochondrial capsule proteins, catalyzed by PHGPx, two-dimensional redox electrophoresis analysis showed homo- and heteropolymers of SMCP and PHGPx, together with other minor components. Adjacent cysteine residues in SMCP peptides are oxidized to cystine by PHGPx. This unusual disulfide is known to drive, by reshuffling oxidative protein folding. On this basis we propose that oxidative polymerization of the mitochondrial capsule is primed by the formation of cystine on SMCP, followed by reshuffling. Occurrence of reshuffling is further supported by the calculated thermodynamic gain of the process. This study suggests a new mechanism where selenium catalysis drives the cross-linking of structural elements of the cytoskeleton via the oxidation of a keratin-associated protein. PMID:16159880

  15. Methods for evaluating the effects of environmental chemicals on human sperm production

    SciTech Connect

    Wyrobek, A.J.

    1982-04-20

    Sperm tests provide a direct and effective way of identifying chemical agents that induce spermatogenic damage in man. Four human sperm tests are available: sperm count, motility, morphology (seminal cytology), and the Y-body test. These sperm tests have numerous advantages over other approaches for assessing spermatogenic damage, and they have already been used to assess the effects of at least 85 different occupational, envionmental, and drug-related chemical exposures. When carefully controlled, seminal cytology appears to be statistically more sensitive than the other human sperm tests and should be considered an integral part of semen analysis when assessing induced spermatogenic damage.

  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. 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 ...

  18. 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 ...

  19. 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.

  20. Boar proacrosin correlation between total proacrosin content and sperm fertilizing capacity.

    PubMed

    Cechová, D; Zelezná, B; Petelíková, J; Pavlok, A

    1984-01-01

    A method of acidic extraction at pH-1.3 was used for quantitative estimation of the proacrosin and acrosin content in boar spermatozoa after various times of storage in four different sperm dilutors. The proacrosin and acrosin contents were correlated with the results of biological tests for sperm penetration in zona-free hamster eggs and sperm survival. The results have shown that proacrosin quantitation should be a convenient biochemical test of sperm fertilizing capacity. PMID:6430126

  1. Nematode sperm maturation triggered by protease involves sperm-secreted serine protease inhibitor (Serpin)

    PubMed Central

    Zhao, Yanmei; Sun, Wei; Zhang, Pan; Chi, Hao; Zhang, Mei-Jun; Song, Chun-Qing; Ma, Xuan; Shang, Yunlong; Wang, Bin; Hu, Youqiao; Hao, Zhiqi; Hühmer, Andreas F.; Meng, Fanxia; L'Hernault, Steven W.; He, Si-Min; Dong, Meng-Qiu; Miao, Long

    2012-01-01

    Spermiogenesis is a series of poorly understood morphological, physiological and biochemical processes that occur during the transition of immotile spermatids into motile, fertilization-competent spermatozoa. Here, we identified a Serpin (serine protease inhibitor) family protein (As_SRP-1) that is secreted from spermatids during nematode Ascaris suum spermiogenesis (also called sperm activation) and we showed that As_SRP-1 has two major functions. First, As_SRP-1 functions in cis to support major sperm protein (MSP)-based cytoskeletal assembly in the spermatid that releases it, thereby facilitating sperm motility acquisition. Second, As_SRP-1 released from an activated sperm inhibits, in trans, the activation of surrounding spermatids by inhibiting vas deferens-derived As_TRY-5, a trypsin-like serine protease necessary for sperm activation. Because vesicular exocytosis is necessary to create fertilization-competent sperm in many animal species, components released during this process might be more important modulators of the physiology and behavior of surrounding sperm than was previously appreciated. PMID:22307610

  2. 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

  3. Sperm capacitation combined with removal of the sperm acrosome and plasma membrane enhances paternal nucleus remodelling and early development of bovine androgenetic embryos.

    PubMed

    Xiao, Yao; Zhang, Hualin; Ahmad, Sibtain; Bai, Liya; Wang, Xiaomin; Huo, Lijun; Zhang, Xin; Li, Wengong; Li, Xiang; Yang, Liguo

    2013-01-01

    The androgenetic embryo is a useful model for functional analysis of the paternal genome during embryogenesis. However, few studies have focused on the factors involved in the suppressed developmental competence of such embryos or why sperm cloning-derived androgenetic embryos fail to develop beyond the morula stage in large domestic animals. To overcome this developmental failure, we tried to improve sperm decondensation, as well as to enhance embryonic development by sperm capacitation and removal of the acrosome and plasma membrane before injection of the spermatozoa. Before injection of the spermatozoa, we quantified the effects of sperm capacitation combined with sperm pretreatment on the acrosome and plasma membrane status. We also evaluated sperm decondensation potential, sperm viability and chromatin integrity. Immunostaining data showed that the sperm acrosome and plasma membrane could be more efficiently removed after capacitation. Dithiothreitol-induced sperm decondensation potential was improved with capacitation and removal of the acrosome and plasma membrane. Although most spermatozoa lost viability after pretreatment, their chromatin remained integrated. The patterns of paternal chromatin remodelling within uncleaved androgenetic embryos and the nucleus morphology of cleaved embryos indicated that capacitation combined with membrane disruption could make injected spermatozoa decondense synchronously not only with each other, but also with the developmental pace of the ooplasm. We successfully produced androgenetic blastocysts, and efficiency increased with sperm pretreatment. In conclusion, sperm decondensation and the early development of androgenetic embryos were enhanced with sperm capacitation and removal of the acrosome and plasma membrane prior to sperm injection. PMID:22950979

  4. 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 ...

  5. 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

  6. 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

  7. Sperm Proteome: What Is on the Horizon?

    PubMed

    Mohanty, Gayatri; Swain, Nirlipta; Samanta, Luna

    2015-06-01

    As the mammalian spermatozoa transcends from the testis to the end of the epididymal tubule, the functionally incompetent spermatozoa acquires its fertilizing capability. Molecular changes in the spermatozoa at the posttesticular level concern qualitative and quantitative modifications of proteins along with their sugar moieties and membranous lipids mostly associated with motility, egg binding, and penetration processes. Proteomic studies have identified numerous sperm-specific proteins, and recent reports have provided a further understanding of their function with respect to male fertility. High-throughput techniques such as mass spectrometry have shown drastic potential for the identification and study of sperm proteins. In fact, compelling evidence has provided that proteins are critically important in cellular remodeling event and that aberrant expression is associated with pronounced defects in sperm function. This review highlights the posttesticular functional transformation in the epididymis and female reproductive tract with due emphasis on proteomics. PMID:25376881

  8. Cryopreservation of domestic animal sperm cells.

    PubMed

    Barbas, J P; Mascarenhas, R D

    2009-02-01

    Sperm cells are the endpoint of male spermatogenesis and have particular anatomic and metabolic features. Sperm cryopreservation and storage currently require liquid nitrogen or ultralow refrigeration methods for long or short term storage, which requires routine maintenance and extensive space requirements. Conserving sperms have several purposes such as artificial reproductive technologies (ART), species conservation and clinical medicine. The combinations of storage temperature, cooling rate, chemical composition of the extender, cryoprotectant concentration, reactive oxygen species (ROS), seminal plasma composition and hygienic control are the key factors that affect the life-span of spermatozoa. Sperm preservation protocols vary among animal species owing to their inherent particularities that change extenders used for refrigeration and freezing. Extenders for freezing sperm cells contain buffers, carbohydrates (glucose, lactose, raffinose, saccharose and trehalose), salts (sodium citrate, citric acid), egg yolk and antibiotics. The use of different cryoprotectants, like trehalose or glycerol, as well as different concentrations of egg yolk and other constituents in semen extenders are being studied in our laboratory. Several cooling rates have been tested to freeze sperm cells. The use of faster rates (15-60 degrees C/min) gives rise to best sperm survivals after freezing-thawing, but more studies are needed to find the adequate cooling rates for each animal species. Sheep and goat males of some native breeds are being used in studies performed in EZN. Semen from those males has been frozen and stored as part of the Portuguese Animal Germplasm Bank. In small ruminants, individual variations in the quality of frozen semen have been observed, suggesting specific differences in sperm susceptibility to freezing methods, particularly obvious in goat males. Best quality frozen semen from small ruminants is being used in cervical artificial insemination studies aiming to increase productive parameters in selected flocks. PMID:18548333

  9. The functional test of complex integrated circuits

    NASA Astrophysics Data System (ADS)

    Bellon, C.

    1984-06-01

    A multilevel automatic generation of test sequences is proposed. The procedure starts by considering the set of functional parameters and the fine structure of the electronic modules composing the circuit. The method may be considered related to automatic software testing or automata identification methods. A functional microprocessor test procedure is described. The method takes into account the synchronous input signals and allows the automatic generation of test programs. A specific high level computer language was developed for that purpose and is described. The 68000 microprocessor was described in that language and its test programs generated. The elements implementing the system are detailed.

  10. Sperm competition in bats.

    PubMed Central

    Hosken, D J

    1997-01-01

    Sperm competition is a widespread phenomenon influencing the evolution of male anatomy, physiology and behaviour. Bats are an ideal group for studying sperm competition. Females store fertile sperm for up to 200 days and the size of social groups varies from single animals to groups of hundreds of thousands. This study examines the relationship between social group size and investment in spermatogenesis across 31 species of microchiropteran bat using new and published data on testis mass and sperm length. In addition to male competition, I examined the effects of female reproductive biology on characteristics of spermatogenesis. Comparative studies indicate that relative testis mass is positively related to sperm competition risk in a wide range of taxa. Social group size may also influence the level of sperm competition, and one of the costs of living in groups may be decreased confidence of paternity. I used comparative analysis of independent contrast (CAIC) to control for phylogeny. Using two possible phylogenies and two measures of social group size, I found a significant positive relationship between social group size and testis mass. There was no relationship between testis mass and the dimension of the female reproductive tract or oestrus duration. Sperm length was not significantly related to body mass or group size, nor was it related to oestrus duration. PMID:9107054

  11. Sperm variation within a single ejaculate affects offspring development in Atlantic salmon

    PubMed Central

    Immler, Simone; Hotzy, Cosima; Alavioon, Ghazal; Petersson, Erik; Arnqvist, Göran

    2014-01-01

    It is generally believed that variation in sperm phenotype within a single ejaculate has no consequences for offspring performance, because sperm phenotypes are thought not to reflect sperm genotypes. We show that variation in individual sperm function within an ejaculate affects the performance of the resulting offspring in the Atlantic salmon Salmo salar. We experimentally manipulated the time between sperm activation and fertilization in order to select for sperm cohorts differing in longevity within single ejaculates of wild caught male salmon. We found that within-ejaculate variation in sperm longevity significantly affected offspring development and hence time until hatching. Whether these effects have a genetic or epigenetic basis needs to be further evaluated. However, our results provide experimental evidence for transgenerational effects of individual sperm function. PMID:24522632

  12. Competition drives cooperation among closely related sperm of deer mice

    E-print Network

    Dean, Matthew D.

    LETTERS Competition drives cooperation among closely related sperm of deer mice Heidi S. Fisher1 of deer mice (genus Peromyscus) form motile aggregations, then we use this system to test predictions with unrelated conspecific sperm. In contrast, in the highly promiscuous deer mouse, Peromyscus maniculatus

  13. Sperm preparation: state-of-the-art—physiological aspects and application of advanced sperm preparation methods

    PubMed Central

    Henkel, Ralf

    2012-01-01

    For assisted reproduction technologies (ART), numerous techniques were developed to isolate spermatozoa capable of fertilizing oocytes. While early methodologies only focused on isolating viable, motile spermatozoa, with progress of ART, particularly intracytoplasmic sperm injection (ICSI), it became clear that these parameters are insufficient for the identification of the most suitable spermatozoon for fertilization. Conventional sperm preparation techniques, namely, swim-up, density gradient centrifugation and glass wool filtration, are not efficient enough to produce sperm populations free of DNA damage, because these techniques are not physiological and not modeled on the stringent sperm selection processes taking place in the female genital tract. These processes only allow one male germ cell out of tens of millions to fuse with the oocyte. Sites of sperm selection in the female genital tract are the cervix, uterus, uterotubal junction, oviduct, cumulus oophorus and the zona pellucida. Newer strategies of sperm preparation are founded on: (i) morphological assessment by means of ‘motile sperm organelle morphological examination (MSOME)' (ii) electrical charge; and (iii) molecular binding characteristics of the sperm cell. Whereas separation methods based on electrical charge take advantage of the sperm's adherence to a test tube surface or separate in an electrophoresis, molecular binding techniques use Annexin V or hyaluronic acid (HA) as substrates. Techniques in this category are magnet-activated cell sorting, Annexin V-activated glass wool filtration, flow cytometry and picked spermatozoa for ICSI (PICSI) from HA-coated dishes and HA-containing media. Future developments may include Raman microspectrometry, confocal light absorption and scattering spectroscopic microscopy and polarization microscopy. PMID:22138904

  14. Exposure of rainbow trout milt to mercury and cadmium alters sperm motility parameters and reproductive success

    Microsoft Academic Search

    Grzegorz J. Dietrich; Mariola Dietrich; R. K. Kowalski; Stefan Dobosz; Halina Karol; Wies?aw Demianowicz; Jan Glogowski

    2010-01-01

    In the current work, seminal plasma was used for the first time as an incubation medium for monitoring short-time exposure effects of sublethal concentrations of mercury and cadmium ions on rainbow trout sperm. Sperm motility parameters (CASA) and hatching rates were used as gamete quality markers. Additionally live\\/dead sperm viability test and comet assay of DNA fragmentation were performed. We

  15. Focus on Determinants of Male Fertility Is sperm evaluation useful in predicting human fertility?

    Microsoft Academic Search

    Sheena E M Lewis

    Traditionally, the diagnosis of male infertility has relied upon microscopic assessment and biochemical assays to determine human semen quality. The conventional parameters given most importance have been the concentration, motility, and morphology of sperm in the ejaculate. Most laboratories also include 'sperm suitability' tests where the subpopulations of sperm more likely to finish the marathon journey to the oocyte are

  16. Higher sperm counts in Southern Sweden compared with Denmark

    Microsoft Academic Search

    J. Richthoff; L. Rylander; L. Hagmar; J. Malm; A. Giwercman

    2002-01-01

    BACKGROUND: A common aetiology behind time-related deterioration of male reproductive function, including decline of sperm concentration and rising incidence of testicular cancer and cryptorchidism, has been suggested. Accordingly, a high incidence of testicular cancer and low sperm counts have been observed in Denmark, while in Finland opposite figures were found. This may be due to genetic factors. The incidence of

  17. A Plant Germline-Specific Integrator of Sperm Specification and Cell Cycle Progression

    Microsoft Academic Search

    Lynette Brownfield; Said Hafidh; Michael Borg; Anna Sidorova; Toshiyuki Mori; David Twell

    2009-01-01

    The unique double fertilisation mechanism in flowering plants depends upon a pair of functional sperm cells. During male gametogenesis, each haploid microspore undergoes an asymmetric division to produce a large, non-germline vegetative cell and a single germ cell that divides once to produce the sperm cell pair. Despite the importance of sperm cells in plant reproduction, relatively little is known

  18. Local anaesthetic effect of antiarrhythmic drugs and human sperm immobilization: mechanism and application of the interrelationship.

    PubMed Central

    Hong, C Y; Chiang, B N

    1984-01-01

    Sperm immobilizing activity of three class I antiarrhythmic drugs, quinidine, procainamide, mexiletine, one adrenoceptor blocking drug, labetalol and one calcium channel blocking drug, diltiazem was measured with a trans-membrane migration method. All these drugs immobilized human sperm. Local anaesthetic effect is the underlying mechanism for these drugs to inhibit sperm motility. Sperm immobilization could be a screening test for the local anaesthetic effect of antiarrhythmic drugs. PMID:6743466

  19. REVIVAL OF MAMMALIAN SPERM AFTER IMMERSION IN LIQUID NITROGEN

    PubMed Central

    Hoagland, Hudson; Pincus, Gregory

    1942-01-01

    1. A wide variety of procedures was used to test the motility of mammalian sperm after plunging them into liquid nitrogen at –195°C. and later rapidly warming them to 35°C. by plunging them into a suitable balanced and isotonic medium. 2. Using seminal fluid sperm from the same human donor, maximal numbers of motile sperm survived vitrification when the samples were (a) very fresh, (b) untreated with plasmolysing solutions, (c) plunged into the refrigerant in the form of a foam. The maximum yield of motile human sperm recoverable from the liquid nitrogen was 50 per cent. Since in this sample only 75 per cent of the sperm were alive before immersion, 67 per cent of the living sperm survived vitrification. 3. Experiments with sperm from 31 rabbits were made with a variety of conditions of pretreatment to obtain maximal yields of recoverable, motile sperm after vitrification by liquid nitrogen. (a) A consistent recoverable yield of about 0.5 per cent was obtained when the untreated suspension of sperm was smeared on cellophane and partially dried in air before immersing in liquid nitrogen. (b) On a few out of many occasions plasmolysis for several minutes with hypertonic Ringer solution gave a recoverable yield of 0.1 per cent as did (c) pretreatment with hypertonic Ringer and butyric acid. PMID:19873277

  20. Intracytoplasmic Sperm Injection (ICSI)

    MedlinePLUS

    ... injection (ICSI) can be done along with in vitro fertilization (IVF) to help fertilize the egg. During ... regardless of the condition of the sperm. • In vitro matured eggs are being used. • Previously frozen eggs ...

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

    E-print Network

    Faucette, Azure

    2012-07-16

    bucks. In the first experiment, testes were harvested from five Alpine bucks at 0, 2, 4, 6, and 8 months of age. Northern and in situ hybridization indicated that the largest change in gene expression occurred during the first 4 months of goat testes...

  2. The frequency of multiple paternity predicts variation in testes size among island populations of house mice.

    PubMed

    Firman, R C; Simmons, L W

    2008-11-01

    Polyandry generates selection on males through sperm competition, which has broad implications for the evolution of ejaculates and male reproductive anatomy. Comparative analyses across species and competitive mating trials within species have suggested that sperm competition can influence the evolution of testes size, sperm production and sperm form and function. Surprisingly, the intraspecific approach of comparing among population variation for investigating the selective potential of sperm competition has rarely been explored. We sampled seven island populations of house mice and determined the frequency of multiple paternity within each population. Applying the frequency of multiple paternity as an index of the risk of sperm competition, we looked for selective responses in male reproductive traits. We found that the risk of sperm competition predicted testes size across the seven island populations of house mice. However, variation in sperm traits was not explained by sperm competition risk. We discuss these findings in relation to sperm competition theory, and other intrinsic and extrinsic factors that might influence ejaculate quality. PMID:18811664

  3. Advantages of digital convergence for functional test

    Microsoft Academic Search

    J. A. Hutchinson

    1998-01-01

    With the ever-increasing complexity and clock rates of today's weapon system electronics, digital functional test plays a critical role in depot test. This is likely to become even more so over the next in years as the next generation of weapon systems are deployed. At the same time that system complexity is increasing, the number of joint weapon system programs

  4. Sperm studies in anesthesiologists

    SciTech Connect

    Wyrobek, A.J. (Lawrence Livermore National Lab., CA); 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.

  5. 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...

  6. Lack of Tyrosylprotein Sulfotransferase-2 Activity Results in Altered Sperm-Egg Interactions and Loss of ADAM3 and ADAM6 in Epididymal Sperm*

    PubMed Central

    Marcello, Matthew R.; Jia, Weitao; Leary, Julie A.; Moore, Kevin L.; Evans, Janice P.

    2011-01-01

    Tyrosine O-sulfation is a post-translational modification catalyzed by two tyrosylprotein sulfotransferases (TPST-1 and TPST-2) in the trans-Golgi network. Tpst2-deficient mice have male infertility, sperm motility defects, and possible abnormalities in sperm-egg membrane interactions. Studies here show that compared with wild-type sperm, fewer Tpst2-null sperm bind to the egg membrane, but more of these bound sperm progress to membrane fusion. Similar outcomes were observed with wild-type sperm treated with the anti-sulfotyrosine antibody PSG2. The increased extent of sperm-egg fusion is not due to a failure of Tpst2-null sperm to trigger establishment of the egg membrane block to polyspermy. Anti-sulfotyrosine staining of sperm showed localization similar to that of IZUMO1, a sperm protein that is essential for gamete fusion, but we detected little to no tyrosine sulfation of IZUMO1 and found that IZUMO1 expression and localization were normal in Tpst2-null sperm. Turning to a discovery-driven approach, we used mass spectrometry to characterize sperm proteins that associated with PSG2. This identified ADAM6, a member of the A disintegrin and A metalloprotease (ADAM) family; members of this protein family are associated with multiple sperm functions. Subsequent studies revealed that Tpst2-null sperm lack ADAM6 and ADAM3. Loss of ADAM3 is strongly associated with male infertility and is observed in knockouts of male germ line-specific endoplasmic reticulum-resident chaperones, raising the possibility that TPST-2 may function in quality control in the secretory pathway. These data suggest that TPST-2-mediated tyrosine O-sulfation participates in regulating the sperm surface proteome or membrane order, ultimately affecting male fertility. PMID:21339297

  7. 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.

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

    E-print Network

    Faucette, Azure

    2012-07-16

    LIST OF FIGURES FIGURE Page 1 Mean total testes weight (g) of goat kids castrated at 0, 2, 4, 6 and 8 months of age... for maintenance of testes differentiation. One of the genes found downstream of SRY is SRY-like HMG (High Mobility Group) box-containing gene-9 (SOX9). The SOX9 gene is responsible for campomelic dysplasia (CD) in humans. Patients with CD are characterized...

  9. 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.

  10. Diisopropyl fluorophosphate labeling of sperm-associated proteinases

    SciTech Connect

    Odem, R.R.; Willand, J.L.; Polakoski, K.L. (Washington Univ. School of Medicine, St. Louis, MO (USA))

    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.

  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. Pulmonary function testing in small laboratory mammals.

    PubMed Central

    O'Neil, J J; Raub, J A

    1984-01-01

    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 animals to measure spirometry (lung volumes), mechanics, distribution of ventilation, gas exchange or control of ventilation. These tests were designed on the basis of similar tests which are used in humans to diagnose and manage patients with lung disease. A major difference is that many of the measurements are performed in anesthetized animals, while human pulmonary function is usually measured in awake cooperating individuals. In addition, the measurement of respiratory events in small animals requires sensitive and rapidly responding equipment, because signals may be small and events can occur quickly. In general, the measurements described provide information on the change in normal lung function which results primarily from structural changes. These tests of pulmonary function can be repetitively and routinely accomplished and the results appear to be highly reproducible. Although some are quite sophisticated, many can be undertaken with relatively inexpensive equipment and provide useful information for toxicological testing. PMID:6434299

  13. 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

  14. Mouse Sperm Membrane Potential Hyperpolarization Is Necessary and Sufficient to Prepare Sperm for the Acrosome Reaction*

    PubMed Central

    De La Vega-Beltran, Jose Luis; Sánchez-Cárdenas, Claudia; Krapf, Darío; Hernandez-González, Enrique O.; Wertheimer, Eva; Treviño, Claudia L.; Visconti, Pablo E.; Darszon, Alberto

    2012-01-01

    Mammalian sperm are unable to fertilize the egg immediately after ejaculation; they acquire this capacity during migration in the female reproductive tract. This maturational process is called capacitation and in mouse sperm it involves a plasma membrane reorganization, extensive changes in the state of protein phosphorylation, increases in intracellular pH (pHi) and Ca2+ ([Ca2+]i), and the appearance of hyperactivated motility. In addition, mouse sperm capacitation is associated with the hyperpolarization of the cell membrane potential. However, the functional role of this process is not known. In this work, to dissect the role of this membrane potential change, hyperpolarization was induced in noncapacitated sperm using either the ENaC inhibitor amiloride, the CFTR agonist genistein or the K+ ionophore valinomycin. In this experimental setting, other capacitation-associated processes such as activation of a cAMP-dependent pathway and the consequent increase in protein tyrosine phosphorylation were not observed. However, hyperpolarization was sufficient to prepare sperm for the acrosome reaction induced either by depolarization with high K+ or by addition of solubilized zona pellucida (sZP). Moreover, K+ and sZP were also able to increase [Ca2+]i in non-capacitated sperm treated with these hyperpolarizing agents but not in untreated cells. On the other hand, in conditions that support capacitation-associated processes blocking hyperpolarization by adding valinomycin and increasing K+ concentrations inhibited the agonist-induced acrosome reaction as well as the increase in [Ca2+]i. Altogether, these results suggest that sperm hyperpolarization by itself is key to enabling mice sperm to undergo the acrosome reaction. PMID:23095755

  15. A Test of the Human Sperm Acrosome Reaction Following lonophore Challenge Relationship to Fertility and Other Seminal Parameters

    Microsoft Academic Search

    JAMES M. CUMMINS; SANDRA M. PEMBER; ANNE M. JEQUIER; JOHN L. YOVICH; PETER E. HARTMANN

    Acrosome reaction capacity was tested on semen samples from 53 fertile and 26 subfertile men. Preparations were divided into two aliquots after 3 or 24 hours of culture. One aliquot received 10 pmoI\\/L calcium lonophore A23187 in dime- thyl sulfoxide (DMSO) and the other received DMSO alone. Ac- rosome reactions were scored on ethanol-permeabilized smears using fluorescein isothiocyanate (FITC)-conjugated Pisum

  16. Sperm counts and sperm sex ratio in male infertility patients.

    PubMed

    Eisenberg, Michael L; Murthy, Lata; Hwang, Kathleen; Lamb, Dolores J; Lipshultz, Larry I

    2012-09-01

    In recent years, investigators have noted a trend toward a declining proportion of male births in many industrialized nations. While men bear the sex-determining chromosome, the role of the female partner as it pertains to fertilization or miscarriage may also alter the gender ratio. We attempted to determine a man's secondary sex ratio (F1 generation) by directly examining the sex chromosomes of his sperm. We examined our male infertility clinic database for all men who had undergone a semen fluorescence in situ hybridization (FISH). Patient demographic and semen parameters were recorded. Chi-squared analysis was used to compare gender ratios (Y chromosomes/total chromosomes). Multivariable logistic regression was used to predict the odds of possessing a Y-bearing sperm after accounting for demographic and semen parameters. A total of 185 men underwent sperm FISH. For the entire cohort, the proportion of Y chromosome-bearing sperm was 51.5%. Men with less than five million motile sperm had a significantly lower proportion of Y chromosome-bearing sperm (50.8%) compared to men with higher sperm counts (51.6%; P=0.02). After multivariable adjustment, a higher sperm concentration, total motile sperm count and semen volume significantly increased the odds of having a Y chromosome-bearing sperm (P<0.01). As a man's sperm production declines, so does the proportion of Y chromosome-bearing sperm. Thus, a man's reproductive potential may predict his ability to sire male offspring. PMID:22842703

  17. Sperm Nuclear Vacuoles in relation to Acrosome Reactions and Sperm Motility

    PubMed Central

    Kawauchi, Yoko; Kato, Tomonori; Tanii, Ichiro; Fuse, Hideki

    2014-01-01

    We investigated sperm nuclear vacuolation in relation to acrosome reactions and the maintenance of sperm motility. Thirty male patients who visited our Male Infertility Clinic were enrolled. These patients underwent conventional semen analyses, Acrobeads tests, and high-magnification observation of the sperm head to evaluate the degree of nuclear vacuolation on the Acrobeads test scoring after 24 hours of incubation. The presence of acrosome reactions was evaluated using the Acrobeads test. The spermatozoa were classified into three groups: (I) those bound to MH61-beads, (II) motile spermatozoa that did not bind to MH61-beads, and (III) immotile spermatozoa that did not bind to MH61-beads. The percentage of spermatozoa with large nuclear vacuoles (%LNV) was compared between the three groups. The degree of sperm nuclear vacuolation was evaluated in 17,992 ejaculated spermatozoa. The mean %LNVs were 2.4% in group I, 5.8% in group II, and 9.8% in group III. These values were significantly different from each other (P < 0.001, paired t-test). There were no correlations between the %LNV values and the Acrobeads scores. In conclusion, the degree of sperm nuclear vacuolation was significantly lower in the acrosome-reacted spermatozoa and spermatozoa with maintained motility, and higher in the immotile spermatozoa that did not bind to MH61-beads. PMID:25097868

  18. 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.

  19. Therapeutic ultrasound as a potential male contraceptive: power, frequency and temperature required to deplete rat testes of meiotic cells and epididymides of sperm determined using a commercially available system

    PubMed Central

    2012-01-01

    Background Studies published in the 1970s by Mostafa S. Fahim and colleagues showed that a short treatment with ultrasound caused the depletion of germ cells and infertility. The goal of the current study was to determine if a commercially available therapeutic ultrasound generator and transducer could be used as the basis for a male contraceptive. Methods Sprague-Dawley rats were anesthetized and their testes were treated with 1 MHz or 3 MHz ultrasound while varying power, duration and temperature of treatment. Results We found that 3 MHz ultrasound delivered with 2.2 Watt per square cm power for fifteen minutes was necessary to deplete spermatocytes and spermatids from the testis and that this treatment significantly reduced epididymal sperm reserves. 3 MHz ultrasound treatment reduced total epididymal sperm count 10-fold lower than the wet-heat control and decreased motile sperm counts 1,000-fold lower than wet-heat alone. The current treatment regimen provided nominally more energy to the treatment chamber than Fahim's originally reported conditions of 1 MHz ultrasound delivered at 1 Watt per square cm for ten minutes. However, the true spatial average intensity, effective radiating area and power output of the transducers used by Fahim were not reported, making a direct comparison impossible. We found that germ cell depletion was most uniform and effective when we rotated the therapeutic transducer to mitigate non-uniformity of the beam field. The lowest sperm count was achieved when the coupling medium (3% saline) was held at 37 degrees C and two consecutive 15-minute treatments of 3 MHz ultrasound at 2.2 Watt per square cm were separated by 2 days. Conclusions The non-invasive nature of ultrasound and its efficacy in reducing sperm count make therapeutic ultrasound a promising candidate for a male contraceptive. However, further studies must be conducted to confirm its efficacy in providing a contraceptive effect, to test the result of repeated use, to verify that the contraceptive effect is reversible and to demonstrate that there are no detrimental, long-term effects from using ultrasound as a method of male contraception. PMID:22289508

  20. 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.

  1. Functional circuit board testing using nanoscale sensors

    Microsoft Academic Search

    R. G. Wright; M. Zgol; D. Adebimpe; L. V. Kirkland

    2003-01-01

    This paper describes research and development efforts in the application of nanoscale sensors to implement an original method for a 'sea-of-needles' kind of bed-of-nails testing for printed circuit boards. This approach performs functional testing that eliminates the need to have a pre-existing model of a circuit board while automating much of the development process. The overall concept involves shrinking the

  2. Porcine Sperm Bind to Specific 6-Sialylated Biantennary Glycans to Form the Oviduct Reservoir1

    PubMed Central

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

    2012-01-01

    ABSTRACT 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

  3. Flow-cytometric analyses of viability biomarkers in pesticide-exposed sperm of three aquatic invertebrates.

    PubMed

    Favret, Karen P; Lynn, John W

    2010-05-01

    Toxicity studies on sperm often use fertilization success as the end point. This type of assay can be affected by sperm density, egg quality, and sperm-egg compatibility. Testing sperm viability biomarkers with flow cytometry is a fast, high-throughput technique for seminal analysis. In this study, we detected sperm viability biomarkers with several fluorescent reporter dyes using flow cytometry in three aquatic invertebrates (Crassostrea virginica, Dreissena polymorpha, and Lytechinus variegatus) after exposure to a pesticide and herbicide. The pesticide, Bayluscide, appeared to affect mitochondrial membrane potential in the sperm of all three species, as measured with MitoTracker Red CMXRos. A decrease in the percentage of sperm stained with SYBR-14 (indicating uncompromised plasma membrane) was observed in C. virginica and D. polymorpha sperm exposed to Bayluscide, but propidium iodide staining (indicating compromised plasma membranes) appeared to be inhibited by Bayluscide. Acrosome-reacted sperm, as measured by FITC-PNA, decreased after Bayluscide exposure in C. virginica and D. polymorpha sperm. The herbicide, Roundup Ready To-Use-Plus, did not affect the overall percentages of sperm stained with MitoTracker but did cause an increase in MitoTracker fluorescence intensity at 16 mg/L in D. polymorpha. Roundup also caused significant decreases in SYBR-14 staining, significant increases in propidium iodide staining, and significant increases in FITC-PNA staining in D. polymorpha sperm. By not having to rely on egg availability and optimal sperm density, sperm toxicity can be more accurately assessed with flow cytometry as being directly correlated to sperm viability rather than the possibility of altered toxicity results due to sperm-to-egg compatibility. PMID:19876686

  4. Pulmonary function tests in beta thalassemia

    Microsoft Academic Search

    Meeta Arora; Jagdish Chandra; J. C. Suri; S. Narayan; A. K. Dutta

    2001-01-01

    To study pulmonary function tests (PFT) in multiple transfusion recipient thalassemics, PFTs were done for 30 thalassemics\\u000a and 20 matched controls. Confirmed cases of thalassemia on regular transfusion therapy were the subject of study. Apart from\\u000a history and physical examination of the thalassemics, serum ferritin estimation and spirometry were done. Parameters studied\\u000a included lung volumes—functional residual capacity (FRC), forced vital

  5. Effects of viscosity on sperm motility studied with optical tweezers

    NASA Astrophysics Data System (ADS)

    Hyun, Nicholas; Chandsawangbhuwana, Charlie; Zhu, Qingyuan; Shi, Linda Z.; Yang-Wong, Collin; Berns, Michael W.

    2012-02-01

    The purpose of this study is to analyze human sperm motility and energetics in media with different viscosities. Multiple experiments were performed to collect motility parameters using customized computer tracking software that measures the curvilinear velocity (VCL) and the minimum laser power (Pesc) necessary to hold an individual sperm in an optical trap. The Pesc was measured by using a 1064 nm Nd:YVO4 continuous wave laser that optically traps motile sperm at a power of 450 mW in the focused trap spot. The VCL was measured frame by frame before trapping. In order to study sperm energetics under different viscous conditions sperm were labeled with the fluorescent dye DiOC6(3) to measure membrane potentials of mitochondria in the sperm midpiece. Fluorescence intensity was measured before and during trapping. The results demonstrate a decrease in VCL but an increase in Pesc with increasing viscosity. Fluorescent intensity is the same regardless of the viscosity level indicating no change in sperm energetics. The results suggest that, under the conditions tested, viscosity physically affects the mechanical properties of sperm motility rather than the chemical pathways associated with energetics.

  6. Bovine sperm plasma membrane proteomics through biotinylation and subcellular enrichment.

    PubMed

    Kasvandik, Sergo; Sillaste, Gerly; Velthut-Meikas, Agne; Mikelsaar, Aavo-Valdur; Hallap, Triin; Padrik, Peeter; Tenson, Tanel; Jaakma, Ülle; Kõks, Sulev; Salumets, Andres

    2015-06-01

    A significant proportion of mammalian fertilization is mediated through the proteomic composition of the sperm surface. These protein constituents can present as biomarkers to control and regulate breeding of agricultural animals. Previous studies have addressed the bovine sperm cell apical plasma membrane (PM) proteome with nitrogen cavitation enrichment. Alternative workflows would enable to expand the compositional data more globally around the entire sperm's surface. We used a cell surface biotin-labeling in combination with differential centrifugation to enrich sperm surface proteins. Using nano-LC MS/MS, 338 proteins were confidently identified in the PM-enriched proteome. Functional categories of sperm-egg interaction, protein turnover, metabolism as well as molecular transport, spermatogenesis, and signal transduction were represented by proteins with high quantitative signal in our study. A highly significant degree of enrichment was found for transmembrane and PM-targeted proteins. Among them, we also report proteins previously not described on bovine sperm (CPQ, CD58, CKLF, CPVL, GLB1L3, and LPCAT2B) of which CPQ and CPVL cell surface localization was further validated. A descriptive overview of the bovine sperm PM integral and peripheral proteins is provided to complement future studies on animal reproduction and its relation to sperm cell surface. All MS data have been deposited in the ProteomeXchange with identifier PXD001096 (http://proteomecentral.proteomexchange.org/dataset/PXD001096). PMID:25603787

  7. In vitro assessment of sperm quality from rams of high and low field fertility.

    PubMed

    Vicente-Fiel, S; Palacín, I; Santolaria, P; Fantova, E; Quintín-Casorrán, F J; Sevilla-Mur, E; Yániz, J L

    2014-04-01

    The aim of the present study was to investigate whether differences in field fertility of rams are reflected in differences in several sperm quality parameters. Ejaculates from 8 adult rams, 4 with high and 4 with low field fertility, were collected weekly using an artificial vagina over 6 consecutive weeks. Analyses of sperm motility by computer-assisted sperm analysis (CASA), membrane integrity by acridine orange-propidium iodide combination and sperm DNA fragmentation using the sperm chromatin dispersion test (SCD) were performed at 0, 3, 6 and 24h of incubation at 37°C. Sperm nuclear morphometry was also determined at 0h by computer-assisted sperm morphometry-fluorescence (CASMA-F). Sperm viability and most CASA sperm motility parameters were higher at 0, 3 and 6h in the high fertility rams. These rams had also a higher sperm nuclear area, perimeter and length (P<0.05) determined by CASMA-F. Significant differences between high and low fertility groups were also found in the dynamics in DNA fragmentation, with significant differences at 6h (14.42±1.40 and 20.27±1.77, respectively, P<0.05) and at 24h (22.32±2.03 and 31.24±2.54, respectively, P<0.01). It was concluded that high and low fertility rams present clear differences in several sperm quality parameters. This opens up the possibility of selection of males for artificial insemination based on sperm quality data. PMID:24602507

  8. Conserved ram seminal plasma proteins bind to the sperm membrane and repair cryopreservation damage.

    PubMed

    Bernardini, A; Hozbor, F; Sanchez, E; Fornés, M W; Alberio, R H; Cesari, A

    2011-08-01

    Whole seminal plasma (SP) enhances the function and fertility of frozen/thawed ram sperm. The objective of the current study was to investigate whether SP proteins capable of binding to molecules from the sperm plasma membrane were conserved among ram breeds, and whether these proteins were sufficient to overcome cryopreservation-induced reductions in sperm quality. Whole ram SP, obtained from rams of various breeds, improved progressive motility of frozen/thawed sperm at all times evaluated (P < 0.05); however, it did not improve total motility (15 min, P = 0.480; 30 min, P = 0.764; and 45 min, P = 0.795). To identify SP proteins responsible for this effect, a new method was developed to retain SP proteins that bound specifically to the sperm membrane by immobilization of sperm membrane proteins. These proteins specifically bound to the sperm surface, especially the acrosomal region. Lactotransferrin, epididymal secretory protein E1, Synaptosomal-associated protein 29, and RSVP-20 were identified (mass spectrometry) in this fraction. The retained SP proteins fraction repaired ultrastructural damage of frozen/thawed sperm and, with the addition of fructose, significantly improved motility of frozen/thawed sperm. We concluded that SP proteins that bound to the sperm membrane were conserved among ram breeds, and that when added to frozen/thawed semen (along with an energy source), they repaired ram sperm damage and enhanced sperm motility. PMID:21601269

  9. 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

  10. 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

  11. Sperm Proteases that May Be Involved in the Initiation of Sperm Motility in the Newt, Cynops pyrrhogaster

    PubMed Central

    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

  12. Modifications of the Test Information Function.

    ERIC Educational Resources Information Center

    Samejima, Fumiko

    Two modification formulas are presented for the test information function in order to provide better measures of local accuracies of the estimation of "theta" when maximum likelihood estimation is used to provide the estimate of ability "theta." A minimum bound of any estimator, biased or unbiased, is considered; and Formula 1 is proposed for the…

  13. Sperm competition in Odonata (Insecta): the evolution of female sperm storage and rivals' sperm displacement

    Microsoft Academic Search

    E. Uh ´ õa; A. Cordero River; E. U. E. T. Forestal

    2003-01-01

    Odonates (dragonflies) are well known for the ability of the males to displace sperm stored in the female's sperm- storage organs during copulation. By this means, copulating males are able to increase their fertilization success. This ability has been used as an example to illustrate a conflict of interests between the sexes in which males have evolved sperm-displacement mechanisms whilst

  14. 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...

  15. 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...

  16. Hanford tanks initiative test facility functions and requirements

    Microsoft Academic Search

    Krieg; Fluor Daniel Hanford

    1997-01-01

    This document presents the functions and requirements for a test facility for testing single-shell tank waste retrieval equipment and systems for the Hanford Tanks Initiative (HTI) project. This effort includes review of previous test facility functions and requirements and conducting a workshop to develop specific functions and requirements for HTI testing needs. Functions and requirements for testing future retrieval systems

  17. 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

  18. Platelet function tests: a comparative review.

    PubMed

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

  19. 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.

  20. Human sperm bound to the zona pellucida have normal nuclear chromatin as assessed by acridine orange fluorescence

    Microsoft Academic Search

    D. Y. Liu; H. W. G. Baker

    2007-01-01

    BACKGROUND: The aim of this study was to determine if only sperm with double stranded DNA could bind to the human zona pellucida (ZP). METHODS: Sperm samples from 124 infertile men with a range of semen abnormalities were studied. Oocytes that had failed to fertilize in IVF or ICSI were used for the sperm-ZP binding test. A group of four

  1. Genetic Influences on Thyroid Function Tests

    Microsoft Academic Search

    Wendy M. van der Deure; Marco Medici; Robin P. Peeters; Theo J. Visser

    \\u000a In this review we will discuss the possible effects of polymorphic variation in the genes important for thyroid hormone synthesis,\\u000a metabolism, and action, on the interindividual variation in thyroid functions tests. The genes involved are summarized in\\u000a the following outline of thyroid hormone production and action, but their role is discussed in detail in other sections (Chaps.\\u000a 1 and 4).

  2. 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

  3. An Automated Preschool Pulmonary Function Test

    PubMed Central

    Budd, Jeffrey R.; Finkelstein, Stanley M.; Warwick, Warren J.

    1981-01-01

    A non-invasive, non-effort dependent pulmonary function test has been created which can be used on preschool subjects. The integration of a mini-computer system with the test procedure allows extensive analysis of flow and gas concentration data. This analysis not only supplies lung volume measurements but also gas mixing efficiency which quantifies the evenness of gas distribution and alveolar efficiency which indicates the extent of ventilation-perfusion inequalities and diffusion abnormalities. The test has been performed on a sample of control subjects and cystic fibrosis patients aged 1 to 23 years old. The results indicate that the measurements are not only sensitive and specific to lung disease but also that they should prove useful for following the extent of lung disease over time.

  4. Sperm and spermatids contain different proteins and bind distinct egg factors.

    PubMed

    Teperek, Marta; Miyamoto, Kei; Simeone, Angela; Feret, Renata; Deery, Michael J; Gurdon, John B; Jullien, Jerome

    2014-01-01

    Spermatozoa are more efficient at supporting normal embryonic development than spermatids, their immature, immediate precursors. This suggests that the sperm acquires the ability to support embryonic development during spermiogenesis (spermatid to sperm maturation). Here, using Xenopus laevis as a model organism, we performed 2-D Fluorescence Difference Gel Electrophoresis (2D-DIGE) and mass spectrometry analysis of differentially expressed proteins between sperm and spermatids in order to identify factors that could be responsible for the efficiency of the sperm to support embryonic development. Furthermore, benefiting from the availability of egg extracts in Xenopus, we also tested whether the chromatin of sperm could attract different egg factors compared to the chromatin of spermatids. Our analysis identified: (1) several proteins which were present exclusively in sperm; but not in spermatid nuclei and (2) numerous egg proteins binding to the sperm (but not to the spermatid chromatin) after incubation in egg extracts. Amongst these factors we identified many chromatin-associated proteins and transcriptional repressors. Presence of transcriptional repressors binding specifically to sperm chromatin could suggest its preparation for the early embryonic cell cycles, during which no transcription is observed and suggests that sperm chromatin has a unique protein composition, which facilitates the recruitment of egg chromatin remodelling factors. It is therefore likely that the acquisition of these sperm-specific factors during spermiogenesis makes the sperm chromatin suitable to interact with the maternal factors and, as a consequence, to support efficient embryonic development. PMID:25244019

  5. Comprehensive profiling of accessible surface glycans of mammalian sperm using a lectin microarray

    PubMed Central

    2014-01-01

    It is well known that cell surface glycans or glycocalyx play important roles in sperm motility, maturation and fertilization. A comprehensive profile of the sperm surface glycans will greatly facilitate both basic research (sperm glycobiology) and clinical studies, such as diagnostics of infertility. As a group of natural glycan binders, lectin is an ideal tool for cell surface glycan profiling. However, because of the lack of effective technology, only a few lectins have been tested for lectin-sperm binding profiles. To address this challenge, we have developed a procedure for high-throughput probing of mammalian sperm with 91 lectins on lectin microarrays. Normal sperm from human, boar, bull, goat and rabbit were collected and analyzed on the lectin microarrays. Positive bindings of a set of ~50 lectins were observed for all the sperm of 5 species, which indicated a wide range of glycans are on the surface of mammalian sperm. Species specific lectin bindings were also observed. Clustering analysis revealed that the distances of the five species according to the lectin binding profiles are consistent with that of the genome sequence based phylogenetic tree except for rabbit. The procedure that we established in this study could be generally applicable for sperm from other species or defect sperm from the same species. We believe the lectin binding profiles of the mammalian sperm that we established in this study are valuable for both basic research and clinical studies. PMID:24629138

  6. Sperm competition effects on sperm production and expenditure in sailfin mollies, Poecilia latipinna

    Microsoft Academic Search

    Andrea S. Aspbury

    2007-01-01

    Sperm competition risk (SCR) models predict that if there is a low probability a male's ejaculate will compete with another male, individuals should invest less sperm in a mating, whereas if the probability of competition is high, males should invest more sperm. Alternatively, models of sperm competition intensity (SCI) predict that increased intensity of sperm competition leads to maximal sperm

  7. 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

  8. The effect of selected staining techniques on bull sperm morphometry.

    PubMed

    Banaszewska, Dorota; Andraszek, Katarzyna; Czubaszek, Magdalena; Biesiada-Drzazga, Barbara

    2015-08-01

    Sperm morphometry has some value as an indicator of reproductive capacity in males. In laboratory practice a variety of slide-staining methods are used during morphological evaluation of semen to predict male fertility. The aim of this study was to determine the effect of staining of semen using four different techniques on the morphometry of the bull sperm cell. The material for the study consisted of semen collected from test bulls of the Black-and-White variety of Holstein-Friesians. The results obtained in the study indicate differences in the dimensions of bull sperm heads when different slide staining techniques were used. The most similar results for sperm head dimensions were obtained in the case of SpermBlue(®) and eosin+gentian violet complex, although statistically significant differences were found between all the staining techniques. Extreme values were noted for the other staining techniques - lowest for the Papanicolaou and highest for silver nitrate, which may indicate more interference in the cell by the reagents used in the staining process. However, silver nitrate staining was best at identifying the structures of the sperm cell. Hence it is difficult to determine which of the staining methods most faithfully reveals the dimensions and shape of the bull sperm. PMID:26149220

  9. 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

  10. Human sperm chromatin epigenetic potential: genomics, proteomics, and male infertility.

    PubMed

    Castillo, Judit; Estanyol, Josep Maria; Ballescá, Josep Lluis; Oliva, Rafael

    2015-01-01

    The classical idea about the function of the mammalian sperm chromatin is that it serves to transmit a highly protected and transcriptionally inactive paternal genome, largely condensed by protamines, to the next generation. In addition, recent sperm chromatin genome-wide dissection studies indicate the presence of a differential distribution of the genes and repetitive sequences in the protamine-condensed and histone-condensed sperm chromatin domains, which could be potentially involved in regulatory roles after fertilization. Interestingly, recent proteomic studies have shown that sperm chromatin contains many additional proteins, in addition to the abundant histones and protamines, with specific modifications and chromatin affinity features which are also delivered to the oocyte. Both gene and protein signatures seem to be altered in infertile patients and, as such, are consistent with the potential involvement of the sperm chromatin landscape in early embryo development. This present work reviews the available information on the composition of the human sperm chromatin and its epigenetic potential, with a particular focus on recent results derived from high-throughput genomic and proteomic studies. As a complement, we provide experimental evidence for the detection of phosphorylations and acetylations in human protamine 1 using a mass spectrometry approach. The available data indicate that the sperm chromatin is much more complex than what it was previously thought, raising the possibility that it could also serve to transmit crucial paternal epigenetic information to the embryo. PMID:25926607

  11. Autocrine regulation of human sperm motility by tachykinins

    PubMed Central

    2010-01-01

    Background We examined the presence and function of tachykinins and the tachykinin-degrading enzymes neprilysin (NEP) and neprilysin-2 (NEP2) in human spermatozoa. Methods Freshly ejaculated semen was collected from forty-eight normozoospermic human donors. We analyzed the expression of substance P, neurokinin A, neurokinin B, hemokinin-1, NEP and NEP2 in sperm cells by reverse-transcriptase polymerase chain reaction (RT-PCR), western blot and immunocytochemistry assays and evaluated the effects of the neprilysin and neprilysin-2 inhibitor phosphoramidon on sperm motility in the absence and presence of tachykinin receptor-selective antagonists. Sperm motility was measured using WHO procedures or computer-assisted sperm analysis (CASA). Results The mRNAs of the genes that encode substance P/neurokinin A (TAC1), neurokinin B (TAC3), hemokinin-1 (TAC4), neprilysin (MME) and neprilysin-2 (MMEL1) were expressed in human sperm. Immunocytochemistry studies revealed that tachykinin and neprilysin proteins were present in spermatozoa and show specific and differential distributions. Phosphoramidon increased sperm progressive motility and its effects were reduced in the presence of the tachykinin receptor antagonists SR140333 (NK1 receptor-selective) and SR48968 (NK2 receptor-selective) but unmodified in the presence of SR142801 (NK3 receptor-selective). Conclusion These data show that tachykinins are present in human spermatozoa and participate in the regulation of sperm motility. Tachykinin activity is regulated, at least in part, by neprilysins. PMID:20796280

  12. Lysozyme activities and immunoglobulin concentrations in seminal plasma and spermatozoa of different teleost species and indications on its significance for sperm function.

    PubMed

    Lahnsteiner, F; Radner, M

    2010-07-15

    The occurrence of lysozyme and immunoglobulin (Ig) in semen of different teleost species (brown trout-Salmo trutta, perch-Perca fluviatilis, burbot-Lota lota) was studied. In all investigated species lysozyme activities (1.13-1.45 U ml(-1)) and Ig concentrations (T-Ig: 1.11-1.61 microg ml(-1), IgG [measured only in brown trout]: 1.49 microg ml(-1)) were detected in seminal plasma. Ig was also found in spermatozoa (T-Ig: 0.234-0.357 microg/g protein, IgG: 0.198 microg ml(-1)) while spermatozoal lysozyme activities were low and fluctuating (0.093-0.164 U/g protein). In Salmo trutta lysozyme activities and immunoglobulin levels were compared between semen samples with high and low sperm motility as motility is an indicator for sperm fertility. Lysozyme activities were higher in seminal plasma of samples with high motility than in those with low motility while seminal plasma and spermatozoal immunoglobulin concentrations (T-Ig, IgG) were increased in samples with low motility in comparison to samples with high motility. Seminal plasma and spermatozoal IgG concentrations and seminal plasma lysozyme activities showed significant correlations with the sperm motility rate and swimming velocity. Moreover, lysozyme improved the viability of spermatozoa in in vitro experiments. Possible physiological meanings of these results are discussed. PMID:20416943

  13. Basic and Clinical Aspects of Sperm Comet Assay

    Microsoft Academic Search

    Luke Simon; Sheena E. M. Lewis

    \\u000a Sperm DNA damage is associated with poorer assisted reproductive treatment (ART) outcomes including reduced fertilization\\u000a rates, embryo quality, and pregnancy rates and higher rates of spontaneous miscarriage and childhood diseases. It shows promise\\u000a as a more robust biomarker of infertility than conventional semen parameters. Among the sperm DNA testing methods, the alkaline\\u000a comet assay is a sensitive, reliable, and powerful

  14. Diagnostic value of sperm DNA fragmentation and sperm high-magnification for predicting outcome of assisted reproduction treatment

    PubMed Central

    López, Gemma; Lafuente, Rafael; Checa, Miguel A; Carreras, Ramón; Brassesco, Mario

    2013-01-01

    Over the last years, major improvements in the field of male infertility diagnosis have been achieved. The aim of this study was to determine the diagnostic usefulness of sperm DNA integrity and sperm vacuolisation for predicting outcome in infertile couples undergoing in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) treatments. A cohort study from 152 infertile couples undergoing sperm DNA fragmentation and high-magnification tests prior to an assisted reproduction treatment was designed. We found that the most predictive cutoff for pregnancy was 25.5% of DNA fragmentation with a negative predictive value of 72.7% (P=0.02). For the degree of vacuolisation, the best predictor of pregnancy was 73.5% of vacuolated sperm grades III+IV with a negative predictive value of 39.4% (P=0.09), which was not statistically significant. In conclusion, sperm DNA fragmentation greater than 25.5% could be associated with higher probability of failure IVF treatment. Regarding the results of the sperm analysis at high magnification, they do not allow us to predict whether or not patients will become pregnant. PMID:23912311

  15. Frozen-thawed rhinoceros sperm exhibit DNA damage shortly after thawing when assessed by the sperm chromatin dispersion assay.

    PubMed

    Portas, T; Johnston, S D; Hermes, R; Arroyo, F; López-Fernadez, C; Bryant, B; Hildebrandt, T B; Göritz, F; Gosalvez, J

    2009-09-15

    This study reports on the successful validation (via in situ nick translation and neutral comet assay) of the equine Sperm-Halomax kit as an appropriate methodology for the assessment of sperm DNA fragmentation in three species of rhinoceros. Rhinoceros sperm nuclei with fragmented DNA (validated using in situ nick translation) were evident as large halos with dispersed DNA fragments, whereas those with nonfragmented DNA displayed small halos of nondispersed DNA within the microgel. There was a high correlation (r) of 0.974 (R(2) value=0.949; P<0.01; n=16) between the respective assessments of the Sperm Chromatin Dispersion test (SCDt) and the neutral comet assay. Application of the SCDt to determine the DNA fragmentation dynamics of rhinoceros (n=6) sperm frozen in liquid nitrogen vapor and incubated postthaw at 37 degrees C for up to 48 h to mimic in vitro conditions in the female reproductive tract, revealed an increase (P=0.001) in DNA damage, as soon as 4h after the start of incubation. Linear regression equations were calculated for all six rhinoceroses over the first 6h of incubation and revealed individual animal variation. Freshly collected and incubated (37 degrees C) rhinoceros (n=3) sperm had no increase in the basal level of DNA fragmentation for up to 48 h, indicating that the cryopreservation of rhinoceros sperm in liquid nitrogen vapor, as used in this study, appeared to result in freeze-thaw DNA damage. PMID:19560805

  16. Diagnostic value of sperm DNA fragmentation and sperm high-magnification for predicting outcome of assisted reproduction treatment.

    PubMed

    López, Gemma; Lafuente, Rafael; Checa, Miguel A; Carreras, Ramón; Brassesco, Mario

    2013-11-01

    Over the last years, major improvements in the field of male infertility diagnosis have been achieved. The aim of this study was to determine the diagnostic usefulness of sperm DNA integrity and sperm vacuolisation for predicting outcome in infertile couples undergoing in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) treatments. A cohort study from 152 infertile couples undergoing sperm DNA fragmentation and high-magnification tests prior to an assisted reproduction treatment was designed. We found that the most predictive cutoff for pregnancy was 25.5% of DNA fragmentation with a negative predictive value of 72.7% (P=0.02). For the degree of vacuolisation, the best predictor of pregnancy was 73.5% of vacuolated sperm grades III+IV with a negative predictive value of 39.4% (P=0.09), which was not statistically significant. In conclusion, sperm DNA fragmentation greater than 25.5% could be associated with higher probability of failure IVF treatment. Regarding the results of the sperm analysis at high magnification, they do not allow us to predict whether or not patients will become pregnant. PMID:23912311

  17. 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.

  18. The role of sperm proteasomes during sperm aster formation and early zygote development: implications for fertilization failure in humans.

    PubMed

    Rawe, Vanesa Y; Díaz, Emilce S; Abdelmassih, Roger; Wójcik, Cezary; Morales, Patricio; Sutovsky, Peter; Chemes, Héctor E

    2008-03-01

    BACKGROUND Sperm aster organization during bovine and human fertilization requires a paternally-derived centriole that must first disengage from the sperm tail connecting-piece. We investigated the participation of the 26S proteasome in this process. METHODS Proteasome localization and enzymatic activity were studied in normal and pathological human spermatozoa by immunocytochemistry and enzyme-substrate assays. The role of proteasomes during bovine zygote development was investigated using a pharmacological proteasome-inhibitor, MG132, and with anti-proteasome antibodies delivered by Streptolysin O-permeabilization or with the Chariot reagent. Human zygotes discarded after ICSI failures (n = 28) were also examined. RESULTS Proteasomes were localized in the sperm acrosome and connecting-piece, as well as in the pronuclei of bovine and human zygotes. Proteasomal enzymatic activities were decreased in defective human spermatozoa. Disrupted sperm aster formation and pronuclear development were found after pharmacological and immunological block of proteasomes in human/bovine spermatozoa and oocytes, as well as in 28 discarded human post-ICSI fertilization failures. CONCLUSIONS Specific proteasome inhibition disrupts sperm aster formation and pronuclear development/apposition in bovine and human zygotes. Human spermatozoa with defective centriolar/pericentriolar structures have decreased proteasomal enzymatic activity. Release of a functional sperm centriole that acts as a zygote microtubule-organizing center probably relies on selective proteasomal proteolysis. These findings suggest an important role of sperm proteasomes in zygotic development. PMID:18089554

  19. Sperm Whale Project

    NSDL National Science Digital Library

    Describes an award-winning project in Homer, Alaska. After a sperm whale carcass washed ashore, a museum-school collaboration implemented a five-year cross-disciplinary marine science education project using major grant funding. Students worked with museum staff, scientists, public agencies and the community to research the whale, preserve, articulate and mount its skeleton. They developed exhibits, school and community education programs interpreting the whale and promoting ocean conservation in the region.

  20. Sperm Chromatin and Environmental Factors

    Microsoft Academic Search

    Aleksander Giwercman

    \\u000a Available experimental and human data show that a number of lifestyle- and environment-related exposures may have negative\\u000a effects on sperm DNA integrity. The extent of the sperm DNA damage seems to differ between different studies. Such a discrepancy\\u000a may partly be due to the use of ­several methods for assessment of sperm DNA integrity, techniques that do not measure exactly

  1. Comprehensive investigation in patients affected by sperm macrocephaly and globozoospermia.

    PubMed

    Chianese, C; Fino, M G; Riera Escamilla, A; López Rodrigo, O; Vinci, S; Guarducci, E; Daguin, F; Muratori, M; Tamburrino, L; Lo Giacco, D; Ars, E; Bassas, L; Costa, M; Pisatauro, V; Noci, I; Coccia, E; Provenzano, A; Ruiz-Castañé, E; Giglio, S; Piomboni, P; Krausz, C

    2015-03-01

    The aim of this study was to provide a comprehensive genetic/phenotypic characterization of subjects suffering infertility owing to sperm macrocephaly (n = 3) or globozoospermia (n = 9) and to investigate whether the patients' genetic status was correlated with the alteration of various sperm parameters. AURKC was sequenced in case of sperm macrocephaly while the DPY19L2 status has been analyzed by multiple approaches including a novel qPCR-based copy number assay in case of globozoospermia. Globozoospermic patients were also analyzed for SPACA1, a novel candidate gene herein tested for the first time in humans. The effect of the patients' genetic status was interrogated by implementing the molecular screening with the characterization of several sperm parameters: (i) routine sperm analysis, integrated with transmission electron microscopy; (ii) sperm fluorescent in situ hybridization (FISH) analysis; (iii) sperm DNA fragmentation (DF) analysis. Moreover, for the first time, we performed microsatellite instability analysis as a marker of genome instability in men with sperm macrocephaly and globozoospermia. Finally, artificial reproductive technology (ART) history has been reported for those patients who underwent the treatment. Macrocephalic patients had an AURKC mutation and >89% tetraploid, highly fragmented spermatozoa. DPY19L2 was mutated in all patients with >80% globozoospermia: the two homozygous deleted men and the compound heterozygous showed the severest phenotype (90-100%). The newly developed qPCR method was fully validated and has the potential of detecting also yet undiscovered deletions. DPY19L2 status is unlikely related to FISH anomalies and DF, although globozoospermic men showed a higher disomy rate and DF compared with internal reference values. No patient was mutated for SPACA1. Our data support the general agreement on the negative correlation between macro/globozoospermia and conventional intracytoplasmic sperm injection outcomes. Microsatellites were stable in all patients analyzed. The comprehensive picture provided on these severe phenotypes causing infertility is of relevance in the management of patients undergoing ART. PMID:25755131

  2. Hanford tanks initiative test facility functions and requirements

    SciTech Connect

    Krieg, S.A., Fluor Daniel Hanford

    1997-03-01

    This document presents the functions and requirements for a test facility for testing single-shell tank waste retrieval equipment and systems for the Hanford Tanks Initiative (HTI) project. This effort includes review of previous test facility functions and requirements and conducting a workshop to develop specific functions and requirements for HTI testing needs. Functions and requirements for testing future retrieval systems that follow HTI are also identified.

  3. A Test for Measuring Gustatory Function

    PubMed Central

    Smutzer, Gregory; Lam, Si; Hastings, Lloyd; Desai, Hetvi; Abarintos, Ray A.; Sobel, Marc; Sayed, Nabil

    2010-01-01

    Objectives The purpose of this study is to determine the usefulness of edible taste strips for measuring human gustatory function. Research Design The physical properties of edible taste strips were examined in order to determine their potential for delivering threshold and suprathreshold amounts of taste stimuli to the oral cavity. Taste strips were then assayed by fluorescence to analyze the uniformity and distribution of bitter tastant in the strips. Finally, taste recognition thresholds for sweet taste were examined in order to determine whether or not taste strips would produce recognition thresholds that were equal to or better than those obtained from aqueous tests. Methodology Edible strips were prepared from pullulan-hydroxypropyl methylcellulose solutions that were dried to a thin film. The maximal amount of a tastant that could be incorporated in a 2.54 × 2.54 cm taste strip was identified by including representative taste stimuli for each class of tastant (sweet, sour, salty, bitter, and umami) during strip formation. Distribution of the bitter tastant quinine hydrochloride in taste strips was assayed by fluorescence emission spectroscopy. The efficacy of taste strips for evaluating human gustatory function was examined by using a single series ascending method of limits protocol. Sucrose taste recognition threshold data from edible strips was then compared to results that were obtained from a standard “sip and spit” recognition threshold test. Results Edible films that formed from a pullulan-hydroxypropyl methylcellulose polymer mixture can be used to prepare clear, thin strips that have essentially no background taste and leave no physical presence after release of tastant. Edible taste strips could uniformly incorporate up to five percent of their composition as tastant. Taste recognition thresholds for sweet taste were over one order of magnitude lower with edible taste strips when compared to an aqueous taste test. Conclusion Edible taste strips are a highly sensitive method for examining taste recognition thresholds in humans. This new means of presenting taste stimuli should have widespread applications for examining human taste function in the lab, in the clinic, or at remote locations. PMID:18528309

  4. [Sperm-antibodies - practical importance in the male fertility disorder].

    PubMed

    Meili, H U; Bandhauer, K

    1976-07-01

    On the basis of abundant statistics it is known that in about 5% of infertile males fertility-inhibiting antibodies are present, which can lead to immobilization or agglutination of the sperm; they can block acrosome activity or become cytotoxically active. The motility of spermatozoon charged with antibodies and partially immobilized or agglutinized is probably not sufficient to penetrate the cervical mucus to reach the egg. Immunological sterility can be suspected in certain situations: infertility in a couple where there is no apparent cause of female infertility; anamnestic or clinical indication of chronic prostatitis, vesiculitis, or epididymitis; spontaneous agglutination or motility reduction in the spermiogram (not demonstrable in all cases); pathological postcoital test by the Sims-Huhner method. Since the last is only 50% reliable, diagnosis of antibodies is dependent on laboratory tests such as: micro-sperm-agglutination test, macro-sperm-agglutination test, sperm immobilization test, hema-agglutination test, and capillary X-ray. As yet there is no satisfactory treatment for this type of male fertility disorder. The only promising results in this area are achieved when the inflamed source for the antigen-antibody reaction is found and removed. Diagnosis of sperm antibodies in male infertility, however, can be the clear indication for heterologic insemination. PMID:947867

  5. Ejaculate Oxidative Stress Is Related with Sperm DNA Fragmentation and Round Cells

    PubMed Central

    Iommiello, Valeria Maria; Albani, Elena; Di Rosa, Alessandra; Marras, Alessandra; Menduni, Francesca; Morreale, Giovanna; Levi, Shanti Lia; Pisano, Benedetta; Levi-Setti, Paolo Emanuele

    2015-01-01

    Oxidative stress (OS) plays an essential role in male infertility aetiology by affecting sperm quality, function, and also the integrity of sperm DNA. The assessment of oxidative stress in semen may be an important tool to improve the evaluation of sperm reproductive capacity. The purpose of this study was the evaluation of any possible relation between the unbalance of oxidative stress caused by superoxide anion in the ejaculate with the presence of sperm DNA fragmentation and high concentration of round cells. 56 semen samples from males from couples suffering from infertility were evaluated according to World Health Organisation (WHO) 2010 guidelines. Oxidative stress levels from N1 (low) to N4 (high) were assessed in ejaculates using oxiSperm; DFI (sperm DNA fragmentation index) as assessed by the SCSA (Sperm Chromatin Structure Assay) was used for evaluation of sperm chromatin integrity. Our data show that high oxidative stress (N3-N4 levels) correlated positively with a DFI ? 30% (P = 0.0379) and round cells ?1.500.000/mL (P = 0.0084). In conclusion, OS increases sperm DNA damage. Thus evaluation of semen OS extent of sperm DNA damage in infertile man could be useful to develop new therapeutic strategies and improve success of assisted reproduction techniques (ART). PMID:25802519

  6. Sperm Development and Motility are Regulated by PP1 Phosphatases in Caenorhabditis elegans

    PubMed Central

    Wu, Jui-ching; Go, Aiza C.; Samson, Mark; Cintra, Thais; Mirsoian, Susan; Wu, Tammy F.; Jow, Margaret M.; Routman, Eric J.; Chu, Diana S.

    2012-01-01

    Sperm from different species have evolved distinctive motility structures, including tubulin-based flagella in mammals and major sperm protein (MSP)-based pseudopods in nematodes. Despite such divergence, we show that sperm-specific PP1 phosphatases, which are required for male fertility in mouse, function in multiple processes in the development and motility of Caenorhabditis elegans amoeboid sperm. We used live-imaging analysis to show the PP1 phosphatases GSP-3 and GSP-4 (GSP-3/4) are required to partition chromosomes during sperm meiosis. Postmeiosis, tracking fluorescently labeled sperm revealed that both male and hermaphrodite sperm lacking GSP-3/4 are immotile. Genetic and in vitro activation assays show lack of GSP-3/4 causes defects in pseudopod development and the rate of pseudopodial treadmilling. Further, GSP-3/4 are required for the localization dynamics of MSP. GSP-3/4 shift localization in concert with MSP from fibrous bodies that sequester MSP at the base of the pseudopod, where directed MSP disassembly facilitates pseudopod contraction. Consistent with a role for GSP-3/4 as a spatial regulator of MSP disassembly, MSP is mislocalized in sperm lacking GSP-3/4. Although a requirement for PP1 phosphatases in nematode and mammalian sperm suggests evolutionary conservation, we show PP1s have independently evolved sperm-specific paralogs in separate lineages. Thus PP1 phosphatases are highly adaptable and employed across a broad range of sexually reproducing species to regulate male fertility. PMID:22042574

  7. Ejaculate oxidative stress is related with sperm DNA fragmentation and round cells.

    PubMed

    Iommiello, Valeria Maria; Albani, Elena; Di Rosa, Alessandra; Marras, Alessandra; Menduni, Francesca; Morreale, Giovanna; Levi, Shanti Lia; Pisano, Benedetta; Levi-Setti, Paolo Emanuele

    2015-01-01

    Oxidative stress (OS) plays an essential role in male infertility aetiology by affecting sperm quality, function, and also the integrity of sperm DNA. The assessment of oxidative stress in semen may be an important tool to improve the evaluation of sperm reproductive capacity. The purpose of this study was the evaluation of any possible relation between the unbalance of oxidative stress caused by superoxide anion in the ejaculate with the presence of sperm DNA fragmentation and high concentration of round cells. 56 semen samples from males from couples suffering from infertility were evaluated according to World Health Organisation (WHO) 2010 guidelines. Oxidative stress levels from N1 (low) to N4 (high) were assessed in ejaculates using oxiSperm; DFI (sperm DNA fragmentation index) as assessed by the SCSA (Sperm Chromatin Structure Assay) was used for evaluation of sperm chromatin integrity. Our data show that high oxidative stress (N3-N4 levels) correlated positively with a DFI ? 30% (P = 0.0379) and round cells ?1.500.000/mL (P = 0.0084). In conclusion, OS increases sperm DNA damage. Thus evaluation of semen OS extent of sperm DNA damage in infertile man could be useful to develop new therapeutic strategies and improve success of assisted reproduction techniques (ART). PMID:25802519

  8. Effects of Kamdhenu Ark and Active Immunization by Gonadotropin Releasing Hormone Conjugate (GnRH-BSA) on Gonadosomatic Indices (GSI) and Sperm Parameters in Male Mus musculus

    PubMed Central

    Ganaie, Javid Ahmad; Gautam, Varsha; Shrivastava, Vinoy Kumar

    2011-01-01

    Background Active immunization against GnRH decreases the secretion of gonadotropins and causes cessation of gonadal function, thereby, inducing infertility. Based on the immunoenhancing activity of Kamdhenu ark (distilled cow urine), this study was performed to evaluate its effects on the gonadosomatic indices (GSI) and sperm parameters in male mice receiving a GnRH contraceptive vaccine. Methods Sixty adult male mice of Parke's strain were divided into three groups of twenty. Group I served as the controls, while group II was immunized by GnRH-BSA conjugate (50/0.2/35 µg/ml/g BW) by four intraperitoneal injections at different intervals on days 1, 30, 60 and 90. However, group III was supplemented daily by oral Kamdhenu ark (100 ppm) along with GnRH-BSA immunizations. The animals were sacrificed after 30, 60, 90 and 120 days and their testis and epididymis were dissected out weighed and semen analysis was performed. Results GSI values, sperm motility, sperm count and sperm morphology in male Mus musculus were decreased significantly in all the experimental groups as compared to the control group (p<0.01). Kamdhenu ark significantly enhanced the effect of GnRH vaccine on the aforesaid parameters especially in 90 and 120 days treated groups (p<0.05). Conclusion The changes witnessed in sperm parameters suggested that the GnRH-BSA immunization suppressed the activities of gonadotropins and testosterone directly through hypothalamo-hypophysial-gonadal axis and indirectly by acting on the testes which may modulate the sperm morphology, sperm count and motility. However, Kamdhenu ark seems to have enhanced these effects because of its immune-modulatory properties too. PMID:23926493

  9. Sperm-mediated gene transfer

    Microsoft Academic Search

    Anthony W. S. Chan; C. Marc Luetjens; Gerald P. Schatten

    2000-01-01

    Since 1989, a new method for the production of transgenic animals has been available, namely sperm- mediated gene transfer (SMGT), based on the intrinsic ability of sperm cells to bind and internalise exogenous DNA molecules and to transfer them into the oocyte at fertilisation. We first described the SMGT procedure in a small animal model, with high efficiency reported in

  10. 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

  11. Spermatangium formation and sperm discharge in the Japanese pygmy squid Idiosepius paradoxus.

    PubMed

    Sato, Noriyosi; Kasugai, Takashi; Munehara, Hiroyuki

    2014-06-01

    In cephalopods, sperm discharge is an important event not only for sperm transfer but also influencing sperm storage capacity of attached spermatangia (everted spermatophores). To investigate sperm discharge from spermatangia and the condition of naturally attached spermatangia in Japanese pygmy squid (Idiosepius paradoxus) we (i) investigated the morphology of spermatophores and spermatangia, and the process of spermatophore evagination and sperm discharge from spermatangia obtained in vitro; (ii) observed spermatangia that were naturally attached to female squids at 6, 12, 18, 24 and 48 h after copulation to investigate alterations in naturally attached spermatangia with time. The spermatophore of I. paradoxus is slender and cylindrical and consists of a sperm mass, a cement body and an ejaculatory apparatus, which is similar to those of loliginid squids. The spermatangium is fishhook-shaped, its distal end being open and narrow. After the spermatangium is formed, the sperm mass gradually moves to the open end of the spermatangium, from where sperm are released. Sperm discharge is a rapid process immediately after the beginning of sperm release, but within 5 min changes to an intermittent release of sperm. Although the volume of residual spermatozoa differed among spermatangia that were naturally attached to a single individual, the probability that spermatangia would be empty increased with time. Most naturally attached spermatangia discharged almost all of their spermatozoa within 24h after copulation, and no spermatangia were attached to females 48 h after copulation. These results suggest that sperm transfer from the spermatangium to the seminal receptacle must occur within 24h, and that the spermatangium functions as a transient sperm storage organ in pygmy squids. PMID:24813852

  12. Sperm donation in Israel.

    PubMed

    Mor-Yosef, S; Schenker, J G

    1995-04-01

    Science and technology in the field of human reproduction present new legal, ethical and religious questions which do not always have immediate answers. The first step in the rapidly developed field of reproductive technology was the use of sperm donation (artificial insemination by donor, AID) and the establishment of sperm banks. The state of Israel faced these problems when the regulations for sperm donation were discussed. The fact that the main holy places for the three monotheistic religions are in Israel directly influences the make-up of the population constituents. Therefore, besides a majority of secular people, a high percentage of the population of Israel is very religious: Jews, Moslems and Christians. Thus any resolution relating to AID should take this demographic combination into account. The practice of AID is opposed by the different monotheistic religions. To avoid the conflict between secular and religious people, and between the different religions' perspectives, the legal problem of AID in Israel was solved not by laws but by regulations which were published by the Ministry of Health. The main idea behind this attitude is that the state and its authorities should not and do not deal with ethical or religious questions. Thus, the decision was left to the couples and to the donors. The regulations address technical requirements, health problems and confidential issues concerning the couple, the donor and the child. In this paper we present the different views relating to these problems as perceived by the different religions, and describe the solution that was accepted by the Israeli Ministry of Health. PMID:7650152

  13. Transforming and Selecting Functional Test Cases for Security Policy Testing Tejeddine Mouelhi, Yves Le Traon

    E-print Network

    Paris-Sud XI, Université de

    Transforming and Selecting Functional Test Cases for Security Policy Testing Tejeddine Mouelhi to reuse and automatically transform existing functional test cases for specifically testing the security, XACML, Or- BAC) and AOP for transforming automatically func- tional test cases into security policy test

  14. 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

  15. Seminal vesicle protein SVS2 is required for sperm survival in the uterus

    PubMed Central

    Kawano, Natsuko; Araki, Naoya; Yoshida, Kaoru; Hibino, Taku; Ohnami, Naoko; Makino, Maako; Kanai, Seiya; Hasuwa, Hidetoshi; Yoshida, Manabu; Miyado, Kenji; Umezawa, Akihiro

    2014-01-01

    In mammals, sperm migrate through the female reproductive tract to reach the egg; however, our understanding of this journey is highly limited. To shed light on this process, we focused on defining the functions of seminal vesicle secretion 2 (SVS2). SVS2?/? male mice produced sperm but were severely subfertile, and formation of a copulatory plug to cover the female genital opening did not occur. Surprisingly, even when artificial insemination was performed with silicon as a substitute for the plug, sperm fertility in the absence of SVS2 remained severely reduced because the sperm were already dead in the uterus. Thus, our results provide evidence that the uterus induces sperm cell death and that SVS2 protects sperm from uterine attack. PMID:24591616

  16. Delayed male maturity is a cost of producing large sperm in Drosophila.

    PubMed Central

    Pitnick, S; Markow, T A; Spicer, G S

    1995-01-01

    Among fruit-fly species of the genus Drosophila there is remarkable variation in sperm length, with some species producing gigantic sperm (e.g., > 10 times total male body length). These flies are also unusual in that males of some species exhibit a prolonged adult nonreproductive phase. We document sperm length, body size, and sex-specific ages of reproductive maturity for 42 species of Drosophila and, after controlling for phylogeny, test hypotheses to explain the variation in rates of sexual maturation. Results suggest that delayed male maturity is a cost of producing long sperm. A possible physiological mechanism to explain the observed relationship is discussed. PMID:7479851

  17. Functional testing philosophies using neural networks

    Microsoft Academic Search

    L. V. Kirkland; R. G. Wright

    1997-01-01

    This paper describes the use of neural networks in combination with algorithmic test programs to aid in improving test efficiency and accuracy, especially in test situations where “bad actor” test programs exist that have difficulty in detecting and isolating Unit Under Test (UUT) failures. The paper will begin with a discussion of the theoretical basis for the use of neural

  18. Formal functional test designs with a test representation language

    NASA Technical Reports Server (NTRS)

    Hops, J. M.

    1993-01-01

    The application of the category-partition method to the test design phase of hardware, software, or system test development is discussed. The method provides a formal framework for reducing the total number of possible test cases to a minimum logical subset for effective testing. An automatic tool and a formal language were developed to implement the method and produce the specification of test cases.

  19. Deletion of murine choline dehydrogenase results in diminished sperm motility

    PubMed Central

    Johnson, Amy R.; Craciunescu, Corneliu N.; Guo, Zhong; Teng, Ya-Wen; Thresher, Randy J.; Blusztajn, Jan K.; Zeisel, Steven H.

    2010-01-01

    Choline dehydrogenase (CHDH) catalyzes the conversion of choline to betaine, an important methyl donor and organic osmolyte. We have previously identified single nucleotide polymorphisms (SNPs) in the human CHDH gene that, when present, seem to alter the activity of the CHDH enzyme. These SNPs occur frequently in humans. We created a Chdh?/? mouse to determine the functional effects of mutations that result in decreased CHDH activity. Chdh deletion did not affect fetal viability or alter growth or survival of these mice. Only one of eleven Chdh?/? males was able to reproduce. Loss of CHDH activity resulted in decreased testicular betaine and increased choline and PCho concentrations. Chdh+/+ and Chdh?/? mice produced comparable amounts of sperm; the impaired fertility was due to diminished sperm motility in the Chdh?/? males. Transmission electron microscopy revealed abnormal mitochondrial morphology in Chdh?/? sperm. ATP content, total mitochondrial dehydrogenase activity and inner mitochondrial membrane polarization were all significantly reduced in sperm from Chdh?/? animals. Mitochondrial changes were also detected in liver, kidney, heart, and testis tissues. We suggest that men who have SNPs in CHDH that decrease the activity of the CHDH enzyme could have decreased sperm motility and fertility.—Johnson, A. R., Craciunescu, C. N., Guo, Z., Teng, Y.-W., Thresher, R. J., Blusztajn, J. K., Zeisel, S. H. Deletion of murine choline dehydrogenase results in diminished sperm motility. PMID:20371614

  20. The alteration of profile analysis to accommodate testing functions

    NASA Technical Reports Server (NTRS)

    Myers, R. H.

    1979-01-01

    The development of a methodology was studied for testing differences among several pilot functions, where the data points represent averages at various frequencies. Topics discussed include: basic assumptions, hypothesis, profile analysis, alteration of profile analysis to accommodate testing functions, test and procedures, and power of tests.

  1. Identification and validation of mouse sperm proteins correlated with epididymal maturation

    PubMed Central

    Ijiri, Takashi W.; Merdiushev, Tanya; Cao, Wenlei; Gerton, George L.

    2012-01-01

    Sperm need to mature in the epididymis to become capable of fertilization. To understand the molecular mechanisms of mouse sperm maturation, we conducted a proteomic analysis using saturation dye labeling to identify proteins of caput and cauda epididymal sperm that exhibited differences in amounts or positions on two-dimensional gels. Of eight caput epididymal sperm-differential proteins, three were molecular chaperones and three were structural proteins. Of nine cauda epididymal sperm-differential proteins, six were enzymes of energy metabolism. To validate these proteins as markers of epididymal maturation, immunoblotting and immunofluorescence analyses were performed. During epididymal transit, heat shock protein 2 was eliminated with the cytoplasmic droplet and smooth muscle ?-actin exhibited reduced fluorescence from the anterior acrosome while the signal intensity of aldolase A increased, especially in the principal piece. Besides these changes, we observed protein spots, such as glutathione S-transferase mu 5 and the E2 component of pyruvate dehydrogenase complex, shifting to more basic isoelectric points, suggesting post-translational changes such dephosphorylation occur during epididymal maturation. We conclude that most caput epididymal sperm-differential proteins contribute to the functional modification of sperm structures and that many cauda epididymal sperm-differential proteins are involved in ATP production that promotes sperm functions such as motility. PMID:21805633

  2. Pulmonary function tests in ulcerative colitis

    PubMed Central

    Amra, Babak; Ataabadi, Ghazal; Emami, Mohamad Hassan; Hassanzadeh, Akbar; Golshan, Mohammad; Soltaninejad, Forogh

    2014-01-01

    Background: Pulmonary impairment in patients suffering ulcerative colitis (UC) has been suggested by several investigators using standard pulmonary function tests (PFTs). This changes in pulmonary function associated with minimal respiratory symptoms have been documented, especially in patients with active disease. The aim of this prospective study was to determine airway resistance and lung volumes in patients with UC who have no respiratory symptoms in comparisons to a healthy control group. Materials and Methods: We evaluated a total of 30 patients with UC by means of spirometry, body plethysmography, and impulse oscillometry. The patients were not complaining of any pulmonary symptoms and did not present any history of previous respiratory diseases. As controls we examined 30 healthy subjects matched for gender, age, and smoking status. The relationship between PFT, lung volume, and airway resistance; and the activity, localization, and duration of the UC disease were analyzed. Results: There was a significant difference between airway resistances (kPa/L/s) measured by body plethysmography in patients with UC and those of the controls (R5hz; 0.60 ± 0.44 vs. 0.39 ± 0.13; P < 0.001) and R20hz (0.37 ± 0.19 vs. 0.29 ± 0.1, P = 0.02). There were no correlation between PFT, airway resistance and site and scoring activity (P > 0.05). Conclusion: Despite the lack of pulmonary symptoms, increased airway resistance was found in UC patients. We also have not found correlation between PFT, lung volume and airway resistance values and scoring of UC activity. PMID:25364358

  3. Male infertility testing: reactive oxygen species and antioxidant capacity.

    PubMed

    Ko, Edmund Y; Sabanegh, Edmund S; Agarwal, Ashok

    2014-12-01

    Reactive oxygen species (ROS) are an integral component of sperm developmental physiology, capacitation, and function. Elevated ROS levels, from processes such as infection or inflammation, can be associated with aberrations of sperm development, function, and fertilizing capacity. We review the impact of ROS on sperm physiology, its place in infertility evaluation, the implications for reproductive outcomes, and antioxidant therapy. Our systematic review of PubMed literature from the last 3 decades focuses on the physiology and etiology of ROS and oxidative stress (OS), evaluation of ROS, and antioxidants. ROS is normally produced physiologically and is used to maintain cellular processes such as sperm maturation, capacitation, and sperm-oocyte interaction. When ROS production exceeds the buffering capacity of antioxidants, OS occurs and can have a negative impact on sperm and fertility. ROS and antioxidant capacity testing can potentially add additional prognostic information to standard laboratory testing for the infertile male, although its role as standard part of an evaluation has yet to be determined. Elevated ROS levels have been implicated with abnormal semen parameters and male infertility, but the impact of ROS on fertilization rates and pregnancy is controversial. This is partly because of the lack of consensus on what type of patients may be suitable for ROS testing and assay standardization. Routine ROS testing for the infertile male is not currently recommended. PMID:25458618

  4. 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.

  5. 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...

  6. DIBROMOACETIC ACID AFFECTS REPRODUCTIVE COMPETENCE AND SPERM QUALITY IN THE MALE RAT

    EPA Science Inventory

    We have recently shown that Dibromoacetic acid (DBAA) alters sperm quality in short duration tests. n this study, male rats were gavaged with 0, 2, 10, 50, 250 mg DBAA/kg/d for up to 49 d. Interim. and terminal measurements of sperm quality & reproductive outcome were made. BAA c...

  7. Sperm motility and fertilizing ability in the Persian sturgeon Acipenser persicus

    E-print Network

    Villefranche sur mer

    Sperm motility and fertilizing ability in the Persian sturgeon Acipenser persicus Sayyed Mohammad_alavi@yahoo.com Abstract The motilityand fertilizing abilityof the Persian stur- geon, Acipenser persicus, spermatozoa were, at dilution rate 1:50 and pH 8.0. To test the fertilizing ability of sperm, two bu¡ered saline solutions were

  8. Shorter sperm confer higher competitive fertilization success.

    PubMed

    García-González, Francisco; Simmons, Leigh W

    2007-04-01

    Spermatozoa exhibit taxonomically widespread patterns of divergent morphological evolution. However, the adaptive significance of variation in sperm morphology remains unclear. In this study we examine the role of natural variation in sperm length on fertilization success in the dung beetle Onthophagus taurus. We conducted sperm competition trials between males that differed in the length of their sperm and determined the paternity of resulting offspring using amplified fragment length polymorphism (AFLP) markers. We also quantified variation in the size and shape of the female's sperm storage organ to determine whether female morphology influenced the competitiveness of different sperm morphologies. We found that fertilization success was biased toward males with relatively shorter sperm, but that selection on sperm length was dependent on female tract morphology; selection was directional for reduced sperm length across most of the spermathecal size range, but stabilizing in females with the smallest spermathecae. Our data provide empirical support for the theory that sperm competition should favor the evolution of numerous tiny sperm. Moreover, because sperm length is both heritable and genetically correlated with condition, our results are consistent with a process by which females can accrue genetic benefits for their offspring from the incitement of sperm competition and/or cryptic female choice, as proposed by the "sexy sperm" and "good sperm" models for the evolution of polyandry. PMID:17439614

  9. 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.

  10. Failed Sperm Development as a Reproductive Isolating Barrier between Species

    PubMed Central

    Wünsch, Lisa K.; Pfennig, Karin S.

    2013-01-01

    SUMMARY Hybrid male sterility is a common reproductive isolating barrier between species. Yet, little is known about the actual developmental causes of this phenomenon, especially in naturally hybridizing species. We sought to evaluate the developmental causes of hybrid male sterility, using spadefoot toads as our study system. Plains spadefoot toads (S. bombifrons) and Mexican spadefoot toads (S. multiplicata) hybridize where they co-occur in the southwestern USA. Hybrids are viable, but hybrid males suffer reduced fertility. We compared testes size and developmental stages of sperm cell maturation between hybrid males and males of each species. We found that testes of hybrid males did not differ in mean size from pure-species males. However, hybrids showed a greater range of within-individual variation in testes size than pure-species males. Moreover, although hybrids produced similar numbers of early stage sperm cells, hybrids produced significantly fewer mature spermatozoids than pure-species males. Interestingly, an introgressed individual produced numbers of live sperm comparable to pure-species males, but the majority of these sperm cells were abnormally shaped and non-motile. These results indicate that hybrid incompatibilities in late sperm development serve as a reproductive isolating barrier between species. The nature of this breakdown highlights the possibilities that hybrid males may vary in fertility and that fertility could possibly be recovered in introgressed males. PMID:24261446

  11. Possible cytoskeletal structures of rainbow trout sperm revealed by electron microscopic observation after detergent extraction

    Microsoft Academic Search

    Maya D. Markova; Ralitsa S. Zhivkova

    2003-01-01

    Despite the considerable research interest in fish sperm ultrastructure, little is known about the functions of different sperm cell components. Our electron microscopic study was aimed at identifying possible tissue-specific cytoskeletal structures in spermatozoa of rainbow trout Oncorhynchus mykiss (Teleostei, Salmoniformes, Salmonidae; formerly Salmo gairdneri). Based on the known resistance of the cytoskeleton to nonionic detergents, we compared the ultrastructure

  12. New staining methods for sperm evaluation estimated by microscopy and flow cytometry

    Microsoft Academic Search

    M. Magistrini; E. Guitton; Y. Levern; J. Cl. Nicolle; M. Vidament; D. Kerboeuf; E. Palmer

    1997-01-01

    New staining methods and automated instruments are now available to evaluate the sperm cell in vitro. Individual compartments of the sperm cell, such as the nucleus and the plasma and acrosomal membranes, may be investigated, as well as the cell function as shown by mitochondria activity and capacitation. Various probes are used and they can be analyzed by direct light

  13. 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

  14. TRY-5 is a sperm-activating protease in Caenorhabditis elegans seminal fluid.

    PubMed

    Smith, Joseph R; Stanfield, Gillian M

    2011-11-01

    Seminal fluid proteins have been shown to play important roles in male reproductive success, but the mechanisms for this regulation remain largely unknown. In Caenorhabditis elegans, sperm differentiate from immature spermatids into mature, motile spermatozoa during a process termed sperm activation. For C. elegans males, sperm activation occurs during insemination of the hermaphrodite and is thought to be mediated by seminal fluid, but the molecular nature of this activity has not been previously identified. Here we show that TRY-5 is a seminal fluid protease that is required in C. elegans for male-mediated sperm activation. We observed that TRY-5::GFP is expressed in the male somatic gonad and is transferred along with sperm to hermaphrodites during mating. In the absence of TRY-5, male seminal fluid loses its potency to transactivate hermaphrodite sperm. However, TRY-5 is not required for either hermaphrodite or male fertility, suggesting that hermaphrodite sperm are normally activated by a distinct hermaphrodite-specific activator to which male sperm are also competent to respond. Within males, TRY-5::GFP localization within the seminal vesicle is antagonized by the protease inhibitor SWM-1. Together, these data suggest that TRY-5 functions as an extracellular activator of C. elegans sperm. The presence of TRY-5 within the seminal fluid couples the timing of sperm activation to that of transfer of sperm into the hermaphrodite uterus, where motility must be rapidly acquired. Our results provide insight into how C. elegans has adopted sex-specific regulation of sperm motility to accommodate its male-hermaphrodite mode of reproduction. PMID:22125495

  15. TRY-5 Is a Sperm-Activating Protease in Caenorhabditis elegans Seminal Fluid

    PubMed Central

    Smith, Joseph R.; Stanfield, Gillian M.

    2011-01-01

    Seminal fluid proteins have been shown to play important roles in male reproductive success, but the mechanisms for this regulation remain largely unknown. In Caenorhabditis elegans, sperm differentiate from immature spermatids into mature, motile spermatozoa during a process termed sperm activation. For C. elegans males, sperm activation occurs during insemination of the hermaphrodite and is thought to be mediated by seminal fluid, but the molecular nature of this activity has not been previously identified. Here we show that TRY-5 is a seminal fluid protease that is required in C. elegans for male-mediated sperm activation. We observed that TRY-5::GFP is expressed in the male somatic gonad and is transferred along with sperm to hermaphrodites during mating. In the absence of TRY-5, male seminal fluid loses its potency to transactivate hermaphrodite sperm. However, TRY-5 is not required for either hermaphrodite or male fertility, suggesting that hermaphrodite sperm are normally activated by a distinct hermaphrodite-specific activator to which male sperm are also competent to respond. Within males, TRY-5::GFP localization within the seminal vesicle is antagonized by the protease inhibitor SWM-1. Together, these data suggest that TRY-5 functions as an extracellular activator of C. elegans sperm. The presence of TRY-5 within the seminal fluid couples the timing of sperm activation to that of transfer of sperm into the hermaphrodite uterus, where motility must be rapidly acquired. Our results provide insight into how C. elegans has adopted sex-specific regulation of sperm motility to accommodate its male-hermaphrodite mode of reproduction. PMID:22125495

  16. Sperm competition and the evolution of male reproductive anatomy in rodents

    PubMed Central

    Ramm, Steven A; Parker, Geoffrey A; Stockley, Paula

    2005-01-01

    Sperm competition is a pervasive selective force in evolution, shaping reproductive anatomy, physiology and behaviour. Here, we present comparative evidence that varying sperm competition levels account for variation in the male reproductive anatomy of rodents, the largest and most diverse mammalian order. We focus on the sperm-producing testes and the accessory reproductive glands, which produce the seminal fluid fraction of the ejaculate. We demonstrate a positive association between relative testis size and the prevalence of within-litter multiple paternity, consistent with previous analyses in which relative testis size has been found to correlate with sperm competition levels inferred from social organization and mating systems. We further demonstrate an association between sperm competition level and the relative size of at least two accessory reproductive glands: the seminal vesicles and anterior prostate. The size of the major product of these glands—the copulatory plug—is also found to vary with sperm competition level. Our findings thus suggest that selection for larger plugs under sperm competition may explain variation in accessory gland size, and highlight the need to consider both sperm and non-sperm components of the male ejaculate in the context of post-copulatory sexual selection. PMID:16024351

  17. No Experimental Evidence for Sneaking in a West African Cichlid Fish with Extremely Long Sperm

    PubMed Central

    Langen, Kathrin; Thünken, Timo; Bakker, Theo C. M.

    2013-01-01

    Alternative reproductive tactics are widespread in fishes, increasing the potential for sperm competition. Sperm competition has enormous impact on both variation in sperm numbers and sperm size. In cichlids, the sperm competition risk is very divergent and longer sperm are usually interpreted as adaptation to sperm competition. Here we examined whether sneaking tactics exist in Pelvicachromis taeniatus, a socially monogamous cichlid with biparental brood care from West Africa. The small testis indicates low gonadal investment which is typical for genetically monogamous species. In contrast, sperm length with up to 85??m is extraordinarily long. We examined the reproductive behaviour of ten groups with a male-biased sex ratio under semi-natural conditions via continuous video recording. We recorded spawning site preferences and correlates of reproductive success and conducted paternity tests using microsatellites. Safe breeding sites that could be successfully defended were preferred. All offspring could be assigned to their parents and no multiple paternities were detected. Body size of spawning pairs predicted their spawning probability and offspring hatching rate suggesting benefits from mating with large individuals. Our study suggests low risk of sperm competition under the given conditions in P. taeniatus and thus first evidence for genetic monogamy in a substrate breeding cichlid. PMID:24386589

  18. 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

  19. 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…

  20. Sperm retrieval for azoospermia and intracytoplasmic sperm injection success rates--a personal overview.

    PubMed

    Silber, Sherman J

    2010-12-01

    It is often questioned whether sperm parameters, including whether retrieved or ejaculated, have any effect on intracytoplasmic sperm injection (ICSI) results. Do severe spermatogenic defects affect embryo quality or pregnancy rate? Further, does it matter in azoospermic patients whether the sperm source is testicular or epididymal? Our studies show there is no significant difference in results with ICSI related to any sperm count parameters either with patient's sperm or even with donor sperm. No matter how poor the sperm count, there was no difference from patients with high sperm counts nor even patients using donor sperm. There is no significant difference between results with epididymal sperm, either fresh or frozen, in comparison to results with ejaculated or donor sperm. However, both pregnancy rate and delivery rate were considerably lower with testicular sperm (testis sperm extraction) than with epididymal sperm (microsurgical epididymal sperm aspiration). This was true for overall results as well as in each category of the female partner's age. It is obvious that in all these cycles the female partner's age was the most important determinant of delivery rate, but testicular sperm always yielded lower results than epididymal sperm. These results show that it is the origin of the sperm rather than the spermatogenic defect that determines success rate with ICSI. PMID:21117935

  1. Effect of ram age on structural and functional competence of frozen-thawed spermatozoa in dairy sheep.

    PubMed

    Lymberopoulos, A G; Tsakmakidis, I A; Khalifa, T A A

    2010-08-01

    The objectives of this study were to investigate the influence of ram age on structural and functional competence of frozen-thawed spermatozoa and to test the hypothesis that increasing number of sperm bound to the zona pellucida in vitro was associated with decreasing in vivo fertility of frozen semen. Rams were allocated into two groups. Each group consisted of five rams aged either 1-2 years (young) or 4-5 years (mature). Three successive ejaculates were collected from each ram using an artificial vagina. Only ejaculates of >or= 2.5 x 10(9) sperm/ml and 80% sperm progressive motility were pooled per ram, diluted with Bioxcell medium and frozen in 0.25 ml straws. The end points of post-thawing semen evaluation were computer-assisted cell motility analysis, sperm capacitation (chlortetracycline assay), simultaneous assessment of plasma membrane integrity, mitochondrial membrane potential and condensation status of nucleus, per-cell analysis of lipid peroxidation using C11-BODIPY(581/591), sperm-hemizona binding (HZB) ability and sperm fertility after laparoscopic insemination of ewes (n = 114) in the progestagen-synchronized oestrus. The results showed that mature rams had significantly lower values of sperm hyperactivated motility and peroxidized sperm, higher percentages of live non-capacitated sperm and sperm cells with intact plasma membrane, functional mitochondria and condensed chromatin, as well as, greater lambing rate and ewe prolificacy. Sperm HZB binding ability was higher (p < 0.05) for young than for mature rams. Significant correlations were found between number of spermatozoa bound to the zona pellucida and semen fertility (r = -0.63 to -0.71). In conclusion, mature rams have better semen quality and in vivo fertility than young rams. Cryocapacitation can be involved in decreasing ram semen fertility as evidenced by the high number of spermatozoa bound to the zona pellucida in vitro. PMID:19055551

  2. Transforming and Selecting Functional Test Cases for Security Policy Testing

    Microsoft Academic Search

    Tejeddine Mouelhi; Yves Le Traon; Benoit Baudry

    2009-01-01

    In this paper, we consider typical applications in which the business logic is separated from the access control logic, implemented in an independent compo- nent, called the Policy Decision Point (PDP). The execution of functions in the business logic should thus include calls to the PDP, which grants or denies the access to the protected resources\\/functionalities of the system, depending

  3. Dephosphorylation of sperm guanylate cyclase during sea urchin fertilization

    SciTech Connect

    Ward, G.E.

    1985-01-01

    When intact Arbacia punctulata spermatozoa are exposed to solubilized egg jelly, the electrophoretic mobility of an abundant sperm flagellar membrane protein changes from an apparent molecular mass of 160 kDa to 150 kDa. A. punctulata spermatozoa can be labeled in vivo with /sup 32/P-labeled cells it was demonstrated that the mobility shift of the 160-kDa protein is due to dephosphorylation. The peptide resact (Cys-Val-Thr-Gly-Ala-Pro-Gly-Cys-Val-Gly-Gly-Gly-Arg-Leu-NH/sub 2/) is the component of egg jelly which is responsible for inducing the dephosphorylation. The 160/150-kdal sperm membrane protein has been purified to homogeneity by affinity chromatography on concanavalin A-agarose, and identified as sperm guanylate cyclase. The enzymatic activity of the guanylate cyclase is tightly coupled to its phosphorylation state. Resact has been shown to act as a potent chemoattractant for A. punctulata spermatozoa. The chemotactic response is concentration-dependent, is abolished by pretreatment of the spermatozoa with resact, and shows an absolute requirement for external calcium. This work represents the first demonstration of animal sperm chemotaxis in response to a precisely-defined molecule of egg origin. The results established a new, biologically meaningful function for resact, and may implicate sperm guanylate cyclase and cGMP in flagellar function and the chemotactic response.

  4. 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

  5. Sperm Trajectories Form Chiral Ribbons

    NASA Astrophysics Data System (ADS)

    Su, Ting-Wei; Choi, Inkyum; Feng, Jiawen; Huang, Kalvin; McLeod, Euan; Ozcan, Aydogan

    2013-04-01

    We report the discovery of an entirely new three-dimensional (3D) swimming pattern observed in human and horse sperms. This motion is in the form of `chiral ribbons', where the planar swing of the sperm head occurs on an osculating plane creating in some cases a helical ribbon and in some others a twisted ribbon. The latter, i.e., the twisted ribbon trajectory, also defines a minimal surface, exhibiting zero mean curvature for all the points on its surface. These chiral ribbon swimming patterns cannot be represented or understood by already known patterns of sperms or other micro-swimmers. The discovery of these unique patterns is enabled by holographic on-chip imaging of >33,700 sperm trajectories at >90-140 frames/sec, which revealed that only ~1.7% of human sperms exhibit chiral ribbons, whereas it increases to ~27.3% for horse sperms. These results might shed more light onto the statistics and biophysics of various micro-swimmers' 3D motion.

  6. Age and gender differences in seven tests of functional mobility

    PubMed Central

    Butler, Annie A; Menant, Jasmine C; Tiedemann, Anne C; Lord, Stephen R

    2009-01-01

    Background The objective of this study was to examine age and gender differences in seven tests of functional mobility. Methods The study included 50 young participants aged 20 to 39 years, and 684 older participants aged 75 to 98 years. Functional mobility measures included the coordinated stability test, the near tandem balance test, the six metre walk test, the sit to stand test with five repetitions, the alternate step test and the stair ascent and descent tests. Results Older participants performed significantly worse than the younger participants in all of the functional mobility tests (p < 0.001), with the older women performing worse than the older men in all of the tests (p < 0.05). Significant correlations were found within the older group among all the functional mobility tests scores (r = 0.24–0.87, p < 0.001), and between functional mobility performance and age (r = 0.14–0.35, p < 0.001). People with arthritis and stroke performed worse than people without these conditions in these tests. Conclusion This study provides a normative database for performance of young and older community-dwelling people in a battery of validated and reliable functional mobility tests. The results confirm age-related differences in functional mobility between young and older adults. PMID:19642991

  7. Sperm-Egg Interaction: Evidence for Boar Sperm Plasma Membrane Receptors for Porcine Zona Pellucida

    NASA Astrophysics Data System (ADS)

    Peterson, Rudolph N.; Russell, Lonnie; Bundman, Donna; Freund, Matthew

    1980-01-01

    Freshly ejaculated, noncapacitated boar sperm bind rapidly and in large numbers to pig egg zona pellucida in vitro. In the present study, the number of sperm bound decreased sharply when sperm motility was lowered by energy poisons or by reducing the temperature. Highly motile sperm from humans, guinea pigs, and rats, added at concentrations ten times higher than control sperm, did not bind to the porcine zona. At the same high concentration, a small number of hamster and bull sperm bound to the zona. Binding of boar sperm to the zona pellucida was blocked almost completely by diluted whole antiserum to sperm plasma membranes and by univalent (Fab) antibody to these membranes. When antibody to sperm plasma membrane was first absorbed with plasma membrane vesicles, sperm binding was not inhibited. These results provide direct evidence for the existence of sperm plasma membrane receptors for the zona pellucida of the pig.

  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. Esophageal function testing: beyond manometry and impedance.

    PubMed

    Mittal, Ravinder K

    2014-10-01

    Manometry and impedance provide only surrogate information regarding longitudinal wall function and are focused on contractile amplitude and lumen content. Ultrasound imaging provides a unique perspective of esophageal function by providing important information regarding longitudinal muscle contraction. Laser Doppler assessment of perfusion may be an important complementary tool to assess abnormal wall blood perfusion as a possible mechanism of pain. PMID:25216911

  10. Calcium Signaling in Sperm: Help from Prostasomes

    NSDL National Science Digital Library

    Dejian Ren (University of Pennsylvania; Department of Biology REV)

    2011-05-17

    Mammalian sperm cells are equipped with sophisticated Ca2+ signaling mechanisms that are fundamental to sperm’s success in fertilization. Unlike most other cells, however, mature sperm generally do not have the luxury of synthesizing new proteins. New evidence indicates that human sperm have a very clever way to solve the conflict between the critical demand for Ca2+ signaling tools and the silence of protein translation. Just before encountering the female reproductive tract, sperm acquire some of the key molecules for Ca2+ signaling from the male reproductive tract itself: prostasomes secreted by the prostate gland.

  11. A Comparison of Statistical Significance Tests for Selecting Equating Functions

    ERIC Educational Resources Information Center

    Moses, Tim

    2009-01-01

    This study compared the accuracies of nine previously proposed statistical significance tests for selecting identity, linear, and equipercentile equating functions in an equivalent groups equating design. The strategies included likelihood ratio tests for the loglinear models of tests' frequency distributions, regression tests, Kolmogorov-Smirnov…

  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. Evolutionary conservation of mammalian sperm proteins associates with overall, not tyrosine, phosphorylation in human spermatozoa.

    PubMed

    Schumacher, Julia; Ramljak, Sanja; Asif, Abdul R; Schaffrath, Michael; Zischler, Hans; Herlyn, Holger

    2013-12-01

    We investigated possible associations between sequence evolution of mammalian sperm proteins and their phosphorylation status in humans. As a reference, spermatozoa from three normozoospermic men were analyzed combining two-dimensional gel electrophoresis, immunoblotting, and mass spectrometry. We identified 99 sperm proteins (thereof 42 newly described) and determined the phosphorylation status for most of them. Sequence evolution was studied across six mammalian species using nonsynonymous/synonymous rate ratios (dN/dS) and amino acid distances. Site-specific purifying selection was assessed employing average ratios of evolutionary rates at phosphorylated versus nonphosphorylated amino acids (?). According to our data, mammalian sperm proteins do not show statistically significant sequence conservation difference, no matter if the human ortholog is a phosphoprotein with or without tyrosine (Y) phosphorylation. In contrast, overall phosphorylation of human sperm proteins, i.e., phosphorylation at serine (S), threonine (T), and/or Y residues, associates with above-average conservation of sequences. Complementary investigations suggest that numerous protein-protein interactants constrain sequence evolution of sperm phosphoproteins. Although our findings reject a special relevance of Y phosphorylation for sperm functioning, they still indicate that overall phosphorylation substantially contributes to proper functioning of sperm proteins. Hence, phosphorylated sperm proteins might be considered as prime candidates for diagnosis and treatment of reduced male fertility. PMID:23919900

  14. Evaluation of sperm DNA damage in bulls by TUNEL assay as a parameter of semen quality.

    PubMed

    Takeda, Kumiko; Uchiyama, Kyoko; Kinukawa, Masashi; Tagami, Takahiro; Kaneda, Masahiro; Watanabe, Shinya

    2015-06-20

    Sperm DNA damage affects the conception rate resulting from human assisted reproduction technology. The objective of this study was to adapt the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to provide a quality parameter for bull semen based on the detection of sperm DNA damage. Fresh semen was collected from two Japanese Black bulls (A, B) several times over the course of a year, and the percentage of TUNEL-positive spermatozoa (sperm TUNEL index) was determined. Individual differences in semen were detected using the sperm TUNEL index in these bulls (P < 0.01). The sperm TUNEL index of cryopreserved semen obtained from test-mated Japanese Black (n = 30, including two bulls with a conception rate lower than 10%) and Holstein (n = 34) bulls were analyzed. The average sperm TUNEL index and conception rate resulting from artificial insemination (AI) were 4.7% and 55.7% for Japanese Black, and 4.9% and 39.5% for Holstein, respectively. A weak negative correlation between sperm TUNEL index and conception rate was observed in Holstein bulls (P < 0.05). Semen samples from six bulls with more than 10% sperm TUNEL index were studied, and these samples showed low sperm viability. However, semen resulting in a very low conception rate did not have a high sperm TUNEL index. Although it would be difficult to predict a low conception rate resulting from AI using the sperm TUNEL index alone, the index can be used as an additional parameter to provide a more comprehensive description of semen quality. PMID:25739957

  15. [A brief protocol for sperm cryopreservation and revival in zebrafish].

    PubMed

    Zheng, Nai-Zhong; Zhang, Bo

    2012-09-01

    Zebrafish is an important vertebrate model organism for the study of embryonic development and the underlying genetic mechanism. Numerous mutants and transgenics have been generated in recent years, long-term and safe storage of these fish lines is of crucial importance for every zebrafish community/lab. Sperm cryopreservation and revival has become a preferred method for this purpose, which provides extra and reliable security as a backup for the cost-effective maintenance of genetic stocks in addition to reducing space demanding for housing large amount of live fish. This is especially critical for invaluable fish lines against accidental loss. Generally, the sperm are obtained by either squeezing the male fish or dissecting out and homogenizing the testes, then they are mixed with the freezing medium before gradually frozen as aliquots in liquid nitrogen. They can be easily revived through in vitro fertilization whenever necessary. This technique was introduced into zebrafish research three decades ago and has gradually become mature and more reliable following the im-provement of many critical factors and steps, including cryoprotectants and conditions for freezing and revival. Base on pioneers' work, our lab has established and improved a simple method for sperm cryopreservation and revival which shows high recovery rate after relatively long storage time. Here we briefly summarize the history and development of the methods for sperm cryopreservation and revival in zebrafish and present a brief protocol for the practice of sperm cryopreservation and revival, which is routinely used in our lab. PMID:23017463

  16. Deletion of murine choline dehydrogenase results in diminished sperm motility.

    PubMed

    Johnson, Amy R; Craciunescu, Corneliu N; Guo, Zhong; Teng, Ya-Wen; Thresher, Randy J; Blusztajn, Jan K; Zeisel, Steven H

    2010-08-01

    Choline dehydrogenase (CHDH) catalyzes the conversion of choline to betaine, an important methyl donor and organic osmolyte. We have previously identified single nucleotide polymorphisms (SNPs) in the human CHDH gene that, when present, seem to alter the activity of the CHDH enzyme. These SNPs occur frequently in humans. We created a Chdh(-/-) mouse to determine the functional effects of mutations that result in decreased CHDH activity. Chdh deletion did not affect fetal viability or alter growth or survival of these mice. Only one of eleven Chdh(-/-) males was able to reproduce. Loss of CHDH activity resulted in decreased testicular betaine and increased choline and PCho concentrations. Chdh(+/+) and Chdh(-/-) mice produced comparable amounts of sperm; the impaired fertility was due to diminished sperm motility in the Chdh(-/-) males. Transmission electron microscopy revealed abnormal mitochondrial morphology in Chdh(-/-) sperm. ATP content, total mitochondrial dehydrogenase activity and inner mitochondrial membrane polarization were all significantly reduced in sperm from Chdh(-/-) animals. Mitochondrial changes were also detected in liver, kidney, heart, and testis tissues. We suggest that men who have SNPs in CHDH that decrease the activity of the CHDH enzyme could have decreased sperm motility and fertility. PMID:20371614

  17. Evolution of protamine genes and changes in sperm head phenotype in rodents.

    PubMed

    Lüke, Lena; Vicens, Alberto; Tourmente, Maximiliano; Roldan, Eduardo R S

    2014-03-01

    Little is known about the genetic basis of evolutionary changes in sperm phenotype. Postcopulatory sexual selection is associated with differences in protamine gene sequences and promoters and is a powerful force acting on sperm form and function, although links between protamine evolution and sperm phenotype are scarce. Protamines are involved in sperm chromatin condensation, and protamine deficiency negatively affects sperm morphology and male fertility, thus suggesting that they are important for sperm design and function. We examined changes in protamine genes and sperm phenotype in rodents to understand the role of sexual selection on protamine evolution and sperm design. We performed a genotype-phenotype association study using root-to-tip dN/dS (nonsynonymous/synonymous substitutions rate ratio) to account for evolutionary rates and phylogenetic generalized least squares analyses to compare genetic and morphometric data. Evolutionary rates of protamine 1 and the protamine 2 domain cleaved off during chromatin condensation correlated with head size and elongation. Protamine 1 exhibited restricted positive selection on some functional sites, which seemed sufficient to preserve its role in head design. The cleaved-protamine 2, whose relaxation is halted by sexual selection, seems to ensure small, elongated heads that would make sperm more competitive. No association existed between mature-protamine 2 and head phenotype, suggesting little involvement during chromatin condensation and a likely role maintaining the condensed state. Our results suggest that evolutionary changes in protamines could be related to complex developmental modifications in the sperm head. This represents an important step toward understanding the role of changes in gene coding sequences in the divergence of germ cell phenotype. PMID:24522148

  18. Variation in Sperm Displacement and Its Association with Accessory Gland Protein Loci in Drosophila Melanogaster

    PubMed Central

    Clark, A. G.; Aguade, M.; Prout, T.; Harshman, L. G.; Langley, C. H.

    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 Drosophilia 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, Glucose dehydrogenase (Gld), and Esterase-6 (Est-6). Acp genes encode proteins that are in some cases known to be transmitted to the female in the seminal fluid and are likely candidates for genes that might mediate the phenomenon of sperm displacement. 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. PMID:7705622

  19. Functional test generation for DMA controllers

    Microsoft Academic Search

    M. Grosso; W. J. H. Pe?rez; D. Ravotto; E. Sanchez; M. S. Reorda; J. V. Medina

    2010-01-01

    Today's SoCs are composed of a high variety of modules, such as microprocessor cores, memories, peripherals, and customized blocks directly related to the targeted application. Testing a peripheral core embedded in a SoC requires two correlated phases: module configuration and module operation. The first one prepares the peripheral on the different operation modes, whereas, the second one is in charge

  20. Male sperm whale ( Physeter macrocephalus ) acoustics in a high-latitude habitat: implications for echolocation and communication

    Microsoft Academic Search

    P. T. Madsen; M. Wahlberg; B. Møhl

    2002-01-01

    Sperm whales (Physeter macrocephalus) are deep-diving predators foraging in meso- and bathypelagic ecosystems off the continental shelves. To investigate the ecophysiological and communicative function of various click types from male sperm whales in a high-latitude habitat, we deployed a large-aperture array of calibrated hydrophones off northern Norway (N69, E15). Data show that sperm whales in this habitat produce three click

  1. Evaluating the CMA Evolution Strategy on Multimodal Test Functions

    Microsoft Academic Search

    Nikolaus Hansen; Stefan Kern

    2004-01-01

    In this paper the performance of the CMA evolution strategy with rank-µ-update and weighted recombination is empirically investi- gated on eight multimodal test functions. In particular the eect of the population size on the performance is investigated. Increasing the pop- ulation size remarkably improves the performance on six of the eight test functions. The optimal population size takes a wide

  2. 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.

  3. [The genetic and epigenetic defect from the sperm for intracytoplasmic sperm injection (ICSI)].

    PubMed

    Ge, Shao-Qin; Kang, Xian-Jiang; Duan, Fei

    2010-04-01

    Intracytoplasmic sperm injection (ICSI) can be applied to treat male infertility patients of oligospermia, asthenospermia, teratospermia, azoospermia and failure of the common in-vitro fertilization (IVF), which may overcome the sperm deficiency and even obtain sperms directly from percutaneous epididymal sperm aspiration (PESA) and testicular sperm extraction (TESE). As for direct injection of a single sperm into an egg, the ICSI disobeys the biological laws of natural insemination, thus leading to high genetic and epigenetic risk for patients owing to genetic and epigenetic defect of sperm. By reviewing the genetic and epigenetic defects of ICSI sperm, as well as related diseases, this article aims at understanding of the risks resulting from the genetic and epigenetic defects of ICSI sperm at a molecular mechanism level. The results show that the quality control of ICSI sperm via detecting its epigenetic factors, such as methylated DNA and acetylated histone, is essential for reducing the genetic and epigenetic risk from ICSI. PMID:20423882

  4. The effects of opiate consumption on serum reproductive hormone levels, sperm parameters, seminal plasma antioxidant capacity and sperm DNA integrity.

    PubMed

    Safarinejad, Mohammad Reza; Asgari, Seyyed Alaeddin; Farshi, Alireza; Ghaedi, Gholamhossein; Kolahi, Ali Asghar; Iravani, Shahrokh; Khoshdel, Ali Reza

    2013-04-01

    We evaluated the effects of opiate consumption on semen quality, sperm function, seminal plasma antioxidant capacity, and sperm DNA integrity. A total of 142 opiate addict men (group 1) were enrolled in the study and 146 healthy age matched male volunteers (group 2) served as controls. Two semen analyses were performed in all participants. Sperm chromatin structure assay (SCSA) was used to identify sperm DNA integrity. The mean±SD sperm concentration in opiate users and in control subjects was 22.2±4.4 and 66.3±8.3 million per ml, respectively (P=0.002). A significant increase in the amount of fragmented DNA was found in opiate consumers compared with that in controls (36.4±3.8% vs. 27.1±2.4%, P=0.004). Significantly decreased levels of catalase-like and superoxide dismutase-like (SOD) activity were observed in group 1 compared with group 2. Opiate consumption has significant adverse effects on semen quality. In cases of unexplained infertility in men, opium consumption should be considered as a possible factor. PMID:23207164

  5. 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.

  6. 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

  7. Pathogenetic transition in the morphology of abnormal sperm in the testes and the caput, corpus, and cauda epididymides of male rats after treatment with 4,6-dinitro- o-cresol

    Microsoft Academic Search

    Ken L. Takahashi; Naofumi Takahashi; Hitoshi Hojo; Maki Kuwahara; Hiroaki Aoyama; Shoji Teramoto

    2006-01-01

    In order to elucidate the pathogenesis of tailless sperm, 4,6-dinitro-o-cresol (DNOC) was administered to Jcl:SD male rats at daily oral doses of 0, 10 or 15mg\\/kg for 5 days. Sperm were collected from the caput, corpus, and cauda epididymides on days 1, 7 and 14 after the last dosing (D1, D7 and D14, respectively), counted and examined morphologically by phase-contrast

  8. Current status of plant products reported to inhibit sperm.

    PubMed

    Farnsworth, N R; Waller, D P

    1982-06-01

    This report reviews research on plant-derived agents that prevent sperm production if taken orally by the male or that incapacitate or kill sperm on contact if used vaginally by the female. It would be of great value to develop fertility inhibitors that are totally selective for reproductive systems and enzymes, and there is a possibility that a plant-derived drug may have this effect. Plants that have been studied for their fertility inhibiting effects in the male include: Aristolochia indica L. (Aristolochiaceae); Azadirachta indica A. Juss (Meliaceae); Balanites roxburghii Planch. (Zygophyllaceae); Calotropis procera (Ait) R.Br. (Asclepiadaceae); Carica papaya L. (Caricaceae); Catharanthus roseus (L.) G. Don (Apocynaceae); Dieffenbachia seguine (Jacquin) Schott. (Araceae); Ecaballium elaterium A. Richard (Cucurbitaceae); Gossypium species (Malvaceae); Hibiscus rosa-sinensis L. (Malvaceae); Hippophae salicifolia D. Don (Elaeagnaceae); Leucaena glauca (L.) Benth. (Leguminosae); Lonicera ciliosa Poir. (Caprifoliaceae); Lupinus termis Forsk. (Leguminosae); Malvaviscus conzattii Greenm. (Malvaceae); Momordica charantia L. (Curcurbitaceae); Ocimum sanctum L. (Labiatae); Prunus emarginata Walp. (Rosaceae); and Withania somnifera (L.) Dunal (Solanaceae). A large number of plants have been randomly selected and screened for spermicidal activity "in vitro" and several seem promising. Those species found to be active and the nature of the active principle(s), when known, are presented in a table as are plant-derived chemical substances of known or partially known structure reported to be spermicidal "in vitro." Plants warrant systematic study as potential sources of sperm-agglutinating compounds. Of 1600 Indian plants tested, 90 showed positive semen coagulating properties. There seems to be a lack of correlation among experimental results obtained by different groups of investigators, between data obtained "in vitro" and "in vivo," and between experimental results and information found in folklore. Factors complicating the adequate assessment of plants affecting male fertility are inadequate numbers of vehicle-treated controls, poor experimental design, problems related to insolubility of crude plant extracts, variation in routes of administration, diversity in reproductive function and control among various laboratory species, and problems in identifying plant names consistently. PMID:12179631

  9. 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

  10. 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

  11. LUNG FUNCTION TESTS IN NORMAL INDIAN CHILDREN

    Microsoft Academic Search

    R. v. Chowgule; V. m. Shetye; J. r. Parmar

    ABSTRACT The forced vital capacity, forced expiratory volume in one second, peak expiratory flow, mid- expiratory flow and ,maximum ,voluntary ,venti- lation was measured in 632 healthy, normal children,from ,Metropolitan ,city of Bombay using,computerized ,spirometer. The children were between age range 6 years to 15 years and belong,to high ,or middle ,and lower socio eco- nomic,status. The pulmonary ,function data

  12. Pancreatic function tests: When to choose, what to use

    Microsoft Academic Search

    Wolfgang G. Boeck; Adler Guido; Thomas M. Gress

    2001-01-01

    Although techniques for high-resolution imaging of the pancreas are constantly being improved, the evaluation of pancreatic\\u000a function remains crucial for the workup of pancreatic diseases. More than 20 direct and indirect tests are available for the\\u000a assessment of pancreatic function. Measurement of fecal elastase-1 is recommended as the most suitable test for the initial\\u000a assessment of pancreatic function. Among other

  13. Adaptation to Sperm Competition in Humans

    Microsoft Academic Search

    Todd K. Shackelford; Aaron T. Goetz

    2007-01-01

    With the recognition, afforded by recent evolutionary science, that female infidelity was a recurrent feature of modern humans' evolutionary history has come the development of a unique area in the study of human mating: sperm competition. A form of male–male postcopulatory competition, sperm competition occurs when the sperm of two or more males concurrently occupy the reproductive tract of a

  14. Testicular Sperm Retrieval in Azoospermic Men

    Microsoft Academic Search

    Peter Pantke; Thorsten Diemer; Marcelo Marconi; Martin Bergmann; Klaus Steger; Hans-Christian Schuppe; Wolfgang Weidner

    2008-01-01

    Context: Sperm retrieval in combination with IVF\\/ICSI is the only medical procedure for an azoospermic man to father a child. Different techniques, especially testicular sperm extraction (TESE), have evolved over time and have dramatically improved the outlook for men with testicular azoospermia. However sperm retrieval rates are associated not only with the operation proposed but especially with a distinct pattern

  15. DNA-fluorometry of mammalian sperm

    Microsoft Academic Search

    Jtirgen Zante; Johannes Schumann; Wolfgang Göhde; Ursula Hacker

    1977-01-01

    The DNA-content of sperm and testicular cells was measured by pulse-cytophotometry with high resolution. From flat sperm symmetric and narrow fluorescence distributions were obtained. Enzymatic treatment with papain or pronase and staining with an ethidiumbromide-mithramycin dye solution generate stoichiometric DNA-staining including that of mature sperm with a coefficient of variation below 2%.

  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. Cryopreservation of Greenshell™ mussel (Perna canaliculus) sperm.

    PubMed

    Adams, Serean L; Smith, John F; Taylor, Jolene; McGowan, Lindsay T; Tervit, H Robin

    2015-01-01

    Cryopreservation is a valuable technique for aquaculture as it enables a library or bank of genetically valuable animals to be maintained in a cost-effective manner. Here, we describe a method to cryopreserve the sperm of the Greenshell™ mussel (Perna canaliculus) and how to use the sperm post-thawing to maximize larval production from thawed sperm in selective breeding. PMID:25428014

  18. 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…

  19. S-allyl cysteine ameliorates the quality of sperm and provides protection from age-related sperm dysfunction and oxidative stress in rats

    PubMed Central

    Takemura, Shigekazu; Ichikawa, Hiroshi; Naito, Yuji; Takagi, Tomohisa; Yoshikawa, Toshikazu; Minamiyama, Yukiko

    2014-01-01

    Reactive oxygen species play a central role in the pathophysiology of the age-related decrease in male fertility. It has been reported that the total protein of DJ-1 was decreased in a proteomic analysis of seminal plasma from asthenozoospermia patients and a DJ-1 protein acts as a sensor of cellular redox homeostasis. Therefore, we evaluated the age-related changes in the ratio of the oxidized/reduced forms of the DJ-1 protein in the epididymis. In addition, the protective effects of S-allyl cysteine (SAC), a potent antioxidant, were evaluated against sperm dysfunction. Male rats aged 15–75 weeks were used to assess age-associated sperm function and oxidative stress. Sperm count increased until 25 weeks, but then decreased at 50 and 75 weeks. The rate of sperm movement at 75 weeks was decreased to approximately 60% of the rate observed at 25 weeks. Expression of DJ-1 decreased, but oxidized-DJ-1 increased with age. In addition, 4-hydroxy-2-nonenal modified proteins in the epididymis increased until 50 weeks of age. The total number and DNA synthetic potential of the sperm increased until 25 weeks, and then decreased. In rats 75 weeks of age, SAC (0.45% diet) attenuated the decrease in the number, motility, and DNA synthesis of sperm and inhibited the oxidized proteins. These results suggest that SAC ameliorates the quality of sperm subjected to age-associated oxidative stress. PMID:25411519

  20. Sperm penetration into cervical mucus in vitro. III. Effect of freezing on estrous bovine cervical mucus

    SciTech Connect

    Lee, W.I.; Gaddum-Rosse, P.; Blandau, R.J.

    1981-08-01

    The influence of the storages period on estrous bovine cervical mucus after it was stored in the freezing compartment of the laboratory refrigerator was evaluated by an in vitro sperm penetration test with human spermatozoa, laser light-scattering, and a spinnbarkeit test. Data obtained from the sperm penetration test were analyzed by a mathematical model that correlates the sperm motility with the sperm transport rate and the penetrability of the mucus. The tests showed that estrous bovine cervical mucus can be stored for up to 4 weeks at -12 degrees C without a change in its physical properties. The results of this study strengthen the suggestion that bovine mucus could be employed as a substitute for human cervical mucus.

  1. Protein disulfide isomerase-P5, down-regulated in the final stage of boar epididymal sperm maturation, catalyzes disulfide formation to inhibit protein function in oxidative refolding of reduced denatured lysozyme

    Microsoft Academic Search

    Kuniko Akama; Tomoe Horikoshi; Atsushi Sugiyama; Satoko Nakahata; Aoi Akitsu; Nobuyoshi Niwa; Atsushi Intoh; Yasutaka Kakui; Michiko Sugaya; Kazuo Takei; Noriaki Imaizumi; Takaya Sato; Rena Matsumoto; Hitoshi Iwahashi; Shin-ichi Kashiwabara; Tadashi Baba; Megumi Nakamura; Tosifusa Toda

    2010-01-01

    In mammalian spermiogenesis, sperm mature during epididymal transit to get fertility. The pig sharing many physiological similarities with humans is considered a promising animal model in medicine. We examined the expression profiles of proteins from boar epididymal caput, corpus, and cauda sperm by two-dimensional gel electrophoresis and peptide mass fingerprinting. Our results indicated that protein disulfide isomerase-P5 (PDI-P5) human homolog

  2. Neurotensin enhances sperm capacitation and acrosome reaction in mice.

    PubMed

    Hiradate, Yuuki; Inoue, Hiroki; Kobayashi, Norio; Shirakata, Yoshiki; Suzuki, Yutaka; Gotoh, Aina; Roh, Sang-Gun; Uchida, Takafumi; Katoh, Kazuo; Yoshida, Manabu; Sato, Eimei; Tanemura, Kentaro

    2014-08-01

    Neurotensin (NT) has multiple functions, ranging from acting as a neurotransmitter to regulating intestinal movement. However, its function in reproductive physiology is unknown. Here, we confirmed the expression and localization of NT receptors (NTR1) in mouse epididymal spermatozoa and investigated the effect of NT on sperm function. Sperm protein tyrosine phosphorylation, one of the indices of sperm capacitation, was facilitated dose-dependently by NT administration. In addition, the acrosome reaction was promoted in capacitated spermatozoa, and addition of a selective antagonist of NTR1 and NTR2 blocked the induction. Furthermore, intracellular calcium mobilization by NT addition was observed. This showed that NT was an accelerator of sperm function via its functional receptors. The presence of NT was confirmed by immunohistochemistry and its localization was observed in epithelia of the uterus and oviduct isthmus and ampulla, which correspond to the fertilization route of spermatozoa. The NT mRNA level in ovulated cumulus cell was remarkably increased by treatment with human chorionic gonadotropin (hCG). Using an in vitro maturation model, we analyzed the effects of FSH, epidermal growth factor (EGF), estradiol, and progesterone in NT production in cumulus cells. We found that FSH and EGF upregulated NT release and mRNA expression. Both FSH- and EGF-induced upregulation were inhibited by U0126, an MAPK kinase inhibitor, indicating that FSH and EGF regulate NT expression via a MAPK-dependent pathway. This evidence suggests that NT can act as a promoter of sperm capacitation and the acrosome reaction in the female reproductive tract. PMID:25031361

  3. [Gender differences on physiological function test].

    PubMed

    Nakagawa, Mikiko

    2015-04-01

    Recently, clinical findings showed sex differences in the manifestation of the cardiovascular diseases. There are gender differences in cardiac electrophysiologic properties and the incidence of various types of arrhythmias. Long QT syndrome is more common in women than in men, whereas the incidence of Brugada syndrome and J-wave syndrome are male predominant. The gender differences in QT interval are not observed before puberty but become obvious after puberty. ST-levels markedly increase after puberty in men but not in women, which suggested that the sex hormones have an important role for the ECG parameters and arrhythmias. In addition, gender differences were reported in age-related changes in echocardiographic parameters such as left ventricular diastolic function. PMID:25936147

  4. Distinct cytoskeletal domains revealed in sperm cells

    PubMed Central

    1984-01-01

    Antibodies against different cytoskeletal proteins were used to study the cytoskeletal organization of human spermatozoa. A positive staining with actin antibodies was seen in both the acrosomal cap region and the principal piece region of the tail. However, no staining was obtained with nitrobenzoxadiazol-phallacidin, suggesting that most of the actin was in the nonpolymerized form. Most of the myosin immunoreactivity was confirmed to a narrow band in the neck region of spermatozoa. Tubulin was located to the entire tail, whereas vimentin was only seen in a discrete band-like structure encircling the sperm head, apparently coinciding with the equatorial segment region. Surface staining of the spermatozoa with fluorochrome-coupled Helix pomatia agglutinin revealed a similar band-like structure that co-distributed with the vimentin- specific staining. Instead, other lectin conjugates used labeled either the acrosomal cap region (peanut and soybean agglutinins), both the acrosomal cap and the postacrosomal region of the head (concanavalin A), or the whole sperm cell surface membrane (wheat germ and lens culinaris agglutinins and ricinus communis agglutinin l). In lectin blotting experiments, the Helix pomatia agglutinin-binding was assigned to a 80,000-mol-wt polypeptide which, together with vimentin, also resisted treatment with Triton X-100. Only the acrosomal cap and the principal piece of the tail were decorated with rabbit and hydridoma antibodies against an immunoanalogue of erythrocyte alpha-spectrin (p230). p230 appeared to be the major calmodulin-binding polypeptide in spermatozoa, as shown by a direct overlay assay of electrophoretic blots of spermatozoa with 125I-calmodulin. The results indicate that spermatozoa have a highly specialized cytoskeletal organization and that the distribution of actin, spectrin, and vimentin can be correlated with distinct surface specializations of the sperm cells. This suggest that cytoskeleton may regulate the maintenance of these surface assemblies and, hence, affect the spermatozoan function. PMID:6381503

  5. Calcium signaling in sperm: help from prostasomes.

    PubMed

    Ren, Dejian

    2011-01-01

    Mammalian sperm cells are equipped with sophisticated Ca2+ signaling mechanisms that are fundamental to sperm's success in fertilization. Unlike most other cells, however, mature sperm generally do not have the luxury of synthesizing new proteins. New evidence indicates that human sperm have a very clever way to solve the conflict between the critical demand for Ca2+ signaling tools and the silence of protein translation. Just before encountering the female reproductive tract, sperm acquire some of the key molecules for Ca2+ signaling from the male reproductive tract itself: prostasomes secreted by the prostate gland. PMID:21586726

  6. Power functional theory for the dynamic test particle limit

    NASA Astrophysics Data System (ADS)

    Brader, Joseph M.; Schmidt, Matthias

    2015-05-01

    For classical Brownian systems both in and out of equilibrium we extend the power functional formalism of Schmidt and Brader (2013 J. Chem. Phys. 138 214101) to mixtures of different types of particles. We apply the framework to develop an exact dynamical test particle theory for the self and distinct parts of the van Hove function, which characterize tagged and collective particle motion. The memory functions that induce non-Markovian dynamics are related to functional derivatives of the excess (over ideal) free power dissipation functional. The method offers an alternative to the recently found nonequilibrium Ornstein–Zernike relation for dynamic pair correlation functions.

  7. Thermosensitive ion channel TRPV1 is endogenously expressed in the sperm of a fresh water teleost fish (Labeo rohita) and regulates sperm motility

    PubMed Central

    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 Ca2+-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

  8. 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

  9. Sperm competition promotes diversity of sperm bundles in Ohomopterus ground beetles

    NASA Astrophysics Data System (ADS)

    Takami, Yasuoki; Sota, Teiji

    2007-07-01

    Diversification of sperm morphology has been investigated in the context of sperm competition, but the adaptive significance of sperm bundles is still unclear. In analyzing 10 taxa of the genus Carabus subgenus Ohomopterus and one related Carabus ground beetles, we found that dimorphic sperm bundles occurred in most species with varied degrees of bimodality, whereas sperm were generally monomorphic. Comparative analyses with phylogenetically independent contrasts revealed that the sizes of large and small sperm bundles evolved more rapidly than, and were not correlated with, the length of sperm, suggesting more intense selection on sperm bundle sizes and their independent responses to different evolutionary forces. The size of large sperm bundles was positively correlated with male genital morphology (pertinent to displacement of rival spermatophores) and postcopulatory guarding duration as well as male body length, suggesting that larger sperm bundles have been favored when the risk of spermatophore displacement is high. Larger sperm bundles may be advantageous because of their ability to migrate more rapidly into the spermatheca. In contrast, no clear association was detected between the small sperm bundle size and mating traits despite its rapid diversification. The present study provides the first record of heteromorphic sperm bundles, the diversity of which may be promoted by sperm competition.

  10. The Sperm Whale, Physeter macrocephalus

    E-print Network

    of the tail- stock. The skin of the trunk is cor- rugated into many series of longitudinal ripples con- tains the spermaceti organ, which is a large reservoir for spermaceti oil. Sperm whales are hunted for this oil as well as for the lower grade oil con- tained in the blubber. The remainder

  11. GENETIC VARIATION IN SPERM PRODUCTION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In boars, the primary determinant of daily sperm production is number of Sertoli cells, which establishes testicular weight. The only breed comparison of fetal testicular development in boars contrasted two diverse breeds, White composite (WC, Landrace-Yorkshire) with Meishan, a Chinese breed that u...

  12. 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

  13. UNIT ROOT TESTING FOR FUNCTIONALS OF LINEAR PROCESSES1

    E-print Network

    Wu, Wei-Biao

    UNIT ROOT TESTING FOR FUNCTIONALS OF LINEAR PROCESSES1 Wei Biao Wu University of Chicago March 17: Wei Biao Wu, Depart- ment of Statistics, University of Chicago, 5734 S. University Avenue, Chicago, IL

  14. Intelligence test performance and the delay function of the ego

    Microsoft Academic Search

    George Spivack; Murray Levine; Herbert Sprigle

    1959-01-01

    The Ss were 123 emotionally disturbed adolescents. Ego delay function correlates with IQ. Intelligence test performance and personality variables can be considered within a single theoretical framework. 18 refs.

  15. An in vivo and in vitro investigation into the effects of alpha- chlorohydrin on sperm motility and correlation with fertility in the Han Wistar rat.

    PubMed

    Woods, J; Garside, D A

    1996-01-01

    Following a recommendation from the International Conference on Harmonisation, pharmaceutical companies are now monitoring possible drug effects on sperm motility in the rat during preclinical safety studies by assessing sperm motility (velocity). However, it is not known precisely how changes in sperm motility relate to fertility. Therefore, the effects of alpha-chlorohydrin on sperm motility were investigated and related to fertility both in vivo and in vitro. alpha-Chlorohydrin was given orally to male rats using a range of doses: 0, 5, 10, and 20 mg/kg for at least 5 consecutive days. Sperm were than assessed for motility using a standard scoring system (operators' observation of sperm) that graded degree of motility (i.e., 0 = i mmotile to 4 = very motile). The results showed a dose-related decrease in sperm motility. The sperm also appeared to move with a "jerky" action. Surprisingly, when this was correlated to fertility, none of the females mated with treated males became pregnant. A dose-related decrease in pregnancy would perhaps have been expected. There was no effect on sperm morphology, and testicular and epididymal pathology were only seen after doses of 20 mg/kg. When sperm from untreated rats were incubated with alpha-chlorohydrin in vitro at concentrations of 0, 0.5, 1.0, and 1.5 mM, sperm motility and motion were similarly affected as observed in vivo. However, the fertilization capacity (in vitro fertilization) of the treated sperm showed a concentration-related reduction in percentage fertilization, and there was also evidence of abnormal embryo development. These findings suggest that the present standard scoring system used in preclinical safety studies is not a comprehensive indicator of sperm function and/or fertility. A better understanding of sperm movement, therefore, is desirable so effects on sperm motility can be related to fertility. PMID:8738556

  16. Branchial cilia and sperm flagella recruit distinct axonemal components.

    PubMed

    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

  17. Simulation analysis of a printed circuit board functional test process 

    E-print Network

    Brinkley, Paul Andrew

    1991-01-01

    SIMULATION ANALYSIS OF A PRINTED CIRCUIT BOARD FUNCTIONAL TEST PROCESS A Thesis by PAUL ANDREW BRINKLEY Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree of MASTER... OF SCIENCE August 1991 Major Subject: Industrial Engineering SIMULATION ANAI. YSIS OF A PRINTED CIRCUIT BOARD FUNCTIONAL TEST PROCESS A Thesis by PAUL ANDREW BRINKLEY Approved as to style and content by: a ph . Disney (Chair of Committee) Thomas E...

  18. A Functional Test Platform for the Community Land Model

    SciTech Connect

    Xu, Yang [ORNL] [ORNL; Thornton, Peter E [ORNL] [ORNL; King, Anthony Wayne [ORNL] [ORNL; Steed, Chad A [ORNL] [ORNL; Gu, Lianhong [ORNL] [ORNL; Schuchart, Joseph [ORNL] [ORNL

    2014-01-01

    A functional test platform is presented to create direct linkages between site measurements and the process-based ecosystem model within the Community Earth System Models (CESM). The platform consists of three major parts: 1) interactive user interfaces, 2) functional test model and 3) observational datasets. It provides much needed integration interfaces for both field experimentalists and ecosystem modelers to improve the model s representation of ecosystem processes within the CESM framework without large software overhead.

  19. Effects of a maximal exercise test on neurocognitive function

    Microsoft Academic Search

    Tracey Covassin; Leigh Weiss; John Powell; Christopher Womack; M. R Lovell

    2007-01-01

    Objective: To examine the effects of a maximal exercise test on cognitive function in recreational athletes.Design: A repeated-measures design was used to compare baseline with post-cognitive function and fatigue symptoms after a maximal exercise test.Setting: Division 1 American Midwestern University, (Michigan State University, Michigan, USA).Participants: 102 male and female recreational athletes.Intervention: Participants in the experimental group (n = 54) were

  20. Lower Bounds on Testing Functions of Low Fourier Degree

    E-print Network

    Hatami, Pooya

    2012-01-01

    We consider the problem of testing whether a Boolean function has Fourier degree $\\leq k$ or it is $\\epsilon$-far from any Boolean function with Fourier degree $\\leq k$, we improve the known lower bound of $\\Omega(k)$ \\cite{BBM11,CGM10}, to $\\Omega(k/\\sqrt{\\epsilon})$. The lower bound uses the recently discovered connections between property testing and communication complexity by Blais \\textit{et. al.} \\cite{BBM11}

  1. 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

  2. Abstract When females mate with more than one male, the ensuing sperm competition leads to the evolution of

    E-print Network

    Rutowski, Ronald L.

    Abstract When females mate with more than one male, the ensuing sperm competition leads). Adequate tests for cryptic fe- male choice can only be designed if male mechanisms of sperm competition competition, usually measured as the proportion of offspring sired by the second of two males to mate

  3. Effect of sperm concentration, medium osmolality and oocyte storage on artificial fertilisation success in a myobatrachid frog (Limnodynastes tasmaniensis)

    Microsoft Academic Search

    D. L. EdwardsA; M. J. Mahony; J. Clulow

    2004-01-01

    The present study optimised artificial fertilisation and oocyte storage conditions in Limnodynastes tasmaniensis (Myobatrachidae). Data on general reproductive biology, the effect of sperm motility and concen- tration, medium osmolality and oocyte storage on artificial fertilisation success are presented. Egg number was most strongly correlated with bodyweight (r = 0.819). Sperm yield was correlated with testes weight (r = 0.827), which

  4. Control of sperm concentration is necessary for standardization of sperm cryopreservation in aquatic species: evidence from sperm agglutination in oysters.

    PubMed

    Dong, Qiaoxiang; Huang, Changjiang; Tiersch, Terrence R

    2007-02-01

    A lack of standardization in sperm cryopreservation of aquatic organisms is one of the main reasons for inconsistency observed among various studies. In particular, there have been few attempts to standardize sperm concentration during procedural optimization. This study was intended to call attention to sperm concentration standardization through research of sperm agglutination in Pacific oysters Crassostrea gigas. Sperm agglutination after thawing is a relatively frequent phenomenon observed for various aquatic species, especially when sub-optimal cryopreservation protocols are used; however, no systematic attempts have been made to explain this phenomenon. The present study evaluated various factors affecting sperm agglutination of thawed samples from diploid and tetraploid Pacific oysters, and is the first detailed report addressing the sperm agglutination phenomenon of thawed samples from any aquatic organism. Agglutination of oyster sperm was classified into six levels with a scale ranging from 0 (homogenous suspension) to 5 (well-developed "noodles"). It was found that agglutination in thawed samples was mainly due to the lack of sufficient cryoprotectant for a specific sperm concentration. Interestingly, high levels of agglutination did not necessarily lead to low fertilization. On the contrary, some sperm cells appeared to gain protection from the formation of peripheral agglutination within 0.5-ml French straws. The exact mechanism of sperm agglutination remains unclear. However, morphological examination of cross sections of the noodles (agglutination level 5) indicated at least two forms of agglutination (formed with and without cryoprotectant) which could be used as a tool to understand the cryopreservation process within the micro-environment of the straw. Furthermore, the fact that the level of sperm agglutination was directly determined by sperm concentration, in addition to the type of cryoprotectant, cryoprotectant concentration, and cooling and thawing methods emphasized the importance of procedural standardization and systematic optimization and integration of protocols involving multiple factors. PMID:17276426

  5. A Use of the Information Function in Tailored Testing

    ERIC Educational Resources Information Center

    Samejima, Fumiko

    1977-01-01

    Several important implications in latent trait theory, with implications for individualized or tailored testing, are pointed out. A way of using the information function in tailored testing in connection with the standard error estimation of the ability level using maximum likelihood estimation is suggested. (Author/JKS)

  6. Understanding Functions without Using the Vertical Line Test

    ERIC Educational Resources Information Center

    Fernandez, Eileen

    2005-01-01

    Study was conducted on how the students were made to think meaningfully and widely within and across the representations in solving and identifying functions in precalculus, without the usage of the vertical line test. It was concluded that the "no vertical line testing" method proved to be easily accessible by all students and also made them…

  7. Differential Item Functioning on Two Tests of EFL Proficiency.

    ERIC Educational Resources Information Center

    Ryan, Katherine E.; Bachman, Lyle F.

    1992-01-01

    The extent to which items from the Test of English as a Foreign Language and the First Certificate in English function differently for test-takers of equal ability from different native language and curricular backgrounds was investigated. Results suggest a need for methods like logistic regression to examine nonuniform differential item…

  8. Effect of Seminal Plasma on Equine Sperm Quality: Preparation and Storage Techniques

    E-print Network

    Whigham, Alison R

    2013-09-03

    to facilitate binding with the oocyte’s zona pellucida for the initiation of the acrosome reaction [11]. Pores on the sperm head are created over the duration of the acrosome reaction to enable enzymes to pass from the sperm to the egg to facilitate further... in the horse found thus far, HSP-7 binds to the oocyte’s zona pellucida to aid in the fertilization capacity of the sperm. Although labeled and identified based on molecular weight, HSP-5 has yet to have its physiological function determined in the stallion...

  9. Effects of genetic captive-breeding protocols on sperm quality and fertility in the white-footed mouse.

    PubMed

    Malo, Aurelio F; Martinez-Pastor, Felipe; Alaks, Glen; Dubach, Jean; Lacy, Robert C

    2010-10-01

    Mice (Peromyscus leucopus noveboracensis) from a captive-breeding program were used to test the effects of three genetic breeding protocols (minimizing mean kinship [MK], random breeding, and selection for docility [DOC]) and inbreeding levels on sperm traits and fertility. Earlier, in generation 8, one DOC replicate went extinct because of poor reproductive success. By generation 10, spermatozoa from DOC mice had more acrosome and midpiece abnormalities, which were shown to be strong determinants of fertility, as well as lower sperm production and resistance to osmotic stress. In addition, determinants of fertility, including male and female components, were assessed in a comprehensive manner. Results showed that the probability (P) of siring litters is determined by sperm number, sperm viability, and midpiece and acrosome abnormalities; that the P of siring one versus two litters is determined by tail abnormalities; and that the total number of offspring is influenced by female size and proportion of normal sperm, showing the relative importance of different sperm traits on fertility. On average, males with 20% normal sperm sired one pup per litter, and males with 70% normal sperm sired eight pups per litter. Interestingly, the proportion of normal sperm was affected by docility but not by relatively low inbreeding. However, inbreeding depression in sperm motility was detected. In the MK group, inbreeding depression not only affected sperm motility but also fertility: An increase in the coefficient of inbreeding (f) of 0.03 reduced sperm motility by 30% and translated into an offspring reduction of three pups in second litters. A genetic load of 48 fecundity equivalents was calculated. PMID:20519695

  10. Effects of L-carnitine and L-acetyl-carnitine on testicular sperm motility and chromatin quality

    PubMed Central

    Aliabadi, Elham; Soleimani Mehranjani, Malek; Borzoei, Zahra; Talaei-Khozani, Tahereh; Mirkhani, Hossein; Tabesh, Hamed

    2012-01-01

    Background: Sperm cells extracted from testes (TESE) have poor chromatin quality and motility. Various substances are used in the laboratory to increase sperm motility and improve the ART outcomes; however, there are few research which considered improving both sperm motility and chromatin quality. Objective: The aim of this investigation was to evaluate the improvement of the testicular sperm motility and chromatin quality exposed to L-carnitine (LC) and L-acetyl-carnitine (LAC), which are normally concentrated in testis and epididymis, compared with Pentoxifylline (PF), which used for sperm motility enhancement in IVF procedures. Materials and Methods: TESE samples from 30 male mice divided into four parts. The sperm samples were added to Ham' F10 (control) or the media contained 1.76mM of LC, LAC or PF), then, the samples were kept in the room temperature for 30, 90 and 180 min. At each time step, sperm motility and chromatin quality were assessed. Chromatin quality was evaluated by chromomycin A3 and aniline blue. Statistical analysis was performed using one way analysis of variance (ANOVA). A p-value less than 0.05 were accepted as a statistically significant difference. Results: The results showed LC, LAC and PF significantly increased the sperm motility. However, sperm chromatin quality only improved significantly by administration of LC and LAC. Conclusion: Administration of LC and LAC to the testicular sperm samples can lead to improve both sperm motility and chromatin quality. It may be because they can mimic in vivo sperm condition during late spermatogenesis. PMID:25242977

  11. The functional standing test development and standardization of a clinical evaluation of standing function

    Microsoft Academic Search

    R. J. Triolo; B. W. B. Reilley; W. Freedman; R. R. Betz

    1992-01-01

    The functional standing test, which quantifies standing performance based on the ability to free the upper extremities from support and balance tasks in order to manipulate the environment, is described. The development of the test was motivated by the need for a new assessment that reflects the clinical usefulness of standing and allows the comparison of functional neuromuscular stimulation (FNS)

  12. Laser radiation and motility patterns of human sperm

    SciTech Connect

    Lenzi, A.; Claroni, F.; Gandini, L.; Lombardo, F.; Barbieri, C.; Lino, A.; Dondero, F. (Univ. of Rome La Sapienza (Italy))

    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.

  13. Symptom Reporting by Functional Dyspeptics During the Water Load Test

    Microsoft Academic Search

    Michael P. Jones; Lorrie M. Roth; Michael D. Crowell

    2005-01-01

    INTRODUCTION:Reporting of symptoms and visceral sensations in functional dyspepsia is subjective and influenced by such factors as somatization and depression. To better understand the role of these factors in symptom reporting, somatization, somatosensory amplification, alexithymia, and depression were evaluated in patients with functional dyspepsia undergoing a water load test.METHODS:Sixty-seven controls and 103 patients with functional dyspepsia underwent a 5-min water

  14. DNA fragmentation in frozen sperm of Equus asinus: Zamorano-Leonés, a breed at risk of extinction.

    PubMed

    Cortés-Gutiérrez, E I; Crespo, F; Gosálvez, A; Dávila-Rodríguez, M I; López-Fernández, C; Gósalvez, J

    2008-05-01

    The dynamics of sperm DNA fragmentation (sDF) and sperm viability were analyzed in frozen-thawed sperm samples of Equus asinus (Zamorano-Leonés), a breed at risk of extinction. Sperm DNA fragmentation was assessed using an adaptation of the sperm chromatin dispersion test developed for stallions in five different frozen samples. Sperm were thawed and incubated at different temperatures (37 degrees C, 25 degrees C, and 4 degrees C) and sDF was assessed at different times and compared. The mean sDF after thawing at the beginning of the experiment was 18.20+/-14.77% and did not differ significantly from the results of a neutral comet assay (22.0+/-19.34%). The tendency in the sDF of all donkeys indicated that sperm DNA is more sensitive to breakage when incubated at 37 degrees C than when incubated at 25 degrees C or 4 degrees C. Interestingly, the tendency was not the same when different animals were compared, and differences in sDF dynamics were established among individuals. sDF correlated negatively with sperm viability in some individuals but not in others. From a conservation perspective, sDF analysis may offer a new way to assess sperm quality in endangered breeds in order to identify and select the best semen samples for artificial reproduction purposes. In particular, we recommend for artificial insemination the use of semen samples with a slow increase in sDF with time after thawing. PMID:18367243

  15. Effect of post-thaw storage time on motility and fertility of cryopreserved beluga sturgeon (Huso huso) sperm.

    PubMed

    Aramli, M S; Nazari, R M; Gharibi, M R

    2015-04-01

    The aim of this study was to test the influence of post-thaw storage time on the duration of sperm motility, percentage of motile sperm, and fertilization and hatching rates of fresh sperm and sperm stored for 0, 30 and 60 min at 4°C post-thawing. After being frozen in liquid nitrogen and then thawed, the percentage of motile sperm and duration of motility were not affected by 30 min of storage at 4°C, whereas a significant decline in these parameters was observed after 60 min of storage. Similarly, fertilization and hatching rates were significantly affected within 60 min of storage at 4°C, and the fertility of frozen-thawed sperm was significantly lower than that of fresh sperm. We conclude that cryopreserved sperm of beluga sturgeon could be stored for 30 min without the loss of sperm quality. This described procedure for beluga sturgeon cryopreservation is reliable and efficient and therefore can be recommended for hatchery practice after scaling up this technique. PMID:25612010

  16. 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

  17. Formal Functional Test Designs: Bridging the Gap Between Test Requirements and Test Specifications

    NASA Technical Reports Server (NTRS)

    Hops, Jonathan

    1993-01-01

    This presentation describes the testing life cycle, the purpose of the test design phase, and test design methods and gives an example application. Also included is a description of Test Representation Language (TRL), a summary of the language, and an example of an application of TRL. A sample test requirement and sample test design are included.

  18. Imbalanced lipid homeostasis in the conditional Dicer1 knockout mouse epididymis causes instability of the sperm membrane.

    PubMed

    Björkgren, Ida; Gylling, Helena; Turunen, Heikki; Huhtaniemi, Ilpo; Strauss, Leena; Poutanen, Matti; Sipilä, Petra

    2015-02-01

    During epididymal sperm maturation, the lipid content of the sperm membrane is modified, which facilitates sperm motility and fertility. However, little is known about the mechanisms regulating the maturation process. By generating a conditional knockout (cKO) of Dicer1 in the proximal part of the mouse epididymis, we studied the role of RNA interference in epididymal functions. The Dicer1 cKO epididymis displayed an altered lipid homeostasis associated with a 0.6-fold reduction in the expression of the gene elongation of very long chain fatty acids-like 2, an enzyme needed for production of long-chain polyunsaturated fatty acids (PUFAs). Furthermore, the expression of several factors involved in cholesterol synthesis was up-regulated. Accordingly, the Dicer1 cKO sperm membrane showed a 0.7-fold decrease in long-chain PUFAs, whereas the amount of cholesterol in acrosome-reacted sperm displayed a 1.7-fold increase. The increased cholesterol:PUFA ratio of the sperm membrane caused breakage of the neck and acrosome region and immotility of sperm. Dicer1 cKO mice sperm also displayed reduced ability to bind to and fertilize the oocyte in vitro. This study thus shows that Dicer1 is critical for lipid synthesis in the epididymis, which directly affects sperm membrane integrity and male fertility. PMID:25366345

  19. 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.

  20. 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

  1. Microfluidic single sperm entrapment and analysis.

    PubMed

    de Wagenaar, B; Berendsen, J T W; Bomer, J G; Olthuis, W; van den Berg, A; Segerink, L I

    2015-03-01

    Selection of healthy spermatozoa is of crucial importance for the success rates of assisted reproduction technologies (ART) such as in vitro fertilization and intra-cytoplasmic sperm injection. Although sperm selection for ART procedures is predominantly based on sperm motility, successful fertilization is not predicted by good motility alone. For example, sperm characteristics such as the acrosome state and DNA integrity have shown significant impact on ART outcome. Although fertilization can be achieved with a single spermatozoon of high quality, current quality assessments are population-based and do not allow investigation of multiple sperm characteristics on a single spermatozoon simultaneously. In order to study sperm cells on the single cell level, we designed and characterized a PDMS microfluidic platform that allows single sperm entrapment. After spatially confining individual sperm cells within microfluidic cell traps, the cell viability, chromosomal content and acrosome state were studied. This platform is suitable for the analysis of individual sperm cells, which could be exploited for (non-invasive) sperm analysis and selection by impedance or Raman spectroscopy. PMID:25578490

  2. State of the Art in Platelet Function Testing

    PubMed Central

    E. Kehrel, Beate; F. Brodde, Martin

    2013-01-01

    Summary Platelets perform many functions in hemostasis but also in other areas of physiology and pathology. Therefore, it is obvious that many different function tests have been developed, each one conceived and standardized for a special purpose. This review will summarize the different fields in which platelet function testing is currently in use; diagnostics of patients with bleeding disorders, monitoring patients’ response to anti-platelet therapy, monitoring in transfusion medicine (blood donors, platelet concentrates, and after transfusion), and monitoring in perioperative medicine to predict bleeding tendency. The second part of the review outlines different methods for platelet function testing, spanning bleeding time, and platelet counting as well as determining platelet adhesion, platelet secretion, platelet aggregation, platelet morphology, platelet signal transduction, platelet procoagulant activity, platelet apoptosis, platelet proteomics, and molecular biology. PMID:23653569

  3. Mouse ICSI with frozen-thawed sperm: the impact of sperm freezing procedure and sperm donor strain.

    PubMed

    Moreira, Pedro N; Jimenéz, Adela; Fernández, Raul; Bury-Madrid, Ninoska; De la Fuente, Julio; Pintado, Belen; Gutiérrez-Adán, Alfonso

    2003-09-01

    Normal mouse offspring can be obtained from oocytes injected with frozen-thawed spermatozoa without cryoprotection, however, embryo development can be affected by sperm freezing procedure and sperm donor strain. In this study we observed that direct contact of mouse spermatozoa with liquid nitrogen did not affect their ability to activate injected oocytes but severely restricted subsequent in vitro embryo development to blastocyst stage. Tris-EDTA buffer and M2 were also shown to be better sperm freezing extenders than DPBS, allowing higher developmental potential. In addition, differences in embryo development obtained by intracytoplasmic sperm injection (ICSI) with frozen-thawed spermatozoa were observed between hybrid sperm donor strains. Frozen-thawed B6D2F1 spermatozoa provided higher embryo development than sperm cells from C57CBAF1. PMID:12874805

  4. 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.

  5. Sperm fertility and skewed paternity during sperm competition in the Australian long-eared bat Nyctophilus geoffroyi (Chiroptera: Vespertilionidae)

    Microsoft Academic Search

    David J. Hosken

    1998-01-01

    Prolonged sperm storage, rare among mammals, is widespread among bats and may promote sperm competition, assuming stored sperm are fertile. However, while sperm storage has been documented in many bat species, there have been few investigations of the fertility of stored sperm. Related to this, and a fundamental question in the study of competition at the gametic level, is the

  6. Availability of sperm examination for male reproductive toxicities in rats treated with boric acid.

    PubMed

    Yoshizaki, H; Izumi, Y; Hirayama, C; Fujimoto, A; Kandori, H; Sugitani, T; Ooshima, Y

    1999-08-01

    Sperm morphological examination, computer-assisted sperm analysis (CASA) and histopathologic examination of the testis and epididymis were performed for male rats treated orally with boric acid for 3 weeks at dosage levels of 50, 150 and 500 mg/kg/day, and treated males were mated with untreated females. None of the males treated with 500 mg/kg/day could impregnate untreated females. The fertility index showed a tendency to decrease in males treated with 150 mg/kg/day. At necropsy, the pre-implantation loss rate in females mated with males treated with 150 mg/kg/day was higher than the control values. Upon epididymal sperm analysis using the CASA system, all parameters including the number of sperm and sperm motions were found to be affected in males treated with 500 mg/kg/day, and the number of sperm, percent motile, velocities and amplitude of lateral head displacement (ALH) were affected in males treated with 150 mg/kg/day. Upon sperm morphological examination, head and tail abnormalities were observed in males treated with 150 and 500 mg/kg/day. In the histopathological examination, atrophy of the seminiferous tubules and multinucleated giant cells in the testes were observed in males treated with 500 mg/kg/day. PMID:10478334

  7. A practical in vitro sperm-egg binding assay that detects subfertile males.

    PubMed

    Barbato, G F; Cramer, P G; Hammerstedt, R H

    1998-03-01

    A series of in vitro assays of sperm-egg binding were developed to identify potentially subfertile roosters. Initial assays used either segments of intact hen's egg perivitelline membrane (PVM) placed on a microscope slide or a heat-solubilized extract of PVM (HS-PVM) dried within a flat-bottomed microwell plate, with bound sperm detected with a DNA-specific stain and epifluorescence microscopy. An automated assay was developed using prestained sulfosuccinimidyl-7-amino-4-methylcoumarin-3-acetate-HS-PVM and enumeration of bound sperm with a fluorometric microwell plate reader. Four populations of chickens differing in fertility were evaluated with the following results: 1) the correlation across lines between in vitro sperm binding and fertility was 0.83 (N = 40; p < 0.0001); 2) correlations with other seminal parameters were low; and 3) the relationship between sperm binding and fertility was not linear, but a threshold plot allowed identification of males with low binding and low fertility. Motile sperm from roosters, turkeys, bulls, humans, mice, rams, and stallions bound in a dose responsive manner. Features of binding were revealed by both scanning and transmission electron microscopy. Use of this assay to cull males whose semen appears normal by traditional modes of analysis but differs in the obligatory trait of sperm-egg binding could be of value to avoid expensive progeny testing. PMID:9565452

  8. Panax ginseng induces the expression of CatSper genes and sperm hyperactivation

    PubMed Central

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

    2014-01-01

    The cation channel of sperm (CatSper) protein family plays important roles in male reproduction and infertility. The four members of this family are expressed exclusively in the testis and are localized differently in sperm. To investigate the effects of Panax ginseng treatment on the expression of CatSper genes and sperm hyperactivation in male mice, sperm motility and CatSper gene expression were assessed using a computer-assisted semen analysis system, a Fluoroskan Ascent microplate fluorometer to assess Ca2+ influx, real-time polymerase chain reaction, Western blotting and immunofluorescence. The results suggested that the Ca2+ levels of sperm cells treated with P. ginseng were increased significantly compared with the normal group. The P. ginseng-treated groups showed increased sperm motility parameters, such as the curvilinear velocity and amplitude of lateral head displacement. Taken together, the data suggest that CatSper messenger ribonucleic acid levels were increased significantly in mouse testes in the P. ginseng-treated group, as was the protein level, with the exception of CatSper2. In conclusion, P. ginseng plays an important role in improving sperm hyperactivation via CatSper gene expression. PMID:24969054

  9. Comparative transcript profiling of gene expression of fresh and frozen-thawed bull sperm.

    PubMed

    Chen, Xiaoli; Wang, Yonggui; Zhu, Huabin; Hao, Haisheng; Zhao, Xueming; Qin, Tong; Wang, Dong

    2015-03-01

    Although frozen semen is widely used commercially in the cattle breeding industry, the resultant pregnancy rate is lower than that produced using fresh semen. Cryodamage is a major problem in semen cryopreservation; it causes changes to sperm transcripts that may influence sperm function and motility. We used suppression subtractive hybridization technology to establish a complementary DNA subtractive library, and combined microarray technology and sequence homology analysis to screen and analyze differentially expressed genes in the library, comparing fresh sperm with the frozen-thawed sperm of nine bulls. Overall, 19 positive differentially expressed unigenes were identified using microarray data and Significance Analysis of Microarrays software (|score (d)| ? 2, fold change > 1, and false discovery rate < 0.05). Of 15 differentially expressed unigenes exhibited high sequence homology (E-value ? 1 × 10(-3)), 12 were upregulated in frozen-thawed sperm, the remaining 3 were upregulated in fresh sperm, and 4 other clones were identified as unknown because of incomplete sequences or because there was no significant sequence homology (E-value > 1E(-03)) and were considered novel genes. The expression of five of these genes-RPL31, PRKCE, PAPSS2, PLP1, and R1G7-was verified by quantitative real-time reverse transcription-polymerase chain reaction. There was a significant differential expression of the RPL31 gene (P < 0.05). Our preliminary results provide an overview of differentially expressed transcripts between fresh and frozen-thawed sperm of Holstein bulls. PMID:25459024

  10. A Powerful Test for Comparing Multiple Regression Functions

    PubMed Central

    Maity, Arnab

    2012-01-01

    In this article, we address the important problem of comparison of two or more population regression functions. Recently, Pardo-Fernández, Van Keilegom and González-Manteiga (2007) developed test statistics for simple nonparametric regression models: Yij = ?j(Zij) + ?j(Zij)?ij, based on empirical distributions of the errors in each population j = 1, … , J. In this paper, we propose a test for equality of the ?j(·) based on the concept of generalized likelihood ratio type statistics. We also generalize our test for other nonparametric regression setups, e.g, nonparametric logistic regression, where the loglikelihood for population j is any general smooth function L{Yj,?j(Zj)}. We describe a resampling procedure to obtain the critical values of the test. In addition, we present a simulation study to evaluate the performance of the proposed test and compare our results to those in Pardo-Fernández et al. (2007). PMID:23074357

  11. Production of transgenic rabbit embryos through intracytoplasmic sperm injection.

    PubMed

    Li, Qiuyan; Hou, Jian; Wang, Sheng; Chen, Yongfu; An, Xiao-Rong

    2010-11-01

    The objective of this study was to test if intracytoplasmic sperm injection (ICSI)-mediated gene transfer was an effective method in the production of transgenic rabbit embryos. Rabbit sperm diluted in different media with various pH were treated by freezing without cryoprotectant, and their ability for DNA uptake was determined. In these experiments using production of transgenic rabbit embryos by ICSI, exogenous genes at three concentrations and of two conformation types were used. The rate of DNA association to the sperm seen by rhodamine-tagged DNA encoding green fluorescent protein (GFP) was 90.0%, 92.7%, 91.0%, 91.7%, and 92.3%, respectively in TCM199, DM, DPBS, CZB, and HCZB media. The DNA attachment to sperm was not affected by media pH within the range of 5.4-9.4 (p > 0.05). Expression of GFP first occurred at the 2-cell stage and continued to blastocyst formation. DNA concentration (between 5, 10, and 20 ng/?l) or conformation (linear and circular) had no effect on the production rate of transgenic embryos. These results indicated that genetically modified rabbit blastocysts can be efficiently produced by ICSI technique. PMID:20663236

  12. 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.

  13. Localization and identification of sumoylated proteins in human sperm: excessive sumoylation is a marker of defective spermatozoa

    PubMed Central

    Vigodner, Margarita; Shrivastava, Vibha; Gutstein, Leah Elisheva; Schneider, Jordana; Nieves, Edward; Goldstein, Marc; Feliciano, Miriam; Callaway, Myrasol

    2013-01-01

    BACKGROUND Sumoylation is a type of post-translational modification that is implicated in the regulation of numerous cellular events. However, its role in the function of human sperm has not yet been characterized. METHODS AND RESULTS In this study, both immunofluorescence and electron microscopy revealed that small ubiquitin-like modifiers (SUMO) SUMO1 and SUMO2/3 were highly enriched in the neck area of human sperm that is associated with the redundant nuclear envelope and were also detectable in the flagella and some head regions. Similar localization patterns of SUMO were also observed in mouse and fly sperm. Nonmotile, two-tailed, curled tailed, misshapen, microcephalic (small head) and aciphalic (no head) sperm exhibited abnormally high levels of sumoylation in their neck and tail regions relative to normal sperm. Numerous sumoylated proteins, ranging from 20 to 260 kDa, were detected via western blotting and identified by mass spectrometry, and 55 SUMO targets that were present specifically in human sperm, and not in the control fraction, corresponded to flagella proteins, proteins involved in the maturation and differentiation of sperm, heat shock proteins and important glycolytic and mitochondrial enzymes. The targets that were identified included proteins with specific functions in germ cells and sperm, such as heat shock-related 70-kDa protein 2, outer dense fiber protein 3, A-kinase anchor proteins 3 and 4, l-lactate dehydrogenase C, sperm protein associated with the nucleus on the X chromosome B/F, valosin-containing protein, seminogelins, histone H4 and ubiquitin. Coimmunoprecipitation experiments confirmed the sumoylation of semenogelin and indicated that some sperm proteins are modified by sumoylation and ubiquitination simultaneously. CONCLUSIONS Numerous proteins are modified by sumoylation in human sperm; excessive sumoylation is a marker of defective spermatozoa. PMID:23077236

  14. Optimization of isolation and storage of sperm cells from pollen of perennial ryegrass ( Lolium perenne L.)

    Microsoft Academic Search

    H. M. van der Maas; M. A. C. M. Zaal; E. R. de Jong; J. L. van Went; F. A. Krens

    1993-01-01

    Summary  A procedure for isolating sperm cells of perennial ryegrass (Lolium perenne L.) was developed. The sperm cells were released from the pollen grains by osmotic shock, with the right combination of pH and osmolality being important for optimal release. Various combinations of vitamins E, C and fetal calf serum were tested with the aim of improving yield and long-term viability,

  15. Effect of increased scrotal temperature on sperm production in normal men

    Microsoft Academic Search

    Christina Wang; Veronica McDonald; Andrew Leung; Laura Superlano; Nancy Berman; Laura Hull; Ronald S. Swerdloff

    1997-01-01

    Objective: To determine whether application of polyester-lined athletic supports to bring the testes closer to the abdomen increases scrotal temperature and decreases sperm production.Design: Prospective clinical study.Setting: University academic medical center.Patient(s): Twenty-one healthy male volunteers.Intervention(s): The study consisted of a pretreatment period of 6 weeks, a treatment phase of 52 weeks, and a recovery phase until return to normal sperm

  16. Post-thaw viability of bull AI-doses with low-sperm numbers

    Microsoft Academic Search

    J. Ballester; A. Johannisson; F. Saravia; M. Håård; H. Gustafsson; D. Bajramovic; H. Rodriguez-Martinez

    2007-01-01

    Use of AI-doses containing low-sperm numbers are increasingly been used to optimise use of elite bulls as well as to accommodate an eventual wider application of sex-sorted semen. Since spermatozoa might, however, suffer from high extension rates, thus compromising fertility, this study evaluated the post-thaw sperm quality of semen from commercial progeny-tested, high-ranked AI-sires whose semen was within acceptable limits

  17. Characteristics of sperm acrosin-like activity of paddlefish ( Polyodon spathula Walbaum)

    Microsoft Academic Search

    Andrzej Ciereszko; Konrad Dabrowski; Steven D. Mims; Jan Glogowski

    2000-01-01

    Spermatozoa of paddlefish and sturgeon fishes (Acipenseriformes), unlike teleost fish, have an acrosome. The objectives of this study were to characterize acrosin-like activity of cryopreserved sperm of paddlefish (Polyodon spathula) and to test and compare stability of paddlefish acrosin-like activity with that of lake sturgeon and bull spermatozoa. Mean acrosin-like activity of cryopreserved paddlefish sperm was 0.372±0.067 ?U\\/106 spermatozoa. This

  18. Clinical testing of otolith function: perceptual thresholds and myogenic potentials.

    PubMed

    Agrawal, Yuri; Bremova, Tatiana; Kremmyda, Olympia; Strupp, Michael; MacNeilage, Paul R

    2013-12-01

    Cervical and ocular vestibular-evoked myogenic potential (cVEMP/oVEMP) tests are widely used clinical tests of otolith function. However, VEMP testing may not be the ideal measure of otolith function given the significant inter-individual variability in responses and given that the stimuli used to elicit VEMPs are not physiological. We therefore evaluated linear motion perceptual threshold testing compared with cVEMP and oVEMP testing as measures of saccular and utricular function, respectively. A multi-axis motion platform was used to measure horizontal (along the inter-aural and naso-occipital axes) and vertical motion perceptual thresholds. These findings were compared with the vibration-evoked oVEMP as a measure of utricular function and sound-evoked cVEMP as a measure of saccular function. We also considered how perceptual threshold and cVEMP/oVEMP testing are each associated with Dizziness Handicap Inventory (DHI) scores. We enrolled 33 patients with bilateral vestibulopathy of different severities and 42 controls to have sufficient variability in otolith function. Subjects with abnormal oVEMP amplitudes had significantly higher (poorer) perceptual thresholds in the inter-aural and naso-occipital axes in age-adjusted analyses; no significant associations were observed for vertical perceptual thresholds and cVEMP amplitudes. Both oVEMP amplitudes and naso-occipital axis perceptual thresholds were significantly associated with DHI scores. These data suggest that horizontal perceptual thresholds and oVEMPs may estimate the same underlying physiological construct: utricular function. PMID:24077672

  19. Sperm interaction with fallopian tube apical membrane enhances sperm motility and delays capacitation

    Microsoft Academic Search

    Shona C. Murray; T. Timothy Smith

    1997-01-01

    Objective: To understand the effect of sperm contact with the apical plasma membrane of tubal epithelial cells on sperm motility, velocity, and capacitation.Design: Prospective, controlled in vitro study.Setting: University medical center.Patient(s): Women of reproductive age undergoing hysterectomy for benign gynecologic indications and normozoospermic donors of proved fertility.Main Outcome Measure(s): Sperm motility as measured manually, velocity as measured by computer-assisted sperm

  20. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... false Hydrogenated sperm oil. 173.275 Section 173...ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED...173.275 Hydrogenated sperm oil. The food additive hydrogenated sperm oil may be safely used in...

  1. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... false Hydrogenated sperm oil. 173.275 Section 173...ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED...173.275 Hydrogenated sperm oil. The food additive hydrogenated sperm oil may be safely used in...

  2. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... false Hydrogenated sperm oil. 173.275 Section 173...ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED...173.275 Hydrogenated sperm oil. The food additive hydrogenated sperm oil may be safely used in...

  3. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... false Hydrogenated sperm oil. 173.275 Section 173...ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED...173.275 Hydrogenated sperm oil. The food additive hydrogenated sperm oil may be safely used in...

  4. 21 CFR 173.275 - Hydrogenated sperm oil.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2009-04-01 true Hydrogenated sperm oil. 173.275 Section 173...ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED...173.275 Hydrogenated sperm oil. The food additive hydrogenated sperm oil may be safely used in...

  5. The Case for an Unregulated Private Sperm Donation Market

    E-print Network

    Acker, Jacqueline M.

    2013-01-01

    Donation? Although sperm donation for human conception datesHuman Services to protect her right to receive privately donated sperm.sperm is an inherent characteristic and an intrinsic part of being a male human, and

  6. 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

  7. Functional assessment of sexual maturity in male macaques (Macaca fascicularis).

    PubMed

    Luetjens, C M; Weinbauer, G F

    2012-08-01

    Selection of suitable criteria for assessing sexual maturity in the male long-tailed macaque (Macaca fascicularis) has yielded conflicting results. The present retrospective work investigates whether the sole presence of sperm in the baseline semen sample unequivocally (i.e. for every animal) hallmarks complete testicular maturation. For 956 animals providing the baseline semen sample, neither age, body weight nor testes volume unequivocally predicted the presence of sperm in that sample, and for 322 animals these parameters failed to predict testicular histology. In contrast, the presence of sperm in the baseline semen sample correlated with mature testis histology at study termination in every single animal (n=197/322). Surprisingly, for the 125/322 animals without sperm in the baseline semen sample, spermatogenesis was also mature in 95 animals. Thus, the mere provision of a semen sample without sperm--implying peripheral reproductive tract maturation--was associated with mature spermatogenesis in approx. 75% of animals. Interestingly, testicular maturation occurred approx. 2 years earlier in Mauritian compared to Asian mainland animals. In conclusion, a single semen sample that contains sperm provides unequivocal evidence for mature spermatogenesis and, thus, is suggested as a functional parameter for sexual maturity assessment in this species. PMID:22579626

  8. Cryopreservation of domestic animal sperm cells

    Microsoft Academic Search

    J. P. Barbas; R. D. Mascarenhas

    2009-01-01

    Sperm cells are the endpoint of male spermatogenesis and have particular anatomic and metabolic features. Sperm cryopreservation\\u000a and storage currently require liquid nitrogen or ultralow refrigeration methods for long or short term storage, which requires\\u000a routine maintenance and extensive space requirements. Conserving sperms have several purposes such as artificial reproductive\\u000a technologies (ART), species conservation and clinical medicine. The combinations of

  9. Postnatal Effects of Sperm Chromatin Damage

    Microsoft Academic Search

    Miriam Pérez-Crespo; Raúl Fernández-González; Miguel Ángel Ramírez; Eva Pericuesta; Alexandra Calle; Alfonso Gutiérrez-Adán

    \\u000a The use of spermatozoa with fragmented DNA has been linked to ­developmental and postnatal effects in animal models. Environmental\\u000a and toxic factors such as radiation, heat stress, air pollution, chemotherapeutic agents, etc. are known to have detrimental\\u000a effects on sperm chromatin. Sperm chromatin damage has also been observed following sperm manipulation techniques (freeze–thawing\\u000a without cryoprotectants, freeze-drying, preincubation under different conditions,

  10. X-y interactions underlie sperm head abnormality in hybrid male house mice.

    PubMed

    Campbell, Polly; Nachman, Michael W

    2014-04-01

    The genetic basis of hybrid male sterility in house mice is complex, highly polygenic, and strongly X linked. Previous work suggested that there might be interactions between the Mus musculus musculus X and the M. m. domesticus Y with a large negative effect on sperm head morphology in hybrid males with an F1 autosomal background. To test this, we introgressed the M. m. domesticus Y onto a M. m. musculus background and measured the change in sperm morphology, testis weight, and sperm count across early backcross generations and in 11th generation backcross males in which the opportunity for X-autosome incompatibilities is effectively eliminated. We found that abnormality in sperm morphology persists in M. m. domesticus Y introgression males, and that this phenotype is rescued by M. m. domesticus introgressions on the X chromosome. In contrast, the severe reductions in testis weight and sperm count that characterize F1 males were eliminated after one generation of backcrossing. These results indicate that X-Y incompatibilities contribute specifically to sperm morphology. In contrast, X-autosome incompatibilities contribute to low testis weight, low sperm count, and sperm morphology. Restoration of normal testis weight and sperm count in first generation backcross males suggests that a small number of complex incompatibilities between loci on the M. m. musculus X and the M. m. domesticus autosomes underlie F1 male sterility. Together, these results provide insight into the genetic architecture of F1 male sterility and help to explain genome-wide patterns of introgression across the house mouse hybrid zone. PMID:24504187

  11. 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

  12. 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.

  13. RNA in situ hybridization makes sperm doesn't make sperm

    E-print Network

    RNA in situ hybridization makes sperm doesn't make sperm #12;spe-47 promoter spe-47 3' UTR structures in developing sperm: the Fibrous Body (FB) Membranous Organelle (MO) complexes. A mutation in spe regions of the protein. #12;Structural alignment of SPE-8 with crystal structures from human c-Abl protein

  14. A Statistical Test for Differential Item Pair Functioning.

    PubMed

    Bechger, Timo M; Maris, Gunter

    2015-06-01

    This paper presents an IRT-based statistical test for differential item functioning (DIF). The test is developed for items conforming to the Rasch (Probabilistic models for some intelligence and attainment tests, The Danish Institute of Educational Research, Copenhagen, 1960) model but we will outline its extension to more complex IRT models. Its difference from the existing procedures is that DIF is defined in terms of the relative difficulties of pairs of items and not in terms of the difficulties of individual items. The argument is that the difficulty of an item is not identified from the observations, whereas the relative difficulties are. This leads to a test that is closely related to Lord's (Applications of item response theory to practical testing problems, Erlbaum, Hillsdale, 1980) test for item DIF albeit with a different and more correct interpretation. Illustrations with real and simulated data are provided. PMID:25223228

  15. 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

  16. Sperm Size Evolution in Drosophila : Inter and Intraspecific Analysis

    Microsoft Academic Search

    Dominique Joly; Abraham Korol; Eviatar Nevo

    2004-01-01

    Numerous reports were devoted to the variation of sperm length in relation to sperm competition amongst species. However, studies on intraspecific variations of sperm size are very scarce and the number of sperm measured, very limited. This paper investigates within-individual, between-individual and between-population variation of sperm length in the two cosmopolitan species, D. simulans and D. melanogaster. Sperm length distributions

  17. Prediction of the in-vitro fertOization (IVF) potential of human spermatozoa using sperm function tests: the effect of the delay between testing and IVF

    Microsoft Academic Search

    Nares Sukcharoen; John Keith; D. Stewart Irvine

    To examine the diagnostic significance of several criteria of semen quality and to determine whether their prognostic value is eroded by the time interval between assessment and the attempt at in-vitro fertilization (IVF) with embryo transfer, 73 couples undergoing IVF and embryo transfer therapy were studied. The ability of human spermatozoa to achieve fertilization in vitro was examined hi relation

  18. Aging of vestibular function evaluated using correlational vestibular autorotation test

    PubMed Central

    Hsieh, Li-Chun; Lin, Hung-Ching; Lee, Guo-She

    2014-01-01

    Background Imbalance from degeneration of vestibular end organs is a common problem in the elderly. However, the decline of vestibular function with aging was revealed in few vestibular function tests such as vestibular autorotation test (VAT). In the current VAT, there are drawbacks of poor test–retest reliability, slippage of the sensor at high-speed rotations, and limited data about the effect of aging. We developed a correlational-VAT (cVAT) system that included a small, light sensor (less than 20 g) with wireless data transmission technique to evaluate the aging of vestibular function. Material and methods We enrolled 53 healthy participants aged between 25 and 75 years and divided them into five age groups. The test conditions were vertical and horizontal head autorotations of frequencies from 0 to 3 Hz with closed eyes or open eyes. The cross-correlation coefficient (CCC) between eye velocity and head velocity was obtained for the head autorotations between 1 Hz and 3 Hz. The mean of the CCCs was used to represent the vestibular function. Results Age was significantly and negatively correlated with the mean CCC for all test conditions, including horizontal or vertical autorotations with open eyes or closed eyes (P<0.05). The mean CCC with open eyes declined significantly at 55–65 years old and the mean CCC with closed eyes declined significantly at 65–75 years old. Conclusion Vestibular function evaluated using mean CCC revealed a decline with age, and the function of visual-vestibulo-ocular reflex declined 10 years earlier than the function of vestibulo-ocular reflex. PMID:25214774

  19. Applications of human sperm parameters for monitoring

    SciTech Connect

    Wyrobek, A.J.

    1985-11-01

    Very few studies of male reproductive hazards in the workplace are being done, and there are several underlying problems slowing the process in this infant field. First, the interaction of many diverse disciplines from management, labor, government, medicine, and science is often required. Outlooks and incentives are generally in conflict from the outset. Second, there is considerable complacency because no workplace agent as toxic as DBCP has been found. Third, there is as yet no requirement to assess the human male reproductive effects of a compound before large groups of men are permitted to work with it. Fourth, there is a poor opinion of the available epidemiological methods as indicators of abnormal reproductive outcome, because they are expensive, prone to confounding factors, and small numbers of exposed men (and pregnancies at risk) limit their sensitivity. Fifth, sperm tests have been criticized for their variability, subjectivity, and lack of standardization. 68 refs., 2 tabs.

  20. [Diagnosis of pancreatic function by the selenomethionine--75Se test].

    PubMed

    Dressler, J; Hör, G; Buttermann, G; Pabst, H W

    1975-02-14

    Dterminations of 75selenium radioactivity and pancreatic enzyme in duodenal aspirate were carried out in 50 patients after i. v. injection of selenomethionine-75Se. After standardized excitation of the pancreas with pancreozymin-secretin, a significant correlation (r = 0.654) between excreted 75Se radioactivity and trypsin production could be established. The test is suitable for the recording of exocrine pancreas function while avoiding expensive enzymatic methods. The combined use of pancreas scintigraphy--using the double radionuclide substraction technique-with the radioselenomethionine test (RSMT) provides comprehensive diagnostic information on the morphology and function of the pancreas. PMID:235085

  1. Testing Odd-Cycle-Freeness in Boolean Functions

    E-print Network

    Bhattacharyya, Arnab; Raghavendra, Prasad; Shapira, Asaf

    2011-01-01

    Call a function f : F_2^n -> {0,1} odd-cycle-free if there are no x_1, ..., x_k in F_2^n with k an odd integer such that f(x_1) = ... = f(x_k) = 1 and x_1 + ... + x_k = 0. We show that one can distinguish odd-cycle-free functions from those eps-far from being odd-cycle-free by making poly(1/eps) queries to an evaluation oracle. To obtain this result, we use connections between basic Fourier analysis and spectral graph theory to show that one can reduce testing odd-cycle-freeness of Boolean functions to testing bipartiteness of dense graphs. Our work forms part of a recent sequence of works that shows connections between testability of properties of Boolean functions and of graph properties. We also prove that there is a canonical tester for odd-cycle-freeness making poly(1/eps) queries, meaning that the testing algorithm operates by picking a random linear subspace of dimension O(log 1/eps) and then checking if the restriction of the function to the subspace is odd-cycle-free or not. The test is analyzed by s...

  2. Simple tests of renal function using creatinine, phenolsulphonphthalein, and pitressin

    PubMed Central

    Healy, J. K.; Edwards, K. D. G.; Whyte, H. M.

    1964-01-01

    The reciprocals of the blood urea nitrogen and plasma creatinine levels, the maximum specific gravity of the urine after vasopressin, and three modified forms of the phenolsulphonphthalein (P.S.P.) excretion test were found to be directly related to the glomerular filtration rate (G.F.R.) in hospital patients. From 34 to 75 patients were studied for each test, and in 21 patients all tests were performed concurrently. The plasma creatinine level and the 15-minute urinary excretion of P.S.P. were found to be the most useful simple tests of renal function and gave sufficiently accurate estimates of total function (G.F.R.) to justify their more extensive use. The G.F.R. (ml./min./1·73 sq. m. of body surface area) could be calculated from each test, using the following equations:— [Formula: see text] G.F.R. = 3·15 × P.S.P.% + 19, where P.S.P.% is the 15-minute urinary excretion of P.S.P., expressed as a percentage of the administered dose. Satisfactory estimates of G.F.R. were also given by the simplified relationships [Formula: see text]. In the presence of impaired renal function more accurate estimates of G.F.R. were obtained from the plasma creatinine and P.S.P. excretion tests (S.D. of estimate 8 and 13 ml./min. respectively). It was thought that these tests could well replace the commonly used blood urea nitrogen estimation as simple tests of renal function. PMID:14207792

  3. Evaluation of upper extremity motor function tests in tetraplegics

    Microsoft Academic Search

    JH van Tuijl; YJM Janssen-Potten; HAM Seelen

    2002-01-01

    Objective: To provide an overview of arm–hand function tests useful in tetraplegic subjects. Considerations for selection of an appropriate test are also provided.Data sources: A Medline literature search was conducted covering the period from 1967 to March 2001. Relevant references cited in the selected papers were also considered, regardless of the year of publication.Study selection: This review was restricted to

  4. Alteration of sperm protein profile induced by cigarette smoking.

    PubMed

    Chen, Xiaohui; Xu, Wangjie; Miao, Maohua; Zhu, Zijue; Dai, Jingbo; Chen, Zhong; Fang, Peng; Wu, Junqing; Nie, Dongsheng; Wang, Lianyun; Wang, Zhaoxia; Qiao, Zhongdong; Shi, Huijuan

    2015-07-01

    Cigarette smoking is associated with lower semen quality, but how cigarette smoking changes the semen quality remains unclear. The aim of this study was to screen the differentially expressed proteins in the sperm of mice with daily exposure to cigarette smoke. The 2D gel electrophoresis (2DE) and mass spectrometry (MS) analyses results showed that the mouse sperm protein profile was altered by cigarette smoking. And 22 of the most abundant proteins that correspond to differentially expressed spots in 2DE gels of the sperm samples were identified. These proteins were classified into different groups based on their functions, such as energy metabolism, reproduction, and structural molecules. Furthermore, the 2DE and MS results of five proteins (Aldoa, ATP5a1, Gpx4, Cs, and Spatc1) were validated by western blot analysis and reverse transcriptase-polymerase chain reaction. Results showed that except Spatc1 the other four proteins showed statistically significant different protein levels between the smoking group and the control group (P < 0.05). The expressions of three genes (Aldoa, Gpx4, and Spatc1) were significantly different (P < 0.05) at transcription level between the smoking group and the control group. In addition, five proteins (Aldoa, ATP5a1, Spatc1, Cs, and Gpx4) in human sperm samples from 30 male smokers and 30 non-smokers were detected by western blot analysis. Two proteins (Aldoa and Cs) that are associated with energy production were found to be significantly altered, suggesting that these proteins may be potential diagnostic markers for evaluation of smoking risk in sperm. Further study of these proteins may provide insight into the pathogenic mechanisms underlying infertility in smoking persons. PMID:26063603

  5. Investigation of the mechanisms by which the molecular chaperone HSPA2 regulates the expression of sperm surface receptors involved in human sperm-oocyte recognition.

    PubMed

    Redgrove, Kate A; Anderson, Amanda L; McLaughlin, Eileen A; O'Bryan, Moira K; Aitken, R John; Nixon, Brett

    2013-03-01

    A unique characteristic of mammalian spermatozoa is that, upon ejaculation, they are unable to recognize and bind to an ovulated oocyte. These functional attributes are only realized following the cells' ascent of the female reproductive tract whereupon they undergo a myriad of biochemical and biophysical changes collectively referred to as 'capacitation'. We have previously shown that this functional transformation is, in part, engineered by the modification of the sperm surface architecture leading to the assembly and/or presentation of multimeric sperm-oocyte receptor complexes. In this study, we have extended our findings through the characterization of one such complex containing arylsulfatase A (ARSA), sperm adhesion molecule 1 (SPAM1) and the molecular chaperone, heat shock 70kDa protein 2 (HSPA2). Through the application of flow cytometry we revealed that this complex undergoes a capacitation-associated translocation to facilitate the repositioning of ARSA to the apical region of the human sperm head, a location compatible with a role in the mediation of sperm-zona pellucida (ZP) interactions. Conversely, SPAM1 appears to reorient away from the sperm surface, possibly reflecting its primary role in cumulus matrix dispersal preceding sperm-ZP recognition. The dramatic relocation of the complex was completely abolished by incubation of capacitating spermatozoa in exogenous cholesterol or broad spectrum protein kinase A (PKA) and tyrosine kinase inhibitors suggesting that it may be driven by alterations in membrane fluidity characteristics and concurrently by the activation of a capacitation-associated signal transduction pathway. Collectively these data afford novel insights into the sub-cellular localization and potential functions of multimeric protein complexes in human spermatozoa. PMID:23247813

  6. Sexual selection and the evolution of sperm quality.

    PubMed

    Fitzpatrick, John L; Lüpold, Stefan

    2014-12-01

    Sperm experience intense and varied selection that dramatically impacts the evolution of sperm quality. Selection acts to ensure that sperm are fertilization-competent and able to overcome the many challenges experienced on their way towards eggs. However, simply being able to fertilize an egg is not enough to ensure male fertility in most species. Owing to the prevalence of female multiple mating throughout the animal kingdom, successful fertilization requires sperm to outcompete rival sperm. In addition, females can actively influence sperm quality, storage or utilization to influence male fertility. This review provides an overview of how these selective forces influence the evolution of sperm quality. After exploring the link between sperm traits and male fertility, we examine how post-mating competition between rival ejaculates influences the evolution of sperm quality. We then describe how complex genetic, social and sexual interactions influence sperm quality, focusing on the importance of seminal fluid and interactions between sperm and the female's reproductive tract. In light of the complexities of selection on sperm traits, greater use of multivariate approaches that incorporate male-male, sperm-sperm and sperm-female interactions to study sperm quality will enhance our understanding of how selection acts on sperm traits and factors influencing male fertility. Because the metric of male reproductive success--fertilization--is the same across the animal kingdom, we argue that information about sperm evolution gained from non-human animals has enormous potential to further our understanding of the factors that impact human fertility. PMID:25323970

  7. Paternal Poly (ADP-ribose) Metabolism Modulates Retention of Inheritable Sperm Histones and Early Embryonic Gene Expression

    PubMed Central

    Leu, N. Adrian; Rao, Shilpa; Li, Fan; Gregory, Brian D.; Zalenskaya, Irina A.; Schultz, Richard M.; Meyer, Ralph G.

    2014-01-01

    To achieve the extreme nuclear condensation necessary for sperm function, most histones are replaced with protamines during spermiogenesis in mammals. Mature sperm retain only a small fraction of nucleosomes, which are, in part, enriched on gene regulatory sequences, and recent findings suggest that these retained histones provide epigenetic information that regulates expression of a subset of genes involved in embryo development after fertilization. We addressed this tantalizing hypothesis by analyzing two mouse models exhibiting abnormal histone positioning in mature sperm due to impaired poly(ADP-ribose) (PAR) metabolism during spermiogenesis and identified altered sperm histone retention in specific gene loci genome-wide using MNase digestion-based enrichment of mononucleosomal DNA. We then set out to determine the extent to which expression of these genes was altered in embryos generated with these sperm. For control sperm, most genes showed some degree of histone association, unexpectedly suggesting that histone retention in sperm genes is not an all-or-none phenomenon and that a small number of histones may remain associated with genes throughout the genome. The amount of retained histones, however, was altered in many loci when PAR metabolism was impaired. To ascertain whether sperm histone association and embryonic gene expression are linked, the transcriptome of individual 2-cell embryos derived from such sperm was determined using microarrays and RNA sequencing. Strikingly, a moderate but statistically significant portion of the genes that were differentially expressed in these embryos also showed different histone retention in the corresponding gene loci in sperm of their fathers. These findings provide new evidence for the existence of a linkage between sperm histone retention and gene expression in the embryo. PMID:24810616

  8. 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.

  9. Test Driving Three 1995 Genetic Algorithms: New Test Functions and Geometric Matching

    E-print Network

    Whitley, Darrell

    . 1 Introduction The use of genetic algorithms as optimization tools is now familiar to a broad rangeTest Driving Three 1995 Genetic Algorithms: New Test Functions and Geometric Matching D. Whitley, R, Colorado 80523 USA (303) 491­5373 whitley@cs.colostate.edu Abstract Genetic algorithms have attracted

  10. Inhibition of sperm capacitation and fertilizing capacity by adjudin is mediated by chloride and its channels in humans

    PubMed Central

    Li, Kun; Ni, Ya; He, Yi; Chen, Wen-Ying; Lu, Jian-Xin; Cheng, C. Yan; Ge, Ren-Shan; Shi, Qi-Xian

    2013-01-01

    STUDY QUESTION Does adjudin disrupt chloride ion (Cl?) ion transport function in human sperm and impede sperm capacitation and fertilizing ability in vitro? SUMMARY ANSWER In this study the results indicate that adjudin is a potent blocker of Cl? channels: disrupting Cl? ion transport function results in a decline in sperm capacitation and fertilizing ability in humans in vitro. WHAT IS KNOWN ALREADY Although our previous studies have demonstrated that adjudin exerts its effect by disrupting sertoli-germ cell adhesion junctions, most notably apical ectoplasmic specialization by targeting testin and actin filament bundles that disrupts the actin-based cytoskeleton in sertoli cells, it remains unclear whether adjudin impedes Cl? ion transport function in the human sperm. STUDY DESIGIN, SIZE AND DURATION Semen samples were obtained from 45 fertile men (aged 25–32). Spermatozoa were isolated from the semen in the human tube fluid (HTF) medium by centrifugation through a discontinuous Percoll gradient, and incubated with adjudin at 10 nM–10 µM and/or other reagents under capacitating conditions for 0–5 h. PARTICIPANTS/MATERIALS, SETTING, METHODS We evaluated the effect of adjudin and different reagents on sperm functions with which they were incubated at 37°C. Sperm motility and hyperactivation were analyzed by a computer-assisted sperm analysis (CASA) system. Sperm capacitation and the acrosome reaction were assessed by chlortetracycline fluorescence staining. Sperm fertilizing ability was evaluated by sperm penetration of zona-free hamster egg assay, and cellular cAMP levels in spermatozoa were quantified by the EIA kit. The proteins tyrosine, serine and threonine phosphorylation in the presence or absence of adjudin were analyzed by means of a immunodetection of spermatozoa, especially, compared the effect of adjudin on sperm hyperactivation and capacitation in the complete HTF medium with the Cl?-deficient HTF medium as well as the various Cl? channel blockers. MAIN RESULTS AND THE ROLE OF CHANCE Adjudin significantly inhibited sperm hyperactivation but not sperm motility. Adjudin-induced inhibition of sperm capacitation was reversible, and it was found to block the rhuZP3?- and progesterone-induced acrosome reaction in a dose-dependent manner. Adjudin also blocked sperm penetration of zona-free hamster eggs, and significantly inhibited both forskolin-activated transmembrane adenylyl cyclase and soluble adenylyl cyclase activities leading to a significant decline in the cellular cAMP levels in human spermatozoa. Adjudin failed to reduce sperm protein tyrosine phosphorylation but it did prevent sperm serine and threonine protein phosphorylation. Interestingly, adjudin was found to exert its inhibitory effects on sperm capacitation and capacitation-associated events only in the complete Cl?-HTF medium but not Cl?-deficient medium, illustrating the likely involvement of Cl?. Adjudin inhibits the fertility capacity of human sperm is mediated by disrupting chloride ion and its transport function. LIMITATIONS, REASONS FOR CAUTION This study has examined the effect of adjudin only on human sperm capacitation and fertilizing ability in vitro and thus has some limitations. Further investigations in vivo are needed to confirm adjudin is a potent male contraceptive. WIDER IMPLICATIONS OF THE FINDINGS Our studies demonstrated that adjudin inhibition of capacitation is reversible and its toxicity is low, opening the door for the examination of adjudin as a mediator of male fertility control. Adjudin may be a safe, efficient and reversible male antifertility agent and applicable to initial clinical trials of adjudin as a male antifertility agent in humans. STUDING FUNDING/COMPETING INTEREST(S) This work was supported by the National Basic Research Program of China (2006CB504002), the Nature Science Foundation of China (Nos. 81000244 and 81170554), Zhejiang Project of Science and Technology (2011C23046), the Nature Science Fund of Zhejian

  11. Particulate and soluble adenylyl cyclases participate in the sperm acrosome reaction.

    PubMed

    Beltrán, Carmen; Vacquier, Victor D; Moy, Gary; Chen, Yanqiu; Buck, Jochen; Levin, Lonny R; Darszon, Alberto

    2007-07-13

    cAMP is important in sea urchin sperm signaling, yet the molecular nature of the adenylyl cyclases (ACs) involved remained unknown. These cells were recently shown to contain an ortholog of the mammalian soluble adenylyl cyclase (sAC). Here, we show that sAC is present in the sperm head and as in mammals is stimulated by bicarbonate. The acrosome reaction (AR), a process essential for fertilization, is influenced by the bicarbonate concentration in seawater. By using functional assays and immunofluorescence techniques we document that sea urchin sperm also express orthologs of multiple isoforms of transmembrane ACs (tmACs). Our findings employing selective inhibitors for each class of AC indicate that both sAC and tmACs participate in the sperm acrosome reaction. PMID:17524362

  12. Effect of Cordyceps militaris supplementation on sperm production, sperm motility and hormones in Sprague-Dawley rats.

    PubMed

    Chang, Ying; Jeng, Kee-Ching; Huang, Kuei-Fen; Lee, Ying-Chung; Hou, Chien-Wei; Chen, Kuan-Hao; Cheng, Fu-Yen; Liao, Jiunn-Wang; Chen, Yuh-Shuen

    2008-01-01

    Cordyceps species have been traditionally used as for the enhancement of sexual function, but its direct evidence is lacking. We investigated the spermatogenic effect of Cordyceps militaris (CM) as supplementation with CM mycelium to 7-week-old male Sprague-Dawley (SD) rats. Ninety rats (30 for each group) were selected to regular diet or diet supplemented with CM mycelium (1% and 5%) for 6 weeks. Epididymal sperm were collected from 6 animals per group at each interval of observation. They were allowed to recover for one week. The quality and quantity of sperm were compared in these rats. The CM supplementation resulted in an increase of serum cordycepin concentration (n = 6, each group) that correlated with treatment time and the cordycepin level was significantly higher (p < 0.05) in 5% group as compared to 1% group at the 5th and 6th week. Epididymal sperm count was enhanced significantly from the control, at the 5th week and peaked at the 6th week in both groups supplemented with CM (each time point, n = 6; p < 0.05) and maintained for 2 weeks after stopping the treatment. Increased serum testosterone and estradiol-17 (E2) concentrations were found in rats with the CM supplementation (p < 0.05), but not other hormones such as follicle stimulating hormone (FSH), luteinizing hormone (LH) or prolactin. Importantly, percentages of motile sperm cells were also enhanced significantly (p < 0.05) paralleled the serum testosterone pattern from the supplement groups as compared to the control group. Taken together, these results indicate that supplementation with CM improves sperm quality and quantity in rats. PMID:19051352

  13. 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.

  14. Sequence-specific packaging of DNA in human sperm chromatin

    Microsoft Academic Search

    J. M. Gatewood; G. R. Cook; R. Balhorn; E. M. Bradbury; C. W. Schmid

    1987-01-01

    The DNA in human sperm chromatin is packaged into nucleoprotamine (approx.85%) and nucleohistone (approx.15%). Whether these two chromatin fractions are sequence-specific subsets of the spermatozoon genome is the question addressed in this report. Sequence-specific packaging would suggest distinct structural and functional roles for nucleohistone and nucleoprotamine in late spermatogenesis or early development or both. After removal of histones with 0.65

  15. Regulation of Protein Tyrosine Phosphorylation in Human Sperm by a Calcium\\/Calmodulin-Dependent Mechanism: Identification of A Kinase Anchor Proteins as Major Substrates for Tyrosine Phosphorylation

    Microsoft Academic Search

    Alfonso Carrera; Jiri Moos; Xiao Ping Ning; George L. Gerton; Jan Tesarik; Gregory S. Kopf; Stuart B. Moss

    1996-01-01

    Signal transduction pathways regulate various aspects of mammalian sperm function. When human sperm were incubated in a medium supporting capacitation, proteins became tyrosine-phosphorylated in a time-dependent manner. This phosphorylation was inhibited by genistein, a protein tyrosine kinase inhibitor. Phosphorylation was also reduced when sperm were incubated either in the presence of increasing concentrations of extracellular Ca2+or in a medium containing

  16. Clinical Factors Associated with Sperm DNA Fragmentation in Male Patients with Infertility

    PubMed Central

    Komiya, Akira; Kato, Tomonori; Kawauchi, Yoko; Watanabe, Akihiko; Fuse, Hideki

    2014-01-01

    Objective. The clinical factors associated with sperm DNA fragmentation (SDF) were investigated in male patients with infertility. Materials and Methods. Fifty-four ejaculates from infertile Japanese males were used. Thirty-three and twenty-one were from the patients with varicoceles and idiopathic causes of infertility, respectively. We performed blood tests, including the serum sex hormone levels, and conventional and computer-assisted semen analyses. The sperm nuclear vacuolization (SNV) was evaluated using a high-magnification microscope. The SDF was evaluated using the sperm chromatin dispersion test (SCDt) to determine the SDF index (SDFI). The SDFI was compared with semen parameters and other clinical variables, including lifestyle factors. Results. The SDFI was 41.3 ± 22.2% (mean ± standard deviation) and did not depend on the cause of infertility. Chronic alcohol use increased the SDFI to 49.6 ± 23.3% compared with 33.9 ± 18.0% in nondrinkers. The SDFI was related to adverse conventional semen parameters and sperm motion characteristics and correlated with the serum FSH level. The SNV showed a tendency to increase with the SDFI. The multivariate analysis revealed that the sperm progressive motility and chronic alcohol use were significant predictors of the SDF. Conclusion. The SCDt should be offered to chronic alcohol users and those with decreased sperm progressive motility. PMID:25165747

  17. Cytochemical evaluation of sperm chromatin and DNA integrity in couples with unexplained recurrent spontaneous abortions.

    PubMed

    Talebi, A R; Vahidi, S; Aflatoonian, A; Ghasemi, N; Ghasemzadeh, J; Firoozabadi, R D; Moein, M R

    2012-05-01

    The aim of this study was to examine the possible relationship between sperm DNA integrity and chromatin packaging evaluated by cytochemical assays, traditional sperm parameters and recurrent spontaneous abortion (RSA) of unknown origin. In this cohort study, 40 couples with a history of RSA and 40 couples with proven fertility were considered as case and control groups respectively. The semen samples of all husbands were analysed for sperm parameters and also sperm chromatin and DNA integrity assessed using cytochemical tests including aniline blue (AB), chromomycin A3 (CMA3), toluidine blue (TB), acridine orange (AOT) and nuclear chromatin stability assay. Among different sperm parameters, only slow motility was significantly different between the two groups. In sperm chromatin evaluations, there were significant differences between the two groups in all of the tests. In addition, the majority of semen samples in RSA patients exhibited upper percentages of abnormal spermatozoa than the cut-off values regarding different cytochemical assays. Our study showed that in the cases of RSA, slow motility had a significant reduction in comparison with controls and also spermatozoa of men from RSA group had less chromatin condensation and poorer DNA integrity than spermatozoa that obtained from fertile men with no history of RSA. PMID:21806662

  18. 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

  19. Inter-rater reliability of the functional standing test

    Microsoft Academic Search

    R. J. Triolo; G. Eisenhower; T. Stabinski; D. Wormser; R. L. Craik

    1994-01-01

    The functional standing test (FST) has been proposed as an evaluation tool to assess the ability to perform one-handed reaching tasks while standing. It is composed of eighteen timed subtests involving manipulation of objects on countertop and simulated kitchen shelves. This study assessed the inter-rater reliability of the FST when administered to adolescents with spinal cord injuries (SCI) standing with

  20. Panning for sperm gold: Isolation and purification of apyrene and eupyrene sperm from lepidopterans.

    PubMed

    Karr, Timothy L; Walters, James R

    2015-08-01

    We describe a simple and straightforward procedure for the purification and separation of apyrene and eupyrene forms of lepidopteran sperm. The procedure is generally applicable to both butterfly and moth species with results varying according to the relative amounts of sperm produced and size of sperm storage organs. The technique relies upon inherent differences between eupyene sperm bundles and free apyrene sperm morphology. These differences allow for separation of the sperm morphs by repeated "panning" of sperm bundles into the center of a plastic dish. The purified eupyrene sperm bundles can then be removed and apyrene sperm collected from the supernatant by centrifugation. Efficacy of the purification process was confirmed by light microscopy and gel electrophoresis of the resulting fractions. Both one- and two-dimensional gel electrophoresis identified significant protein differences between the fractions further suggesting that the panning procedure effectively separated eurpyrene from apyrene sperm. The panning procedure should provide a convenient and accessible technique for further studies of sperm biology in lepidopterans. PMID:26141489

  1. Sperm proteomics reveals intensified selection on mouse sperm membrane and acrosome genes.

    PubMed

    Dorus, Steve; Wasbrough, Elizabeth R; Busby, Jennifer; Wilkin, Elaine C; Karr, Timothy L

    2010-06-01

    Spermatozoa are a focal point for the impact of sexual selection due to sperm competition and sperm-female interactions in a wide range of sexually reproducing organisms. In-depth molecular investigation of the ramifications of these selective regimes has been limited due to a lack of information concerning the molecular composition of sperm. In this study, we utilize three previously published proteomic data sets in conjunction with our whole mouse sperm proteomic analysis to delineate cellular regions of sperm most impacted by positive selection. Interspecific analysis reveals robust evolutionary acceleration of sperm cell membrane genes (which include genes encoding acrosomal and sperm cell surface proteins) relative to other sperm genes, and evidence for positive selection in approximately 22% of sperm cell membrane components was obtained using maximum likelihood models. The selective forces driving the accelerated evolution of these membrane proteins may occur at a number of locations during sperm development, maturation, and transit through the female reproductive tract where the sperm cell membrane and eventually the acrosome are exposed to the extracellular milieu and available for direct cell-cell interactions. PMID:20080865

  2. A Comparative Analysis of the Morphology and Evolution of Permanent Sperm Depletion in Spiders

    PubMed Central

    2011-01-01

    Once thought to be energetically cheap and easy to produce, empirical work has shown that sperm is a costly and limited resource for males. In some spider species, there is behavioral evidence that sperm are permanently depleted after a single mating. This extreme degree of mating investment appears to co-occur with other reproductive strategies common to spiders, e.g. genital mutilation and sexual cannibalism. Here we corroborate that sperm depletion in the golden orb-web spider Nephila clavipes is permanent by uncovering its mechanistic basis using light and electron microscopy. In addition, we use a phylogeny-based statistical analysis to test the evolutionary relationships between permanent sperm depletion (PSD) and other reproductive strategies in spiders. Male testes do not produce sperm during adulthood, which is unusual in spiders. Instead, spermatogenesis is nearly synchronous and ends before the maturation molt. Testis size decreases as males approach their maturation molt and reaches its lowest point after sperm is transferred into the male copulatory organs (pedipalps). As a consequence, the amount of sperm available to males for mating is limited to the sperm contained in the pedipalps, and once it is used, males lose their ability to fertilize eggs. Our data suggest that PSD has evolved independently at least three times within web-building spiders and is significantly correlated with the evolution of other mating strategies that limit males to monogamy, including genital mutilation and sexual cannibalism. We conclude that PSD may be an energy-saving adaptation in species where males are limited to monogamy. This could be particularly important in web-building spiders where extreme sexual size dimorphism results in large, sedentary females and small, searching males who rarely feed as adults and are vulnerable to starvation. Future work will explore possible energetic benefits and the evolutionary lability of PSD relative to other mate-limiting reproductive behaviors. PMID:21264312

  3. Examining the Relationship between Differential Item Functioning and Differential Test Functioning

    ERIC Educational Resources Information Center

    Pae, Tae-Il; Park, Gi-Pyo

    2006-01-01

    The present study utilized both the IRT-LR (item response theory likelihood ratio) and a series of CFA (confirmatory factor analysis) multi-sample analyses to systematically examine the relationships between DIF (differential item functioning) and DTF (differential test functioning) with a random sample of 15 000 Korean examinees. Specifically,…

  4. Control of the low voltage-activated calcium channel of mouse sperm by egg ZP3 and by membrane hyperpolarization during capacitation

    PubMed Central

    Arnoult, Christophe; Kazam, Imrana G.; Visconti, Pablo E.; Kopf, Gregory S.; Villaz, Michel; Florman, Harvey M.

    1999-01-01

    Sperm adhesion to egg zonae pellucidae initiates sperm acrosome reactions, an exocytotic event that is an early step during fertilization. Previously, it was suggested that zona pellucida-evoked Ca2+ entry into sperm through low voltage-activated Ca2+ channels is an essential step in acrosome reactions, based on the inhibitory effects of Ca2+ channel antagonists. However, analysis of this channel is limited by the inability to apply electrophysiological methods directly to sperm. In this report, optical methods of determining membrane potential and internal Ca2+ levels were used to demonstrate that (i) contact with zonae pellucidae activates a transient Ca2+ response in sperm that has a time course and antagonist sensitivity anticipated of low voltage-activated Ca2+ channels; (ii) these channels are unavailable for opening in uncapacitated sperm because of voltage-dependent, steady state inactivation; (iii) membrane hyperpolarization during sperm capacitation is sufficient to recruit channels into a closed state, from which they are available for opening during fertilization; and (iv) channel conductance state may be a factor in determines the efficacy with which channel antagonists inhibit fertilization. This study provides evidence for the activation of sperm Ca2+ channels during gamete adhesion and offers a mechanism that may account for aspects of the regulation of sperm fertility during capacitation through the control of channel availability. Finally, these results suggest that channel conductance state may be a central feature in the design of channel antagonists that inhibit sperm function. PMID:10359785

  5. Advanced Techniques in Pulmonary Function Test Analysis Interpretation and Diagnosis

    PubMed Central

    Gildea, T.J.; Bell, C. William

    1980-01-01

    The Pulmonary Functions Analysis and Diagnostic System is an advanced clinical processing system developed for use at the Pulmonary Division, Department of Medicine at the University of Nebraska Medical Center. The system generates comparative results and diagnostic impressions for a variety of routine and specialized pulmonary functions test data. Routine evaluation deals with static lung volumes, breathing mechanics, diffusing capacity, and blood gases while specialized tests include lung compliance studies, small airways dysfunction studies and dead space to tidal volume ratios. Output includes tabular results of normal vs. observed values, clinical impressions and commentary and, where indicated, a diagnostic impression. A number of pulmonary physiological and state variables are entered or sampled (A to D) with periodic status reports generated for the test supervisor. Among the various physiological variables sampled are respiratory frequency, minute ventilation, oxygen consumption, carbon dioxide production, and arterial oxygen saturation.

  6. Can platelet function tests predict the clinical efficacy of aspirin?

    PubMed

    Haubelt, Hannelore; Anders, Christof; Hellstern, Peter

    2005-01-01

    "Aspirin resistance" and "aspirin nonresponsiveness" are terms used both to describe both the failure of aspirin to protect subgroups of individuals from severe vascular events and to evoke an appropriate inhibition of platelet function. Several studies utilizing a broad range of platelet function tests have shown that some subgroups of individuals exhibit a reduced or completely missing antiplatelet response to aspirin. The clinical significance of aspirin nonresponsiveness for the prediction of clinical endpoints remains, however, to be determined. Thus far, only three prospective clinical trials have demonstrated a possible relationship between aspirin nonresponsiveness and subsequent vascular events. Most platelet function tests used in respective clinical studies cannot be reliably performed in clinical routine and are not interchangeable for monitoring antiplatelet treatment. There is a need for a simple and reliable assay for predicting the clinical efficacy of antiplatelet therapy. Recent data demonstrate that none of the currently developed assays, including the PFA-100 system, are presently able to accomplish these objectives. PMID:16149017

  7. A derived transfer of functions and the implicit association test.

    PubMed

    O'Toole, Catriona; Barnes-Holmes, Dermot; Smyth, Sinead

    2007-09-01

    Participants were trained in a series of interrelated conditional discriminations that aimed to establish four 4-member equivalence classes (i.e., A1-B1-C1-D1, A2-B2-C2-D2, A3-B3-C3-D3, A4-B4-C4-D4). During this training, the four A stimuli (i.e., A1, A2, A3, and A4) were compounded with pictures containing positive or negative evaluative functions (A1/A2 negative & A3/A4 positive). The transfer of evaluative functions to directly and indirectly related members of the equivalence classes (i.e., B, C, and D stimuli) was measured using an Implicit Association Test (IAT). During consistent test blocks, participants were required to press the same response key for target words that were related to those A stimuli that possessed similar evaluative functions (A1/A2-left key & A3/A4-right key). During inconsistent test blocks, target words that were related to those A stimuli with different evaluative functions were assigned to the same response key (A1/A4-left key & A2/A3-right key). Results showed that all 8 participants, who passed a matching-to-sample equivalence test following the IAT, responded more rapidly on consistent relative to inconsistent test blocks. This typical IAT effect was not observed for those participants who did not pass the equivalence test. The results suggest that the IAT effect may arise from formally untested derived relations, and supports the argument that such relations could provide a valid behavioral model of semantic categories in natural language. PMID:17970419

  8. 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

  9. Evaluation of a bedside test of utricular function – the bucket test – in older individuals

    PubMed Central

    SUN, DANIEL Q.; ZUNIGA, M. GERALDINE; DAVALOS-BICHARA, MARCELA; CAREY, JOHN P.; AGRAWAL, YURI

    2015-01-01

    Conclusion As a bedside test of subjective visual vertical (SVV), the ‘bucket test’ has a role as a viable and cost-effective clinical test of unilateral utricular dysfunction in older individuals. Objective To investigate whether the bucket test as a test of the SVV is a valid bedside test of utricular function in older individuals. Methods This was a diagnostic validation study at a tertiary academic medical center. Vestibular function was evaluated using sound-evoked cervical and tap-evoked ocular vestibular evoked myogenic potential (cVEMP and oVEMP, respectively) asymmetry ratios, the bucket test of SVV, and the Dizziness Handicap Index (DHI), in 51 older individuals aged 70–95 years. Results Bucket test scores are correlated in both magnitude and direction with utricle-selective tap-evoked oVEMP asymmetry ratios, but not with sound-evoked cVEMP asymmetry ratios, which are saccule-selective, or with the DHI. Receiver operating characteristics analysis suggests that the bucket test is more specific than sensitive for utricular dysfunction, and a bucket test SVV score of 2° may maximize diagnostic yield relative to the currently accepted score of 3. PMID:24460151

  10. Segmentation of white rat sperm image

    NASA Astrophysics Data System (ADS)

    Bai, Weiguo; Liu, Jianguo; Chen, Guoyuan

    2011-11-01

    The segmentation of sperm image exerts a profound influence in the analysis of sperm morphology, which plays a significant role in the research of animals' infertility and reproduction. To overcome the microscope image's properties of low contrast and highly polluted noise, and to get better segmentation results of sperm image, this paper presents a multi-scale gradient operator combined with a multi-structuring element for the micro-spermatozoa image of white rat, as the multi-scale gradient operator can smooth the noise of an image, while the multi-structuring element can retain more shape details of the sperms. Then, we use the Otsu method to segment the modified gradient image whose gray scale processed is strong in sperms and weak in the background, converting it into a binary sperm image. As the obtained binary image owns impurities that are not similar with sperms in the shape, we choose a form factor to filter those objects whose form factor value is larger than the select critical value, and retain those objects whose not. And then, we can get the final binary image of the segmented sperms. The experiment shows this method's great advantage in the segmentation of the micro-spermatozoa image.

  11. Geometric Morphometrics of Rodent Sperm Head Shape

    PubMed Central

    Varea Sánchez, María; Bastir, Markus; Roldan, Eduardo R. S.

    2013-01-01

    Mammalian spermatozoa, particularly those of rodent species, are extremely complex cells and differ greatly in form and dimensions. Thus, characterization of sperm size and, particularly, sperm shape represents a major challenge. No consensus exists on a method to objectively assess size and shape of spermatozoa. In this study we apply the principles of geometric morphometrics to analyze rodent sperm head morphology and compare them with two traditional morphometry methods, that is, measurements of linear dimensions and dimensions-derived parameters calculated using formulae employed in sperm morphometry assessments. Our results show that geometric morphometrics clearly identifies shape differences among rodent spermatozoa. It is also capable of discriminating between size and shape and to analyze these two variables separately. Thus, it provides an accurate method to assess sperm head shape. Furthermore, it can identify which sperm morphology traits differ between species, such as the protrusion or retraction of the base of the head, the orientation and relative position of the site of flagellum insertion, the degree of curvature of the hook, and other distinct anatomical features and appendices. We envisage that the use of geometric morphometrics may have a major impact on future studies focused on the characterization of sperm head formation, diversity of sperm head shape among species (and underlying evolutionary forces), the effects of reprotoxicants on changes in cell shape, and phenotyping of genetically-modified individuals. PMID:24312234

  12. Comparative Evolutionary Psychology of Sperm Competition

    Microsoft Academic Search

    Todd K. Shackelford; Aaron T. Goetz

    2006-01-01

    A comparative evolutionary psychological perspective predicts that species that recurrently faced similar adaptive problems may have evolved similar psychological mechanisms to solve these problems. Sperm competition provides an arena in which to assess the heuristic value of such a comparative evolutionary perspective. The sperm competition that results from female infidelity and polyandry presents a similar class of adaptive problems for

  13. Wonders of the Seas: Sperm Whales

    NSDL National Science Digital Library

    Overview of sperm whale biology, adaptations, and behaviors. Basic introduction to anatomy, diet, echolocation, diving, whale oil and other products derived from sperm whales, whaling industry and impacts, and conservation measures. Accessible text, illustrated with excellent photographs. A NSTA SciLinks selected site.

  14. Hydrodynamics of Sperm Cells near Surfaces

    PubMed Central

    Elgeti, Jens; Kaupp, U. Benjamin; Gompper, Gerhard

    2010-01-01

    Sperm are propelled by an actively beating tail, and display a wide variety of swimming patterns. When confined between two parallel walls, sperm swim either in circles or on curvilinear trajectories close to the walls. We employ mesoscale hydrodynamics simulations in combination with a mechanical sperm model to study the swimming behavior near walls. The simulations show that sperm become captured at the wall due to the hydrodynamic flow fields which are generated by the flagellar beat. The circular trajectories are determined by the chiral asymmetry of the sperm shape. For strong (weak) chirality, sperm swim in tight (wide) circles, with the beating plane of the flagellum oriented perpendicular (parallel) to the wall. For comparison, we also perform simulations based on a local anisotropic friction of the flagellum. In this resistive force approximation, surface adhesion and circular swimming patterns are obtained as well. However, the adhesion mechanism is now due to steric repulsion, and the orientation of the beating plane is different. Our model provides a theoretical framework that explains several distinct swimming behaviors of sperm near and far from a wall. Moreover, the model suggests a mechanism by which sperm navigate in a chemical gradient via a change of their shape. PMID:20712984

  15. Operating Characteristics of Executive Functioning Tests Following Traumatic Brain Injury

    PubMed Central

    Demery, Jason A.; Larson, Michael J.; Dixit, Neha K.; Bauerand, Russell M.; Perlstein, William M.

    2010-01-01

    The primary purposes of this study were to determine if controls, mild, and moderate/severe traumatic brain injury (TBI) patients performed differently on a battery of executive functioning (EF) tests, and to identify the operating characteristics of EF tests in this population. Participants consisted of 46 brain injured individuals and 24 healthy controls. All participants completed an extensive battery of EF tests. Results showed that mild TBI participants performed worse than controls on the Trail Making Test Part B, and that moderate/severe TBI participants consistently performed worse than either group on a variety of EF measures. Tests of EF exhibited a wide range of operating characteristics, suggesting that some EF tests are better than others in identifying TBI-related neurocognitive impairment. Predictive values were better for individuals with moderate/severe TBI than mild TBI. Overall, the Digit Span Backward Test showed the best positive predictive power in differentiating TBI. Our results provide useful data that may guide test selection in evaluating EF in patients with traumatic brain injury. PMID:21069617

  16. Sperm ubiquitination in epididymal feline semen.

    PubMed

    Vernocchi, Valentina; Morselli, Maria Giorgia; Varesi, Sara; Nonnis, Simona; Maffioli, Elisa; Negri, Armando; Tedeschi, Gabriella; Luvoni, Gaia Cecilia

    2014-09-01

    Ubiquitin is a 8.5-kDa peptide that tags other proteins for proteasomal degradation. It has been proposed that ubiquitination might be responsible for the elimination of defective spermatozoa during transit through the epididymis in humans and cattle, but its exact biological function in seminal plasma has not yet been clarified. In the domestic cat (Felis catus), the percentage of immature, unviable, and abnormal spermatozoa decreases during the epididymal transit, indicating the existence of a mechanism that removes defective spermatozoa. Magnetic cell separation techniques, based on the use of magnetic beads coated with anti-ubiquitin antibodies, may allow the selective capture of ubiquitinated spermatozoa from semen, thus contributing to the identification of a potential correlation between semen quality and ubiquitination process. Moreover, the selective identification of all the ubiquitinated proteins in different epididymal regions could give a better understanding of the ubiquitin role in feline sperm maturation. The aims of this study were as follows: (1) to verify the possibility of separating ubiquitinated spermatozoa with magnetic ubiquitin beads and identify the morphological and acrosomal differences between whole sample and unbound gametes, (2) to characterize all the ubiquitinated proteins in spermatozoa retrieved in the three epididymal regions by a proteomic approach. The data indicated the presence of ubiquitinated proteins in cat epididymal semen. However, a correlation between abnormal and ubiquitinated spermatozoa has not been found, and ubiquitin cannot be considered as a biomarker of quality of epididymal feline spermatozoa. To the author's knowledge, this is the first identification of all the ubiquitinated proteins of cat spermatozoa collected from different epididymal regions. The proteomic pattern allows a further characterization of cat epididymal semen and represents a contribute to a better understanding of the ubiquitin role in feline sperm maturation. PMID:24999010

  17. Testing the Link between Functional Diversity and Ecosystem Functioning in a Minnesota Grassland Experiment

    PubMed Central

    Butterfield, Bradley J.; Reich, Peter B.

    2012-01-01

    The functional diversity of a community can influence ecosystem functioning and reflects assembly processes. The large number of disparate metrics used to quantify functional diversity reflects the range of attributes underlying this concept, generally summarized as functional richness, functional evenness, and functional divergence. However, in practice, we know very little about which attributes drive which ecosystem functions, due to a lack of field-based tests. Here we test the association between eight leading functional diversity metrics (Rao’s Q, FD, FDis, FEve, FDiv, convex hull volume, and species and functional group richness) that emphasize different attributes of functional diversity, plus 11 extensions of these existing metrics that incorporate heterogeneous species abundances and trait variation. We assess the relationships among these metrics and compare their performances for predicting three key ecosystem functions (above- and belowground biomass and light capture) within a long-term grassland biodiversity experiment. Many metrics were highly correlated, although unique information was captured in FEve, FDiv, and dendrogram-based measures (FD) that were adjusted by abundance. FD adjusted by abundance outperformed all other metrics in predicting both above- and belowground biomass, although several others also performed well (e.g. Rao’s Q, FDis, FDiv). More generally, trait-based richness metrics and hybrid metrics incorporating multiple diversity attributes outperformed evenness metrics and single-attribute metrics, results that were not changed when combinations of metrics were explored. For light capture, species richness alone was the best predictor, suggesting that traits for canopy architecture would be necessary to improve predictions. Our study provides a comprehensive test linking different attributes of functional diversity with ecosystem function for a grassland system. PMID:23300787

  18. How should we test for nonsevere heritable platelet function disorders?

    PubMed

    Norman, J E; Westbury, S K; Jones, M L; Mumford, A D

    2014-06-01

    Heritable platelet function disorders (HPFD) are a heterogeneous group of bleeding disorders with diverse clinical and laboratory characteristics. In contrast to the severe phenotype disorders, Glanzmann thrombasthenia and Bernard-Soulier syndrome, most nonsevere HPFD are incompletely characterized. This is a consequence of the poor standardization of diagnostic tests and of the lack of consensus about diagnostic criteria for the different subgroups of nonsevere HPFD. Distinguishing patients who have a nonsevere HPFD from those who do not is an essential first step in diagnosis which may be aided by bleeding assessment tools and screening tests such as the Platelet Function Analyser-100. However, high diagnostic accuracy can only be achieved with both light transmission aggregation (LTA) and secretion tests, for which streamlined agonist panels may be of similar utility to extended panels. Standardization of the methodology of these tests and quality assurance are essential for robust diagnosis. Identification of which platelet pathway is defective in patients with nonsevere HPFD is also usually possible with LTA and secretion tests. This strategy also sometimes enables exact diagnosis by implicating a single candidate protein and gene. Next-generation sequencing may offer a rapid approach to diagnosis of nonsevere HPFD, although rigorous strategies must be adopted to distinguish causative gene defects from bystander variations. PMID:24750679

  19. 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

  20. Response of isolated sperm plasma membranes from sea urchin to egg jelly.

    PubMed

    Darszon, A; Gould, M; De De La Torre, L; Vargas, I

    1984-11-01

    The acrosome reaction in sea urchin sperm is induced by a glycoprotein jelly surrounding the egg and is accompanied by changes in ion permeability of sperm plasma membrane. In an attempt to learn what membrane components are involved in the response to jelly, we have begun to reassemble sperm membrane components into artificial membranes and assay for permeability changes mimicking those that occur in sperm. Jelly in sea water at concentrations that induce the acrosome reaction did not significantly change 45Ca2+ uptake of sonicated unilamellar vesicles made with soybean lipid only (ratio jelly:control uptake = 1.08 +/- 0.36 SD, n = 21). Experiments with pure lipid planar bilayers made with soybean lipid or a lipid extract from sperm and held at various voltages, also did not reveal substantial permeability changes at comparable jelly concentrations. Thus, jelly by itself does not change the conductance of a pure lipid bilayer. In contrast, significant (P----0.0005, t test for two sample means) 45Ca2+ uptake was observed with vesicles made by cosonicating soybean phospholipids and Strongylocentrotus purpuratus sperm membranes isolated by the method of Cross, N. L. [1983, J. Cell Sci. 59, 13-25] (ratio jelly: control uptake = 1.51 +/- 0.75, n = 20, 16 positive out of 20 experiments). The calcium uptake response of the mixed vesicles was also species-specific: it did not occur with jelly from Arbacia punctulata (ratio Arbacia jelly: control = 1.18 +/- 0.51; ratio Strongylocentrotus jelly: control = 1.71 +/- 0.97, n = 10; P----0.025, paired t statistic). Vesicles made with soybean lipid and an octyl glucoside extract of sperm membranes also responded to jelly with increased 45Ca2+ uptake. Our results indicate that we have the starting conditions to isolate and characterize the sperm membrane components that participate in the egg jelly induced permeability changes. PMID:6436025

  1. Effect of different extenders and storage temperatures on sperm viability of liquid ram semen.

    PubMed

    Paulenz, Heiko; Söderquist, Lennart; Pérez-Pé, Rosaura; Berg, Kjell Andersen

    2002-01-15

    Semen was collected with an artificial vagina from four adult rams. The ejaculates were pooled and diluted, using a split-sample technique, in four different extenders: one for milk (Mi), one for sodium citrate (Na), and two for Tris-based extenders (T1 and T2) including egg yolk. Thereafter, the diluted semen was stored at 5 and 20 degrees C, 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-osmotic resistance test (ORT) and a fluorophore staining (SYBR-14 and propidium iodide) technique. We evaluated acrosomal status with Spermac and capacitation status with Chlortetracycline (CTC assay). All sperm viability parameters were influenced by storage time and extender, while sperm motility was the only evaluated parameter that was influenced by the interaction between extender and temperature. Semen that was diluted and stored in the commercially available Tris-based extender (T2) maintained sperm motility for a longer period of time, and acrosome and membrane integrity was higher during storage for up to 30 h as compared to the other extenders independent of storage temperature. In general, however, storage of ram semen at 5 degrees C seemed to influence sperm viability parameters less than storage at 20 degrees C. In conclusion, the results of the present study indicate that Tris-based extenders, especially T2, preserved sperm viability better than both the sodium citrate- and the milk-based extender did when liquid ram semen was stored up to 30 h at 5 and 20 degrees C. Whether the differences found between the extenders will be reflected in the fertility results after AI is yet unknown and needs to be further studied. PMID:11991386

  2. Boar sperm storage capacity of BTS and Androhep Plus: viability, motility, capacitation, and tyrosine phosphorylation.

    PubMed

    Dubé, Charlotte; Beaulieu, Martin; Reyes-Moreno, Carlos; Guillemette, Christine; Bailey, Janice L

    2004-09-01

    Androhep Plus, a long-term extender (up to 7 days) and Beltsville Thawing Solution (BTS), a short-term extender (up to 3 days), are commonly used for liquid storage of porcine semen. To test the hypothesis that modifications in sperm viability, motility, chlortetracycline (CTC) fluorescence patterns, and protein tyrosine phosphorylation occur during semen storage in extenders, we compared these end points at different periods of storage in either Androhep Plus or BTS. Sperm from five boars were assessed daily over 12 days of storage (n = 5 ejaculates from different boars). Viability was not different (P < 0.05 between extenders, except on Day 2, when Androhep Plus maintained better viability. Differences in the percentage of motile (total) sperm due to extender were evident on Days 2, 4, 5, and 6, when Androhep Plus was superior to BTS (P < 0.05). The percentages of progressively motile sperm also differed, with Androhep Plus supporting higher rates on Days 2, 4, 5, 7, 8, 9, 10, and 11 (P < 0.05). The CTC fluorescence pattern distribution differed due to extender as early as Day 2; storage in Androhep Plus induced higher levels of pattern B sperm (P < 0.05) than storage in BTS. A tyrosine-phosphorylated protein of Mr 21,000 appeared after 10 days in sperm incubated in BTS, and was identified as a phospholipid hydroperoxide glutathione peroxidase. Therefore, modifications in viability, motility, CTC fluorescence patterns, and sperm protein tyrosine phosphorylation were apparent during sperm storage in extenders; these may affect the fertilizing capacity of the semen. PMID:15251239

  3. Bicarbonate stimulated phospholipid scrambling induces cholesterol redistribution and enables cholesterol depletion in the sperm plasma membrane.

    PubMed

    Flesch, F M; Brouwers, J F; Nievelstein, P F; Verkleij, A J; van Golde, L M; Colenbrander, B; Gadella, B M

    2001-10-01

    Mammalian sperm cells are activated prior to fertilization by high bicarbonate levels, which facilitate lipoprotein-mediated cholesterol efflux. The role of bicarbonate and cholesterol acceptors on the cholesterol organization in the sperm plasma membrane was tested. Bicarbonate induced an albumin-independent change in lipid architecture that was detectable by an increase in merocyanine staining (due to protein kinase A-mediated phospholipid scrambling). The response was limited to a subpopulation of viable sperm cells that were sorted from the non-responding subpopulation by flow cytometry. The responding cells had reduced cholesterol levels (30% reduction) compared with non-responding cells. The subpopulation differences were caused by variable efficiencies in epididymal maturation as judged by cell morphology. Membrane cholesterol organization was observed with filipin, which labeled the entire sperm surface of non-stimulated and non-responding cells, but labeled only the apical surface area of bicarbonate-responding cells. Addition of albumin caused cholesterol efflux, but only in bicarbonate-responding cells that exhibited virtually no filipin labeling in the sperm head area. Albumin had no effect on other lipid components, and no affinity for cholesterol in the absence of bicarbonate. Therefore, bicarbonate induces first a lateral redistribution in the low cholesterol containing spermatozoa, which in turn facilitates cholesterol extraction by albumin. A model is proposed in which phospholipid scrambling induces the formation of an apical membrane raft in the sperm head surface that enables albumin mediated efflux of cholesterol. PMID:11682613

  4. 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

  5. SEPT12 mutations cause male infertility with defective sperm annulus.

    PubMed

    Kuo, Yung-Che; Lin, Ying-Hung; Chen, Hau-Inh; Wang, Ya-Yun; Chiou, Yu-Wei; Lin, Hsi-Hui; Pan, Hsien-An; Wu, Ching-Ming; Su, Shih-Ming; Hsu, Chao-Chin; Kuo, Pao-Lin

    2012-04-01

    Septins are members of the GTPase superfamily, which has been implicated in diverse cellular functions including cytokinesis and morphogenesis. Septin 12 (SEPT12) is a testis-specific gene critical for the terminal differentiation of male germ cells. We report the identification of two missense SEPT12 mutations, c.266C>T/p.Thr89Met and c.589G>A/p.Asp197Asn, in infertile men. Both mutations are located inside the GTPase domain and may alter the protein structure as suggested by in silico modeling. The p.Thr89Met mutation significantly reduced guanosine-5'-triphosphate (GTP) hydrolytic activity, and the p.Asp197Asn mutation (SEPT12(D197N)) interfered with GTP binding. Both mutant SEPT12 proteins restricted the filament formation of the wild-type SEPT12 in a dose-dependent manner. The patient carrying SEPT12(D197N) presented with oligoasthenozoospermia, whereas the SEPT12(T89M) patient had asthenoteratozoospermia. The characteristic sperm pathology of the SEPT12(D197N) patient included defective annulus with bent tail and loss of SEPT12 from the annulus of abnormal sperm. Our finding suggests loss-of-function mutations in SEPT12 disrupted sperm structural integrity by perturbing septin filament formation. PMID:22275165

  6. Cytoplasmic incompatibility and sperm cyst infection in different Drosophila-Wolbachia associations.

    PubMed

    Veneti, Zoe; Clark, Michael E; Zabalou, Sofia; Karr, Timothy L; Savakis, Charalambos; Bourtzis, Kostas

    2003-06-01

    Wolbachia are a group of maternally transmitted obligatory intracellular alpha-proteobacteria that infect a wide range of arthropod and nematode species. Wolbachia infection in Drosophila in most cases is associated with the induction of cytoplasmic incompatibility (CI), manifested as embryonic lethality of offspring in a cross between infected males and uninfected females. While the molecular basis of CI is still unknown, it has been suggested that two bacterial functions are involved: mod (for modification) modifies the sperm during spermatogenesis and resc (for rescue) acts in the female germline and/or in early embryos, neutralizing the modification. There is considerable variation in the level of incompatibility in different Wolbachia/host interactions. We examine the relationship between the levels of CI in a number of naturally infected and transinfected Drosophila hosts and the percentage of Wolbachia-infected sperm cysts. Our results indicate the presence of two main groups of Drosophila-Wolbachia associations: group I, which exhibits a positive correlation between CI levels and the percentage of infected sperm cysts (mod(+) phenotype), and group II, which does not express CI (mod(-) phenotype) irrespective of the infection status of the sperm cysts. Group II can be further divided into two subgroups: The first one contains associations with high numbers of heavily Wolbachia-infected sperm cysts while in the second one, Wolbachia is rarely detected in sperm cysts, being mostly present in somatic cells. We conclude that there are three requirements for the expression of CI in a host-Wolbachia association: (a) Wolbachia has to be able to modify sperm (mod(+) genotype), (b) Wolbachia has to infect sperm cysts, and (c) Wolbachia has to be harbored by a permissive host. PMID:12807775

  7. Effects of repeated electroejaculations on kinematic sperm subpopulations and quality markers of Mexican creole goats.

    PubMed

    Vázquez, A J F; Cedillo, M J; Quezada, V J; Rivas, A C; Morales, E C L; Ayala, E M E; Hernández, M J; González, R A; Aragón, M A

    2015-03-01

    Here we show the effects of repeated electroejaculation (EE) on mean values of motility, mitochondrial functionality, and expression of active caspases on goat sperm obtained by EE. Evaluations were done using CASA and flow cytometry. A strategy for identification of kinematic sperm subpopulations, when individual data of sperm are not provided by the CASA system, is provided. Fifty semen samples, five of each of ten adult creole goats, were obtained by electroejaculation. Mean values of total motility, progressive motility and flow cytometry evaluations were compared among EEs. Relationships among mean values of variables were investigated using Spearman correlation and principal component analysis (PCA). For identification of kinematic sperm subpopulations, PCA followed by hierarchical clustering was applied on data of the intervals provided automatically by the CASA system. Total motility does no change after repeated EE. Mean values of motility parameters and molecular markers were unrelated in multivariate space, but bivariate correlations were found. Values in upper and lower intervals defined clearly the sperm subpopulations, which had motility parameters changing over time. Taken together, our results show that repeated EE does not affect mean values of total motility, that molecular markers are not related with motility parameters, and that it is possible to identify kinematic sperm subpopulations when individual data, of motility parameters, are not provided by the CASA system. PMID:25600145

  8. Fertilization induced changes in sea urchin sperm: mitochondrial deformation and phosphatidylserine exposure.

    PubMed

    Kazama, Makoto; Asami, Kouichi; Hino, Akiya

    2006-10-01

    This study demonstrates that the single mitochondrion of the sea urchin sperm undergoes a shape change at fertilization that is linked to respiration. The mitochondrion swells and shifts to the lateral side of the sperm head on contact with the homologous egg jelly or egg surface; Mg(2+)- or Na(+)-free seawater or respiratory inhibitors also induce this change. During the mitochondrial deformation, the sperm decreases the rate of oxygen consumption and their redox-state of cytochromes is disrupted b-c(1)/c. Simultaneously, the adenine nucleotides content changes precipitously. This suggests that mitochondrial morphology is strongly associated with respiratory activities in the sea urchin sperm. These changes in mitochondrial morphology and function are similar to the mitochondrial changes in apoptotic cells such as swelling, decrease in its membrane potential, and release of cytochrome c. In apoptotic cells, the exposure of phosphatidylserine from the inner to outer leaflet of the plasma membrane is one of prominence phenomena. This change was visualized by staining the sea urchin sperm with Annexin V-Fluorescein. It is possible that mitochondrial deformation is an initial sign of sperm destruction, which like as apoptotic cells. PMID:16865719

  9. SNAREs in Mammalian Sperm: Possible Implications for Fertilization

    E-print Network

    Wessel, Gary M.

    sperm (human, rhesus monkey, bull, hamster, mouse). Sperm SNAREs are sloughed off during the acrosomeSNAREs 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

  10. Effect of method and clinician on stallion sperm morphology evaluation

    E-print Network

    Watson, Craig A.

    , the World Health Organization (WHO) recommends the use of Papanicolaou stain for evaluation of human sperm morphology [3]. Several studies from laboratories evaluating human sperm described variations in spermEffect of method and clinician on stallion sperm morphology evaluation Leonardo F.C. Brito ,1

  11. Treating ram sperm with cholesterol-loaded cyclodextrins improves cryosurvival

    Microsoft Academic Search

    Eva Mocé; Phillip H. Purdy; James K. Graham

    2010-01-01

    Acceptable fertility using cryopreserved ram sperm is currently only achieved using laparoscopic intrauterine insemination. Improving the cryosurvival of ram sperm may permit greater fertility rates using more practical techniques. This study was conducted to determine if treating ram sperm with six different cyclodextrins pre-loaded with cholesterol (CLC), prior to cryopreservation increases sperm cryosurvival and if this technology can be used

  12. 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...

  13. Boar sperm quality in lines of pigs selected for either ovulation rate or uterine capacity.

    PubMed

    Freking, B A; Purdy, P H; Spiller, S F; Welsh, C S; Blackburn, H D

    2012-08-01

    Selection for 11 generations in swine for ovulation rate (OR) or uterine capacity (UC) resulted in significant changes in component traits of litter size. Our objective was to conserve the unique germplasm for the future and to characterize sperm quality as a correlated response to the selection criterion imposed compared with an unselected control line (CO). Boars representing genetic diversity available in all 3 lines were produced in 2 farrowing seasons. Season 1 was born in September 2005 and was sampled for semen characteristics in October 2006. Season 2 was born in March 2006 and was sampled for semen characteristics in February and March 2007. Each boar (n = 60) was collected twice. The sperm-rich fraction was obtained, and volume and concentration of sperm cells were measured to estimate total sperm production. Each ejaculate was extended 1:3 (vol/vol) with Androhep Plus (Minitube, Verona, WI) and was packed for shipping to the National Animal Germplasm Program laboratory for processing into frozen straws. Semen quality was measured by computer-assisted semen analysis at 3 semen processing points: fresh (FR), 24 h after extender added (E), and postthaw (PT). A mixed model ANOVA was applied to the data. Fixed effects of farrowing season, line, and 2-way interactions were fitted. The random effect of boar (n = 60) within farrowing season and line was used to test line differences. Sperm concentration was not different (P = 0.18) among the lines (0.594 × 10(9), 0.691 × 10(9), and 0.676 × 10(9) cells/mL for CO, OR, and UC lines, respectively). However, significance (P = 0.04) was detected for the volume of the sperm-rich fraction, greatest for OR (86.4 mL), intermediate for UC (75.5 mL), and least for CO (70.2 mL). Line differences were thus detected (P = 0.02) for total sperm production per ejaculate, greatest for OR (54.9 × 10(9)), intermediate for UC (48.7 × 10(9)), and least for CO (40.5 × 10(9)). A larger percentage of progressively motile sperm and greater estimates of sperm velocity only at processing point E (P < 0.01) were detected in favor of CO. Estimates of motility, velocity, and other parameters of sperm movement measured on E processing points were positively correlated with the same estimates obtained PT, but the magnitude was low to moderate (r range -0.03 to 0.23). Thus, selection for component traits of female reproduction had a favorable effect on total sperm production of boars. PMID:22896731

  14. Boar sperm quality in lines of pigs selected for either ovulation rate or uterine capacity.

    PubMed

    Freking, B A; Purdy, P H; Spiller, S F; Welsh, C S; Blackburn, H D

    2012-02-10

    Selection for 11 generations in swine for ovulation rate (OR) or uterine capacity (UC) resulted in significant changes in component traits of litter size. Our objective was to conserve the unique germplasm for the future and to characterize sperm quality as a correlated response to the selection criterion imposed compared to an unselected control line (CO). Boars representing genetic diversity available in all 3 lines were produced in 2 farrowing seasons. Season 1 was born in September 2005 and sampled for semen characteristics in October 2006. Season 2 was born in March 2006 and sampled for semen characteristics in February and March 2007. Each boar (n = 60) was collected twice. The sperm-rich fraction was obtained and volume and concentration of sperm cells were measured to estimate total sperm production. Each ejaculate was extended 1:3 vol/vol with Androhep Plus (Minitube, Verona, WI) and packed for shipping to the National Animal Germplasm Program laboratory for processing into frozen straws. Semen quality was measured by computer-assisted semen analysis at 3 semen processing points: fresh (FR), 24 h after extender added (E), and post-thaw (PT). A mixed model analysis of variance was applied to the data. Fixed effects of farrowing season, line and 2-way interactions were fitted. Random effect of boar (n = 60) within farrowing season and line was used to test line differences. Sperm concentration was not different (P = 0.18) among the lines (0.594 × 10(9), 0.691 × 10(9), and 0.676 × 10(9) cells/mL) for CO, OR, and UC lines, respectively. However, significance (P = 0.04) was detected for volume of the sperm-rich fraction, greatest for OR (86.4 mL), intermediate for UC (75.5 mL), and least for CO (70.2 mL). Line differences were thus detected (P = 0.02) for total sperm production per ejaculate, greatest for OR (54.9 × 10(9)), intermediate for UC (48.7 × 10(9)), and least for CO (40.5 × 10(9)). A larger percentage of progressively motile sperm and greater estimates of sperm velocity only at processing point E (P < 0.01) were detected in favor of CO. Estimates of motility, velocity, and other parameters of sperm movement measured on E processing points were positively correlated with the same estimates obtained PT but the magnitude was low to moderate (r range -0.03 to 0.23). Thus, selection for component traits of female reproduction had a favorable effect on total sperm production of boars. PMID:22328726

  15. Estimating and testing of functional data with restrictions 

    E-print Network

    Lee, Sang Han

    2009-05-15

    INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1.1 Motivation and problem . . . . . . . . . . . . . . . . . . . 1 1.2 Pre-processing of the data . . . . . . . . . . . . . . . . . . 3 II FUNCTIONAL ANALYSIS ON PANIC DATA . . . . . . . . . . 7... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 13 Paired NL-SL, extracted trend: Pointwise t-test on differences . . . . 22 14 Mean curves for N+L (line) and S+L (dotted line) after pre- processing. The left panel is for traces after subtraction of com- ponents at 4sec and less, the right panel...

  16. 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 ...

  17. Pheochromocytoma with Markedly Abnormal Liver Function Tests and Severe Leukocytosis

    PubMed Central

    Eun, Chai Ryoung; Ahn, Jae Hee; Seo, Ji A

    2014-01-01

    Pheochromocytoma is a rare neuroendocrine tumor arising from the medulla of the adrenal glands, which causes an overproduction of catecholamines. The common symptoms are headache, palpitations, and sweating; however, various other clinical manifestations might also be present. Accurate diagnosis of pheochromocytoma is important because surgical treatment is usually successful, and associated clinical problems are reversible if treated early. A 49-year-old man with a history of uncontrolled hypertension and diabetes mellitus presented with chest pain, fever, and sweating. His liver function tests and white blood cell counts were markedly increased and his echocardiography results suggested stress-induced cardiomyopathy. His abdominal computed tomography showed a 5×5-cm-sized tumor in the left adrenal gland, and laboratory tests confirmed catecholamine overproduction. After surgical resection of the left adrenal gland, his liver function tests and white blood cell counts normalized, and echocardiography showed normal cardiac function. Moreover, his previous antihypertensive regimen was deescalated, and his previously uncontrolled blood glucose levels normalized without medication. PMID:24741459

  18. Reproductive functions of the ageing male

    Microsoft Academic Search

    Bianca Kuhnert; Eberhard Nieschlag

    2004-01-01

    Delayed childbearing is a common phenomenon in industrialized countries. This review focuses on age-associated alterations of male fertility and genetic risks. Semen volume, sperm motility and sperm morphology decrease with age, whereas the data concerning sperm concentration are conflicting. The age-related changes of semen parameters reflect the histological modifications which are found to varying degrees in individual testes. Men aged

  19. SLC6 family transporter SNF-10 is required for protease-mediated activation of sperm motility in C. elegans.

    PubMed

    Fenker, Kristin E; Hansen, Angela A; Chong, Conrad A; Jud, Molly C; Duffy, Brittany A; Norton, J Paul; Hansen, Jody M; Stanfield, Gillian M

    2014-09-01

    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 ameboid sperm of Caenorhabditis 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

  20. Lung function tests in clinical decision-making.

    PubMed

    Puente Maestú, Luis; García de Pedro, Julia

    2012-05-01

    In this article, we review the utility of the most common lung function tests (spirometry, reversibility test, peak expiratory flow, lung volumes, maximal respiratory pressure, carbon monoxide transference, arterial blood gas, 6-minute walk test and desaturation with exercise and ergospirometry) related to the most frequent pathologies (dyspnea of undetermined origin, chronic cough, asthma, COPD, neuromuscular diseases, interstitial diseases, pulmonary vascular diseases, pre-operative evaluation and disability evaluation). Our analysis has been developed from the perspective of decision-making, clinical interpretation or aspects that the physician should take into account with their use. Consequently, the paper does not deal with aspects of quality, technique or equipment, with the exception of when regarding costs as we believe that this is an important element in the decision-making process. The document is extensively supported by references from the literature. PMID:22364671

  1. Hypertonic saline test for the investigation of posterior pituitary function.

    PubMed

    Mohn, A; Acerini, C L; Cheetham, T D; Lightman, S L; Dunger, D B

    1998-11-01

    The hypertonic saline test is a useful technique for distinguishing partial diabetes insipidus from psychogenic polydipsia, and for the diagnosis of complex disorders of osmoreceptor and posterior pituitary function. However, there is little information concerning its use in childhood. The experience of using this test in five children (11 months to 18 years) who presented diagnostic problems is reported. In two patients, in whom water deprivation tests were equivocal or impractical, an inappropriately low antidiuretic hormone (ADH) concentration (< 1 pmol/l) was demonstrated in the presence of an adequate osmotic stimulus (plasma osmolality > 295 mosmol/kg). In two children--one presenting with adipsic hypernatraemia and the other with hyponatraemia complicating desmopressin treatment of partial diabetes insipidus--defects of osmoreceptor function were identified. Confirming a diagnosis of idiopathic syndrome of inappropriate ADH secretion (SIADH) was possible in a patient with no other evidence of pituitary dysfunction. The hypertonic saline test was well tolerated, easy to perform, and diagnostic in all cases. PMID:10193258

  2. Hypertonic saline test for the investigation of posterior pituitary function

    PubMed Central

    Mohn, A.; Acerini, C.; Cheetham, T.; Lightman, S.; Dunger, D.

    1998-01-01

    The hypertonic saline test is a useful technique for distinguishing partial diabetes insipidus from psychogenic polydipsia, and for the diagnosis of complex disorders of osmoreceptor and posterior pituitary function. However, there is little information concerning its use in childhood. The experience of using this test in five children (11 months to 18 years) who presented diagnostic problems is reported. In two patients, in whom water deprivation tests were equivocal or impractical, an inappropriately low antidiuretic hormone (ADH) concentration (< 1 pmol/l) was demonstrated in the presence of an adequate osmotic stimulus (plasma osmolality > 295 mosmol/kg). In two children—one presenting with adipsic hypernatraemia and the other with hyponatraemia complicating desmopressin treatment of partial diabetes insipidus—defects of osmoreceptor function were identified. Confirming a diagnosis of idiopathic syndrome of inappropriate ADH secretion (SIADH) was possible in a patient with no other evidence of pituitary dysfunction. The hypertonic saline test was well tolerated, easy to perform, and diagnostic in all cases.?? PMID:10193258

  3. Towards a statistical test for functional connectivity dynamics.

    PubMed

    Zalesky, Andrew; Breakspear, Michael

    2015-07-01

    Sliding-window correlation is an emerging method for mapping time-resolved, resting-state functional connectivity. To avoid mapping spurious connectivity fluctuations (false positives), Leonardi and Van De Ville recently recommended choosing a window length exceeding the longest wavelength composing the BOLD signal, usually assumed to be ~100s. Here, we provide further statistical support for this rule of thumb. However, we demonstrate that non-stationary fluctuations in functional connectivity can in theory be detected with much shorter window lengths (e.g. 40s), while maintaining nominal control of false positives. We find that statistical power is near-maximal for window lengths chosen according to Leonardi and Van De Ville's rule of thumb. Furthermore, we lay some foundations for a parametric test to identify non-stationary fluctuations in functional connectivity, also noting limitations of the sinusoidal model upon which our work, and the work of Leonardi and Van De Ville, is based. Most notably, our analytical results pertain to covariances, as does our statistical test, whereas functional connectivity is more commonly measured using correlations. PMID:25818688

  4. Morphometric characterization and classification of alpaca sperm heads using the Sperm-Class Analyzer ® computer-assisted system

    Microsoft Academic Search

    P Buend??a; C Soler; F Paolicchi; G Gago; B Urquieta; F Pérez-Sánchez; E Bustos-Obregón

    2002-01-01

    Sperm morphology has been identified as one characteristic which can be useful in the prediction of sperm fertility, therefore, we hope that this study aimed at establishing standardized morphological criteria might serve in future studies dealing with the search for sperm parameters which facilitate an estimation of sperm quality. For this purpose, ejaculates from fertile alpacas were used to evaluate

  5. Redox regulation of mammalian sperm capacitation.

    PubMed

    O'Flaherty, Cristian

    2015-01-01

    Capacitation is a series of morphological and metabolic changes necessary for the spermatozoon to achieve fertilizing ability. One of the earlier happenings during mammalian sperm capacitation is the production of reactive oxygen species (ROS) that will trigger and regulate a series of events including protein phosphorylation, in a time-dependent fashion. The identity of the sperm oxidase responsible for the production of ROS involved in capacitation is still elusive, and several candidates are discussed in this review. Interestingly, ROS-induced ROS formation has been described during human sperm capacitation. Redox signaling during capacitation is associated with changes in thiol groups of proteins located on the plasma membrane and subcellular compartments of the spermatozoon. Both, oxidation of thiols forming disulfide bridges and the increase on thiol content are necessary to regulate different sperm proteins associated with capacitation. Reducing equivalents such as NADH and NADPH are necessary to support capacitation in many species including humans. Lactate dehydrogenase, glucose-6-phospohate dehydrogenase, and isocitrate dehydrogenase are responsible in supplying NAD (P) H for sperm capacitation. Peroxiredoxins (PRDXs) are newly described enzymes with antioxidant properties that can protect mammalian spermatozoa; however, they are also candidates for assuring the regulation of redox signaling required for sperm capacitation. The dysregulation of PRDXs and of enzymes needed for their reactivation such as thioredoxin/thioredoxin reductase system and glutathione-S-transferases impairs sperm motility, capacitation, and promotes DNA damage in spermatozoa leading to male infertility. PMID:25926608

  6. Comparative profiling of the sperm proteome.

    PubMed

    Holland, Ashling; Ohlendieck, Kay

    2015-02-01

    The highly complex and species-selective mechanism of fertilization is a central theme of developmental biology. Gametogenesis, sperm activation, and egg-sperm recognition are fundamental biological processes, warranting detailed studies into the molecular composition of gametes. Biological MS has been instrumental for the comprehensive itemizing of gamete proteomes. The protein constellation of sperm cells and its subcellular structures has been established for a variety of animal species. Spermatogenesis and the crucial activation of sperm cells as a prerequisite of successful fertilization and physiological adaptations to external stressors was investigated using proteomics, as well as the underlying mechanisms of male infertility with respect to proteome-wide alterations. This review outlines recent achievements of sperm proteomics and exemplifies the usefulness of gel-based surveys by outlining the comparative analysis of abnormal spermatozoa in globozoospermia. Besides label-free MS techniques and cell-based labeling methodology, high-resolution fluorescence 2DE has been shown to be highly suitable as a proteomic biomarker discovery tool in sperm protein research. The appropriateness of novel protein markers for improving our understanding of normal spermatogenesis and sperm activation versus the molecular pathogenesis of male infertility will be discussed. New biomarker candidates might be useful to improve diagnostic, prognostic, and therapeutic aspects of infertility. PMID:24909132

  7. Sperm protein 17 is an oncofetal antigen: a lesson from a murine model.

    PubMed

    Arnaboldi, F; Menon, A; Menegola, E; Di Renzo, F; Mirandola, L; Grizzi, F; Figueroa, J A; Cobos, E; Jenkins, M; Barajon, I; Chiriva-Internati, Maurizio

    2014-10-01

    Sperm protein 17 (Sp17) was originally identified in the flagellum of spermatozoa and subsequently included in the subfamily of tumor-associated antigens known as cancer-testes antigens (CTA). Sp17 has been associated with the motility and migratory capacity in tumor cells, representing a link between gene expression patterns in germinal and tumor cells of different histological origins. Here we review the relevance of Sp17 expression in the mouse embryo and cancerous tissues, and present additional data demonstrating Sp17 complex expression pattern in this murine model. The expression of Sp17 in embryonic as well as adult neoplastic cells, but not normal tissues, suggests this protein should be considered an "oncofetal antigen." Further investigations are necessary to elucidate the mechanisms and functional significance of Sp17 aberrant expression in human adult cells and its implication in the pathobiology of cancer. PMID:24811209

  8. Effects of ocean warming and acidification on fertilization in the Antarctic echinoid Sterechinus neumayeri across a range of sperm concentrations.

    PubMed

    Ho, M A; Price, C; King, C K; Virtue, P; Byrne, M

    2013-09-01

    The gametes of marine invertebrates are being spawned into an ocean that is simultaneously warming and decreasing in pH. Predicting the potential for interactive effects of these stressors on fertilization is difficult, especially for stenothermal polar invertebrates adapted to fertilization in cold, viscous water and, when decreased sperm availability may be an additional stressor. The impact of increased temperature (2-4 °C above ambient) and decreased pH (0.2-0.4 pH units below ambient) on fertilization in the Antarctic echinoid Sterechinus neumayeri across a range of sperm concentrations was investigated in cross-factorial experiments in context with near future ocean change projections. The high temperature treatment (+4 °C) was also used to assess thermal tolerance. Gametes from multiple males and females in replicate experiments were used to reflect the multiple spawner scenario in nature. For fertilization at low sperm density we tested three hypotheses, 1) increased temperature enhances fertilization success, 2) low pH reduces fertilization and, 3) due to the cold stenothermal physiology of S. neumayeri, temperature would be the more significant stressor. Temperature and sperm levels had a significant effect on fertilization, but decreased pH did not affect fertilization. Warming enhanced fertilization at the lowest sperm concentration tested likely through stimulation of sperm motility and reduced water viscosity. Our results indicate that fertilization in S. neumayeri, even at low sperm levels potentially found in nature, is resilient to near-future ocean warming and acidification. PMID:23948149

  9. Trypan blue/giemsa staining to assess sperm membrane integrity in salernitano stallions and its relationship to pregnancy rates.

    PubMed

    Serafini, R; Longobardi, V; Spadetta, M; Neri, D; Ariota, B; Gasparrini, B; Di Palo, R

    2014-02-01

    Aim of this study was to test the reliability of Trypan blue/Giemsa staining to evaluate sperm membrane integrity, acrosomal intactness and morphology in stallion to verify whether it could be applied in vitro as useful tool for sperm fertilizing ability. Fertility data on inseminated mares were collected to evaluate the relationship of sperm quality to pregnancy rates. Forty-one ejaculates were collected from 3 stallions of Salernitano Horse Breed and evaluated for gross appearance, volume, visual motility and membrane integrity with Trypan blue/Giemsa staining and thirty-five mares were inseminated during the breeding season from April to July. Differences among stallions were found in volume, sperm concentration (p < 0.05) and visual motility (p < 0.01). A decrease in sperm motility, concentration (p < 0.05) and total sperm number was found in June-July (p < 0.01). Live sperm with intact acrosome (LSIA) and proximal droplets (PD) were lower (p < 0.01) in June-July, while acrosome reacted sperm (ARS) percentage increased (p < 0.05). No fertility differences were found among stallions with an average fertility per cycle of 44.6% and a pregnancy rate of 68.6%. Higher percentages of LSIA were found in the ejaculates used to inseminate mares that became pregnant vs those used in mares not pregnant (p < 0.05). The significance of LSIA as test variable to verify the reliability of Trypan blue/Giemsa staining was confirmed by Receiver operating characteristic ROC analysis and the sensitivity of the test was 85% at a cut-off value of 48% LSIA. Trypan blue-Giemsa showed to be an accurate method that can be applied on field to evaluate sperm membrane integrity and to identify poor-quality ejaculates. PMID:23889365

  10. Association between sperm DNA integrity and seminal plasma antioxidant levels in health workers occupationally exposed to ionizing radiation.

    PubMed

    Kumar, Dayanidhi; Salian, Sujith Raj; Kalthur, Guruprasad; Uppangala, Shubhashree; Kumari, Sandhya; Challapalli, Srinivas; Chandraguthi, Shrinidhi Gururajarao; Jain, Navya; Krishnamurthy, Hanumanthappa; Kumar, Pratap; Adiga, Satish Kumar

    2014-07-01

    There is a paucity of data regarding the association between occupational radiation exposure and risk to human fertility. Recently, we provided the first evidence on altered sperm functional characteristics, DNA damage and hypermethylation in radiation health workers. However, there is no report elucidating the association between seminal plasma antioxidants and sperm chromatin integrity in occupationally exposed subjects. Here, we assessed the seminal plasma antioxidants and lipid peroxidation level in 83 men who were occupationally exposed to ionizing radiation and then correlated with the sperm chromatin integrity. Flow cytometry based sperm chromatin integrity assay revealed a significant decline in ?t value in the exposed group in comparison to the non-exposed group (P<0.0001). Similarly, both total and reduced glutathione levels and total antioxidant capacity in the seminal plasma were significantly higher in exposed group than the non-exposed group (P<0.01, 0.001 and 0.0001, respectively). However, superoxide dismutase level and malondialdehyde level, which is an indicator of lipid peroxidation in the seminal plasma, did not differ significantly between two groups. The total antioxidant capacity (TAC) and GSH level exhibited a positive correlation with sperm DNA integrity in exposed subjects. To conclude, this study distinctly shows that altered sperm chromatin integrity in radiation health workers is associated with increase in seminal plasma antioxidant level. Further, the increased seminal plasma GSH and TAC could be an adaptive measure to tackle the oxidative stress to protect genetic and functional sperm deformities in radiation health workers. PMID:24834825

  11. Comparative analysis of macaque and human sperm proteomes: Insights into sperm competition.

    PubMed

    Zhou, Tao; Wang, Gaigai; Chen, Minjian; Zhang, Mianqiu; Guo, Yueshuai; Yu, Chunmei; Zhou, Zuomin; Si, Wei; Sha, Jiahao; Guo, Xuejiang

    2015-05-01

    Male macaques produce faster sperm than male humans due to a higher pressure of sperm competition in macaques. To explore the molecular basis of this biological difference, we firstly constructed macaque and human sperm proteomes using LC-MS/MS. We then detected the positively selected genes specifically on the branch of macaque based on branch-site likelihood method. We identified 197 positively selected genes specifically on the branch of macaque that are unselected in corresponding human orthologs. These genes are highly associated with mitochondria and axoneme that directly drive sperm motility. We further compared the ultrastructural differences of the midpiece between macaque and human sperms to provide evidence for our findings using transmission electron microscopy. In conclusion, our results provide potential molecular targets for explaining the different phenotypes under sperm competition between macaques and humans, and also provide resources for the analysis of male fertility. PMID:25545774

  12. Relationship of seminal reactive nitrogen and oxygen species and total antioxidant capacity with sperm DNA fragmentation in infertile couples with normal and abnormal sperm parameters.

    PubMed

    Khosravi, F; Valojerdi, M R; Amanlou, M; Karimian, L; Abolhassani, F

    2012-11-01

    The objective of this study was to investigate the association between the amount of superoxide anion, peroxynitrite as oxidative stress (OS) markers and total antioxidant capacity (TAC) with sperm DNA fragmentation in infertile men with abnormal semen parameters. Semen samples were obtained from 102 infertile couples and divided into groups with normal and abnormal semen parameters according to the World Health Organization (WHO). Peroxynitrite and superoxide anions were detected using spectrofluorometric assays combined with 2,7 dicholorofluorescein (DCF)-DA and 4-chloro-7-nitrobenzo-2-oxa -1, 3-diazole (NBD-CL). Colorimetric assay was used for evaluation of TAC, while DNA fragmentation was studied by using sperm chromatin dispersion test. Superoxide anion, peroxynitrite and DNA fragmentation were significantly higher in infertile couples with abnormal semen parameters as compared to infertile couples with normal semen (P < 0.01). TAC was significantly lower in infertile men with abnormal semen parameters (P < 0.01). There was also a significant positive correlation between OS markers with sperm DNA fragmentation (r = 0.59, P < 0.01 and r = 0.67, P < 0.01, respectively). We have found that imbalance between superoxide anion and peroxynitrite with antioxidant capacity in infertile men with abnormal sperm parameters is associated with higher sperm DNA fragmentation. PMID:23126684

  13. Effects of oral antioxidant treatment upon the dynamics of human sperm DNA fragmentation and subpopulations of sperm with highly degraded DNA.

    PubMed

    Abad, C; Amengual, M J; Gosálvez, J; Coward, K; Hannaoui, N; Benet, J; García-Peiró, A; Prats, J

    2013-06-01

    The primary aim of this study was to determine the effect of oral antioxidant treatment (1500 mg of l-Carnitine; 60 mg of vitamin C; 20 mg of coenzyme Q10; 10 mg of vitamin E; 10 mg of zinc; 200 ?g of vitamin B9; 50 ?g of selenium; 1 ?g of vitamin B12) during a time period of 3 months upon the dynamics of sperm DNA fragmentation following varying periods of sperm storage (0 h, 2 h, 6 h, 8 h and 24 h) at 37 °C in a cohort of 20 infertile patients diagnosed with asthenoteratozoospermia. A secondary objective was to use the sperm chromatin dispersion test (SCD) to study antioxidant effects upon a specific subpopulation of highly DNA degraded sperm (DDS). Semen parameters and pregnancy rate (PR) were also determined. Results showed a significant improvement of DNA integrity at all incubation points (P < 0.01). The proportion of DDS was also significantly reduced (P < 0.05). Semen analysis data showed a significant increase in concentration, motility, vitality and morphology parameters. Our results suggest that antioxidant treatment improves sperm quality not only in terms of key seminal parameters and basal DNA damage, but also helps to maintain DNA integrity. Prior administration of antioxidants could therefore promote better outcomes following assisted reproductive techniques. PMID:22943406

  14. Comparative Analysis of Mafriwal (Bos taurus × Bos indicus) and Kedah Kelantan (Bos indicus) Sperm Proteome Identifies Sperm Proteins Potentially Responsible for Higher Fertility in a Tropical Climate

    PubMed Central

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

    2013-01-01

    The fertility of zebu cattle (Bos indicus) is higher than that of the European purebred (Bos taurus) and crossbred (Bos taurus × Bos indicus) cattle in tropical areas. To identify proteins related to the higher thermo-tolerance and fertility of Zebu cattle, this study was undertaken to identify differences in sperm proteome between the high fertile Malaysian indigenous zebu cattle (Kedah Kelantan) and the sub-fertile crossbred cattle (Mafriwal). Frozen semen from three high performance bulls from each breed were processed to obtain live and pure sperm. Sperm proteins were then extracted, and two-dimensional gel electrophoresis performed to compare proteome profiles. Gel image analysis identified protein spots of interest which were then identified by liquid chromatography mass spectrometry quadrupole time-of-flight (LC MS/MS Q-TOF). STRING network analysis predicted interactions between at least 20 of the identified proteins. Among the identified proteins, a number of motility and energy related proteins were present in greater abundance in Kedah Kelantan. Sperm motility evaluation by Computer Assisted Semen Analysis (CASA) confirmed significantly higher motility in Kedah Kelantan. While results from this study do identify proteins that may be responsible for the higher fertility of Kedah Kelantan, functional characterization of these proteins is warranted to reinforce our understanding of their roles in sperm fertility. PMID:23903046

  15. Human sperm molecular anatomy: the enzyme 5?-reductase (SRD5A) is present in the sperm and may be involved in the varicocele-related infertility.

    PubMed

    Aquila, Saveria; Montanaro, Daniela; Guido, Carmela; Santoro, Marta; Perrotta, Ida; Gervasi, Serena; De Amicis, Francesca; Lanzino, Marilena

    2015-07-01

    The most common cause of male infertility is the testicular varicocele, a condition that impairs production and decreases quality of sperm. Male fertility also strictly depends on androgens acting through their own receptor. The enzyme 5?-reductase (SRD5A) is involved in the conversion of testosterone to 5?-dihydrotestosterone, both required for the development and maintenance of male reproductive function. Here, we evaluated, by western blotting analysis, the presence of SRD5A in human ejaculated spermatozoa and evidenced differences in sperm SRD5A content between healthy donors and varicocele-affected patients. Additionally, SRD5A sperm ultrastructural localization was also assessed by transmission electron microscopy and immunogold assay. We evidenced that SRD5A enzyme is present in the human spermatozoa and that its cellular content is lowered in sperm samples from varicocele patients compared to healthy subjects. The presence of SRD5A in human ejaculated spermatozoa highlights the potential role of this enzyme in sperm physiopathology suggesting that the decrease in its content, by affecting the conversion of testosterone into 5?-dihydrotestosterone, may be an important additional mechanism involved in the harmful effect of varicocele in male fertility. PMID:25850410

  16. Automated optical function testing of diffractive beam shapers

    NASA Astrophysics Data System (ADS)

    Lizotte, Todd E.; Ohar, Orest P.

    2006-08-01

    Function testing of diffractive optics is a straight forward procedure when dealing with single elements or discrete systems of optics on an optical bread board. As the volumes of a particular diffractive optical device increase to production levels approaching hundreds to thousands of optics, the testing task can quickly become arduous and mind-numbing. At this level of production, data collection and quality monitoring is required to close the feedback loop to the manufacturing process. In this paper, a description of a prototype semi-automated diffractive optics testing platform used to measure performance, map quality and confirm working distances of a two element diffractive beam shaper system will be given. The testing system architecture and operation will be outlined and each subsystem will be described including software. A wafer map, consisting of data from an array of beam shapers, representing performance and etch depth from each diffractive optic within the wafer will be presented. An example of how data is compiled from a tested wafer and then interlaced with external sources of topographic data for ease of analysis and interrogation will be explained. Methods for beam analysis will be disclosed including multiple profilometer techniques. Further information will be shown on how the data from the system can be interpreted to identify defects on specific optics, manufacturing defects and non-linearity effects due to grayscale lithography and etch processes.

  17. Yolk protein is expressed in the insect testis and interacts with sperm

    PubMed Central

    Bebas, Piotr; Kotwica, Joanna; Joachimiak, Ewa; Giebultowicz, Jadwiga M

    2008-01-01

    Background Male and female gametes follow diverse developmental pathways dictated by their distinct roles in fertilization. While oocytes of oviparous animals accumulate yolk in the cytoplasm, spermatozoa slough off most of their cytoplasm in the process of individualization. Mammalian spermatozoa released from the testis undergo extensive modifications in the seminal ducts involving a variety of glycoproteins. Ultrastructural studies suggest that glycoproteins are involved in sperm maturation in insects; however, their characterization at the molecular level is lacking. We reported previously that the circadian clock controls sperm release and maturation in several insect species. In the moth, Spodoptera littoralis, the secretion of glycoproteins into the seminal fluid occurs in a daily rhythmic pattern. The purpose of this study was to characterize seminal fluid glycoproteins in this species and elucidate their role in the process of sperm maturation. Results We collected seminal fluid proteins from males before and after daily sperm release. These samples were separated by 2-D gel electrophoresis, and gels were treated with a glycoprotein-detecting probe. We observed a group of abundant glycoproteins in the sample collected after sperm release, which was absent in the sample collected before sperm release. Sequencing of these glycoproteins by mass spectroscopy revealed peptides bearing homology with components of yolk, which is known to accumulate in developing oocytes. This unexpected result was confirmed by Western blotting demonstrating that seminal fluid contains protein immunoreactive to antibody against yolk protein YP2 produced in the follicle cells surrounding developing oocytes. We cloned the fragment of yp2 cDNA from S. littoralis and determined that it is expressed in both ovaries and testes. yp2 mRNA and YP2 protein were detected in the somatic cyst cells enveloping sperm inside the testis. During the period of sperm release, YP2 protein appears in the seminal fluid and forms an external coat on spermatozoa. Conclusion One of the yolk protein precursors YP2, which in females accumulate in the oocytes to provision developing embryos, appears to have a second male-specific role. It is produced in the testes and released into the seminal fluid where it interacts with sperm. These data reveal unexpected common factor in the maturation of insect eggs and sperm. PMID:18549506

  18. Tactic-specific differences in seminal fluid influence sperm performance

    PubMed Central

    Locatello, Lisa; Poli, Federica; Rasotto, Maria B.

    2013-01-01

    Seminal fluid often makes up a large part of an ejaculate, yet most empirical and theoretical studies on sperm competition have focused on how sperm characteristics (number and quality) affect fertilization success. However, seminal fluid influences own sperm performance and may potentially influence the outcome of sperm competition, by also affecting that of rivals. As a consequence males may be expected to allocate their investment in both sperm and seminal fluid in relation to the potential level of competition. Grass goby (Zosterisessor ophiocephalus) is an external fertilizer with guard-sneaker mating tactics, where sperm competition risk varies according to the tactic adopted. Here, we experimentally manipulated grass goby ejaculates by separately combining sperm and seminal fluid from territorial and sneaker males. While sperm of sneaker and territorial males did not differ in their performance when they interacted with their own seminal fluid only, sperm of sneakers increased their velocity and fertilization rate in the presence of territorial males' seminal fluid. By contrast, sneaker males' seminal fluid had a detrimental effect on the performance of territorial males' sperm. Sperm velocity was unaffected by the seminal fluid of males employing the same tactic, suggesting that seminal fluid's effect on rival-tactic sperm is not based on a self/non-self recognition mechanism. Our findings show that cross interactions of sperm and seminal fluid may influence the fertilization success of competing ejaculates with males investing in both sperm and seminal fluid in response to sperm competition risk. PMID:23363633

  19. Long sperm fertilize more eggs in a bird

    PubMed Central

    Bennison, Clair; Hemmings, Nicola; Slate, Jon; Birkhead, Tim

    2015-01-01

    Sperm competition, in which the ejaculates of multiple males compete to fertilize a female's ova, results in strong selection on sperm traits. Although sperm size and swimming velocity are known to independently affect fertilization success in certain species, exploring the relationship between sperm length, swimming velocity and fertilization success still remains a challenge. Here, we use the zebra finch (Taeniopygia guttata), where sperm size influences sperm swimming velocity, to determine the effect of sperm total length on fertilization success. Sperm competition experiments, in which pairs of males whose sperm differed only in length and swimming speed, revealed that males producing long sperm were more successful in terms of (i) the number of sperm reaching the ova and (ii) fertilizing those ova. Our results reveal that although sperm length is the main factor determining the outcome of sperm competition, complex interactions between male and female reproductive traits may also be important. The mechanisms underlying these interactions are poorly understood, but we suggest that differences in sperm storage and utilization by females may contribute to the outcome of sperm competition. PMID:25621327

  20. Sperm retrieval for intra-cytoplasmic sperm injection in non-obstructive azoospermia

    Microsoft Academic Search

    G. M. COLPI; G. PIEDIFERRO; F. NERVA; D. GIACCHETTA; E. M. COLPI; E. PIATTI

    2005-01-01

    Surgical testicular sperm retrieval for intra-cy- toplasmic sperm injection (ICSI) purposes is the only possibility of biological fathering in case of non-obstructive azoospermia (NOA). Successful retrieval only correlates with his- tology, not with FSH values or testicular vol- ume. Concurrent AZFa and AZFb microdele- tions predict unsuccessful recovery. Testicular sperm extraction (TESE) (mean of successful re- trievals in literature: 52.7%)