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Semen analysis and sperm function testing  

PubMed Central

Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both developing and developed countries. Other optional tests or sophisticated technologies have not been widely applied. The current review addresses important changes in the analysis of semen as described in the new World Health Organization (WHO) manual for semen analysis. The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality. Apart from the above mentioned changes, the initial evaluation and handling methods remain, in most instances, the same as in previous editions. Furthermore, the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology. WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995. The external quality control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training. This review reports on current sperm functional tests, such as the induced acrosome reaction, and sperm–zona pellucida binding assays, as well as the impact of sperm quality in terms of DNA integrity, and the relationship of sperm function tests to sperm morphology.

Franken, Daniel R; Oehninger, Sergio



Generation of functional eggs and sperm from cryopreserved whole testes.  


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

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



Generation of functional eggs and sperm from cryopreserved whole testes  

PubMed Central

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.

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



Spermatogonial stem cell transplantation into Rhesus testes regenerates spermatogenesis producing functional sperm  

PubMed Central

Summary Spermatogonial stem cells (SSCs) maintain spermatogenesis throughout a man’s life and may have application for treating some cases of male infertility, including those caused by chemotherapy before puberty. We performed autologous and allogeneic SSC transplantations into the testes of 18 adult and 5 prepubertal recipient macaques that were rendered infertile with alkylating chemotherapy. After autologous transplant, the donor genotype from lentivirus-marked SSCs was evident in the ejaculated sperm of 9/12 adult and 3/5 prepubertal recipients after they reached maturity. Allogeneic transplant led to donor-recipient chimerism in sperm from 2/6 adult recipients. Ejaculated sperm from one recipient transplanted with allogeneic donor SSCs were injected into 85 rhesus oocytes via intracytoplasmic sperm injection. Eighty-one oocytes were fertilized, producing embryos ranging from 4-cell to blastocyst with donor paternal origin confirmed in 7/81 embryos. This demonstration of functional donor spermatogenesis following SSC transplantation in primates is an important milestone for informed clinical translation.

Hermann, Brian P.; Sukhwani, Meena; Winkler, Felicity; Pascarella, Julia N.; Peters, Karen A.; Sheng, Yi; Valli, Hanna; Rodriguez, Mario; Ezzelarab, Mohamed; Dargo, Gina; Peterson, Kim; Masterson, Keith; Ramsey, Cathy; Ward, Thea; Lienesch, Maura; Volk, Angie; Cooper, David K.; Thomson, Angus W.; Kiss, Joseph E.; Penedo, Maria Cecilia T.; Schatten, Gerald P.; Mitalipov, Shoukhrat; Orwig, Kyle E.



Functional nonequivalence of sperm in Drosophila pseudoobscura.  


We report on a form of sperm polymorphism, termed polymegaly, that occurs in species of the Drosophila obscura group. Individual males of species in this group characteristically produce more than one discrete length of nucleated, motile sperm. Hypotheses suggested to explain the evolutionary significance of sperm polymorphism have been either nonadaptive or adaptive, with the latter focusing on sperm competition or nutrient provisioning. These hypotheses assume all sperm types fertilize eggs; however, no data have been gathered to test this assumption. We found that two size classes of sperm are produced and transferred to females in approximately equal numbers by the male; only long sperm persist in significant numbers in female sperm storage organs. Furthermore, we used a DNA-specific dye (bisbenzimide) and sperm-specific antibodies to ask if both sperm types fertilize eggs in Drosophila pseudoobscura. Confocal microscopy and immunofluorescent analyses of fertilized eggs using anti-sperm polyclonal antisera demonstrated that only long sperm participate in fertilization. These data falsify those hypotheses in which all sperm types are assumed to be functionally equivalent (fertilize eggs). Any remaining or new hypotheses for the evolutionary significance of polymegaly must incorporate these findings. Several new areas of research are suggested. PMID:7972038

Snook, R R; Markow, T A; Karr, T L



Mitochondria functionality and sperm quality.  


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 Ca(2)(+) stores, although their role in signalling is still unclear. PMID:23901129

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



The Sperm Chromatin Dispersion Test: A Simple Method for the Determination of Sperm DNA Fragmentation  

Microsoft Academic Search

Sperm DNA fragmentation is being increasingly rec- ognized as an important cause of infertility. We herein describe the Sperm Chromatin Dispersion (SCD) test, a novel assay for sperm DNA fragmentation in semen. The SCD test is based on the principle that sperm with fragmented DNA fail to produce the characteristic halo of dispersed DNA loops that is observed in sperm




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


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

Ramu, Sivakumar; Jeyendran, Rajasingam S



Sperm fate and function in reproductive isolation in Drosophila.  


Central to the study of speciation is the identification of the isolating mechanisms that reduce gene flow. Of special interest are those isolating mechanisms that are the earliest to arise. Sperm fate and function play a major role in two distinct types of reproductive isolation in the genus Drosophila. First, postcopulatory-prezygotic (PCPZ) incompatibilities between the male ejaculate and a heterospecific female reproductive tract can result in a reduction in sperm fertilisation success. Although PCPZ incompatibilities have, until recently, received little attention, overwhelming diversity in reproductive morphology and biochemistry indicates that they may play an important role in speciation in this genus. The second type of isolating mechanism that affects sperm occurs in the testes of hybrid males produced by heterospecific matings. These individuals often suffer from dysfunction in spermatogenesis, resulting in the production of aberrant sperm. Hybrid male sterility and hybrid inviability are examples of postzygotic (PZ) reproductive isolation. The observation that hybrid sterility is pervasive among males of all taxonomic groups is known as Haldane's rule. Here we discuss both the evolutionary origins, and functional causes of both PCPZ incompatibilities, which affects sperm fate and function in females, and one type of PZ incompatibility, hybrid male sterility, or sperm dysfunction in hybrid males. Although these two mechanisms of isolation are quite distinct, they are similarly caused by breakdowns in epistatic interactions which occur in the encounter between two divergent genomes. Molecular, cytological, and empirical data are discussed, as is relevant evolutionary theory. PMID:17644960

Markow, Therese Ann; Reed, Laura K; Kelleher, Erin S



Assessment of sperm survival and functional membrane integrity of the six-banded armadillo (Euphractus sexcinctus).  


The objective was to evaluate sperm survival in the six-banded armadillo, using a thermoresistance test, and to compare sugar solutions with varying osmolarities to analyze the integrity of the functional sperm plasma membrane in this species. Twelve ejaculates were obtained from four mature males by electroejaculation and evaluated for sperm motility, vigor, live sperm, and morphology. Sperm survival was evaluated during a thermoresistance test at 34 °C (the body temperature of this species). The functional integrity of the plasma membrane was evaluated by means of the hypo-osmotic swelling test (HOST), using solutions of varying osmolarities (0, 50, 100, and 150 mOsm/L). During the thermoresistance test, at each evaluation, there was a reduction (P < 0.05) in mean values for sperm motility, sperm vigor, and percentage of live sperm (no movement was observed at 360 min). Sperm survival varied among individual armadillos (P < 0.05). In two individuals, sperm vigor was significantly enhanced when semen was diluted in Tris extender. The response of armadillo sperm to the HOST varied among individuals (P < 0.05). On average, maximal values (P < 0.05) of reactive sperm (59%) were detected with 50 mOsm/L solution; furthermore, this concentration had the largest significant positive correlation (r = 0.84) to live sperm percentage. In conclusion, six-banded armadillos had significant individual variation with regard to sperm survival in a thermoresistance test at 34 °C; in some individuals, sperm survived until 360 min. The use of a 50 mOsm/L fructose solution was recommended for conducting a HOST in this species. PMID:21565395

Santos, E A A; Sousa, P C; Dias, C E V; Castelo, T S; Peixoto, G C X; Lima, G L; Ricarte, A R F; Simão, B R; Freitas, C I A; Silva, A R



Xenoestrogenic chemicals effectively alter sperm functional behavior in mice.  


Xenoestrogenic compounds (XCs) can disrupt endogenous hormone function and affect sperm function by binding to receptors on sperm membrane. Albeit spermatozoa are potentially a useful model for screening estrogenic activities of endocrine disruptors, high-quality in vitro test system that examination of the XCs effects on sperm function is required. The objective of this study was to compare the effects of XCs (genistein and 4-tert-octylphenol) to those of steroids (estrogen and progesterone) and heparin on in vitro capacitation and acrosome reaction (AR) in mouse spermatozoa. Mouse spermatozoa were incubated with various concentrations (0.001-100 ?M) of each chemical for 15 or 30 min, and then capacitation and AR were assessed using chlortetracycline. All chemicals studied effectively alter capacitation and/or AR in mouse spermatozoa with different manner. Therefore, we believed that our system will provide a good in vitro model system to characterize the physiological effect of XCs especially when compared with steroids. PMID:22036770

Park, Yoo-Jin; Mohamed, El-Sayed A; Kwon, Woo-Sung; You, Young-Ah; Ryu, Buom-Yong; Pang, Myung-Geol



The sperm chromatin dispersion test: a simple method for the determination of sperm DNA fragmentation.  


Sperm DNA fragmentation is being increasingly recognized as an important cause of infertility. We herein describe the Sperm Chromatin Dispersion (SCD) test, a novel assay for sperm DNA fragmentation in semen. The SCD test is based on the principle that sperm with fragmented DNA fail to produce the characteristic halo of dispersed DNA loops that is observed in sperm with non-fragmented DNA, following acid denaturation and removal of nuclear proteins. This was confirmed by the analysis of DNA fragmentation using the specific DNA Breakage Detection-Fluorescence In Situ Hybridization (DBD-FISH) assay, which allows the detection of DNA breaks in lysed sperm nuclei. Sperm suspensions either prepared from semen or isolated from semen by gradient centrifugation were embedded in an agarose microgel on slides and treated with 0.08 N HCl and lysing solutions containing 0.8 M dithiothreitol (DTT), 1% sodium dodecyl sulfate (SDS), and 2 M NaCl. Then, the slides were sequentially stained with DAPI (4',6-diamidino-2-phenylindole) and/or the Diff-Quik reagent, and the percentages of sperm with nondispersed and dispersed chromatin loops were monitored by fluorescence and brightfield microscopy, respectively. The results indicate that all sperm with nondispersed chromatin displayed DNA fragmentation, as measured by DBD-FISH. Conversely, all sperm with dispersed chromatin had very low to undetectable DBD-FISH labeling. SCD test values were significantly higher in patients being screened for infertility than in normozoospermic sperm donors who had participated in a donor insemination program. The coefficient of variation obtained using 2 different observers, either by digital image analysis (DIA) or by brightfield microscopy scoring, was less than 3%. In conclusion, the SCD test is a simple, accurate, highly reproducible, and inexpensive method for the analysis of sperm DNA fragmentation in semen and processed sperm. Therefore, the SCD test could potentially be used as a routine test for the screening of sperm DNA fragmentation in the andrology laboratory. PMID:12514084

Fernández, Jose Luis; Muriel, Lourdes; Rivero, Maria Teresa; Goyanes, Vicente; Vazquez, Rosana; Alvarez, Juan G


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

PubMed Central

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

Dowling, Damian K.; Simmons, Leigh W.



Protective function of alpha-lipoic acid on sperm motility and mitochondrial function during goat sperm-mediated gene transfer  

Microsoft Academic Search

Sperm-mediated gene transfer is a promising technology to generate transgenic animals, but the incubation of the sperm cell with target gene plasmids often impairs the sperm motility and mitochondrial function. Alpha-lipoic acid is one of the beneficial antioxidants, which has been used to protect many types of cells from impairment of their function. The objective of this study was to

Huiming Ma; Fusheng Quan; Dongmei Chen; Yanling Zheng; Bowei Zhang; Yongsheng Wang; Yong Zhang



The effects of increased testicular temperature on testis-specific isoform of Na+/K+ -ATPase in sperm and its role in spermatogenesis and sperm function.  


Impaired testicular thermoregulation is commonly implicated in abnormal spermatogenesis and impaired sperm function in animals and humans, with outcomes ranging from subclinical infertility to sterility. Bovine testes must be maintained 4-5 °C below body-core temperature for normal spermatogenesis. The effects of elevated testicular temperature have been extensively studied in cattle using a scrotal insulation model, which results in abnormal spermatogenesis and impaired sperm morphology and function. Using this model and proteomic approaches, we compared normal and abnormal sperm (from the same bulls) to elucidate the molecular basis of impaired function. We identified a cohort of sperm functional proteins differentially expressed between normal vs abnormal sperm, including a testis-specific isoform of Na(+) /K(+) -ATPase. In addition to its role as a sodium pump regulating sperm motility, Na(+) /K(+) -ATPase is also involved as a signalling molecule during sperm capacitation. In conclusion, because of its involvement in regulation of sperm function, this protein has potential as a fertility marker. Furthermore, comparing normal vs abnormal sperm (induced by scrotal insulation) is a useful model for identifying proteins regulating sperm function. PMID:22827367

Thundathil, J C; Rajamanickam, G D; Kastelic, J P; Newton, L D



Functional Nonequivalence of Sperm in Drosophila pseudoobscura  

Microsoft Academic Search

We report on a form of sperm polymorphism, termed polymegaly, that occurs in species of the Drosophila obscura group. Individual males of species in this group characteristically produce more than one discrete length of nucleated, motile sperm. Hypotheses suggested to explain the evolutionary significance of sperm polymorphism have been either nonadaptive or adaptive, with the latter focusing on sperm competition

Rhonda R. Snook; Therese A. Markow; Timothy L. Karr



Sperm competition: linking form to function  

PubMed Central

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.



Development of a Sperm Cell Toxicity Test for Marine Waters.  

National Technical Information Service (NTIS)

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

P. A. Dinnel Q. J. Stober S. C. Crumley R. E. Nakatani



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

Code of Federal Regulations, 2010 CFR

...2009-04-01 false Seminal fluid (sperm) immunological test system. 866...Systems § 866.5800 Seminal fluid (sperm) immunological test system. (a) Identification. A seminal fluid (sperm) immunological test system...



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

Code of Federal Regulations, 2010 CFR

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



Sperm DNA tests as useful adjuncts to semen analysis.  


Male infertility has traditionally been diagnosed by microscopic assessment of concentration, motility and morphology of sperm in the ejaculate. Most laboratories use sperm isolated by various methods such as density gradient centrifugation to enrich for subpopulations of sperm believed to have greater fertilization potential. These tests are essential to provide the fundamental information on which clinicians base their initial diagnosis. However, in the clinical setting, tests with superior prognostic value are needed. Tests showing much promise are those determining sperm DNA integrity, particularly the Comet, TUNEL, and Sperm Chromatin Structure assays. Sperm nuclear DNA fragmentation has been positively correlated with lower fertilization rates in IVF, impaired implantation rates, an increased incidence of abortion and disease in offspring, including childhood cancer. The mitochondrial genome of sperm has also been shown to be a sensitive marker of sperm health. Although the usefulness of these tests is recognized, insufficient resources have been available to develop standardized tests and protocols that could lead to universally accepted clinical thresholds. Associated with the lack of useful prognostic tests is the lack of improvement in assisted conception success rates despite thirty years of worldwide use. International collaborations should be initiated to develop agreed protocols and establish clinical thresholds. PMID:18570047

Lewis, Sheena E M; Agbaje, Ishola; Alvarez, Juan


Increased Aneuploidy Rate in Sperm With Fragmented DNA as Determined by the Sperm Chromatin Dispersion (SCD) Test and FISH Analysis  

Microsoft Academic Search

Previous studies suggest that sperm DNA fragmen- tation may be associated with aneuploidy. However, currently available tests have not made it possible to simultaneously perform DNA fragmentation and chromosomal analyses on the same sperm cell. The recently introduced sperm chromatin dispersion (SCD) test allows users to determine this relationship. Semen samples from 16 males, including 4 fertile donors, 7 normozoospermic,




Functional evidence that a recently evolved Drosophila sperm-specific gene boosts sperm competition  

PubMed Central

In many species, both morphological and molecular traits related to sex and reproduction evolve faster in males than in females. Ultimately, rapid male evolution relies on the acquisition of genetic variation associated with differential reproductive success. Many newly evolved genes are associated with novel functions that might enhance male fitness. However, functional evidence of the adaptive role of recently originated genes in males is still lacking. The Sperm dynein intermediate chain multigene family, which encodes a Sperm dynein intermediate chain presumably involved in sperm motility, originated from complex genetic rearrangements in the lineage that leads to Drosophila melanogaster within the last 5.4 million years since its split from Drosophila simulans. We deleted all the members of this multigene family resident on the X chromosome of D. melanogaster by chromosome engineering and found that, although the deletion does not result in a reduction of progeny number, it impairs the competence of the sperm in the presence of sperm from wild-type males. Therefore, the Sperm dynein intermediate chain multigene family contributes to the differential reproductive success among males and illustrates precisely how quickly a new gene function can be incorporated into the genetic network of a species.

Yeh, Shu-Dan; Do, Tiffanie; Chan, Carolus; Cordova, Adriana; Carranza, Francisco; Yamamoto, Eugene A.; Abbassi, Mashya; Gandasetiawan, Kania A.; Librado, Pablo; Damia, Elisabetta; Dimitri, Patrizio; Rozas, Julio; Hartl, Daniel L.; Roote, John; Ranz, Jose M.



Sperm mitochondria of patients with normal sperm motility and with asthenozoospermia: morphological and functional study.  


Studies were performed on ejaculated human spermatozoa (32 subjects with normal sperm motility and 25 subjects with low sperm motility). Morphology of sperm midpiece was evaluated in light, fluorescent and transmission or scanning electron microscope. Changes in mitochondrial membrane potential (delta(psi)m) and mass of mitochondria were analysed by flow cytometry using mitochondrial specific probes JC-1 and Mito Tracker Green FM. Moreover, oxidoreductive capability of sperm mitochondria was assessed using cytochemical reaction for NADH-dependent dehydrogenases. In flow cytometry analysis of JC-1-stained spermatozoa, two asthenozoospermic subpopulations were distinguished: patients with a high percentage (76 +/- 11%, 13 subjects) and patients with a low percentage (29 +/- 14%,12 subjects) of spermatozoa with functional-polarized mitochondria with high delta(psi)m. Our microscopic investigations of spermatozoa of seven asthenozoospermic patients reveal that the deformed and unusually thickened sperm midpieces (50-70% of cells), occasionally with persistent cytoplasmic droplet, contain supernumerary mitochondria with normal substructure, full oxidoreductive capability and high delta(psi)m. The midpiece deformations cause nonprogressive movement or immotility. They can also appear in smaller number of spermatozoa (5-35% of cells) in patients with normal sperm motility. Moreover, in three cases of asthenozoospermia midpiece malformations were accompanied by abnormal morphology of outer dense fibers and axoneme. The cytochemical, fluorescence and SEM studies showed the absence of midpieces in many (60-80%) spermatozoa in some other cases of asthenozoospermia. The morphological observations corresponded with flow cytometry analysis of Mito Tracker Green FM-stained spermatozoa. Our results suggest that in some cases of asthenozoospermia the sperm mitochondria can be functionally active and display high delta(psi)m in large number of cells. The results may suggest that asthenozoospermia does not necessarily result from energetic disturbances of sperm mitochondria. The low sperm motility may be associated with deformations of the mitochondrial sheath containing functional mitochondria. The combination of fluorescence microscopy and flow cytometry with electron microscopic investigations is a sensitive, precise and comprehensive examination which helps discover sperm abnormalities responsible for asthenozoospermia. PMID:13678331

Piasecka, Ma?gorzata; Kawiak, Jerzy



Evaluation of DNA fragmentation in llama (Lama glama) sperm using the sperm chromatin dispersion test.  


The integrity of sperm chromatin is now viewed as an important factor in male fertility and in early embryonic development. The objectives of this study were: (1) adapt the simple and inexpensive sperm chromatin dispersion (SCD) test to evaluate DNA fragmentation in llama sperm and establish the halo patterns observed in this species, (2) determine an effective and reliable positive control for this technique and (3) evaluate correlation between the SCD test and the toluidine blue (TB) stain. To adapt the SCD test, three different mercaptoethanol (ME) concentrations were assayed (2.5%, 5% and 10% ME). To determine an effective positive control, three treatments (incubation at 100 °C for 30 min, incubation with 0.3 M NaOH for 30 min at room temperature and exposure to UV light for 2h) were assayed. The concentration selected to use in the SCD test was 5% ME, because it produced the largest halo while still conserving the structure of the core. Four DNA dispersion patterns were clearly observed: (I) nuclei with large DNA dispersion halos; (II) nuclei with medium halos; (III) nuclei with very small halos and (IV) nuclei with no halo. All treatments used as positive controls were effective in producing DNA fragmentation. A high correlation (r=0.84, P=0.03) was observed between spermatozoa without halos and TB positive cells. To conclude, SCD patterns in llama sperm have been established as well as a repeatable positive control for the assay. The SCD test and TB stain are simple and inexpensive techniques that can be used to evaluate DNA damage in llama sperm. PMID:22437148

Carretero, M I; Lombardo, D; Arraztoa, C C; Giuliano, S M; Gambarotta, M C; Neild, D M



Not All Sperm Are Equal: Functional Mitochondria Characterize a Subpopulation of Human Sperm with Better Fertilization Potential  

Microsoft Academic Search

Human sperm samples are very heterogeneous and include a low amount of truly functional gametes. Distinct strategies have been developed to characterize and isolate this specific subpopulation. In this study we have used fluorescence microscopy and fluorescence-activated cell sorting to determine if mitochondrial function, as assessed using mitochondrial-sensitive probes, could be employed as a criterion to obtain more functional sperm

Ana Paula Sousa; Alexandra Amaral; Marta Baptista; Renata Tavares; Pedro Caballero Campo; Pedro Caballero Peregrín; Albertina Freitas; Artur Paiva; Teresa Almeida-Santos; João Ramalho-Santos; Hugh Clarke



Establishment of a practical sperm penetration test for bovine semen.  


Objective: Several methods for evaluating semen quality have been developed in addition to traditional semen analysis. Most of these methods are very complex and require expensive technical equipment as well as special knowledge to be performed. The aim of this study was to establish an easy and cost-effective sperm penetration test for bovine semen using artificial media. Additionally a standard procedure for test performance should be figured out. Material and methods: In the first part of the study four different capillary tubes or pipettes were evaluated in order to determine the most appropriate one to use in the penetration test. In the second part of the study the tubes were filled with different compositions of polyacrylamide gel and subsequently incubated at 37.5 °C for 30 minutes. Sperm penetration was then evaluated using a phase contrast microscope, measuring the penetration distance of the first, the first motile and the five vanguard motile spermatozoa as well as determining the number of sperm cells cumulated per low power field (LPF) at three distinct distances in the tube. Results: Non-heparinised haematocrit capillary tubes were the most appropriate tubes for performance of sperm penetration test as 100% of used capillaries were evaluable after incubation and size of the capillaries allowed a simple handling and good visualisation of migrated spermatozoa. Best results were achieved with 1.9% polyacrylamide gel containing phosphate-buffered saline. Measuring the penetration distance of the vanguard motile sperm cell can be recommended for test evaluation. Conclusion: A sperm penetration test with 1.9% polyacrylamide using non-heparinised capillary tubes was established. Results showed high reproducibility, the procedure is simple to carry out and only requires a phase contrast microscope. PMID:24126998

Becher, A C; Failing, K; Kauffold, J; Wehrend, A



Standardizing the methodology of sperm cell test with Paracentrotus lividus.  


The sperm cell toxicity test with Paracentrotus lividus is widely used in Italy to assess the quality of complex aqueous matrices, but at present there is a shortage of standardized methodologies for the bioassay performance. In this work several critical points were considered, in order to verify the reliability of this bioassay and to improve its standardization. In particular, we have studied the differences in EC50 values at different sperm: egg ratios and with different quantities of gametes; the influence of sperm and eggs on fertilization rate and the influence of different recipients materials (plastic or glass). At the same time, it was evaluated if the obtained EC50 values were in accordance with those reported in literature. Experiments performed yielded EC50 values for copper ranging from 16.54 microg/l to 69.59 microg/l, with a total mean value +/- SD of 39.79 microg/l +/- 11.17. These data fit with those found in literature for other echinoid species. According to the same authors, poor fertilization was probably due to the effects on sperm and not on eggs, and moreover the linear regression analysis performed on each experiment suggested that the best amount of eggs to use in the test was 1000. Results from the comparison between the performance of the test in plastic or glass tubes showed an higher success in fertilization rate using borosilicate glass tubes. PMID:16738765

Lera, S; Macchia, S; Pellegrini, D



Biogenesis of sperm perinuclear theca and its role in sperm functional competence and fertilization.  


The perinuclear theca is a condensed cytosolic protein layer that surrounds the mammalian sperm nucleus except in the region of tail implantation. It is resistant to non-ionic detergent extraction and makes up most of the cytosol of the sperm head. The perinuclear theca can be divided both structurally and compositionally into subacrosomal and postacrosomal regions. Most of the sequence-identities of investigated perinuclear theca proteins have been unexpected, revealing novel proteins as well as isoforms of somatic and conventionally nuclear proteins. Their characterizations have led us to propose that the perinuclear theca can be regionalized into two functional categories: the subacrosomal layer, involved in acrosomal assembly, and the postacrosomal sheath, involved in sperm-egg interactions during fertilization such as egg activation. Immunocytochemical investigation of the origins of well-characterized perinuclear theca proteins have led us to propose that: the subacrosomal layer is assembled relatively early in spermiogenesis from cytosolic proteins that "piggy-back" their way to the forming perinuclear theca on the periphery of acrosomic vesicles, while the postacrosomal sheath is assembled later, from cytosolic proteins that are transported up the microtubular manchette as it descends over the caudal half of the elongating spermatid nucleus. In this review data collected on resident perinuclear theca proteins, SubH2Bv, RAB2, PAWP and the four core somatic histones, is used to substantiate these hypotheses. PMID:19883945

Oko, Richard; Sutovsky, Peter



Sperm adhesion molecules: structure and function.  


Fertilisation is a unique event in which the morphologically disparate gametes recognise, bind and fuse with each other. This event follows a highly regulated schedule of biochemical interactions, in which molecules are involved that mediate cell adhesion, signal transduction and the initiation of metabolic pathways. A plethora of molecules has been found on the male gamete and with regard to the different protein structures it is almost impossible to overlook the structures involved. Even more, carbohydrate structures cause an additional diversity with regard to the generation of surface structures. In this communication we try to elucidate the structures of proteins that have been known so far. We have focussed on spermadhesins, the zonadhesin, proacrosin and the PH-20 antigen. The variety of structures and also the common features among them as well as the presence of redundant systems are attributable to the evolutionary force of intraspecific sperm competition. This evolutionary force is assumed to be also responsible for the species selectivity observed in these adhesion molecules, which explains the preferential binding of gametes in a homologous system. PMID:11114590

Jansen, S; Ekhlasi-Hundrieser, M; Töpfer-Petersen, E



Ultraviolet damages sperm mitochondrial function and membrane integrity in the sea urchin Anthocidaris crassispina.  


Effects of ultraviolet A (UVA) and ultraviolet B (UVB) on mitochondrial function and membrane integrity of sea urchin sperm were investigated using flow cytometry and fluorescent probes. Both UVA and UVB impaired sperm mitochondrial function in a dose-dependent manner. Covariance analysis further showed that the slopes of change in mitochondrial function in relation to UVA and UVB were significantly different, suggesting that the modes of action were different. UVA did not affect membrane integrity, while membrane integrity showed a linear reduction with increasing UVB doses. Sperm mitochondria function showed significant positive correlations with sperm motility and subsequent fertilization success. Overall, our results showed that both UVA and UVB could decrease sperm motility and fertilization success through impairment of mitochondrial function, whereas UVB alone could cause additional damage through impairing the functional integrity of sperm membrane. Mitochondrial function of sperm may also offer a reliable ecotoxicological biomarker for predicting fertilization success in urchins. PMID:15814310

Lu, X Y; Wu, R S S



The heparin-glutathione test: an alternative to the hypo-osmotic swelling test to select viable sperm for intracytoplasmic sperm injection  

Microsoft Academic Search

Objective: To evaluate the heparin-glutathione test (HEGLUT) for the selection of viable sperm for intracytoplasmic sperm injection (ICSI).Design: A prospective study.Setting: Department of Pediatrics, Obstetrics and Gynecology, University of Valencia and Instituto Valenciano de Infertilidad.Patient(s): Semen samples from healthy donors and patients with infertility.Intervention(s): Sperm samples were kept in culture for different periods in Ham’s F-10 medium supplemented or not

Francisco J Vendrell; Carmen Rubio; Juan J Tar??n



[Cyclic nucleotide-gated channels and sperm function].  


The cyclic nucleotide-gated (CNG) channel is a nonselective cation channel and one of the main entrances of Ca2+ influxion into cells. CNG channels are opened by direct binding of cyclic nucleotides. Six different genes encode the CNG protein, 4 A subunits and 2 B subunits. The activity of CNG channels can be regulated by Ca2+/Ca(2+)-binding proteins (CaM) and phosphorylation/dephosphorylation. Recently, extensive attention has been drawn to the researches on CNG channels in the reproductive system, and many studies show that CNG channels play a pivotal role in sperm motility, capacitation and acrosome reaction. This article focuses on the relationship of CNG channels with sperm function. PMID:23700737

Xu, Jian-Hua; Du, Jing



K+ and Cl? Channels and Transporters in Sperm Function  

PubMed Central

To succeed in fertilization, spermatozoa must decode environmental cues which require a set of ion channels. Recent findings have revealed that K+ and Cl? channels participate in some of the main sperm functions. This work reviews the evidence indicating the involvement of K+ and Cl? channels in motility, maturation, and the acrosome reaction, and the advancement in identifying their molecular identity and modes of regulation. Improving our insight on how these channels operate will strengthen our ability to surmount some infertility problems, improve animal breeding, preserve biodiversity, and develop selective and secure male contraceptives.

Santi, C.M.; Orta, G.; Salkoff, L.



Mechanisms of sperm competition: testing the fair raffle.  


Sperm competition is a major force of sexual selection, but its implications for mating system and life-history evolution are just beginning to be understood. Of particular importance is understanding the mechanisms of sperm competition. Models have been developed to determine if sperm competition operates in a fair raffle process, whereby each sperm from competing males has an equal chance of fertilizing a female's ova, or if it operates in a loaded raffle process, whereby one male's sperm has a fertilization advantage. These models require data on relative sperm and offspring (paternity) numbers of competing males. Here we develop a model based on maximum-likelihood methods for differentiating between the fair and loaded raffle processes. The model calculates the relative competitiveness of two males' sperm (loadings) as well as the economy of scale (nonlinear returns to sperm number). Previous models implicitly assumed that there is no economy of scale, which may not be the case when there is cooperation or interference among sperm from a given male. We demonstrate that our model has superior power-in some instances more than double-than previous models. We apply our model to an example of sperm competition in the guppy (Poecilia reticulata) and show that the system may be characterized by a loaded raffle attributable to effects of second male precedence. PMID:15446436

Neff, Bryan D; Wahl, Lindi M



Effect of polyvinylpyrrolidone on sperm function and early embryonic development following intracytoplasmic sperm injection in human assisted reproduction.  


The objective here was to review the effects of polyvinylpyrrolidone (PVP) upon sperm function and embryonic development in humans. PVP has been used successfully in intracytoplasmic sperm injection (ICSI) to facilitate the handling and immobilization of sperm for both domestic animals and humans. In our previous reports, PVP solution exists locally in embryos injected during the early developmental period, and also exerts influence over the developmental capacity of such embryos. In other reports, PVP causes significant damage to sperm membranes that can be detected by transmission electron microscopy, and has been associated with chromosomal abnormalities in pregnancy derived from ICSI embryos. In some Japanese clinics, PVP-free media has been used for sperm immobilization in order to optimise safety. Consequently, it is strongly suggested that the success rate of fertilization and clinical pregnancy could be improved by using PVP-free solution for human ICSI. In conclusion, our interpretation of the available data is to perform ICSI without PVP or select a lower concentration of PVP solution in order to reduce safety for pregnancy and children born via ICSI. PMID:23483084

Kato, Yoku; Nagao, Yoshikazu



Measurement of epididymal sperm motility as a test variable in the rat  

Microsoft Academic Search

Several environmental contaminants, notably dibromochloropropane (Whorton et al. 1977) and kepone (Taylor et al. 1978; Cannon et al. 1978) have been implicated in sperm deficiencies among occupationally exposed males. These incidents emphasize the need for adequate testing of chemicals for effects on the male reproductive system. Although important in clinical diagnosis, the evaluation of sperm motility has not been used

Ralph E. Linder; Lillian F. Strader; W. Keith McElroy



Seasonal functional relevance of sperm characteristics in equine spermatozoa.  


A group of stallions with different reproductive indexes were used to study seasonal variations in sperm quality (Equus caballus). Semen samples were collected from late September to July and analyzed according to four seasonal periods: late September-December, January-March, late March-May, and June-July. Parameters monitored included sperm concentration, sperm motility, sperm morphology, sperm viability, acrosomal status, plasma membrane stability, and sperm mitochondrial membrane potential. Overall, seminal parameters monitored are affected mostly by time period, followed by animal and lastly by fertility, stressing the importance of individual variations in out-bred animal models. The analysis of multiple ejaculates from the same animals showed clear seasonal-based differences (P<0.05) with poor semen quality in winter and a noticeable improvement in sperm quality with increasing photoperiod. Better semen quality was observed between late March and May. Interactions between month period, animal, and fertility were evident (P<0.05) for sperm concentration, head and tail sperm anomalies, and acrosomal integrity. Thus, it may be advisable to adjust the use of stallion semen according to seasonal variations. PMID:20083300

Gamboa, S; Rodrigues, A S; Henriques, L; Batista, C; Ramalho-Santos, J



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

Code of Federal Regulations, 2013 CFR




Sperm wars and the evolution of male fertility.  


Females frequently mate with several males, whose sperm then compete to fertilize available ova. Sperm competition represents a potent selective force that is expected to shape male expenditure on the ejaculate. Here, we review empirical data that illustrate the evolutionary consequences of sperm competition. Sperm competition favors the evolution of increased testes size and sperm production. In some species, males appear capable of adjusting the number of sperm ejaculated, depending on the perceived levels of sperm competition. Selection is also expected to act on sperm form and function, although the evidence for this remains equivocal. Comparative studies suggest that sperm length and swimming speed may increase in response to selection from sperm competition. However, the mechanisms driving this pattern remain unclear. Evidence that sperm length influences sperm swimming speed is mixed and fertilization trials performed across a broad range of species demonstrate inconsistent relationships between sperm form and function. This ambiguity may in part reflect the important role that seminal fluid proteins (sfps) play in affecting sperm function. There is good evidence that sfps are subject to selection from sperm competition, and recent work is pointing to an ability of males to adjust their seminal fluid chemistry in response to sperm competition from rival males. We argue that future research must consider sperm and seminal fluid components of the ejaculate as a functional unity. Research at the genomic level will identify the genes that ultimately control male fertility. PMID:22984191

Simmons, Leigh W; Fitzpatrick, John L



Evaluation of Sperm Tests as Indicators of Germ-Cell Damage in Men Exposed to Chemical or Physical Agents.  

National Technical Information Service (NTIS)

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 ex...

A. J. Wyrobek G. Watchmaker L. Gordon



Cooling and freezing of boar spermatozoa: Supplementation of the freezing media with reduced glutathione preserves sperm function  

Microsoft Academic Search

In this study, we evaluated the effects of glutathione\\u000a (L-\\\\?-glutamyl-L-cysteinylglycine; GSH) supplementation of the\\u000a freezing extender on semen parameters during the cooling (2 hours at\\u000a 5\\\\&DEG; C) and freezing phases of the cryopreservation process to\\u000a compensate for the decrease in GSH content observed during sperm\\u000a freezing. To fully address these questions, we incorporated a new set\\u000a of functional sperm tests.




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

PubMed Central

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

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




EPA Science Inventory

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


Phosphoglycerate Kinase 2 (PGK2) Is Essential for Sperm Function and Male Fertility in Mice1  

PubMed Central

Phosphoglycerate kinase 2 (PGK2), an isozyme that catalyzes the first ATP-generating step in the glycolytic pathway, is encoded by an autosomal retrogene that is expressed only during spermatogenesis. It replaces the ubiquitously expressed phosphoglycerate kinase 1 (PGK1) isozyme following repression of Pgk1 transcription by meiotic sex chromosome inactivation during meiotic prophase and by postmeiotic sex chromatin during spermiogenesis. The targeted disruption of Pgk2 by homologous recombination eliminates PGK activity in sperm and severely impairs male fertility, but does not block spermatogenesis. Mating behavior, reproductive organ weights (testis, excurrent ducts, and seminal vesicles), testis histology, sperm counts, and sperm ultrastructure were indistinguishable between Pgk2?/? and wild-type mice. However, sperm motility and ATP levels were markedly reduced in males lacking PGK2. These defects in sperm function were slightly less severe than observed in males lacking glyceraldehyde-3-phosphate dehydrogenase, spermatogenic (GAPDHS), the isozyme that catalyzes the step preceding PGK2 in the sperm glycolytic pathway. Unlike Gapdhs?/? males, the Pgk2?/? males also sired occasional pups. Alternative pathways that bypass the PGK step of glycolysis exist. We determined that one of these bypass enzymes, acylphosphatase, is active in mouse sperm, perhaps contributing to phenotypic differences between mice lacking GAPDHS or PGK2. This study determined that PGK2 is not required for the completion of spermatogenesis, but is essential for sperm motility and male fertility. In addition to confirming the importance of the glycolytic pathway for sperm function, distinctive phenotypic characteristics of Pgk2?/? mice may provide further insights into the regulation of sperm metabolism.

Danshina, Polina V.; Geyer, Christopher B.; Dai, Qunsheng; Goulding, Eugenia H.; Willis, William D.; Kitto, G. Barrie; McCarrey, John R.; Eddy, E.M.; O'Brien, Deborah A.




EPA Science Inventory

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


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

SciTech Connect

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.

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



Chemical Inhibition of Rabbit Sperm Cell Motility in Toxicological Testing.  

National Technical Information Service (NTIS)

The use of motile rabbit spermatozoa to assess the potential cytotoxic effect of chemicals was investigated. A suspension of motile sperm cells in a defined medium was incubated at 37 C under an atmosphere of 5% Carbon Dioxide 8% diatomic oxygen and 87% d...

D. Burnett R. J. Young



Comparative Evaluation of Three Rapid Marine Toxicity Tests: Sea Urchin Early Embryo Growth Test, Sea Urchin Sperm Cell Toxicity Test and Microtox.  

National Technical Information Service (NTIS)

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 (EC5...

D. Nacci E. Jackim R. Walsh



Identification and Characterization of Functional Nongenomic Progesterone Receptors on Human Sperm Membrane  

Microsoft Academic Search

The presence of functional nongenomic progesterone (P) receptors in human spermatozoa has been investigated by equilibrium binding studies in intact spermatozoa, ligand blot and Western blot analysis of sperm lysates, as well as determination of the effects of the steroid on sperm intracellular Ca21 concentrations. Binding experiments were performed using progesterone-11a-glucuronide-(125I)iodotyra- mine as tracer. Computer analysis of competition curves using




Toxicity of titanium oxide nanoparticles causes functionality and DNA damage in buffalo (Bubalus bubalis) sperm in vitro.  


The present study has examined the effect of different concentrations (1 ?g/ml, 10 ?g/ml and 100 ?g/ml) of titanium oxide (TiO(2)) nanoparticles (NPs) (<100 nm) on viability, membrane integrity, capacitation status and DNA integrity of buffalo spermatozoa. Characterization of NPs was done by the transmission electron microscopy (TEM) and dynamic light scattering (DLS). Sperm chromatin dispersion (SCD) test and acridine orange test (AOT) were employed to detect DNA fragmentation in sperm treated with NPs. There was significant (p < 0.05) decrease in cell viability and membrane integrity (assessed by enzyme leakage) at 6 h of incubation with NPs. However, significant (p < 0.05) increase in sperm capacitation was observed for TiO(2) NP albeit at lower concentrations. In DNA fragmentation assay, there was dose-dependent increase in the DNA fragmentation (r = 0.96). Ultrathin cross-sections revealed TiO(2) NPs inside head and plasma membrane of the buffalo spermatozoa as assessed by TEM. These studies suggest that TiO(2) NPs may have cytotoxic effect on buffalo spermatozoa by affecting sperm functionality and causing high amount of DNA fragmentations. PMID:23064765

Pawar, Kamlesh; Kaul, Gautam



The effects of deletions of the mouse Y chromosome long arm on sperm function--intracytoplasmic sperm injection (ICSI)-based analysis.  


In mouse and man, Y chromosome deletions are frequently associated with spermatogenic defects. XY(Tdy)(m1)qdelSry males have an extensive Yq deletion that almost completely abolishes the expression of two gene families, Ssty and Sly, located within the male-specific region of the mouse Y long arm. These males exhibit severe sperm defects and sterility. XY(RIII)qdel males have a smaller interstitial Yq deletion, removing approximately two thirds of Ssty/Sly gene copies, and display an increased incidence of mild sperm head anomalies with impairment of fertility and an intriguing distortion in the sex ratio of offspring in favor of females. Here we used intracytoplasmic sperm injection (ICSI) to investigate the functional capacity of sperm from these Yq deletion males. Any selection related to the ability of sperm to fertilize in vitro is removed by ICSI, and we obtained two generations of live offspring from the infertile males. Genotyping of ICSI-derived offspring revealed that the Y(Tdym1)qdel deletion does not interfere with production of Y chromosome-bearing gametes, as judged from the frequency of Y chromosome transmission to the offspring. ICSI results for XY(RIII)qdel males also indicate that there is no deficiency of Y sperm production in this genotype, although the data show an excess of females following in vitro fertilization and natural mating. Our findings suggest that 1) Yq deletions in mice do not bias the primary sex ratio and 2) Y(RIII)qdel spermatozoa have poorer fertilizing ability than their X-bearing counterparts. Thus, a normal complement of the Ssty and/or Sly gene families on mouse Yq appears necessary for normal sperm function. Summary: ICSI was successfully used to reproduce infertile mice with Yq deletions, and the analysis of sperm function in obtained offspring demonstrated that gene families located within the deletion interval are necessary for normal sperm function. PMID:16354792

Ward, Monika A; Burgoyne, Paul S



Sperm aster in rabbit zygotes: its structure and function  

Microsoft Academic Search

Microscope observations of rabbit zygotes demonstrate that a sperm aster forms in association with the male pronucleus approximately 1 h postinsemination and consists of two regions. One, the centrosphere, contains a dense aggregation of cisternae of smooth endoplasmic reticulum and microtubules. The second consists of fascicles of microtubules which emanate from the centrosphere. Fertilized rabbit eggs were cultured in medium




Effects of antiepileptic drugs on reproductive endocrine function, sexual function and sperm parameters in Chinese Han men with epilepsy.  


The effects of the antiepileptic drugs sodium valproate (VPA) and levetiracetam (LEV) on reproductive endocrine function, sexual function, and spermatozoa were explored, together with their possible etiological mechanisms, in Chinese Han men with epilepsy. Following VPA treatment (n=32), luteinizing hormone and follicle-stimulating hormone levels were significantly lower than in controls (n=30). The bioactive testosterone/luteinizing hormone ratio and the prolactin level were significantly elevated in the VPA treatment group. There were no significant differences in these hormones between the LEV treatment (n=20) and control groups. The rates of sperm morphologic abnormality (head, body, and tail) were significantly higher in the VPA treatment group than the control group but did not differ significantly between the LEV treatment and control groups. The sperm motility rate was significantly lower in the VPA treatment group (grade A sperm motility rate <25%, grade A+B sperm motility rate <50%) than in controls, as well as in the LEV treatment group (grade A sperm motility rate <25%). Patients in the VPA and LEV treatment groups had lower scores on questions 1, 2 and 3 of a simplified International Index of Erectile Function Scale than controls, but no significant difference on questions 4 or 5. The total International Index of Erectile Function Scale scores were significantly lower in the VPA and LEV treatment groups. We conclude that treatment with VPA adversely affects reproductive endocrine function, sperm parameters and sexual function to varying degrees in Chinese men with epilepsy. PMID:23938015

Xiaotian, Xu; Hengzhong, Zhang; Yao, Xu; Zhipan, Zhao; Daoliang, Xu; Yumei, Wu



Sperm competition, sperm numbers and sperm quality in muroid rodents.  


Sperm competition favors increases in relative testes mass and production efficiency, and changes in sperm phenotype that result in faster swimming speeds. However, little is known about its effects on traits that contribute to determine the quality of a whole ejaculate (i.e., proportion of motile, viable, morphologically normal and acrosome intact sperm) and that are key determinants of fertilization success. Two competing hypotheses lead to alternative predictions: (a) sperm quantity and quality traits co-evolve under sperm competition because they play complementary roles in determining ejaculate's competitive ability, or (b) energetic constraints force trade-offs between traits depending on their relevance in providing a competitive advantage. We examined relationships between sperm competition levels, sperm quantity, and traits that determine ejaculate quality, in a comparative study of 18 rodent species using phylogenetically controlled analyses. Total sperm numbers were positively correlated to proportions of normal sperm, acrosome integrity and motile sperm; the latter three were also significantly related among themselves, suggesting no trade-offs between traits. In addition, testes mass corrected for body mass (i.e., relative testes mass), showed a strong association with sperm numbers, and positive significant associations with all sperm traits that determine ejaculate quality with the exception of live sperm. An "overall sperm quality" parameter obtained by principal component analysis (which explained 85% of the variance) was more strongly associated with relative testes mass than any individual quality trait. Overall sperm quality was as strongly associated with relative testes mass as sperm numbers. Thus, sperm quality traits improve under sperm competition in an integrated manner suggesting that a combination of all traits is what makes ejaculates more competitive. In evolutionary terms this implies that a complex network of genetic and developmental pathways underlying processes of sperm formation, maturation, transport in the female reproductive tract, and preparation for fertilization must all evolve in concert. PMID:21464956

Gómez Montoto, Laura; Magaña, Concepción; Tourmente, Maximiliano; Martín-Coello, Juan; Crespo, Cristina; Luque-Larena, Juan José; Gomendio, Montserrat; Roldan, Eduardo R S




EPA Science Inventory

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


Tripeptidyl peptidase II regulates sperm function by modulating intracellular Ca(2+) stores via the ryanodine receptor.  


Recent studies have identified Ca(2+) stores in sperm cells; however, it is not clear whether these Ca(2+) stores are functional and how they are mobilized. Here, in vitro and in vivo, we determined that tripeptidyl peptidase II antagonists strongly activated the cAMP/PKA signaling pathway that drives sperm capacitation-associated protein tyrosine phosphorylation. We demonstrated that in the absence of Ca(2+), TPIII antagonists elevated the intracellular Ca(2+) levels in sperm, resulting in a marked improvement in sperm movement, capacitation, acrosome reaction, and the in vitro fertilizing ability. This antagonist-induced release of intracellular Ca(2+) could be blocked by the inhibitors of ryanodine receptors (RyRs) which are the main intracellular Ca(2+) channels responsible for releasing stored Ca(2+). Consistent with these results, indirect immunofluorescence assay using anti-RyR antibodies further validated the presence of RyR3 in the acrosomal region of mature sperm. Thus, TPPII can regulate sperm maturation by modulating intracellular Ca(2+) stores via the type 3 RyR. PMID:23818952

Zhou, Yuchuan; Ru, Yanfei; Wang, Chunmei; Wang, Shoulin; Zhou, Zuomin; Zhang, Yonglian



Lipid rafts function in Ca2+ signaling responsible for activation of sperm motility and chemotaxis in the ascidian Ciona intestinalis.  


Lipid rafts are specialized membrane microdomains that function as signaling platforms across plasma membranes of many animal and plant cells. Although there are several studies implicating the role of lipid rafts in capacitation of mammalian sperm, the function of these structures in sperm motility activation and chemotaxis remains unknown. In the ascidian Ciona intestinalis, egg-derived sperm activating- and attracting-factor (SAAF) induces both activation of sperm motility and sperm chemotaxis to the egg. Here we found that a lipid raft disrupter, methyl-?-cyclodextrin (MCD), inhibited both SAAF-induced sperm motility activation and chemotaxis. MCD inhibited both SAAF-promoted synthesis of intracellular cyclic AMP and sperm motility induced by ionophore-mediated Ca(2+) entry, but not that induced by valinomycin-mediated hyperpolarization. Ca(2+)-imaging revealed that lipid raft disruption inhibited Ca(2+) influx upon activation of sperm motility. The Ca(2+)-activated adenylyl cyclase was clearly inhibited by MCD in isolated lipid rafts. The results suggest that sperm lipid rafts function in signaling upstream of cAMP synthesis, most likely in SAAF-induced Ca(2+) influx, and are required for Ca(2+)-dependent pathways underlying activation and chemotaxis in Ciona sperm. PMID:21887722

Zhu, Lihong; Inaba, Kazuo



Sperm Donation  


... for infectious diseases, such as HIV. If all of your test results come back negative, your frozen sample will be thawed and sperm quantity, quality and movement will be evaluated again. Sperm samples from some men are more susceptible to damage during the freezing ...


Clinical and Consumer Trial Performance of a Sensitive Immunodiagnostic Home Test That Qualitatively Detects Low Concentrations of Sperm Following Vasectomy  

PubMed Central

Purpose Compliance with post-vasectomy semen analysis could be improved with the availability of a simple, rapid and accurate home test. SpermCheck Vasectomy®, a highly sensitive lateral flow immunochromatographic diagnostic device, was designed to detect extreme oligospermia or azoospermia in men after vasectomy. We report the results of clinical and consumer testing of SpermCheck. Materials and Methods A prospective, noncomparative observational study assessed the ability of SpermCheck Vasectomy to predict post-vasectomy sperm counts obtained using a hemacytometer procedure based on standard World Health Organization methodology. Consumer studies evaluated ease of use. Results A cohort of 144 post-vasectomy semen samples was tested in the clinical trial. SpermCheck was 96% accurate in predicting whether sperm counts were greater or less than a threshold of 250,000 sperm per ml, a level associated with little or no risk of pregnancy. Sensitivity was 93% (95% CI 79% to 98%) and specificity was 97% (91% to 99%). The positive predictive value of the test was 93% (79% to 98%), and most importantly the negative predictive value was 97% (91% to 99%). The test gave a positive result 100% of the time at sperm concentrations of 385,000/ml or greater. Consumer studies with 109 lay volunteers showed that SpermCheck was easy to use. Volunteers obtained the correct or expected test result in every case and the correct response rate on a 20 question survey about the test was 97%. Conclusions SpermCheck Vasectomy, a simple and reliable immunodiagnostic test that can provide evidence of vasectomy success or failure, offers a useful alternative to improve compliance with post-vasectomy sperm monitoring. It is currently the only Food and Drug Administration approved test for this purpose.

Klotz, Kenneth L.; Coppola, Michael A.; Labrecque, Michel; Brugh, Victor M.; Ramsey, Kim; Kim, Kyung-ah; Conaway, Mark R.; Howards, Stuart S.; Flickinger, Charles J.; Herr, John C.



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

PubMed Central

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

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



High-resolution mapping of sperm function defects in the t complex fourth inversion.  


Structural variants of the mouse Chr 17-specific t complex, known as t haplotypes, express factors that alter the ability of sperm to carry out their roles in the normal fertilization process. In previous studies of males carrying heterospecific combinations of the t complex, we discovered a unique M. spretus/t haplotype phenotype of male sterility. In additional studies with mice carrying a series of M. spretus-M. m. domesticus recombinant Chr 17 homologs and a complete t haplotype (S-+/t), we monitored physiological aspects of sperm function to map a locus (Hst6) responsible for expression of the t-specific "curlicue" sperm flagellar curvature phenotype to 1 cM within the fourth inversion of the t complex. In the present report, we quantitatively analyze the in vitro capability of sperm from mice with similar S-+/t Chr 17 genotypes to fertilize zona pellucida-free mouse eggs. The results identify a locus, Stop1, mapping distal to Pim1, with acute effects on the ability of sperm to penetrate the oolemma. The data suggest that Stop1 is a complex locus consisting of at least two genetic elements, a proximal one overlapping the Hst6 locus, and another, distal to the Hst6 locus. Further quantitative analyses of the "curlicue" phenotype produced by sperm derived from these same animals indicate that expression of this chronic flagellar curvature phenotype also derives from at least two elements, both mapping within the Hst6 locus. Thus, these studies provide higher resolution mapping of the molecular basis of t haplotype-specific sperm dysfunction emanating from In(17)4. PMID:9745038

Redkar, A A; Olds-Clarke, P; Dugan, L M; Pilder, S H



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

PubMed Central

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

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



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

SciTech Connect

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

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



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

Microsoft Academic Search

Flow cytometry is now a recognized methodology within animal spermatology, and has moved from being a research tool to become routine in the assessment of animal semen destined to breeding. The availability of ‘bench-top’ flow cytometers and of newer and versatile markers for cell structure and function had allowed the instrumentation to measure more sperm parameters, from viability to reactiveness

Anders Johannisson; Margareta Wallgren; Szabolcs Nagy; Amanda Pimenta Siqueira; Heriberto Rodriguez-Martinez



Effects of Sperm Conjugation and Dissociation on Sperm Viability In Vitro  

PubMed Central

Sperm conjugation is an unusual variation in sperm behavior where two or more spermatozoa physically unite for motility or transport through the female reproductive tract. Conjugation has frequently been interpreted as sperm cooperation, including reproductive altruism, with some sperm advancing their siblings toward the site of fertilization while ostensibly forfeiting their own ability to fertilize through damage incurred during conjugate break-up. Conversely, conjugation has been proposed to protect sensitive regions of spermatozoa from spermicidal conditions within the female reproductive tract. We investigated the possibility of dissociation-induced sperm mortality and tested for a protective function of conjugation using the paired sperm of the diving beetle, Graphoderus liberus. Sperm conjugates were mechanically dissociated and exposed to potentially damaging tissue extracts of the female reproductive tract and somatic tissue. We found no significant difference in viability between paired sperm and dissociated, single sperm. The results further indicate that the reproductive tract of female G. liberus might not be spermicidal and conjugation is not protective of sperm viability when damaging conditions do exist. Our results support the interpretation that, at least in some taxa, sperm conjugation is neither protective nor damaging to sperm viability.

Higginson, Dawn M.; Henn, Kali R. H.



Acrosome reaction of sperm in the mud crab Scylla serrata as a sensitive toxicity test for metal exposures.  


In order to test the sensitivity of the sperm cell of the mud crab Scylla serrata to heavy metals, the toxic effects of Ag+, Cd2+, Cu2+, and Zn2+ on the acrosome reaction (AR) were studied by artificially inducing the AR of sperm exposed to heavy metals, counting the AR rates by light microscopy, and observing structural changes in sperm by transmission electron microscopy. The AR in S. serrata occurs at two stages. The first stage (ARI) is the eversion of the subacrosomal material. The second stage (ARII) is the ejection of the acrosomal filament. The results showed the EC50 values of the AR based on (ARI + ARII)% for Ag+, Cd2+, Cu2+, and Zn2+ were 10.02, 2.14, 13.69, and 2.21 microg/L, and the EC50 values based on ARII % of Ag+, Cd2+, Cu2+, and Zn2+ were 1.96, 0.20, 1.46, and 0.34 microg/L. The order of toxicity is Cd2+ > Zn2+ > Cu2+ > Ag+ based on the percentage of reacted sperm at the second stage. Sperm cells exposed to heavy metals showed an increased rate of swelling, shape irregularities, and the acrosomal filament of some sperm cells was, crooked, ruptured, and even dissolved. The AR of the sperm cell from S. serrata is more sensitive to the tested heavy metals compared to sea urchin sperm cell toxicity tests. PMID:19399549

Zhang, Ziping; Cheng, Hua; Wang, Yilei; Wang, Shuhong; Xie, Fangjing; Li, Shaojin



A Common Mutation in DEFB126 Causes Impaired Sperm Function and Subfertility  

PubMed Central

A glycosylated polypeptide, ?-defensin 126 (DEFB126), derived from the epididymis and adsorbed onto the sperm surface, has been implicated in immunoprotection and efficient movement of sperm in mucosal fluids of the female reproductive tract. Here, we report a sequence variant in DEFB126 that has a 2-nucleotide deletion in the open reading frame, which generates a non-stop mRNA. The allele frequency of this variant sequence is high in both a European (0.47) and a Chinese (0.45) population cohort. Binding of the Agaricus bisporus lectin to the sperm surface glycocalyx was significantly lower in men with the homozygous variant (del/del) genotype than in those with either a del/wt or wt/wt genotype, suggesting an altered sperm glycocalyx with fewer O-linked oligosaccharides in del/del men. Moreover, sperm from the del/del donors exhibited an 84% reduction in the rate of penetration of a hyaluronic acid (HA) gel, a surrogate for cervical mucus, compared to the other genotypes. This reduction in sperm performance in HA gels was not a result of decreased progressive motility (average curvilinear velocity) or morphological deficits. However, DEFB126 genotype and lectin binding were highly correlated with performance in the penetration assays. In a prospective cohort study of newly married couples who were trying to conceive by natural means, couples were less likely to become pregnant and took longer to achieve a live birth if the male partner was homozygous for the variant sequence. This common sequence variation in DEFB126, and its apparent cause of impaired reproductive function, provides an opportunity to better understand, clinically evaluate, and possibly treat human infertility.

Tollner, Theodore L.; Venners, Scott A.; Hollox, Edward J.; Yudin, Ashley I.; Liu, Xue; Tang, Genfu; Xing, Houxun; Kays, Robert J.; Lau, Tsang; Overstreet, James W.; Xu, Xiping; Bevins, Charles L.; Cherr, Gary N.



The relationship between sperm morphology and chromatin integrity in the koala (Phascolarctos cinereus) as assessed by the Sperm Chromatin Dispersion test (SCDt).  


Koala (Phascolarctos cinereus) sperm nuclei show a tendency to swell after cryopreservation, but it is uncertain whether this phenomenon is associated with DNA fragmentation. In this study, we validated a modified version of the sperm chromatin dispersion test (SCDt) for use with koala spermatozoa, which is the first use of the test for a marsupial. Cryopreserved spermatozoa (multiple straws) from a single koala were used to explore the relationship between sperm morphology, viability, chromatin dispersion, and DNA fragmentation. A SCDt prototype kit (Sperm Halomax) was specifically developed for koala spermatozoa with the use of a lysing solution that did not contain dithiothreitol. DNA fragmentation of lysed and nonlysed spermatozoa was examined in microgel slides and validated by means of in situ nick translation (ISNT). The SCDt was then applied to the analysis of extended and frozen-thawed semen samples of 3 different koalas. Spermatozoa were classified into 3 distinct koala sperm morphotypes (KSMs) after the SCDt: 1) KSM-1, rod-shaped cells with no halo of DNA; 2) KSM-2, rounded nuclei with various degrees of halo formation about a dense chromatin core; and 3) KSM-3, rod-shaped or rounded nuclei consisting of an inner chromatin core but with large dispersed halos of stellar chromatin. Although ISNT after the SCDt did not label KSM-1, both KSM-2 and KSM-3 stained positively for DNA fragmentation. ISNT was not able to differentiate between KSM-2 and KSM-3. Although application of the SCDt to the spermatozoa of another 3 koalas showed no difference in the percentage of the 3 sperm morphotypes found between extended and frozen-thawed semen, thawed spermatozoa incubated at 35 degrees C for 2 hours showed an increase in the incidence of KSM-3 and a corresponding decrease in KSM-2. We propose that KSM-1 and KSM-2 represent nuclei that show either no, or only limited, sperm DNA fragmentation, respectively. It is likely that the halos formed around KSM-2 are from DNA that is damaged as part of the normal processing of the spermatozoa and is a consequence of the lack of cysteine residues and associated stabilizing disulfide bonds in marsupial sperm DNA. "True" sperm DNA damage is most likely associated with KSM-3, which shows a massive dispersion of chromatin similar to that described in other species. A model of koala sperm chromatin structure is proposed to explain the behavior of the sperm nuclei after the SCDt. Further studies are required to determine whether DNA damage found in KSM-2 is indicative of single-stranded DNA breakage associated with an inherent lack of cysteine residues in marsupial sperm chromatin. Conversely, it will also be important to establish whether KSM-3 is caused by an increased incidence of double-stranded DNA breakage and whether this abnormality is correlated with impaired fertility as it is in other species. PMID:17609294

Johnston, Stephen D; López-Fernández, Carmen; Gosálbez, Altea; Zee, Yengpeng; Holt, William V; Allen, Camryn; Gosálvez, Jaime



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

Microsoft Academic Search

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

R. Robin Baker; Mark A. Bellis


Integrated and independent evolution of heteromorphic sperm types  

PubMed Central

Sperm are a simple cell type with few components, yet they exhibit tremendous between-species morphological variation in those components thought to reflect selection in different fertilization environments. However, within a species, sperm components are expected to be selected to be functionally integrated for optimal fertilization of eggs. Here, we take advantage of within-species variation in sperm form and function to test whether sperm components are functionally and genetically integrated both within and between sperm morphologies using a quantitative genetics approach. Drosophila pseudoobscura males produce two sperm types with different functions but which positively interact together in the same fertilization environment; the long eusperm fertilizes eggs and the short parasperm appear to protect eusperm from a hostile female reproductive tract. Our analysis found that all sperm traits were heritable, but short sperm components exhibited evolvabilities 10 times that of long sperm components. Genetic correlations indicated functional integration within, but not between, sperm morphs. These results suggest that sperm, despite sharing a common developmental process, can become developmentally and functionally non-integrated, evolving into separate modules with the potential for rapid and independent responses to selection.

Moore, Allen J.; Bacigalupe, Leonardo D.; Snook, Rhonda R.



Evidence that anandamide-signaling regulates human sperm functions required for fertilization.  


Ejaculated mammalian sperm require several hours exposure to secretions in female reproductive tracts, or incubation in appropriate culture medium in vitro, before acquiring the capacity to fertilize eggs. Arachidonylethanolamide (AEA), also known as anandamide, is a novel lipid-signal molecule that is an endogenous agonist (endocannabinoid) for cannabinoid receptors. We now report that AEA is present in human seminal plasma, mid-cycle oviductal fluid, and follicular fluid analyzed by high-performance liquid chromatography/mass spectrometry. Sperm are sequentially exposed to these reproductive fluids as they move from the vagina to the site of fertilization in the oviduct. Specific binding of the potent cannabinoid agonist [(3)H]CP-55,940 to human sperm was saturable (K(D) 9.71 +/- 1.04 nM), suggesting that they express cannabinoid receptors. R-methanandamide [AM-356], a potent and metabolically stable AEA analog, and (-)delta(9) tetrahydrocannabinol (THC), the major psychoactive constituent of Cannabis, modulated capacitation and fertilizing potential of human sperm in vitro. AM-356 elicited biphasic effects on the incidence of hyperactivated sperm motility (HA) between 1 and 6 hr of incubation: at (2.5 nM) it inhibited HA, while at (0.25 nM) it stimulated HA. Both AM-356 and THC inhibited morphological alterations over acrosomal caps between 2 and 6 hr (IC(50) 5.9 +/- 0.6 pM and 3.5 +/- 1.5 nM, respectively). Sperm fertilizing capacity, measured in the Hemizona Assay, was reduced 50% by (1 nM) AM-356. These findings suggest that AEA-signaling may regulate sperm functions required for fertilization in human reproductive tracts, and imply that smoking of marijuana could impact these processes. This study has potential medical and public policy ramifications because of the incidence of marijuana abuse by adults in our society, previously documented reproductive effects of marijuana, and the ongoing debate about medicinal use of marijuana and cannabinoids. PMID:12237954

Schuel, Herbert; Burkman, Lani J; Lippes, Jack; Crickard, Kent; Mahony, Mary C; Giuffrida, Andrea; Picone, Robert P; Makriyannis, Alexandros



Kinematic study on the effect of pH on bull sperm function.  


Since the mammalian spermatozoa became capable of motion, during the epididymal transit, the spermatozoon swims in a liquid medium and it is completely dependent on the environmental conditions. Some reports have suggested an influence of pH on sperm kinetic characteristics, but no study has objectively described how motility changes in a different environmental pH. In this study, we evaluated the effect of different environmental pHs (5.5, 6, 6.5, 7, 7.5, 8, and 8.5) on kinetic parameters, sperm viability, mitochondrial activity, and sperm morphology of bull semen immediately and 1h after dilution. The results showed higher values for sperm motility characteristics, viability, and mitochondrial activity at pH 7 and 7.5. Values of pH lower than 6.5 and higher than 8 resulted in suboptimal motility, with a decrease in most parameters. At pH 8 and 8.5, a discrepancy between viability and total and progressive motility was found, with a significant amount of spermatozoa that were live but immotile. This reduction seemed related to a decrease in mitochondrial activity, possibly due to the increase in pH. The flow cytometric evaluation of sperm viability assessed by calcein AM was very consistent with the amount of spermatozoa with membrane integrity, evaluated in fluorescence by propidium iodide/SYBR-14 stain. Thus, the calcein AM stain could be used as viability stain instead the classic propidium iodide/SYBR-14 stain because this could allow the addiction of other functional stains without a overlapping of the fluorescent signal in the flow cytometer. PMID:23238050

Contri, Alberto; Gloria, Alessia; Robbe, Domenico; Valorz, Claudio; Wegher, Laura; Carluccio, Augusto



Poor Centrosomal Function of Cat Testicular Spermatozoa Impairs Embryo Development In Vitro after Intracytoplasmic Sperm Injection1  

PubMed Central

In the domestic cat, morula-blastocyst formation in vitro is compromised after intracytoplasmic sperm injection (ICSI) with testicular compared to ejaculated spermatozoa. The aim of this study was to determine the cellular basis of the lower developmental potential of testicular spermatozoa. Specifically, we examined the influence of sperm DNA fragmentation (evaluated by TUNEL assay) and centrosomal function (assessed by sperm aster formation after ICSI) on first-cleavage timing, developmental rate, and morula-blastocyst formation. Because the incidences of DNA fragmentation were not different between testicular and ejaculated sperm suspensions, DNA integrity was not the origin of the reduced developmental potential of testicular spermatozoa. After ICSI, proportions of fertilized and cleaved oocytes were similar and not influenced by sperm source. However, observations made at 5 h post-activation clearly demonstrated that 1) zygotes generally contained a large sperm aster after ICSI with ejaculated spermatozoa, a phenomenon never observed with testicular spermatozoa, and 2) proportions of zygotes with short or absent sperm asters were higher after ICSI with testicular spermatozoa than using ejaculated spermatozoa. The poor pattern of aster formation arose from the testicular sperm centrosome, which contributed to a delayed first cleavage, a slower developmental rate, and a reduced formation of morulae and blastocysts compared to ejaculated spermatozoa. When a testicular sperm centrosome was replaced by a centrosome from an ejaculated spermatozoon, kinetics of first cell cycle as well as embryo development quality significantly improved and were comparable to data from ejaculated spermatozoa. Results demonstrate for the first time in mammals that maturity of the cat sperm centrosome (likely via epididymal transit) contributes to an enhanced ability of the spermatozoon to produce embryos that develop normally to the morula and blastocyst stages.

Comizzoli, Pierre; Wildt, David E.; Pukazhenthi, Budhan S.



Sperm Chromatin  

PubMed Central

Sperm are remarkably complex cells with a singularly important mission: to deliver paternal DNA and its associated factors to the oocyte to start a new life. The integrity of sperm DNA is a keystone of reproductive success, which includes fertilization and embryonic development. In addition, the significance in these processes of proteins that associate with sperm DNA is increasingly being appreciated. In this review, we highlight proteomic studies that have identified sperm chromatin proteins with fertility roles that have been validated by molecular studies in model organisms or correlations in the clinic. Up to 50% of male-factor infertility cases in the clinic have no known cause and therefore no direct treatment. In-depth study of the molecular basis of infertility has great potential to inform the development of sensitive diagnostic tools and effective therapies that will address this incongruity. Because sperm rely on testis-specific protein isoforms and post-translational modifications for their development and function, sperm-specific processes are ideal for proteomic explorations that can bridge the research lab and fertility clinic.

Wu, Tammy F.; Chu, Diana S.



Pulmonary function tests  


PFTs; Spirometry; Spirogram; Lung function tests ... Pulmonary function tests are done to: Diagnose certain types of lung disease (such as asthma , bronchitis , and emphysema) Find the cause of shortness of breath Measure whether exposure ...


Thyroid Function Tests  


... Services NATIONAL INSTITUTES OF HEALTH What is the thyroid gland? The thyroid is a small, butterfly-shaped ... and their release into the bloodstream. Why are thyroid function tests performed? Thyroid function tests are used ...


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


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

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



Response to capacitating stimuli indicates extender-related differences in boar sperm function.  


Spermatozoa, especially those of the porcine species, are highly susceptible to in vitro chilling and ageing. Extenders are continuously developed to protect boar spermatozoa from chilling injury. New semen extenders and other modified preservation strategies require sensitive testing for essential sperm functions. The key process on the pathway of fertilization is capacitation. The aim of the present study was to examine whether the specific response to capacitating stimuli is sensitive enough to indicate different preservation capacities of extenders during hypothermic storage of boar spermatozoa. Semen was diluted in Beltsville Thawing Solution (BTS) and Androstar Plus and kept for 3 h at 22°C or stored at 17°C, 10°C, and 5°C. Semen was analyzed at 24 and 96 h of storage. Motility and membrane integrity remained at high levels, except for lower values when stored in BTS at 5°C. Washed subsamples were incubated in capacitating medium (Tyrode) and control medium and were assessed for intracellular calcium concentration and integrity of plasma membranes using a flow cytometer. On the basis of the loss of low-calcium live cells in a kinetic approach, the specific response to capacitation stimuli was determined. There was a higher loss of response in semen stored hypothermically in the standard extender BTS compared to Androstar Plus. Assessment of the extent of phospholipid disorder under capacitating and control conditions by use of merocyanine staining did not reveal any significant extender-related differences. A field insemination trial with 778 sows was performed to relate in vitro results to fertility. Fertility parameters did not differ in semen stored up to 48 h at 10°C in Androstar Plus compared to controls stored at 17°C in BTS. In conclusion, assessment of specific reactivity to capacitating stimuli appears to be a sensitive tool for detection of extender-dependent alterations in functionality of chilled boar spermatozoa. PMID:23989873

Schmid, S; Henning, H; Petrunkina, A M; Weitze, K F; Waberski, D



The stochastic dance of circling sperm cells: sperm chemotaxis in the plane  

NASA Astrophysics Data System (ADS)

Biological systems such as single cells must function in the presence of fluctuations. It has been shown in a two-dimensional experimental setup that sea urchin sperm cells move toward a source of chemoattractant along planar trochoidal swimming paths, i.e. drifting circles. In these experiments, a pronounced variability of the swimming paths is observed. We present a theoretical description of sperm chemotaxis in two dimensions which takes fluctuations into account. We derive a coarse-grained theory of stochastic sperm swimming paths in a concentration field of chemoattractant. Fluctuations enter as multiplicative noise in the equations for the sperm swimming path. We discuss the stochastic properties of sperm swimming and predict a concentration-dependence of the effective diffusion constant of sperm swimming which could be tested in experiments.

Friedrich, B. M.; Jülicher, F.



Reproductive effects of two neonicotinoid insecticides on mouse sperm function and early embryonic development in vitro.  


Acetamiprid (ACE) and imidacloprid (IMI) are two major members in the family of neonicotinoid pesticides, which are synthesized with a higher selectivity to insects. The present study determined and compared in vitro effects of ACE, IMI and nicotine on mammalian reproduction by using an integrated testing strategy for reproductive toxicology, which covered sperm quality, sperm penetration into oocytes and preimplantation embryonic development. Direct chemical exposure (500 µM or 5 mM) on spermatozoa during capacitation was performed, and in vitro fertilization (IVF) process, zygotes and 2-cell embryos were respectively incubated with chemical-supplemented medium until blastocyst formation to evaluate the reproductive toxicity of these chemicals and monitor the stages mainly affected. Generally, treatment of 500 µM or 5 mM chemicals for 30 min did not change sperm motility and DNA integrity significantly but the fertilization ability in in vitro fertilization (IVF) process, indicating that IVF process could detect and distinguish subtle effect of spermatozoa exposed to different chemicals. Culture experiment in the presence of chemicals in medium showed that fertilization process and zygotes are adversely affected by direct exposure of chemicals (P<0.05), in an order of nicotine>IMI>ACE, whereas developmental progression of 2-cell stage embryos was similar to controls (P>0.05). These findings unveiled the hazardous effects of neonicotinoid pesticides exposure on mammalian sperm fertilization ability as well as embryonic development, raising the concerns that neonicotinoid pesticides may pose reproductive risks on human reproductive health, especially in professional populations. PMID:23922925

Gu, Yi-Hua; Li, Yan; Huang, Xue-Feng; Zheng, Ju-Fen; Yang, Jun; Diao, Hua; Yuan, Yao; Xu, Yan; Liu, Miao; Shi, Hui-Juan; Xu, Wen-Ping



Ovarian fluid mediates the temporal decline in sperm viability in a fish with sperm storage.  


A loss of sperm viability and functionality during sperm transfer and storage within the female reproductive tract can have important fitness implications by disrupting fertilization and impairing offspring development and survival. Consequently, mechanisms that mitigate the temporal decline in sperm function are likely to be important targets of selection. In many species, ovarian fluid is known to regulate and maintain sperm quality. In this paper, we use the guppy Poecilia reticulata, a highly polyandrous freshwater fish exhibiting internal fertilization and sperm storage, to determine whether ovarian fluid (OF) influences the decline in sperm viability (the proportion of live sperm in the ejaculate) over time and whether any observed effects depend on male sexual ornamentation. To address these questions we used a paired experimental design in which ejaculates from individual males were tested in vitro both in presence and absence of OF. Our results revealed that the temporal decline in sperm viability was significantly reduced in the presence of OF compared to a saline control. This finding raises the intriguing possibility that OF may play a role in mediating the decline in sperm quality due to the deleterious effects of sperm ageing, although other possible explanations for this observation are discussed. Interestingly, we also show that the age-related decline in sperm viability was contingent on male sexual ornamentation; males with relatively high levels of iridescence (indicating higher sexual attractiveness) exhibited a more pronounced decline in sperm viability over time than their less ornamented counterparts. This latter finding offers possible insights into the functional basis for the previously observed trade-off between these key components of pre- and postcopulatory sexual selection. PMID:23691216

Gasparini, Clelia; Evans, Jonathan P



Andrology Lab Corner* The Clinical Implementation of Sperm Chromosome Aneuploidy Testing: Pitfalls and Promises  

Microsoft Academic Search

Severe male infertility has been shown to be associated with improper meiotic recombination and elevated sperm chromosome aneuploidy. Elevated sperm aneuploidy increases the risk of embryo lethality or fetal anomalies. Although difficulties in interpreting aneuploidy data still exist, advances in fluorescent in situ hybridization (FISH) technology have facilitated the study of sperm from patients with severe spermatogenesis defects, which has



Pulmonary Function Tests  


Patient Information Series AMERICAN THORACIC SOCIETY Lung function tests can be used to: Compare your lung function with known ? ? standards that show how well your lungs should be working. Measure the ...


Sperm-peritoneal fluid incubation test: influence of a GnRH agonist treatment.  


Objectives of our work were to determine whether the presence of endometriosis, stage and the activity of the disease influence the sperm-peritoneal fluid (PF) incubation test and, possibly, whether a GnRH agonist (GnRHa) treatment in these patients can reduce the PF toxicity toward sperms. Preovulatory PFs from 18 patients with endometriosis (11 stage I-II rAFS and/stage III-IV; 10 with active lesions and 8 with inactive disease), and from 15 women with unexplained infertility were collected and frozen at the time of diagnostic laparoscopy. The ten patients with active endometriosis were treated by 3 months ovarian desensitization by a GnRH a (Decapeptyl 3.75 mg, Ipsen, Milan Italy) followed by a controlled ovarian hyperstimulation with a purified FSH (Metrodin, Serono, Rome, Italy) and HCG 10.000 UI (Profasi, Serono) when appropriate. In these patients, immediately before performing intrauterine and/or intraperitoneal insemination, a small aliquot of PF was collected transvaginally checking the absence of oocytes. Five hundreds suspension for one hour. PFs inducing a decrease in sperm motility greater than 20% were defined "toxic". The prevalence of toxic PF was not statistically greater in endometriosis (8/18) than in unexplained infertility (3/15) and was not dependent upon the stage of the disease, while it was significantly (rho = 0.01) more frequent in patients with active endometriosis (7/10) than other women (4/23). After a three months GnRHa treatment of these patients the rate of toxic PFs was significantly (p = 0.02) decreased (1 out of 10 treated cases).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7660716

Zullo, F; Corea, D; Torano, P; Placco, C; Tropea, G; Mastrantonio, P


Pulmonary Function Tests  

PubMed Central

Pulmonary function tests are valuable investigations in the management of patients with suspected or previously diagnosed respiratory disease. They aid diagnosis, help monitor response to treatment and can guide decisions regarding further treatment and intervention. The interpretation of pulmonary functions tests requires knowledge of respiratory physiology. In this review we describe investigations routinely used and discuss their clinical implications.

Ranu, Harpreet; Wilde, Michael; Madden, Brendan



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


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

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



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

PubMed Central

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

Henkel, Ralf R



Identifying environmental risk to male reproductive function by occupational sperm studies: logistics and design options.  

PubMed Central

Malfunction of the male reproductive system might be a sensitive marker of environmental hazards, the effects of which may extend beyond reproductive function. The testis is more vulnerable to heat and ionising radiation than any other organ of the body and several xenobiotics are known to disrupt spermatogenesis after low level exposure. Studies of environmental impact on human health are often most informative and accurate when carried out in the workplace where exposures can be high and easy to document. Semen analysis provides readily obtainable information on testicular function. The main advantages in comparison with functional measures such as fertility rates and time taken to conceive are the possibilities to examine men independently of marriage and pregnancy, to find changes of fecundity with different exposures within the same person and to detect adverse effects when no alteration of fertility is yet taking place. In the implementation of an occupational sperm study considerable attention must be paid to logistic issues. A mobile laboratory unit for initial semen preparation and processing may in some situations increase worker compliance and the quality of sperm cell motility. The cross sectional design which has been used in almost all male reproductive studies so far has several severe limitations including selection bias because of differential participation, difficulties in defining a suitable reference group, and lack of information about the time dimension of the cause-effect relation. The longitudinal design deals adequately with most of these constraints. Semen samples are collected before, during, and possibly after exposure to the risk factor of interest and causal inferences are based upon change of semen variables within a man over time rather than upon differences between men. The logistics of the longitudinal study may benefit from pre-employment health examinations to enrol newly hired workers and require fewer participants to obtain comparable statistical power. In conclusion, andrological methods and epidemiological designs are available for the implementation of valid studies concerned with environmental impact on human testicular function. Occupational sperm studies should probably not be the first choice when the objective is initial screening of environmental impact on fertility but should be implemented when their is a need to corroborate or refuse earlier evidence that specific exposures have impact on testicular function. Images p514-a

Bonde, J P; Giwercman, A; Ernst, E



Prenatal testing in ICSI pregnancies: incidence of chromosomal anomalies in 1586 karyotypes and relation to sperm parameters  

Microsoft Academic Search

BACKGROUND: Prenatal testing was offered in all pregnancies obtained after ICSI with ejaculated or non- ejaculated sperm as part of the evaluation of the safety of ICSI. METHODS: Between 1990 and 2001, a chorionic villus sampling (CVS) or amniocentesis was offered for multiple or singleton pregnancies respectively during a genetic counselling session for all couples applying for ICSI. ICSI was

Maryse Bonduelle; Elvire Van Assche; Hubert Joris; Kathelijn Keymolen; Paul Devroey; AndreVan Steirteghem; Inge Liebaers


Obesity and Clomiphene Challenge Test as Predictors of Outcome of in vitro Fertilization and Intracytoplasmic Sperm Injection  

Microsoft Academic Search

One hundred and sixty-two consecutive patients undergoing in vitro fertilization (IVF) or IVF\\/intracytoplasmic sperm injection (ICSI) were studied to determine the effect of obesity on the outcome of this treatment and to evaluate the prognostic value of the Clomiphene Challenge Test (CCT) in controlled ovarian hyperstimulation. In this prospective clinical study, we assessed the mean number of stimulation days, the

Esther C. A. M. van Swieten; Loes van der Leeuw-Harmsen; Erik A. Badings



Infant Pulmonary Function Testing  

PubMed Central

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.

Davis, Stephanie D.



The CIL-1 phosphoinositide 5-phosphatase regulates ciliary localization of the TRP polycystins and sperm function in C. elegans  

PubMed Central

Summary Background C. elegans male sexual behaviors include chemotaxis and response to hermaphrodites, backing/turning, vulva location, spicule insertion and sperm transfer, culminating in cross fertilization of hermaphrodite oocytes with male sperm. The LOV-1 and PKD-2 transient receptor potential polycystin (TRPP) complex localizes to ciliated endings of C. elegans male-specific sensory neurons and mediates several aspects of male mating behavior. TRPP complex ciliary localization and sensory function is evolutionarily conserved. A genetic screen for C. elegans mutants with PKD-2 ciliary localization (Cil) defects led to the isolation of a mutation in the cil-1 gene. Results Here, we report that a phosphoinositide (PI) 5-phosphatase CIL-1 regulates TRPP complex ciliary receptor localization and sperm activation. cil-1 does not regulate the localization of other ciliary proteins, including intraflagellar transport (IFT) components, sensory receptors, or other TRP channels in different cell types. Rather, cil-1 specifically controls TRPP complex trafficking in male-specific sensory neurons and does so in a cell autonomous fashion. In these cells, cil-1 is required for normal PI(3)P distribution, indicating that a balance between PI(3,5)P2 and PI(3)P is important for TRPP localization. cil-1 mutants are infertile due to sperm activation and motility defects. In sperm, the CIL-1 5-phosphatase and a wortmannin sensitive PI 3-kinase act antagonistically to regulate the conversion of sessile spermatids into motile spermatozoa, implicating PI(3,4,5)P3 signaling in nematode sperm activation. Conclusion Our studies identify the CIL-1 5-phosphatase as key regulator of PI metabolism in cell types that are important in several aspects of male reproductive biology.

Bae, Young-Kyung; Kim, Eunsoo; L'Hernault, Steven W.; Barr, Maureen M.



Inhibition of Rabbit Sperm Cell Hyperactivated Motility by Metallic Ions in Toxicological Testing.  

National Technical Information Service (NTIS)

Rabbit sperm cells developed hyperactivated motility when incubated in medium N. Hyperactivated but not normal progressive motility was inhibited by lead and cadmium, two metals implicated in fertility dysfunction. A metal of military interest, Cr+6, had ...

R. J. Young D. H. Heitkamp



Measurement of Epididymal Sperm Motility as a Test Variable in the Rat.  

National Technical Information Service (NTIS)

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

R. E. Linder L. F. Strader W. K. McElroy



Infertile Men With Varicocele Show a High Relative Proportion of Sperm Cells With Intense Nuclear Damage Level, Evidenced by the Sperm Chromatin Dispersion Test  

Microsoft Academic Search

The frequency of sperm cells with fragmented DNA was studied in a group of 18 infertile patients with varicocele and compared with those obtained in a group of 51 normozoospermic patients, 103 patients with abnormal standard semen parameters, and 22 fertile men. The spermatozoa were processed to discriminate different levels of DNA fragmentation using the HalospermT kit, an improved Sperm



Molecular and functional characterization of voltage-gated sodium channels in human sperm  

Microsoft Academic Search

BACKGROUND: We have investigated the expression of voltage-gated sodium channels in human spermatozoa and characterized their role in sperm motility. METHODS: Freshly ejaculated semen was collected from thirty normozoospermic human donors, with each donor supplying 2 different samples. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence techniques were used to detect the mRNAs and proteins of interest. Sperm motility was measured

Francisco M Pinto; Cristina G Ravina; Manuel Fernández-Sánchez; Manuel Gallardo-Castro; Antonio Cejudo-Román; Luz Candenas



Immune Activation Reduces Sperm Quality in the Great Tit  

PubMed Central

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.

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



Calibrating Ultraviolet Irradiation of Fish Sperm  

Microsoft Academic Search

Milt collected from common carp, Cyprinus carpio, and bluegill, Lepomis macrochirus, was irradiated with ultraviolet (UV) light to evaluate the effects on sperm viability as a function of sperm concentration and UV dose. The optical density of milt samples was determined using UV spectrophotometry. Sperm viability was estimated, based on motility and fertilization of eggs following UV radiation. Sperm samples

Michael D. Porter



Sperm-associated antigen-17 gene is essential for motile cilia function and neonatal survival.  


Primary ciliary dyskinesia (PCD), resulting from defects in cilia assembly or motility, is caused by mutations in a number of genes encoding axonemal proteins. PCD phenotypes are variable, and include recurrent respiratory tract infections, bronchiectasis, hydrocephaly, situs inversus, and male infertility. We generated knockout mice for the sperm-associated antigen-17 (Spag17) gene, which encodes a central pair (CP) protein present in the axonemes of cells with "9 + 2" motile cilia or flagella. The targeting of Spag17 resulted in a severe phenotype characterized by immotile nasal and tracheal cilia, reduced clearance of nasal mucus, profound respiratory distress associated with lung fluid accumulation and disruption of the alveolar epithelium, cerebral ventricular expansion consistent with emerging hydrocephalus, failure to suckle, and neonatal demise within 12 hours of birth. Ultrastructural analysis revealed the loss of one CP microtubule in approximately one quarter of tracheal cilia axonemes, an absence of a C1 microtubule projection, and other less frequent CP structural abnormalities. SPAG6 and SPAG16 (CP proteins that interact with SPAG17) were increased in tracheal tissue from SPAG17-deficient mice. We conclude that Spag17 plays a critical role in the function and structure of motile cilia, and that neonatal lethality is likely explained by impaired airway mucociliary clearance. PMID:23418344

Teves, Maria Eugenia; Zhang, Zhibing; Costanzo, Richard M; Henderson, Scott C; Corwin, Frank D; Zweit, Jamal; Sundaresan, Gobalakrishnan; Subler, Mark; Salloum, Fadi N; Rubin, Bruce K; Strauss, Jerome F



Sperm Competition Selects for Sperm Quantity and Quality in the Australian Maluridae  

PubMed Central

When ejaculates from rival males compete for fertilization, there is strong selection for sperm traits that enhance fertilization success. Sperm quantity is one such trait, and numerous studies have demonstrated a positive association between sperm competition and both testes size and the number of sperm available for copulations. Sperm competition is also thought to favor increases in sperm quality and changes in testicular morphology that lead to increased sperm production. However, in contrast to sperm quantity, these hypotheses have received considerably less empirical support and remain somewhat controversial. In a comparative study using the Australian Maluridae (fairy-wrens, emu-wrens, grasswrens), we tested whether increasing levels of sperm competition were associated with increases in both sperm quantity and quality, as well as an increase in the relative amount of seminiferous tubule tissue contained within the testes. After controlling for phylogeny, we found positive associations between sperm competition and sperm numbers, both in sperm reserves and in ejaculate samples. Additionally, as sperm competition level increased, the proportion of testicular spermatogenic tissue also increased, suggesting that sperm competition selects for greater sperm production per unit of testicular tissue. Finally, we also found that sperm competition level was positively associated with multiple sperm quality traits, including the proportion of motile sperm in ejaculates and the proportion of both viable and morphologically normal sperm in sperm reserves. These results suggest multiple ejaculate traits, as well as aspects of testicular morphology, have evolved in response to sperm competition in the Australian Maluridae. Furthermore, our findings emphasize the importance of post-copulatory sexual selection as an evolutionary force shaping macroevolutionary differences in sperm phenotype.

Rowe, Melissah; Pruett-Jones, Stephen



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

NASA Astrophysics Data System (ADS)

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.

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



Role of the epididymis in sperm competition.  


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

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



Sperm Chromatin: An Overview  

Microsoft Academic Search

\\u000a The dramatic changes in the structure and function of sperm chromatin that occur during spermatogenesis have continued to\\u000a intrigue researchers for more than a century. In addition to wanting to understand how these changes in chromatin organization\\u000a affect genome function, many of the studies conducted in placental mammals have been driven by a desire to understand the\\u000a relationship between sperm

Rod Balhorn


Cysteine-rich secretory protein 4 is an inhibitor of transient receptor potential M8 with a role in establishing sperm function  

PubMed Central

The cysteine-rich secretory proteins (CRISPs) are a group of four proteins in the mouse that are expressed abundantly in the male reproductive tract, and to a lesser extent in other tissues. Analysis of reptile CRISPs and mouse CRISP2 has shown that CRISPs can regulate cellular homeostasis via ion channels. With the exception of the ability of CRISP2 to regulate ryanodine receptors, the in vivo targets of mammalian CRISPs function are unknown. In this study, we have characterized the ion channel regulatory activity of epididymal CRISP4 using electrophysiology, cell assays, and mouse models. Through patch-clamping of testicular sperm, the CRISP4 CRISP domain was shown to inhibit the transient receptor potential (TRP) ion channel TRPM8. These data were confirmed using a stably transfected CHO cell line. TRPM8 is a major cold receptor in the body, but is found in other tissues, including the testis and on the tail and head of mouse and human sperm. Functional assays using sperm from wild-type mice showed that TRPM8 activation significantly reduced the number of sperm undergoing the progesterone-induced acrosome reaction following capacitation, and that this response was reversed by the coaddition of CRISP4. In accordance, sperm from Crisp4 null mice had a compromised ability to undergo to the progesterone-induced acrosome reaction. Collectively, these data identify CRISP4 as an endogenous regulator of TRPM8 with a role in normal sperm function.

Gibbs, Gerard M.; Orta, Gerardo; Reddy, Thulasimala; Koppers, Adam J.; Martinez-Lopez, Pablo; Luis de la Vega-Beltran, Jose; Lo, Jennifer C. Y.; Veldhuis, Nicholas; Jamsai, Duangporn; McIntyre, Peter; Darszon, Alberto; O'Bryan, Moira K.



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

Microsoft Academic Search

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

R. Robin Baker; Mark A. Bellis


Pattern of sperm transfer in redback spiders: implications for sperm competition and male sacrifice  

Microsoft Academic Search

Many sperm competition studies have identified copulation duration as an important predictor of paternity. This result is often interpreted as a sperm transfer effect--it is assumed that sperm transfer is limited by copulation duration. Here we test the assumption of duration-dependent sperm transfer in the Australian redback spider, Latrodectus hasselti, in which a correlation between copulation duration and paternity has

Lindsay S. E. Snow; Maydianne C. B. Andrade



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

Microsoft Academic Search

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

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



The inhibition of the human sperm phosphatidylinosytol 3-kinase by LY294002 does not interfere with sperm/oocyte interaction.  


It has recently been reported that the selective inhibition of phosphatidylinosytol 3-kinase (PI3K) enhances human sperm motility. However, little information exists on a possible role of PI3K in other sperm functions involved in the fertilization process. In this study, we investigated whether LY294002 could affect human sperm ability to fuse with oocytes, by means of the hamster egg penetration test (HEPT). The effect on acrosome reactions (AR) and on sperm/zona pellucida (ZP) binding was also evaluated. The pre-incubation with scalar doses of LY294002 (0.1, 1 and 10 microm) did not interfere with sperm ability to fuse with oocytes either in the conventional version of the HEPT or in the version enhanced with progesterone (P). No interference with the stimulatory effect on AR exerted by P or mannose-bovine serum albumin (mannose-BSA) was revealed. Finally, LY294002 had no effect on sperm/ZP binding. These results indicate that the inhibition of PI3K by LY294002 does not interfere with sperm interaction with oocytes. This is noteworthy in the view of a possible clinical use of LY294002 as an in vitro stimulator of the sperm motility of asthenozoospermic patients for assisted reproduction techniques. PMID:16480410

Barbonetti, A; Zugaro, A; Sciarretta, F; Santucci, R; Necozione, S; Ruvolo, G; Francavilla, S; Francavilla, F



Kidney function tests  


... chap 8. Oh MS. Evaluation of renal function, water, electrolytes and acid-base balance. In: McPherson RA, Pincus MR, eds. Henry's Clinical Diagnosis and Management by Laboratory Methods . 22nd ed. Philadelphia, Pa: Saunders ...


Functions and Vertical Line Test  

NSDL National Science Digital Library

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.



Properties of a Novel pH-dependent Ca2+ Permeation Pathway Present in Male Germ Cells with Possible Roles in Spermatogenesis and Mature Sperm Function  

PubMed Central

Rises of intracellular Ca2+ ([Ca2+]i) are key signals for cell division, differentiation, and maturation. Similarly, they are likely to be important for the unique processes of meiosis and spermatogenesis, carried out exclusively by male germ cells. In addition, elevations of [Ca2+]i and intracellular pH (pHi) in mature sperm trigger at least two events obligatory for fertilization: capacitation and acrosome reaction. Evidence implicates the activity of Ca2+ channels modulated by pHi in the origin of these Ca2+ elevations, but their nature remains unexplored, in part because work in individual spermatozoa are hampered by formidable experimental difficulties. Recently, late spermatogenic cells have emerged as a model system for studying aspects relevant for sperm physiology, such as plasmalemmal ion fluxes. Here we describe the first study on the influence of controlled intracellular alkalinization on [Ca2+]i on identified spermatogenic cells from mouse adult testes. In BCECF [(2?,7?)-bis(carboxymethyl)- (5,6)-carboxyfluorescein]-AM-loaded spermatogenic cells, a brief (30–60 s) application of 25 mM NH4Cl increased pHi by ?1.3 U from a resting pHi ?6.65. A steady pHi plateau was maintained during NH4Cl application, with little or no rebound acidification. In fura-2-AM-loaded cells, alkalinization induced a biphasic response composed of an initial [Ca2+]i drop followed by a two- to threefold rise. Maneuvers that inhibit either Ca2+ influx or intracellular Ca2+ release demonstrated that the majority of the Ca2+ rise results from plasma membrane Ca2+ influx, although a small component likely to result from intracellular Ca2+ release was occasionally observed. Ca2+ transients potentiated with repeated NH4Cl applications, gradually obliterating the initial [Ca2+]i drop. The pH-sensitive Ca2+ permeation pathway allows the passage of other divalents (Sr2+, Ba2+, and Mn2+) and is blocked by inorganic Ca2+ channel blockers (Ni2+ and Cd2+), but not by the organic blocker nifedipine. The magnitude of these Ca2+ transients increased as maturation advanced, with the largest responses being recorded in testicular sperm. By extrapolation, these findings suggest that the pH-dependent Ca2+ influx pathway could play significant roles in mature sperm physiology. Its pharmacology and ion selectivity suggests that it corresponds to an ion channel different from the voltage-gated T-type Ca2+ channel also present in spermatogenic cells. We postulate that the Ca2+ permeation pathway regulated by pHi, if present in mature sperm, may be responsible for the dihydropyridine-insensitive Ca2+ influx required for initiating the acrosome reaction and perhaps other important sperm functions.

Santi, Celia M.; Santos, Teresa; Hernandez-Cruz, Arturo; Darszon, Alberto



Sperm preparation for ART.  


The onset of clinical assisted reproduction, a quarter of a century ago, required the isolation of motile spermatozoa. As the indication of assisted reproduction shifted from mere gynaecological indications to andrological indications during the years, this urged andrological research to understand the physiology of male germ cell better and develop more sophisticated techniques to separate functional spermatozoa from those that are immotile, have poor morphology or are not capable to fertilize oocytes. Initially, starting from simple washing of spermatozoa, separation techniques, based on different principles like migration, filtration or density gradient centrifugation evolved. The most simple and cheapest is the conventional swim-up procedure. A more sophisticated and most gentle migration method is migration-sedimentation. However, its yield is relatively small and the technique is therefore normally only limited to ejaculates with a high number of motile spermatozoa. Recently, however, the method was also successfully used to isolate spermatozoa for intracytoplasmic sperm injection (ICSI). Sperm separation methods that yield a higher number of motile spermatozoa are glass wool filtration or density gradient centrifugation with different media. Since Percoll as a density medium was removed from the market in 1996 for clinical use in the human because of its risk of contamination with endotoxins, other media like IxaPrep, Nycodenz, SilSelect, PureSperm or Isolate were developed in order to replace Percoll. Today, an array of different methods is available and the selection depends on the quality of the ejaculates, which also includes production of reactive oxygen species (ROS) by spermatozoa and leukocytes. Ejaculates with ROS production should not be separated by means of conventional swim-up, as this can severely damage the spermatozoa. In order to protect the male germ cells from the influence of ROS and to stimulate their motility to increase the yield, a number of substances can be added to the ejaculate or the separation medium. Caffeine, pentoxifylline and 2-deoxyadenosine are substances that were used to stimulate motility. Recent approaches to stimulate spermatozoa include bicarbonate, metal chelators or platelet-activating factor (PAF). While the use of PAF already resulted in pregnancies in intrauterine insemination, the suitability of the other substances for the clinical use still needs to be tested. Finally, the isolation of functional spermatozoa from highly viscous ejaculates is a special challenge and can be performed enzymatically to liquefy the ejaculate. The older method, by which the ejaculate is forcefully aspirated through a narrow-gauge needle, should be abandoned as it can severely damage spermatozoa, thus resulting in immotile sperm. PMID:14617368

Henkel, Ralf R; Schill, Wolf-Bernhard



Sperm preparation for ART  

PubMed Central

The onset of clinical assisted reproduction, a quarter of a century ago, required the isolation of motile spermatozoa. As the indication of assisted reproduction shifted from mere gynaecological indications to andrological indications during the years, this urged andrological research to understand the physiology of male germ cell better and develop more sophisticated techniques to separate functional spermatozoa from those that are immotile, have poor morphology or are not capable to fertilize oocytes. Initially, starting from simple washing of spermatozoa, separation techniques, based on different principles like migration, filtration or density gradient centrifugation evolved. The most simple and cheapest is the conventional swim-up procedure. A more sophisticated and most gentle migration method is migration-sedimentation. However, its yield is relatively small and the technique is therefore normally only limited to ejaculates with a high number of motile spermatozoa. Recently, however, the method was also successfully used to isolate spermatozoa for intracytoplasmic sperm injection (ICSI). Sperm separation methods that yield a higher number of motile spermatozoa are glass wool filtration or density gradient centrifugation with different media. Since Percoll® as a density medium was removed from the market in 1996 for clinical use in the human because of its risk of contamination with endotoxins, other media like IxaPrep®, Nycodenz, SilSelect®, PureSperm® or Isolate® were developed in order to replace Percoll®. Today, an array of different methods is available and the selection depends on the quality of the ejaculates, which also includes production of reactive oxygen species (ROS) by spermatozoa and leukocytes. Ejaculates with ROS production should not be separated by means of conventional swim-up, as this can severely damage the spermatozoa. In order to protect the male germ cells from the influence of ROS and to stimulate their motility to increase the yield, a number of substances can be added to the ejaculate or the separation medium. Caffeine, pentoxifylline and 2-deoxyadenosine are substances that were used to stimulate motility. Recent approaches to stimulate spermatozoa include bicarbonate, metal chelators or platelet-activating factor (PAF). While the use of PAF already resulted in pregnancies in intrauterine insemination, the suitability of the other substances for the clinical use still needs to be tested. Finally, the isolation of functional spermatozoa from highly viscous ejaculates is a special challenge and can be performed enzymatically to liquefy the ejaculate. The older method, by which the ejaculate is forcefully aspirated through a narrow-gauge needle, should be abandoned as it can severely damage spermatozoa, thus resulting in immotile sperm.

Henkel, Ralf R; Schill, Wolf-Bernhard



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

PubMed Central

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

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



Functional human sperm capacitation requires both bicarbonate-dependent PKA activation and down-regulation of Ser/Thr phosphatases by Src family kinases.  


In all mammalian species studied so far, sperm capacitation correlates with an increase in protein tyrosine (Tyr) phosphorylation mediated by a bicarbonate-dependent cAMP/protein kinase A (PKA) pathway. Recent studies in mice revealed, however, that a Src family kinase (SFK)-induced inactivation of serine/threonine (Ser/Thr) phosphatases is also involved in the signaling pathways leading to Tyr phosphorylation. In view of these observations and with the aim of getting a better understanding of the signaling pathways involved in human sperm capacitation, in the present work we investigated the involvement of both the cAMP/PKA and SFK/phosphatase pathways in relation to the capacitation state of the cells. For this purpose, different signaling events and sperm functional parameters were analyzed as a function of capacitation time. Results revealed a very early bicarbonate-dependent activation of PKA indicated by the rapid (1 min) increase in both phospho-PKA substrates and cAMP levels (P < 0.05). However, a complete pattern of Tyr phosphorylation was detected only after 6-h incubation at which time sperm exhibited the ability to undergo the acrosome reaction (AR) and to penetrate zona-free hamster oocytes. Sperm capacitated in the presence of the SFK inhibitor SKI606 showed a decrease in both PKA substrate and Tyr phosphorylation levels, which was overcome by exposure of sperm to the Ser/Thr phosphatase inhibitor okadaic acid (OA). However, OA was unable to induce phosphorylation when sperm were incubated under PKA-inhibitory conditions (i.e. in the absence of bicarbonate or in the presence of PKA inhibitor). Moreover, the increase in PKA activity by exposure to a cAMP analog and a phosphodiesterase inhibitor did not overcome the inhibition produced by SKI606. Whereas the presence of SKI606 during capacitation produced a negative effect (P < 0.05) on sperm motility, progesterone-induced AR and fertilizing ability, none of these inhibitions were observed when sperm were exposed to SKI606 and OA. Interestingly, different concentrations of inhibitors were required to modulate human and mouse capacitation revealing the species specificity of the molecular mechanisms underlying this process. In conclusion, our results describe for the first time the involvement of both PKA activation and Ser/Thr phosphatase down-regulation in functional human sperm capacitation and provide convincing evidence that early PKA-dependent phosphorylation is the convergent regulatory point between these two signaling pathways. PMID:23630234

Battistone, M A; Da Ros, V G; Salicioni, A M; Navarrete, F A; Krapf, D; Visconti, P E; Cuasnicú, P S



Correlation between nitric oxide levels, the hypo-osmotic swelling test for sperm membranes and semen analysis in patients with varicocele.  


Abstract Objective. Nitric oxide (NO) is a well-known oxidative stress agent that directly inhibits mitochondrial respiration and the synthesis of DNA. A case-control study of the concentration of NO in infertile patients with varicocele versus car-accident controls was performed. The concentration of NO in infertile patients with varicocele, and its correlation with the sperm fertility test named the hypo-osmotic swelling (HOS) test for sperm membranes and semen analysis, was also examined. Material and methods. A spectrophotometric method was used to measure the concentration of NO in infertile subjects, and the HOS test was carried out according to Jeyendran's original protocol. NO values from left testicular vein and left cubital vein of 19 infertile patients with left-sided varicocele were compared to 15 patients in the control group who underwent explorative laparoscopies after car accidents. Semen analysis and the HOS test were performed only in the left-sided varicocele group and compared to the NO outcomes. Results.The results indicate that the concentration of NO was significantly higher in the testicular vein than in the peripheral veins of patients with varicocele, as well as being significantly higher than in the testicular vein of control patients. An increased NO concentration in the testicular vein, higher sperm count and higher sperm motility in infertile patients with varicocele followed negative or suspicious findings of the HOS test for sperm membranes. There was also a significant correlation between NO concentration and sperm count in patients with varicocele. Conclusions. Further studies with more patients, related to NO concentrations and the HOS test are needed to verify these results. PMID:23373534

Pajovic, Bogdan; Radojevic, Nemanja; Terzic, Natasa; Dimitrovski, Antonio



Mammalian Sperm Fertility Related Proteins  

PubMed Central

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.

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



Pulmonary Function Testing in Children  


... can be compared to the results of other children of the same sex, age, and height, like the standard ranges on ... P3-P4, 2009. ATS Patient Education Series © 2009 American Thoracic Society Pulmonary Function Testing in Children Pulmonary function tests (PFT’s) measure how well your ...


Human Spermatozoa Contain Multiple Targets for Protein S-Nitrosylation: An Alternative Mechanism of the Modulation of Sperm Function by Nitric Oxide?  

PubMed Central

Nitric oxide (NO) enhances human sperm motility and capacitation associated with increased protein phosphorylation. NO activates soluble guanylyl cyclase, but can also modify protein function covalently via S-nitrosylation of cysteine. Remarkably, this mechanism remains unexplored in sperm although they depend on post-translational protein modification to achieve changes in function required for fertilisation. Our objective was to identify targets for S-nitrosylation in human sperm. Spermatozoa were incubated with NO donors and S-nitrosylated proteins were identified using the biotin switch assay and a proteomic approach using tandem mass spectrometry. 240 S-nitrosylated proteins were detected in sperm incubated with S-nitrosoglutathione. Minimal levels were observed in glutathione or untreated samples. Proteins identified consistently based on multiple peptides included established targets for S-nitrosylation in other cells e.g. tubulin,, glutathione-S-transferase and heat shock proteins but also novel targets including A-kinase anchoring protein (AKAP) types 3 and 4, voltage-dependent anion-selective channel protein 3 and semenogelin 1 and 2. In situ localisation revealed S-nitrosylated targets on the post-acrosomal region of the head and throughout the flagellum. Potential targets for S-nitrosylation in human sperm include physiologically significant proteins not previously reported in other cells. Their identification will provide novel insight into the mechanism of action of NO in spermatozoa.

Lefievre, Linda; Chen, Yongjian; Conner, Sarah J; Scott, Joanna L; Publicover, Steve J; Ford, W Christopher L; Barratt, Christopher LR



Clinical aspects of sperm DNA fragmentation detection and male infertility  

Microsoft Academic Search

Over the past 25 years, various methods have been developed to measure sperm DNA strand breaks in situ. Currently, there are four major tests of sperm DNA fragmentation, including the Comet, Tunel, sperm chromatin structure assay (SCSA) and the acridine orange test (AOT). The Comet assay is a light microscope technique where the sperm cells are mixed with melted agarose

Donald P. Evenson; Regina Wixon



Methods of sperm vitality assessment.  


Sperm vitality is a reflection of the proportion of live, membrane-intact spermatozoa determined by either dye exclusion or osmoregulatory capacity under hypo-osmotic conditions. In this chapter we address the two most common methods of sperm vitality assessment: eosin-nigrosin staining and the hypo-osmotic swelling test, both utilized in clinical Andrology laboratories. PMID:22992899

Moskovtsev, Sergey I; Librach, Clifford L



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


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

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



Sperm storage in caecilian amphibians  

PubMed Central

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.



Cytometric analysis of shape and DNA content in mammalian sperm  

SciTech Connect

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.

Gledhill, B.L.



Maturational changes in the survivability and fertility of fowl sperm during their passage through the male reproductive tract.  


The objective of this study was to examine whether domestic fowl (Gallus domesticus) sperm undergo maturation in their capacity for survival and fertilization in the male reproductive tract. Sperm collected from the testis, epididymis and the proximal, middle and distal vas deferens were simultaneously stored in vitro in minimum essential medium (MEM) at 39°C for 0, 3 and 6h, and at 4°C for 24 and 48h. Sperm membrane integrity was measured using the dual fluorescent stain SYBR-14/propidium iodide (PI). Aliquots of sperm from the various sites were subjected to artificial insemination (AI) into the uteri of hens to assess the duration of sperm survival in the oviduct and to determine the fertility status of the sperm. Testicular sperm exhibited a very low capacity to survive under in vitro liquid storage conditions, irrespective of the storage temperature used, and in the oviduct, and they had a low ability to fertilize the ovum. On the contrary, sperm from the distal vas deferens had a higher survival rate during in vitro storage periods, a longer life span in the oviduct, and high fertility. Survival and fertilizing capacity of the sperm recovered from the testes increased gradually (P<0.05) from the testes to the distal vas deferens. In conclusion, we suggest that fowl sperm may undergo functional maturation through a process of gradual changes in their survival and fertilization capacities during their passage through the successive parts of the male reproductive tract. PMID:21996579

Ahammad, Muslah U; Nishino, C; Tatemoto, H; Okura, N; Kawamoto, Y; Okamoto, S; Nakada, T



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


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

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



[Cytological, morphological and functional changes of Russian sturgeons's (Acipenser guldenshtadti) sperm cells after cryopreservation].  


The article presents some experimental results concerning the problem of genetic conservation of valuable and endangered animal species. Biotechnical development of the cryopreservation of spermatic cells for the purpose of their extended storage at low and ultra low temperatures is a priority line of the investigations in the topical area and implies creation of animal genomes cryobanks. It is known that water crystals of plasma and the cell itself often cause the spermatholysis in the course of freezing. That is why the search for the substances with cryoprotective properties continues up to the present moment. In this study, we investigated cryoprotective properties of glycerin, dimethyl sulfoxide and heparin in different proportions using spermatic fluid of sturgeon Acipenser guldenshtadti (Brandt). These results obtained were compared with the cryoprotectors of well-known composition. In addition, freezing and storage of the spermatic fluid were executed in two variations: at low and ultralow temperatures. Cold tolerance of the sperm cells was estimated by the sperm mobility and by light microscopy morphological analysis of the cells. It was shown to be linked to the composition of cryoprotectors. The heaviest violations were observed under addition of osmotic active substances, some inorganic compounds among them. Principle of our modification is based on that we used fluid substances: glycerin, dimethyl sulfoxide, heparin eliminating water as a solvent. The results obtained showed that our modified cryoprotectors and the standard cryoprotectors had equal efficiency. The article also presents the data on the spermatic fluid freezing at low temperature. They are important in case of modern freezing systems. PMID:20058814

Zemkov, G V; Akimochkina, T N



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


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

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



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


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. PMID:19308348

Evans, Jonathan P



Clomiphene citrate challenge test in the assessment of ovarian reserve before controlled ovarian hyperstimulation for intracytoplasmic sperm injection.  


The objective of this study is to evaluate the performance of clomiphene citrate (CC) challenge test to predict diminished ovarian reserve before controlled ovarian hyperstimulation for intracytoplasmic sperm injection (ICSI). The 198 women who underwent the CC challenge test fulfilled the following criteria; over 35 years of age, removal of one ovary or previous ovarian surgery, the presence of ovarian endometrioma or previous poor response to ovarian hyperstimulation. Of the patients tested, 141 were found to have a normal CC challenge test while 57 had an abnormal result. The cancellation rate of the cycle with a poor response was significantly higher in women with an abnormal test (36.8%) than in those with a normal test (19.8%) (P < 0.05). The sensitivity of CC test for cycle cancellation was found to be 43% with a specificity of 76%, positive and negative predictive values of 37 and 80%, respectively. The estradiol values on hCG day, the number of retrieved oocytes and metaphase II oocytes and the rate of transfer cycles were significantly lower in females with an abnormal test. Women with normal test results had higher pregnancy rates per embryo transfer than those with abnormal test results (21.5 vs. 13.3%) and the predictive value of an abnormal test for failing to conceive was 93% (53/57) with a sensitivity of 31%, specificity of 84% and negative predictive value of 15.6%. Of 57 women with an abnormal test result, 25 (43.8%) were abnormal due only to an elevated day 10 or 11 value of FSH, which could not be detected using only basal FSH screening. In this group, the cancellation rate (48 vs. 19.8%, P < 0.01), the rate of transfer cycles (48 vs. 72.3%, P < 0.05) and the mean number of retrieved oocytes (4.9 +/- 2.5 vs. 6.4 +/- 3.1, P < 0.01) were all significantly different from normal test group. Although the rate of pregnancies per started cycle (8 vs. 15.6%) did not show a statistically significant difference, this is most probably due to the low number of patients. In conclusion, an abnormal CC challenge test is a good predictor of diminished ovarian reserve and it is better than a basal FSH concentration on day 3. It provides valuable information for both patients as to their chances of achieving a pregnancy and also for the medical team deciding on options for stimulation protocols. PMID:9228501

Kahraman, S; Vicdan, K; I?ik, A Z; Ozgün, O D; Alaybeyo?lu, L; Polat, G; Bibero?lu, K



Persistence and intergenerational transmission of differentially expressed genes in the testes of intracytoplasmic sperm injection conceived mice*  

PubMed Central

Intracytoplasmic sperm injection (ICSI) is commonly used to solve male infertility problems. Previous studies showed that early environmental exposure of an embryo may influence postnatal development. To detect whether ICSI operations affect the reproductive health of a male or his offspring, we established assisted reproductive technologies (ART) conceived mouse models, and analyzed gene expression profiles in the testes of both ICSI and naturally conceived (NC) newborn F1 mice using micro-array analysis. Among the differentially expressed genes, we focused on the expression of eight male reproduction-related genes. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was used to analyze the expression of these genes in the testes of both adult and old F1 generation mice and adult F2 generation mice. Our results showed that down-regulated and somatic cell-expressed genes in newborn mice retained their differential expression patterns in adult and old F1 generation individuals, implying the persistence and fetal origin of the alteration in the expression of these genes. The intergenerational transmission of differential gene expression was observed, but most changes tended to be reduced in adult F2 generations. Controlled ovarian hyperstimulation (COH) and in vitro fertilization (IVF) mice models were added to explore the precise factors contributing to the differences in ICSI offspring. The data demonstrated that superovulation, in vitro culture, and mechanical stimulation involved in ICSI had a cumulative effect on the differential expression of these male reproductive genes.

Wang, Li-ya; Wang, Ning; Le, Fang; Li, Lei; Li, Le-jun; Liu, Xiao-zhen; Zheng, Ying-ming; Lou, Hang-ying; Xu, Xiang-rong; Zhu, Xiao-ming; Zhu, Yi-min; Huang, He-feng; Jin, Fan



Automated sperm head morphology analyzer for open-source software  

Microsoft Academic Search

Sperm head morphology has been identified as a characteristic that can be used to predict a male's semen quality. In the present study, we have developed an automated sperm head morphology analysis (ASMA) plug-in for open-source ImageJ software (http:\\/\\/\\/ij\\/). We describe the plug-in's functionality, and confirm its validity for sperm head morphology analysis using fish sperm. Sperm head morphological measurements

I. A. E. Butts; M. A. R. Ward; M. K. Litvak; T. E. Pitcher; S. M. H. Alavi; E. A. Trippel; R. M. Rideout



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

Microsoft Academic Search

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

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



Comparison of human cervical mucus and artificial sperm penetration media.  


The cervical mucus penetration tests aid research and determine the clinical importance of positive sperm antibody tests. Limited availability and variability of human cervical mucus have instigated the search for mucus substitutes for these tests. This study compares sperm migration in cervical mucus with that in artificial media including hyaluronate solution, egg white and albumin Tyrode solution. Results were quantified by measuring the migration distance (the maximum distance of capillary migration from a semen reservoir by spermatozoa after 1 h) and the sperm concentration at half the migration distance. The mean of both measures for cervical mucus and hyaluronate solution were equivalent [4.4 +/- 1.1 (SD) versus 4.3 +/- 1.0 cm and 118 +/- 51 versus 111 +/- 44x10(3)/ml], and higher than in egg white and albumin Tyrode solution. Antisperm antibodies impaired sperm penetration in cervical mucus and hyaluronate solution in a similar manner (r = 0.92). These results suggest that hyaluronate solution sufficiently resembles human cervical mucus in terms of penetrability that it may be used as a substitute for mucus in capillary tube tests of sperm function. The higher penetrability of cervical mucus and hyaluronate solution is probably related to a channelling effect due to their polymeric structure. PMID:10548628

Tang, S; Garrett, C; Baker, H W



Sperm competition and maternal effects differentially influence testis and sperm size in Callosobruchus maculatus.  


The evolutionary factors affecting testis size are well documented, with sperm competition being of major importance. However, the factors affecting sperm length are not well understood; there are no clear theoretical predictions and the empirical evidence is inconsistent. Recently, maternal effects have been implicated in sperm length variation, a finding that may offer insights into its evolution. We investigated potential proximate and microevolutionary factors influencing testis and sperm size in the bruchid beetle Callosobruchus maculatus using a combined approach of an artificial evolution experiment over 90 generations and an environmental effects study. We found that while polyandry seems to select for larger testes, it had no detectable effect on sperm length. Furthermore, population density, a proximate indicator of sperm competition risk, was not significantly associated with sperm length or testis size variation. However, there were strong maternal effects influencing sperm length. PMID:19309491

Gay, L; Hosken, D J; Vasudev, R; Tregenza, T; Eady, P E



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

SciTech Connect

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

Dolan, C E



Mating behavior and the evolution of sperm design  

PubMed Central

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.

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



Aquaporin-9 immunohistochemistry in varicocele testes as a consequence of hypoxia in the sperm production site.  


Aquaporin-9 (AQP-9) regulates tissue hydration by promoting transmembrane exchanges of both water and solutes, such as lactate. The latter is a key metabolite of primary spermatocytes and of maturing haploid germ cells (h-GCs). The present investigation was aimed at immunolocalising human AQP-9 in both normal and varicocele testes. Histology and immmunocytochemistry were investigated in archival biopsies from 20 varicocele testes and in eight unaffected ones. AQP-9 immunostaining was performed using a rabbit antibody, and either focal or diffuse cell membrane labelling was recorded. Varicocele testes showed disarranged tubular compartments, with sloughing h-GCs, tissue hyperhydration, spermiogenesis failure and fibrosis. AQP-9 immunohistology of the control testes showed a diffuse cell membrane staining of the primary spermatocytes and h-GCs, without any positive reaction of spermatogonia and Sertoli cells. AQP-9 cell expression in the varicocele testes was focal or lacking in both adluminal and sloughing GCs. AQP-9 expression occurs in normal human testis, at cell membrane of primary spermatocytes and h-GCs, suggesting a possible role of AQP-9 in the water and lactate transport from Sertoli cells to GCs. AQP-9 is focal or lacking in adolescent varicocele testes, and this suggests AQP-9 to be downregulated in such testicular disorder, leading to lactate deprivation with subsequent hypospermatogenesis. PMID:21219380

Arena, S; Arena, F; Maisano, D; Di Benedetto, V; Romeo, C; Nicòtina, P A



Sperm characteristics and ultrastructure of testes of rats after long-term treatment with the methanol subfraction of Carica papaya seeds  

PubMed Central

The contraceptive efficacy of Carica papaya seeds after short-term evaluation has been well established. We have examined the safety and mechanism of contraception in rats after long-term treatment with the methanol subfraction (MSF) of C. papaya seeds. The test substance was administered orally to the male albino rats (n = 40) at 50 mg per kg body weight each day for 360 days. Control animals (n = 40) received olive oil as a vehicle. Recovery was assessed up to 120 days after treatment withdrawal. Sperm parameters, serum testosterone levels, fertility, histology and ultrastructure of the testis, haematology and serum clinical chemistry were evaluated to establish the safety and efficacy of the test substance. Safety of long-term treatment was evidenced by unaltered health status, organ weight, haematology and clinical chemistry, and by an increase in body weight. The mechanism of contraception was shown by reduction in nuclear and cytoplasmic volume, normal nuclear characteristics and vacuolization in the cytoplasmic organelles of the Sertoli cells, as well as nuclear degeneration in spermatocytes and spermatids indicating disturbed spermatogenesis. Leydig cells were normal. Initial effects were observed in Sertoli cells at 60 days of treatment. Spermatocytes and spermatids were affected after 120–240 days of treatment. A significant decline in sperm count and viability, total inhibition of sperm motility, increased numbers of sperm abnormalities, normal serum testosterone levels and 100% sterility were evident after 60 days of treatment. All the altered parameters, including percent fertility, were restored to control level 120 days after treatment withdrawal. It is concluded that the MSF is safe for long-term treatment and the mechanism of contraception is shown by its effect on spermatid differentiation in the testis, possibly mediated by the Sertoli cell factors.

Manivannan, Boomi; Mittal, Ruchi; Goyal, Shipra; Ansari, Abdul S.; Lohiya, Nirmal K.



Mating system evolution in sperm-heteromorphic Drosophila.  


In Drosophila species of the obscura group, males exhibit sperm-heteromorphism, simultaneously producing both long sperm, capable of fertilization, and short sperm that are not. The production of multiple sperm types calls into question whether mating system correlates, such as sperm length and number trade-offs and female remating behavior, are the same as previously described in sperm-monomorphic systems. We examine three obscura group species, D. pseudoobscura, D. persimilis, and D. affinis that differ significantly in the lengths of their long fertilizing sperm, to test predictions about the relationship between sperm length and four mating system characters: male age at sexual maturity; sperm number; female remating; and male reproductive output. In D. affinis, where males produce the longest fertilizing sperm, their sexual maturity is delayed and they produce fewer long sperm compared to the other two species, as predicted if long sperm are costly to produce. Female D. affinis, although they receive fewer sperm than females of the other two species, do not remate more frequently or produce fewer progeny from a single mating. Different responses between sperm-heteromorphic and sperm-monomorphic systems underscore the complex nature of the coevolution between male and female mating system characters. PMID:11472758

Snook, R R.; Markow, T A.



14 CFR 35.40 - Functional test.  

Code of Federal Regulations, 2013 CFR

... AIRCRAFT AIRWORTHINESS STANDARDS: PROPELLERS Tests and Inspections § 35.40 Functional test. The variable-pitch propeller system must be subjected to the applicable...functional tests of this section. The same propeller system used in the endurance...



Permutation Tests Using Estimated Distribution Functions  

Microsoft Academic Search

In this article we develop permutation tests for estimated distribution functions. The tests are formed by averaging a functional of estimated distribution functions that are calculated from independent sampling units, where the units may be a single response, a set of repeated responses, or a censored response. We study primarily two functionals—the difference in means functional and the Mann-Whitney functional,

Michael P. Fay; Joanna H. Shih



Bridging the gap between chemistry, physiology, and evolution: quantifying the functionality of sperm whale myoglobin mutants.  


This work merges a large set of previously reported thermochemical data for myoglobin (Mb) mutants with a physiological model of O(2)-transport and -storage. The model allows a quantification of the functional proficiency of myoglobin (Mb) mutants under various physiological conditions, i.e. O(2)-consumption rate resembling workload, O(2) partial pressure resembling hypoxic stress, muscle cell size, and Mb concentration, resembling different organism-specific and compensatory variables. We find that O(2)-storage and -transport are distinct functions that rank mutants and wild type differently depending on O(2) partial pressure. Specifically, the wild type is near-optimal for storage at all conditions, but for transport only at severely hypoxic conditions. At normoxic conditions, low-affinity mutants are in fact better O(2)-transporters because they still have empty sites for O(2), giving rise to a larger [MbO(2)] gradient (more varying saturation curve). The distributions of functionality reveal that many mutants are near-neutral with respect to function, whereas only a few are strongly affected, and the variation in functionality increases dramatically at lower O(2) pressure. These results together show that conserved residues in wild type (WT) Mb were fixated under a selection pressure of low P(O2). PMID:21903173

Dasmeh, Pouria; Kepp, Kasper P



Using genetic algorithms to generate test plans for functionality testing  

Microsoft Academic Search

Like in other fields, computer products (applications, hardware, etc.), before being marketed, require some level of testing to verify whether they meet their design and functional specifications -- called functionality test. The general process of performing functionality test consists in the production of a test plan that is then executed by humans or by automated software tools. The main difficulty

Francisca Emanuelle Vieira; Francisco Martins; Rafael Silva; Ronaldo Menezes; Márcio Braga



Preservation of Mouse Sperm by Convective Drying and Storing in 3-O-Methyl-D-Glucose  

PubMed Central

With the fast advancement in the genetics and bio-medical fields, the vast number of valuable transgenic and rare genetic mouse models need to be preserved. Preservation of mouse sperm by convective drying and subsequent storing at above freezing temperatures could dramatically reduce the cost and facilitate shipping. Mouse sperm were convectively dried under nitrogen gas in the Na-EGTA solution containing 100 mmol/L 3-O-methyl-D-glucose and stored in LiCl sorption jars (Relative Humidity, RH, 12%) at 4°C and 22°C for up to one year. The functionality of these sperm samples after storage was tested by intracytoplasmic injection into mouse oocytes. The percentages of blastocysts produced from sperm stored at 4°C for 1, 2, 3, 6, and 12 months were 62.6%, 53.4%, 39.6%, 33.3%, and 30.4%, respectively, while those stored at 22°C for 1, 2, and 3 months were 28.8%, 26.6%, and 12.2%, respectively. Transfer of 38 two- to four-cell embryos from sperm stored at 4°C for 1 year produced two live pups while 59 two- to four-cell embryos from sperm stored at 22°C for 3 months also produced two live pups. Although all the pups looked healthy at 3 weeks of age, normality of offspring produced using convectively dried sperm needs further investigation. The percentages of blastocyst from sperm stored in the higher relative humidity conditions of NaBr and MgCl2 jars and driest condition of P2O5 jars at 4°C and 22°C were all lower. A simple method of mouse sperm preservation is demonstrated. Three-O-methyl-D-glucose, a metabolically inactive derivative of glucose, offers significant protection for dried mouse sperm at above freezing temperatures without the need for poration of cell membrane.

Liu, Jie; Lee, Gloria Y.; Lawitts, Joel A.; Toner, Mehmet; Biggers, John D.



Female presence influences sperm velocity in the guppy.  


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

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



Sperm viability - Determination of sperm viability using fluorescence microscopy  

Technology Transfer Automated Retrieval System (TEKTRAN)

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


Alterations to the bull sperm surface proteins that bind sperm to oviductal epithelium.  


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

Hung, Pei-hsuan; Suarez, Susan S



Lactate Dehydrogenase C and Energy Metabolism in Mouse Sperm1  

PubMed Central

We demonstrated previously that disruption of the germ cell-specific lactate dehydrogenase C gene (Ldhc) led to male infertility due to defects in sperm function, including a rapid decline in sperm ATP levels, a decrease in progressive motility, and a failure to develop hyperactivated motility. We hypothesized that lack of LDHC disrupts glycolysis by feedback inhibition, either by causing a defect in renewal of the NAD+ cofactor essential for activity of glyceraldehyde 3-phosphate dehydrogenase, sperm (GAPDHS), or an accumulation of pyruvate. To test these hypotheses, nuclear magnetic resonance analysis was used to follow the utilization of labeled substrates in real time. We found that in sperm lacking LDHC, glucose consumption was disrupted, but the NAD:NADH ratio and pyruvate levels were unchanged, and pyruvate was rapidly metabolized to lactate. Moreover, the metabolic disorder induced by treatment with the lactate dehydrogenase (LDH) inhibitor sodium oxamate was different from that caused by lack of LDHC. This supported our earlier conclusion that LDHA, an LDH isozyme present in the principal piece of the flagellum, is responsible for the residual LDH activity in sperm lacking LDHC, but suggested that LDHC has an additional role in the maintenance of energy metabolism in sperm. By coimmunoprecipitation coupled with mass spectrometry, we identified 27 proteins associated with LDHC. A majority of these proteins are implicated in ATP synthesis, utilization, transport, and/or sequestration. This led us to hypothesize that in addition to its role in glycolysis, LDHC is part of a complex involved in ATP homeostasis that is disrupted in sperm lacking LDHC.

Odet, Fanny; Gabel, Scott A.; Williams, Jason; London, Robert E.; Goldberg, Erwin; Eddy, Edward M.



Expression, localization and functions in acrosome reaction and sperm motility of Ca(V)3.1 and Ca(V)3.2 channels in sperm cells: an evaluation from Ca(V)3.1 and Ca(V)3.2 deficient mice.  


In spermatozoa, voltage-dependent calcium channels (VDCC) have been involved in different cellular functions like acrosome reaction (AR) and sperm motility. Multiple types of VDCC are present and their relative contribution is still a matter of debate. Based mostly on pharmacological studies, low-voltage-activated calcium channels (LVA-CC), responsible of the inward current in spermatocytes, were described as essential for AR in sperm. The development of Ca(V)3.1 or Ca(V)3.2 null mice provided the opportunity to evaluate the involvement of such LVA-CC in AR and sperm motility, independently of pharmacological tools. The inward current was fully abolished in spermatogenic cells from Ca(V)3.2 deficient mice. This current is thus only due to Ca(V)3.2 channels. We showed that Ca(V)3.2 channels were maintained in sperm by Western-blot and immunohistochemistry experiments. Calcium imaging experiments revealed that calcium influx in response to KCl was reduced in Ca(V)3.2 null sperm in comparison to control cells, demonstrating that Ca(V)3.2 channels were functional. On the other hand, no difference was noticed in calcium signaling induced by zona pellucida. Moreover, neither biochemical nor functional experiments, suggested the presence of Ca(V)3.1 channels in sperm. Despite the Ca(V)3.2 channels contribution in KCl-induced calcium influx, the reproduction parameters remained intact in Ca(V)3.2 deficient mice. These data demonstrate that in sperm, besides Ca(V)3.2 channels, other types of VDCC are activated during the voltage-dependent calcium influx of AR, these channels likely belonging to high-voltage activated Ca(2+) channels family. The conclusion is that voltage-dependent calcium influx during AR is due to the opening of redundant families of calcium channels. PMID:17450521

Escoffier, Jessica; Boisseau, Sylvie; Serres, Catherine; Chen, Chien-Chang; Kim, Daesoo; Stamboulian, Séverine; Shin, Hee-Sup; Campbell, Kevin P; De Waard, Michel; Arnoult, Christophe



Cryopreservation of rooster sperm.  


Successful cryopreservation of sperm requires: 1) selection of proper diluent; 2) selection of the best cryoprotectant; 3) determination of freezing and thawing rates for optimum retention of fertilization potential; and 4) removal of any materials deleterious to fertility (e.g., glycerol) before insemination. An economically useful process must allow recovery of sperm with sufficient fertilization capacity to enable maximum use of any given superior male. A series of experiments tested a novel semen freezing container (BioPore CryoCell container) having physical characteristics that permit reproducible freezing and thawing plus facile removal of glycerol from the sample after processing. Experiments tested the effect of: a) residual glycerol; b) initial glycerol concentrations on retention of fertility when samples were frozen and thawed at 6 C/min; c) Beltsville Poultry Semen Extender and Minnesota A buffers used during the dialysis procedure; and d) dialysis time. Respectively, the results were: a) .8% (vol/vol) reduced fertility by 5 to 10%; b) 12% glycerol was superior to 10% and 8% glycerol; c) no difference was observed between the two buffers; and d) 90 and 120 min were both superior to 60 min. Numerous pools of rooster sperm cryopreserved in CryoCell containers and dialyzed after thawing in a prototype BioStore environmental control chamber for 90 or 120 min resulted in a mean fertility of 55.6%. This mean fertility of frozen-thawed sperm was based on 3,263 eggs laid by 400 hens on Days +1 through 9 after inseminations on Days -1, 2, and 5. It is likely that broiler stocks might have lower fertility than that obtained from the Barred Plymouth Rock males and the Single Comb White Leghorn females used in these studies. Nevertheless, the procedure described is the first to consistently result in > 50% fertilized eggs as a result of conventional intravaginal insemination (< 200 x 10(6) sperm in 100 microL extender) of sperm processed after thawing by a procedure amenable to the scaleup required for commercial applications. PMID:8502616

Buss, E G



"OMICS" of Human Sperm: Profiling Protein Phosphatases.  


Abstract Phosphorylation is a major regulatory mechanism in eukaryotic cells performed by the concerted actions of kinases and phosphatases (PPs). Protein phosphorylation has long been relevant to sperm physiology, from acquisition of motility in the epididymis to capacitation in the female reproductive tract. While the precise kinases involved in the regulation of sperm phosphorylation have been studied for decades, the PPs have only recently received research interest. Tyrosine phosphorylation was first implicated in the regulation of several sperm-related functions, from capacitation to oocyte binding. Only afterwards, in 1996, the inhibition of the serine/threonine-PP phosphoprotein phosphatase 1 (PPP1) by okadaic acid and calyculin-A was shown to initiate motility in caput epididymal sperm. Today, the current mechanisms of sperm motility acquisition based on PPP1 and its regulators are still far from being fully understood. PPP1CC2, specifically expressed in mammalian sperm, has been considered to be the only sperm-specific serine/threonine-PP, while other PPP1 isoforms were thought to be absent from sperm. This article examines the "Omics" of human sperm, and reports, for the first time, the identification of three new serine/threonine-protein PPs, PPP1CB, PPP4C, and PPP6C, in human sperm, together with two tyrosine-PPs, MKP1 and PTP1C. We specifically localized in sperm PPP1CB and PPP1CC2 from the PPP1 subfamily, and PPP2CA, PPP4C, and PPP6C from the PPP2 subfamily of the serine/threonine-PPs. A semi-quantitative analysis was performed to determine the various PPs' differential expression in sperm head and tail. These findings contribute to a comprehensive understanding of human sperm PPs, and warrant further research for their clinical and therapeutic significance. PMID:23895272

Fardilha, Margarida; Ferreira, Mónica; Pelech, Steven; Vieira, Sandra; Rebelo, Sandra; Korrodi-Gregorio, Luís; Sousa, Mário; Barros, Alberto; Silva, Vladimiro; da Cruz E Silva, Odete A B; da Cruz E Silva, Edgar F



The sperm penetration assay for the assessment of fertilization capacity.  


The sperm penetration assay, or zona-free hamster oocyte penetration assay, is utilized to measure the ability of sperm to undergo capacitation, fuse with the egg membrane, and decondense the sperm head within the cytoplasm of the oocyte, resulting in the formation of the male pronucleus. The test is scored by calculation of the percentage of ova that are penetrated (the original assay developed) or the average number of sperm penetrations per ovum (the sperm capacitation index of the optimized assay). The sperm penetration assay identifies those couples that will have a high likelihood of success with in vitro fertilization. PMID:22992907

Hwang, Kathleen; Lamb, Dolores J



Cytometry of mammalian sperm  

SciTech Connect

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

Gledhill, B.L.



Evaluation of abnormal liver function tests  

PubMed Central

Interpretation of abnormalities in liver function tests is a common problem faced by clinicians. This has become more common with the introduction of automated routine laboratory testing. Not all persons with one or more abnormalities in these tests actually have liver disease. The various biochemical tests, their pathophysiology, and an approach to the interpretation of abnormal liver function tests are discussed in this review.

Limdi, J; Hyde, G



How nematode sperm crawl  

Microsoft Academic Search

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

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



Technology Transfer Automated Retrieval System (TEKTRAN)

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


Chromatin Organization in Sperm May Be the Major Functional Consequence of Base Composition Variation in the Human Genome  

Microsoft Academic Search

Chromatin in sperm is different from that in other cells, with most of the genome packaged by protamines not nucleosomes. Nucleosomes are, however, retained at some genomic sites, where they have the potential to transmit paternal epigenetic information. It is not understood how this retention is specified. Here we show that base composition is the major determinant of nucleosome retention

Tanya Vavouri; Ben Lehner



Colourful male guppies produce faster and more viable sperm.  


In guppies (Poecilia reticulata) precopulatory sexual selection (via female choice) and post-copulatory selection (via sperm competition) both favour males with relatively high levels of carotenoid (orange) pigmentation, suggesting that colourful males produce more competitive ejaculates. Here we test whether there is a positive association between male orange pigmentation and sperm quality. Our analysis of sperm quality focused on sperm swimming speeds (using CASA: computer-assisted sperm analysis to estimate three parameters of sperm velocity in vitro), sperm viability (proportion of live sperm per stripped ejaculate) and sperm lengths. We found that males with relatively large areas of orange pigmentation had significantly faster and more viable sperm than their less ornamented counterparts, suggesting a possible link between dietary carotenoid intake and sperm quality. By contrast, we found no relationship between sperm length (head length and total sperm length) and male phenotype. These findings, in conjunction with previous work showing that highly ornamented male guppies sire higher quality offspring, suggest that female preference for colourful males and sperm competition work in concert to favour intrinsically higher quality males. PMID:16910988

Locatello, L; Rasotto, M B; Evans, J P; Pilastro, A



Multiple mating in the ant Cataglyphis cursor: testing the sperm limitation and the diploid male load hypotheses  

Microsoft Academic Search

.  Multiple mating (i.e., polyandry) by queens in social Hymenoptera is expected to weaken social cohesion since it lowers within-colony relatedness,\\u000a and hence, indirect fitness benefits from kin selection. Yet, there are many species where queens mate multiply. Several hypotheses\\u000a have been put forward to explain the evolution and maintenance of polyandry. Here,we investigated the ‘sperm limitation’ and\\u000a the ‘diploid male

M. Pearcy; I. Timmermans; D. Allard; S. Aron



The nuclear status of human sperm cells  

Microsoft Academic Search

In the last decade, and in particular since the development of in vitro fertilization techniques, the nuclear status of human sperm cells has shown to be a key parameter in the assessment of male fertility. The shape and condensed state of the mature sperm nucleus are determined by structural and functional events that occur during spermiogenesis. This paper reviews essential

Jean-Pierre Dadoune



Mass-Specific Metabolic Rate and Sperm Competition Determine Sperm Size in Marsupial Mammals  

PubMed Central

Two complementary hypotheses have been proposed to explain variation in sperm size. The first proposes that post-copulatory sexual selection favors an increase in sperm size because it enhances sperm swimming speed, which is an important determinant of fertilization success in competitive contexts. The second hypothesis proposes that mass-specific metabolic rate acts as a constraint, because large animals with low mass-specific metabolic rates will not be able to process resources at the rates needed to produce large sperm. This constraint is expected to be particularly pronounced among mammals, given that this group contains some of the largest species on Earth. We tested these hypotheses among marsupials, a group in which mass-specific metabolic rates are roughly 30% lower than those of eutherian mammals of similar size, leading to the expectation that metabolic rate should be a major constraint. Our findings support both hypotheses because levels of sperm competition are associated with increases in sperm size, but low mass-specific metabolic rate constrains sperm size among large species. We also found that the relationship between sperm size and mass-specific metabolic rate is steeper among marsupials and shallower among eutherian mammals. This finding has two implications: marsupials respond to changes in mass-specific metabolic rate by modifying sperm length to a greater extent, suggesting that they are more constrained by metabolic rate. In addition, for any given mass-specific metabolic rate, marsupials produce longer sperm. We suggest that this is the consequence of marsupials diverting resources away from sperm numbers and into sperm size, due to their efficient sperm transport along the female tract and the existence of mechanisms to protect sperm.

Tourmente, Maximiliano; Gomendio, Montserrat; Roldan, Eduardo R. S.




EPA Science Inventory

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


Microdissection testicular sperm extraction: an update  

PubMed Central

Patients with non-obstructive azoospermia (NOA) were once considered to be infertile with few treatment options due to the absence of sperm in the ejaculate. In the last two decades, the advent of intracytoplasmic sperm injection (ICSI), and the application of various testicular sperm retrieval techniques, including fine needle aspiration (FNA), conventional testicular sperm extraction (TESE) and microdissection testicular sperm extraction (micro-TESE) have revolutionized treatment in this group of men. Because most men with NOA will have isolated regions of spermatogenesis within the testis, studies have illustrated that sperm can be retrieved in most men with NOA, including Klinefelter's syndrome (KS), prior history of chemotherapy and cryptorchidism. Micro-TESE, when compared with conventional TESE has a higher sperm retrieval rate (SRR) with fewer postoperative complications and negative effects on testicular function. In this article, we will compare the efficacy of the different procedures of sperm extraction, discuss the medical treatment and the role of testosterone optimization in men with NOA and describe the micro-TESE surgical technique. Furthermore, we will update our overall experience to allow counseling on the prognosis of sperm retrieval for the specific subsets of NOA.

Dabaja, Ali A; Schlegel, Peter N



Sperm competition and sex allocation in simultaneous hermaphrodites  

Microsoft Academic Search

Summary Sex allocation theory is developed for hermaphrodites having frequent copulations and long-term sperm storage. Provided the sperm displacement mechanisms are similar to those known in insects, the ESS allocation to sperm versus eggs satisfies a rather simple rule. There are no data to test this rule, as yet.

Eric L. Charnov



Capacitation and the acrosome reaction in equine sperm  

Microsoft Academic Search

During sexual reproduction, the sperm and oocyte must fuse before the production of a diploid zygote can proceed. In mammals such as equids, fusion depends critically on complex changes in the plasma membrane of the sperm and, not surprisingly, this membrane differs markedly from that of somatic cells. After leaving the testes, sperm cease to synthesize plasma membrane lipids or

B. M. Gadella; R. Rathi; J. F. H. M. Brouwers; T. A. E. Stout; B. Colenbrander



Wolbachia infection reduces sperm competitive ability in an insect  

Microsoft Academic Search

The maternally inherited bacterium Wolbachia pipientis imposes significant fitness costs on its hosts. One such cost is decreased sperm production resulting in reduced fertility of male Drosophila simulans infected with cytoplasmic incompatibility (CI) inducing Wolbachia. We tested the hypothesis that Wolbachia infection affects sperm competitive ability and found that Wolbachia infection is indeed associated with reduced success in sperm competition

Fleur E. Champion de Crespigny; Nina Wedell



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


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

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



Immunolocalization of humanin in human sperm and testis.  


We have discovered, by immunocytochemistry and immunoelectronmicroscopy, that humanin (HN) is expressed in human ejaculated sperm and testis. In sperm, the HN immunolabeling pattern depends on sperm morphology; in particular, HN is mainly localized in the midpiece of sperm in semen samples with normal morphology and in cytoplasmic residues and entire tail in those with abnormal morphology. We also found HN in the cytoplasm and nucleus of spermatocytes and spermatids and in experimentally uncoiled chromatin of mature ejaculated sperm. Because it has been established that HN has antiapoptotic properties, it is reasonably hypothesized that HN may play an important role in preventing apoptosis in human sperm and testis. Thus, the examination of the HN localization in normal and abnormal sperm could be proposed as an auxiliary test to better define sperm quality. PMID:20542501

Moretti, Elena; Giannerini, Valentina; Rossini, Lara; Matsuoka, Masaaki; Trabalzini, Lorenza; Collodel, Giulia



Lung function testing — useless in ventilated newborns?  

Microsoft Academic Search

Several methods have been used for lung function testing in the ventilated newborn. The interest in the field has been stimulated by the recent appearance of commercially available equipment for assessment of mechanical parameters and of functional residual capacity in this group. Nevertheless, lung function testing is rarely used as a clinical routine, even such simple variables as tidal volume

Ola Hjalmarson



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 ...


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 ...


Two types of assays for detecting frog sperm chemoattraction.  


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

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



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

Microsoft Academic Search

Sexual reproduction requires the fusion of sperm cell and oocyte during fertilization to produce the diploid zygote. In mammals complex changes in the plasma membrane of the sperm cell are involved in this process. Sperm cells have unusual membranes compared to those of somatic cells. After leaving the testes, sperm cells cease plasma membrane lipid and protein synthesis, and vesicle

Frits M Flesch; Barend M Gadella



Biparental Inheritance of ?-Tubulin during Human Fertilization: Molecular Reconstitution of Functional Zygotic Centrosomes in Inseminated Human Oocytes and in Cell-free Extracts Nucleated by Human Sperm  

PubMed Central

Human sperm centrosome reconstitution and the parental contributions to the zygotic centrosome are examined in mammalian zygotes and after exposure of spermatozoa to Xenopus laevis cell-free extracts. The presence and inheritance of the conserved centrosomal constituents ?-tubulin, centrin, and MPM-2 (which detects phosphorylated epitopes) are traced, as is the sperm microtubule-nucleating capability on reconstituted centrosomes. ?-Tubulin is biparentally inherited in humans (maternal >> than paternal): Western blots detect the presence of paternal ?-tubulin. Recruitment of maternal ?-tubulin to the sperm centrosome occurs after sperm incorporation in vivo or exposure to cell-free extract, especially after sperm “priming” induced by disulfide bond reduction. Centrin is found in the proximal sperm centrosomal region, demonstrates expected calcium sensitivity, but appears absent from the zygotic centrosome after sperm incorporation or exposure to extracts. Sperm centrosome phosphorylation is detected after exposure of primed sperm to egg extracts as well as during the early stages of sperm incorporation after fertilization. Finally, centrosome reconstitution in cell-free extracts permits sperm aster microtubule assembly in vitro. Collectively, these results support a model of a blended zygotic centrosome composed of maternal constituents attracted to an introduced paternal template after insemination.

Simerly, Calvin; Zoran, Sara S.; Payne, Chris; Dominko, Tanja; Sutovsky, Peter; Navara, Christopher S.; Salisbury, Jeffery L.; Schatten, Gerald



Pulmonary Function Testing in Small Laboratory Mammals.  

National Technical Information Service (NTIS)

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 whi...

J. J. O'Neil J. A. Raub



Flow Cytometry of Sperm.  

National Technical Information Service (NTIS)

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 chromoso...

B. L. Gledhill



Normal spermatogenesis and sperm function in a subject affected by cerebellar ataxia due to congenital vitamin E deficiency.  


Vitamin E possesses potent beneficial effects on mammalian spermatogenesis and sperm quality. Subjects affected by cerebellar ataxia due to congenital isolated vitamin E deficiency (AVED) show vitamin E deficiency caused by a selective impaired gastrointestinal absorption of vitamin E for a mutation in the gene for ?-tocopherol transfer protein leading to impairment of vitamin E absorption and decreased vitamin E plasma levels. Here, we present a 34-year-old male patient with AVED showing normal seminal parameters and normal gonadotrophins, testosterone and inhibin B plasma levels. The normal standard seminal parameters of this patient with AVED possibly question the role of vitamin E in human spermatogenesis. PMID:23445347

Rossato, M; Mariotti, C



Sperm chromatin structure assay (SCSA®).  


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

Evenson, Donald P



Flow cytometry of sperm  

SciTech Connect

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.

Gledhill, B.L.



Contraceptive vaccines targeting sperm.  


Overpopulation is a global problem of significant magnitude, with grave implications for the future. Development of new contraceptives is necessary, as existing forms of birth control are unavailable, impractical and/or too expensive for many individuals due to sociological, financial or educational limitations. Immunocontraception and, in particular, the targeting of antibodies to sperm-specific antigens implicated in sperm-egg binding and fertilisation offers an attractive approach to control fertility. Sperm-specific antibodies may impair fertility by inhibiting sperm motility, by reducing penetration of the cervical mucus by sperm, or by interfering in sperm capacitation or the acrosome reaction; alternatively, antisperm antibodies may invoke the complement cascade, resulting in sperm lysis. The antibodies raised against sperm-specific antigens have proved to be extremely effective at reducing sperm-egg interactions in vitro; fertility trials in subhuman primates will eventually be needed to prove the effectiveness of the sperm antigens in terms of contraceptive efficacy before trials in humans can be justified. In addition, existing and emerging strategies (such as sperm proteomics, the determination of molecular and structural details of sperm proteins, and the modelling of protein-ligand interactions using X-ray and/or NMR structures to name a few) are expected to provide the experimental foundation for the design of small molecule inhibitors with antifertility effects. The technology underpinning vaccine development is constantly being developed and the introduction of DNA/RNA vaccines is certain to impact upon the field of immunocontraception. PMID:15833075

Suri, Anil



Sperm competition in bats  

Microsoft Academic Search

SUMMARY 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

D. J. Hosken



Functional Performance Testing in Athletes with Functional Ankle Instability  

PubMed Central

Purpose To determine if functional performance deficits are present in athletes with functional ankle instability (FAI) compared to healthy athletes using various functional performance tests. Methods Sixty two athletes (mean age-21.7±1.8years; height-168.2±9.1cm; weight-63.8±11.0kg) participated in this case control study. Athletes were divided into two groups: athletes with FAI (FAI group, n=31) and healthy athletes (Non-FAI group, n=31). The FAI group was further divided into two subgroups: FAI with giving way (FAI-GW), FAI with no giving way (FAI-NGW). Functional performance was assessed with the single-limb hopping test, figure-of-8 hop test, side-hop test, single-limb hurdle test, square hop test and single hop test. Results Significant differences (P<0.05) were observed for all the functional performance tests (FPTs) except the single hop test between FAI and Non-FAI groups; between FAI-GW, FAI-NGW and Non-FAI groups. Additionally, the involved limb performed significantly worse (P<0.05) than the contra-lateral uninvolved limb of the FAI-GW group for the above-mentioned FPTs. Conclusion Significant functional performance deficits were observed in the FAI group in all tests except single hop test with greater deficits observed in the FAI-GW group. Hence, these tests can be used to determine the presence of FAI. However no deficits were identified for the test involving sagittal plane functional activities suggesting that this test can not be used as a criterion to discriminate individuals with FAI. It was further ascertained that functional performance was not affected by limb dominance.

Sharma, Nidhi; Sharma, Archna; Singh Sandhu, Jaspal



Applied physiology: lung function testing in children  

Microsoft Academic Search

Recent developments in the techniques available to measure lung function in children have greatly improved our understanding of lung development in both health and disease. In everyday clinical practice, lung function tests can provide objective information about the severity and progression of disease and the response to treatment. Although spirometry remains the most useful and widely used test, newer techniques

Jonathan M. Couriel; Frances Child



Snail sperm production characteristics vary with sperm competition risk  

PubMed Central

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

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



Cohomologies of spaces of Schwartz test functions  

NASA Astrophysics Data System (ADS)

We calculate cohomologies of the Schwartz algebras D and S, which are basic spaces (test-function spaces) of the distribution theory. In the process, we find special cohomologies of the quotient algebras E/D and M/S (E andMare also test-function spaces), which are quite unusual from the standpoint of the standard functional analysis and are interesting for theoretical and mathematical physics.

Zharinov, V. V.



The ability of sperm selection techniques to remove single- or double-strand DNA damage  

PubMed Central

A wide variety of techniques for the preparation of sperm are currently available, of which the most commonly employed are density–gradient centrifugation (DGC) and swim-up (SUP). To date, these methods appear to be effective in selecting functional sperm for assisted reproduction techniques (ART), but they may have negative effects on sperm DNA. In this study, the ability of these semen processing techniques to eliminate spermatozoa containing single- and double-strand DNA damage was assessed by the two-tailed comet assay and the sperm chromatin dispersion test in 157 semen samples from patients seeking assisted reproduction treatment. Our results indicated that SUP and DGC are equally efficient in eliminating spermatozoa containing double-strand DNA damage and sperm with highly damaged (degraded) DNA, as characterized by the presence of both single- and double-strand DNA breaks. However, DGC is more efficient than SUP in selecting spermatozoa that are free from single-strand DNA damage. Future studies should characterise the importance of the various types of DNA damage and examine the sperm processing protocols used in each laboratory to determine their ability to eliminate DNA damage and hence, prevent the potential transmission of genetic mutations via ART.

Enciso, Maria; Iglesias, Miriam; Galan, Isabel; Sarasa, Jonas; Gosalvez, Antonio; Gosalvez, Jaime



Effects of testing on learning of functions.  


Is learning of a complex functional relationship enhanced by trying to predict what output will go with a given input, as compared to studying an input-output pair? We examined learning of a bilinear function and transfer to new items outside the trained range. Subjects either saw the input-output pairs (study-only condition) or attempted to guess the output and then saw the pair (test/study condition). The total study times were equated, and motivation was enhanced with a monetary bonus. Performance was markedly better for the test/study condition, both within the trained range and in the transfer test. This benefit of testing during training was observed on a criterial test administered shortly after training. Testing has long been shown to enhance the explicit learning and retention of verbal material; our present findings reveal a novel domain for which testing can also be advantageous-that is, function learning. PMID:21630113

Kang, Sean H K; McDaniel, Mark A; Pashler, Harold



Accelerated Functional Testing of Digital Microfluidic Biochips  

Microsoft Academic Search

Structural testing of digital microfluidic biochips targets the detection of physical defects, but it does not guarantee robust execution of target bioassays or the integrity of assay outcomes. Functional testing is needed to detect fluidic malfunctions. Such tests ensure, whether or not, the elementary fluidic operations, such as droplet transportation, mixing, incubation, and splitting are reliably executed on the microfluidic

Debasis Mitra; Sarmishtha Ghoshal; Hafizur Rahaman; Bhargab B. Bhattacharya; D. D. Majumder; K. Chakrabarty



Accelerated Functional Testing of Digital Microfluidic Biochips  

Microsoft Academic Search

Structural testing of digital microfluidic biochips targets the detection of physical defects, but it does not guarantee robust execution of target bioassays or the integrity of assay outcomes. Functional testing is needed to detect fluidic malfunctions. Such tests ensure whether or not, the elementary fluidic operations, such as droplet transportation, mixing, incubation, and splitting are reliably executed on the microfluidic

Debasis Mitra; Sarmishtha Ghoshal; Hafizur Rahaman; Bhargab B. Bhattacharya; D. D. Majumder; K. Chakrabarty



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

Microsoft Academic Search

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

Nina Wedell; Matthew Gage; Geoffrey Parker


Decreasing Epididymal Sperm Reserves Enhances the Detection of Ethoxyethanol-Induced Spermatotoxicity.  

National Technical Information Service (NTIS)

Current test strategies for assessing male reproductive toxicity may be inadequate for estimating risk in humans. High levels of sperm production and existence of large epididymal sperm reserves in most test species may impede the detection of spermatoxic...

M. E. Hurtt H. Zenick



ACS CCD Functional Test for HRC  

NASA Astrophysics Data System (ADS)

During the ACS Repair {ACS-R} conducted during Hubble Servicing Mission 4 {SM4}, astronauts will install a CCD Electronics Box Replacement {CEB-R} and Low-Voltage Power Supply Replacement {LVPS-R}. A rudimentary aliveness/functional test {AT/FT} will be conducted on-orbit during SM4. Shortly after SM4, the comprehensive CCD function test {FT} summarized here will be conducted as part of SMOV. This program is modeled after the original CCD functional test {HST program 9005, PI Mark Clampin} conducted following the initial installation of ACS during Hubble Servicing Mission 3B. This HRC-only proposal has a companion WFC proposal 11369.;

Mutchler, Max



Binomial test statistics using Psi functions  

SciTech Connect

For the negative binomial model (probability generating function (p + 1 - pt){sup -k}) a logarithmic derivative is the Psi function difference {psi}(k + x) - {psi}(k); this and its derivatives lead to a test statistic to decide on the validity of a specified model. The test statistic uses a data base so there exists a comparison available between theory and application. Note that the test function is not dominated by outliers. Applications to (i) Fisher's tick data, (ii) accidents data, (iii) Weldon's dice data are included.

Bowman, Kimiko o [ORNL



Collection and freezing of epididymal stallion sperm.  


The ability to harvest and preserve epididymal sperm from a stallion after simple elective castration, a catastrophic injury, or severe acute illness and subsequent death has been realized, allowing for the preservation of genetics that would have been lost otherwise.Currently, the care taken to collect the testes and epididymides properly, coupled with proper packaging and shipping, could make the greatest contribution to salvaging viable sperm. As advances in assisted reproductive techniques continue, more offspring may be obtained from stored epididymal sperm from valuable stallions. PMID:17129795

Bruemmer, Jason E



Functional Testing of Semiconductor Random Access Memories  

Microsoft Academic Search

This paper presents an overview of the problem of testing semiconductor random access memories (RAMs). An important aspect of this test procedure is the detection of permanent faults that cause the memory to function incorrectly. Functional-level fault models are very useful for describing a wide variety of RAM faults. Several fault models are &scussed throughout the paper, including the stuck-at-0\\/1

Magdy S. Abadir; Hassan K. Reghbati



Autonomic Function Tests: Some Clinical Applications  

PubMed Central

Modern autonomic function tests can non-invasively evaluate the severity and distribution of autonomic failure. They have sufficient sensitivity to detect even subclinical dysautonomia. Standard laboratory testing evaluates cardiovagal, sudomotor and adrenergic autonomic functions. Cardiovagal function is typically evaluated by testing heart rate response to deep breathing at a defined rate and to the Valsalva maneuver. Sudomotor function can be evaluated with the quantitative sudomotor axon reflex test and the thermoregulatory sweat test. Adrenergic function is evaluated by the blood pressure and heart rate responses to the Valsalva maneuver and to head-up tilt. Tests are useful in defining the presence of autonomic failure, their natural history, and response to treatment. They can also define patterns of dysautonomia that are useful in helping the clinician diagnose certain autonomic conditions. For example, the tests are useful in the diagnosis of the autonomic neuropathies and distal small fiber neuropathy. The autonomic neuropathies (such as those due to diabetes or amyloidosis) are characterized by severe generalized autonomic failure. Distal small fiber neuropathy is characterized by an absence of autonomic failure except for distal sudomotor failure. Selective autonomic failure (which only one system is affected) can be diagnosed by autonomic testing. An example is chronic idiopathic anhidrosis, where only sudomotor function is affected. Among the synucleinopathies, autonomic function tests can distinguish Parkinson's disease (PD) from multiple system atrophy (MSA). There is a gradation of autonomic failure. PD is characterized by mild autonomic failure and a length-dependent pattern of sudomotor involvement. MSA and pure autonomic failure have severe generalized autonomic failure while DLB is intermediate.

Tomalia, Victoria A.; Park, Ki-Jong



Autonomic function tests: some clinical applications.  


Modern autonomic function tests can non-invasively evaluate the severity and distribution of autonomic failure. They have sufficient sensitivity to detect even subclinical dysautonomia. Standard laboratory testing evaluates cardiovagal, sudomotor and adrenergic autonomic functions. Cardiovagal function is typically evaluated by testing heart rate response to deep breathing at a defined rate and to the Valsalva maneuver. Sudomotor function can be evaluated with the quantitative sudomotor axon reflex test and the thermoregulatory sweat test. Adrenergic function is evaluated by the blood pressure and heart rate responses to the Valsalva maneuver and to head-up tilt. Tests are useful in defining the presence of autonomic failure, their natural history, and response to treatment. They can also define patterns of dysautonomia that are useful in helping the clinician diagnose certain autonomic conditions. For example, the tests are useful in the diagnosis of the autonomic neuropathies and distal small fiber neuropathy. The autonomic neuropathies (such as those due to diabetes or amyloidosis) are characterized by severe generalized autonomic failure. Distal small fiber neuropathy is characterized by an absence of autonomic failure except for distal sudomotor failure. Selective autonomic failure (which only one system is affected) can be diagnosed by autonomic testing. An example is chronic idiopathic anhidrosis, where only sudomotor function is affected. Among the synucleinopathies, autonomic function tests can distinguish Parkinson's disease (PD) from multiple system atrophy (MSA). There is a gradation of autonomic failure. PD is characterized by mild autonomic failure and a length-dependent pattern of sudomotor involvement. MSA and pure autonomic failure have severe generalized autonomic failure while DLB is intermediate. PMID:23346153

Low, Phillip A; Tomalia, Victoria A; Park, Ki-Jong



Use of Fluorescent Dyes for Readily Recognizing Sperm Damage  

PubMed Central

Sperm is produced by the testis and mature in the epididymis. For having a successful conception, the fertilizing sperm should have functional competent membranes, intact acrosome, functional mitochondria and an intact haploid genome. The effects of genetic and environmental factors result in sperm vulnerability to damage in the process of spermatogenesis and maturation. In recent years, the feasibility of detecting sperm damage is enhanced through the advances in technologies like fluoscerent staining techniques assisted with fluorescence microscope, flow cytometry and computer analysis systems. Fluoscerent staining techniques involve the use of fluorescent dyes, either directly or indirectly for binding them with some ingredients of sperm and evaluating the damage of the structure or function of the sperm, i.e. membrane, acrosome, mitochondria, chromosome or DNA.

Farah, Omar Ibrahim; Cuiling, Li; Jiaojiao, Wang; Huiping, Zhang



A quantitative test of upper extremity function  

Microsoft Academic Search

IN A number of clinical situations, there is a need for a simple semi-quantitative test of upper extremity function. Measurement of function is of particular importance in evaluating the results of hand surgery and upper extremity prosthesis, in following the development of hand deformities in rheumatoid arthritis and neurological diseases, and in judging the effectiveness of different forms of therapy.





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...


Effect of tulsi (Ocimum Sanctum Linn.) on sperm count and reproductive hormones in male albino rabbits  

PubMed Central

Fresh leaves of Ocimum Sanctum (OS) were used to study its effect on male reproductive function (sperm count and reproductive hormones) in male albino rabbits. Animals in the test group received supplementation of 2 g of fresh leaves of OS per rabbit for 30 days, while the control group was maintained on normal diet for the same duration. Sperm count and hormonal estimation [testosterone, follicle stimulating hormone (FSH), and luteinizing hormone (LH)] were done in serum samples of both groups and compared. A significant decrease was noted in the sperm count in test group rabbits. Serum testosterone levels showed marked increase while FSH and LH levels were significantly reduced in OS-treated rabbits. The results suggest the potential use of OS as an effective male contraceptive agent.

Sethi, Jyoti; Yadav, Mridul; Sood, Sushma; Dahiya, Kiran; Singh, Veena



Sperm-activating peptide induces asymmetric flagellar bending in sea urchin sperm.  


Speract, a sperm-activating peptide (SAP) from sea urchin eggs, induces various sperm responses including a transient increase in the intracellular Ca2+ concentration. However, it has not been clarified how speract modulates sperm motility and whether it functions as a chemoattractant. To confirm the effect of speract on sperm motility, we observed the flagellar bending response to speract in sperm of Hemicentrotus pulcherrimus, in experiments using caged speract and a lighting system for a microscope newly developed with a power LED. We found that speract induces increases in curvature of swimming paths and changes flagellar bending shape to asymmetric. These facts show that speract directly regulates flagellar motility, and suggest that speract-induced increases in intracellular Ca2+ concentration play an actual role in regulation of the flagellar movement. PMID:15795491

Shiba, Kogiku; Ohmuro, Junko; Mogami, Yoshihiro; Nishigaki, Takuya; Wood, Christopher D; Darszon, Alberto; Tatsu, Yoshiro; Yumoto, Noboru; Baba, Shoji A



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.  


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

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



The Full Function Test Explosive Generator  

SciTech Connect

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.

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



Ion channels in sperm motility and capacitation.  


Spermatozoa depend upon ion channels to rapidly exchange information with the outside world and to fertilise the egg. These efficient ion transporters participate in many of the most important sperm processes, such as motility and capacitation. It is well known that sperm swimming is regulated by [Ca2+]i. In the sea urchin sperm speract, a decapeptide isolated from egg outer envelope, induces changes in intracellular Ca2+ ([Ca2+]i), Na+, cAMP and cGMP, membrane potential (Em) and pH (pHi). Photoactivation of a speract analogue induces Ca2+ fluctuations that generate turns that are followed by straighter swimming paths. A fast component of the [Ca2+], increase that most likely occurs through voltage dependent Ca2+ channels (Ca(v)s) is essential for these turns. The Ca(v)s involved are modulated by the Em changes triggered by speract. On the other hand, mammalian sperm gain the ability to fertilise the egg after undergoing a series of physiological changes in the female tract. This maturational process, known as capacitation, encompasses increases in [Ca2+]i and pHi, as well as an Em hyperpolarization in mouse sperm. Our electrophysiological, immunological and molecular-biological experiments indicate that inwardly rectifying K+ channels regulated by ATP (KATP channels) and epithelial Na+ channels (ENaCs) are functionally present in mouse spermatogenic cells and sperm. Notably, pharmacological experiments indicate that the opening of KATP channels and closure of ENaCs may contribute to the hyperpolarization that accompanies mouse sperm capacitation. Remarkably, both in the sea urchin sperm speract response and in the mouse sperm capacitation, Em hyperpolarization seems necessary to remove inactivation from Ca(v) channels so they can then open. PMID:17644965

Darszon, A; Treviño, C L; Wood, C; Galindo, B; Rodríguez-Miranda, E; Acevedo, J J; Hernandez-González, E O; Beltrán, C; Martínez-López, P; Nishigaki, T



PhysioChemical Characteristics of Seminal Plasma and Development of Media and Methods for the Cryopreservation of European eel Sperm  

Microsoft Academic Search

The high sperm density, together with the short spermatozoa swimming time, makes European eel sperm manipulation and assessment for quality difficult. Two diluting media (K15 and K30) previously designed for Japanese eel sperm were tested. After 24 h, European eel sperm showed significant reduction in the percentage of motile spermatozoa after activation and different motility parameters (VAP, angular velocity; VCL, curvilinear

J. F. Asturiano; L. Pérez; D. L. Garzón; F. Marco-Jiménez; D. S. Peñaranda; J. S. Vicente; M. Jover



Effects of molybdenum on sperm quality and testis oxidative stress.  


In order to investigate the effects of molybdenum (Mo) on sperm parameters and testicular oxidative stress, the ICR strain of adult mice were exposed to different doses of molybdenum for a sub-acute toxicity test. Compared to the control, our results showed that the sperm parameters, including the epididymis index, sperm motility, sperm count, and morphology, increased by a moderate dose of Mo (25?mg/L), but were negatively affected at high doses (? 100?mg/L). In addition, the changes of sperm parameters were accompanied with changes of the superoxide dismutase (SOD) activities, the glutathione peroxidase (GPx) activities, and the malondialdehyde (MDA) levels in testes. In conclusion, Mo affects the sperm quality through regulating the testicular oxidative stress in a complex manner. PMID:23651332

Zhai, Xiao-Wei; Zhang, Yu-Ling; Qi, Qiao; Bai, Yu; Chen, Xiao-Li; Jin, Li-Jun; Ma, Xue-Gang; Shu, Run-Zhe; Yang, Zi-Jun; Liu, Feng-Jun



Novel System for the Co-Culture of Epididymal Epithelial Cells and Sperm from Adult Rats.  

National Technical Information Service (NTIS)

To study interactions which occur between the epididymal epithelial cells and sperm within the epididymis during sperm maturation, a specialized co-culture system capable of supporting the differentiated function of these cell types must be utilized. A mu...

G. R. Klinefelter



ACS CCD functional test for WFC  

NASA Astrophysics Data System (ADS)

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

Mutchler, Max



Towards a physiological role for cytochrome P450 aromatase in ejaculated human sperm  

Microsoft Academic Search

BACKGROUND: Advances in the definition of the function and the mechanism of estrogen action in different tis- sues have come from human and animal models of estrogen insufficiency. Recently we have demonstrated that aromatase is present and biologically active in human ejaculated sperm, suggesting that autonomous estradiol sperm production may influence sperm functions. In the present study we investigate a

Saveria Aquila; Diego Sisci; Mariaelena Gentile; Amalia Carpino; Emilia Middea; Stefania Catalano; Vittoria Rago



The testis anion transporter TAT1 (SLC26A8) physically and functionally interacts with the cystic fibrosis transmembrane conductance regulator channel: a potential role during sperm capacitation.  


The Slc26 gene family encodes several conserved anion transporters implicated in human genetic disorders, including Pendred syndrome, diastrophic dysplasia and congenital chloride diarrhea. We previously characterized the TAT1 (testis anion transporter 1; SLC26A8) protein specifically expressed in male germ cells and mature sperm and showed that in the mouse, deletion of Tat1 caused male sterility due to a lack of sperm motility, impaired sperm capacitation and structural defects of the flagella. Ca(2+), Cl(-) and HCO(3)(-) influxes trigger sperm capacitation events required for oocyte fertilization; these events include the intracellular rise of cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA)-dependent protein phosphorylation. The cystic fibrosis transmembrane conductance regulator (CFTR) is expressed in mature sperm and has been shown to contribute to Cl(-) and HCO(3)(-) movements during capacitation. Furthermore, several members of the SLC26 family have been described to form complexes with CFTR, resulting in the reciprocal regulation of their activities. We show here that TAT1 and CFTR physically interact and that in Xenopus laevis oocytes and in CHO-K1 cells, TAT1 expression strongly stimulates CFTR activity. Consistent with this, we show that Tat1 inactivation in mouse sperm results in deregulation of the intracellular cAMP content, preventing the activation of PKA-dependent downstream phosphorylation cascades essential for sperm activation. These various results suggest that TAT1 and CFTR may form a molecular complex involved in the regulation of Cl(-) and HCO(3)(-) fluxes during sperm capacitation. In humans, mutations in CFTR and/or TAT1 may therefore be causes of asthenozoospermia and low fertilizing capacity of sperm. PMID:22121115

Rode, Baptiste; Dirami, Thassadite; Bakouh, Naziha; Rizk-Rabin, Marthe; Norez, Caroline; Lhuillier, Pierre; Lorès, Patrick; Jollivet, Mathilde; Melin, Patricia; Zvetkova, Ilona; Bienvenu, Thierry; Becq, Frédéric; Planelles, Gabrielle; Edelman, Aleksander; Gacon, Gérard; Touré, Aminata



Dynamics of sperm transfer in the ant Leptothorax gredleri  

NASA Astrophysics Data System (ADS)

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.

Oppelt, Angelika; Heinze, Jürgen



Improved methodology for a sea urchin sperm cell bioassay for marine waters  

Microsoft Academic Search

A simple sperm\\/fertilization bioassay, primarily using sea urchin (and sand dollar) gametes, was improved to yield a quick, sensitive, and cost-effective procedure for measuring toxicity in marine waters. Standard sperm bioassays are conducted by exposing sperm cells to test solutions for 60 min prior to addition of eggs to the test solution for fertilization. Reduced fertilization success (as indicated by

Paul A. Dinnel; Jeanne M. Link; Quentin J. Stober



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


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

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



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

USGS Publications Warehouse

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

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



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

SciTech Connect

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.

Wyrobek, A.J.



The endocannabinoid system in bull sperm and bovine oviductal epithelium: role of anandamide in sperm-oviduct interaction.  


Anandamide binds to cannabinoid receptors and plays several central and peripheral functions. The aim of this work was to study the possible role for this endocannabinoid in controlling sperm-oviduct interaction in mammals. We observed that bull sperm and bovine oviductal epithelial cells express cannabinoid receptors, CB1 and CB2, and fatty acid amide hydrolase, the enzyme that controls intracellular anandamide levels. A quantitative assay to determine whether anandamide was involved in bovine sperm-oviduct interaction was developed. R(+)-methanandamide, a non-hydrolysable anandamide analog, inhibited sperm binding to and induced sperm release from oviductal epithelia. Selective CB1 antagonists (SR141716A or AM251) completely blocked R(+)-methanandamide effects. However, SR144528, a selective CB2 antagonist, did not exert any effect, indicating that only CB1 was involved in R(+)-methanandamide effect. This effect was not caused by inhibition of the sperm progressive motility or by induction of the acrosome reaction. Overall, our findings indicate for the first time that the endocannabinoid system is present in bovine sperm and oviductal epithelium and that anandamide modulates the sperm-oviduct interaction, by inhibition of sperm binding and induction of sperm release from oviductal epithelial cells, probably by activating CB1 receptors. PMID:19042982

Gervasi, María Gracia; Rapanelli, Maximiliano; Ribeiro, María Laura; Farina, Mariana; Billi, Silvia; Franchi, Ana María; Perez Martinez, Silvina



Identification and characterization of highly expressed proteins in sperm cells of the marine mussel Mytilus edulis.  


Proteomic analysis on sperm has been restricted to only a few model organisms. We present here a 2DE PAGE proteome map of sperm cells from a nonmodel organism, the marine mussel Mytilus edulis, a free-spawning marine invertebrate with external fertilization. Ninety-six protein spots showing high expression were selected and of these 77 were successfully identified by nESI-MS analysis. Many of the identifications are relevant to sperm cell physiology and mtDNA functioning. The results and proteomics approach used are discussed in relation to their potential for advancing understanding of the unusual system of mtDNA inheritance described in Mytilus spp., and for the testing of evolutionary hypotheses pertaining to the role of fertilization in the speciation process. PMID:22623270

Diz, Angel P; Dudley, Edward; Skibinski, David O F



Instrumented testing of functional knee braces  

Microsoft Academic Search

The ability of seven functional knee braces to control anterior tibial displacement in three severely lax ACL deficient knees using two instrumented testing devices was studied. Some braces were statistically shown to be much better in this regard than others, but not all data obtained was statistically significant. This material should aid one in determining which braces offer the greatest

Charles Beck; David Drez; John Young; W. Dilworth Cannon; Mary Lou Stone



Detection of dilute sperm samples using photoacoustic flowmetry  

NASA Astrophysics Data System (ADS)

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

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



[Human sperm chromatin damage and its determination: an update].  


Human sperm chromatin damage is one of the common reasons for reduced male reproductive capacity, which is influenced by genetics, environment, lifestyle and other factors, as well as associated with male infertility and habitual abortion. Along with deeper insights into the structure and function of sperm chromatin, improvement and promotion of detection technology for its integrity, and wider application of assisted reproductive technology, sperm DNA damage has been recognized as an important new indicator for the evaluation of sperm quality, and has a great clinical significance in the assessment of male fertility and selection of assisted reproductive technology. PMID:23757973

Zhao, Kai; Xiong, Cheng-Liang



Cryopreservation of domestic animal sperm cells.  


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

Barbas, J P; Mascarenhas, R D



Targeted disruption of the Akap4 gene causes defects in sperm flagellum and motility.  


A-kinase anchoring proteins (AKAPs) tether cyclic AMP-dependent protein kinases and thereby localize phosphorylation of target proteins and initiation of signal-transduction processes triggered by cyclic AMP. AKAPs can also be scaffolds for kinases and phosphatases and form macromolecular complexes with other proteins involved in signal transduction. Akap4 is transcribed only in the postmeiotic phase of spermatogenesis and encodes the most abundant protein in the fibrous sheath, a novel cytoskeletal structure present in the principal piece of the sperm flagellum. Previous studies indicated that cyclic AMP-dependent signaling processes are important in the regulation of sperm motility, and gene targeting was used here to test the hypothesis that AKAP4 is a scaffold for protein complexes involved in regulating flagellar function. Sperm numbers were not reduced in male mice lacking AKAP4, but sperm failed to show progressive motility and male mice were infertile. The fibrous sheath anlagen formed, but the definitive fibrous sheath did not develop, the flagellum was shortened, and proteins usually associated with the fibrous sheath were absent or substantially reduced in amount. However, the other cytoskeletal components of the flagellum were present and appeared fully developed. We conclude that AKAP4 is a scaffold protein required for the organization and integrity of the fibrous sheath and that effective sperm motility is lost in the absence of AKAP4 because signal transduction and glycolytic enzymes fail to become associated with the fibrous sheath. PMID:12167408

Miki, Kiyoshi; Willis, William D; Brown, Paula R; Goulding, Eugenia H; Fulcher, Kerry D; Eddy, Edward M



The Sperm Penetration Assay for the Assessment of Fertilization Capacity  

PubMed Central

Summary The sperm penetration assay, or zona-free hamster oocyte penetration assay is utilized to measure the ability of sperm to undergo capacitation, fuse with the egg membrane and decondense the sperm head within the cytoplasm of the oocyte resulting in the formation of the male pronucleus. The test is scored by calculation the percentage of ova that are penetrated or the average number of sperm penetrations per ovum. It has been used to identify those couples who will have a high likelihood of success with in vitro fertilization.

Hwang, Kathleen; Lamb, Dolores J.



An update on sperm retrieval techniques for azoospermic males  

PubMed Central

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.

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



Ion channels: Key elements in sea urchin sperm physiology  

NASA Astrophysics Data System (ADS)

Ion channels are deeply involved in sea urchin sperm activation, motility, chemotaxis and in the acrosome reaction. Unraveling ion channel function and regulation in sperm behavior has required a combination of complementary approaches since spermatozoa are very tiny cells. Planar bilayer and patch clamp techniques have allowed us to detect, for the first time, the activity of single channels in the plasma membrane of these cells. Unlike intact sperm, swollen sperm can be much more easily patch clamped and single channel activity recorded. These techniques, together with studies of membrane potential, intracellular Ca2+ and pH in whole sperm, have established the presence of K+, Ca2+, and Cl- channels in this cell. The strategies developed to study sea urchin sperm channels are applicable to mammalian spermatozoa. We recently detected a Ca2+ channel resembling one found in S. purpuratus sperm in planar bilayers containing mouse sperm plasma membranes. The presence of this Ca2+ channel in such diverse species suggests it is important in sperm function.

Darszon, Alberto; de de Latorre, Lucia; Vargas, Irma; Liévano, Arturo; Beltrán, Carmen; Santi, Celia; Labarca, Pedro; Zapata, Otilia



Sperm competition and the evolution of sperm design in mammals  

PubMed Central

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



Family of sperm associated antigens: relevance in sperm-egg interaction and immunocontraception.  


Overpopulation is a global problem of significant magnitude, with grave implications for the future. Development of new contraceptives is necessary, since current forms of birth control are unavailable, impractical and/or too expensive to many individuals due to sociological, financial, or educational limitations. A novel contraceptive strategy that is receiving considerable attention is that of immunocontraception. The targeting of antibodies to gamete-specific antigens implicated in sperm function, sperm-egg binding and fertilization offers an attractive approach to the growing global problem of over population. The sermatozoon has proteins that are unique, cell specific, immunogenic and accessible to antibodies. Immunological interaction with such molecules can cause block of sperm binding to the oocyte and thus fertilization. Modern biotechnologies (such as sperm proteomics, the determination of molecular and structural details of sperm proteins, and the modelling of protein-ligand interaction using X-ray and/or NMR structures to name a few) are trying to make intervention into the domain of human reproduction possible through the development of a variety of new methods and products to control fertility. The present article highlights the various sperm associated antigens involved in various aspects of sperm-egg interaction. PMID:17566289

Suri, A



Formation of primary sperm conjugates in a haplogyne spider (Caponiidae, Araneae) with remarks on the evolution of sperm conjugation in spiders.  


Sperm conjugation, where two or more sperm are physically united, is a rare but widespread pheno-menon across the animal kingdom. One group well known for its different types of sperm conjugation are spiders. Particularly, haplogyne spiders show a high diversity of sperm traits. Besides individual cleistospermia, primary (synspermia) and secondary (coenospermia, "spermatophore") sperm conjugation occurs. However, the evolution of sperm conjugates and sperm is not understood in this group. Here, we look at how sperm are transferred in Caponiidae (Haplogynae) in pursuit of additional information about the evolution of sperm transfer forms in spiders. Additionally, we investigated the male reproductive system and spermatozoa using light- and transmission electron-microscopy and provide a 3D reconstruction of individual as of well as conjugated spermatozoa. Mature spermatozoa are characterized by an extremely elongated, helical nucleus resulting in the longest spider sperm known to date. At the end of spermiogenesis, synspermia are formed by complete fusion of four spermatids. Thus, synspermia might have evolved early within ecribellate Haplogynae. The fused sperm cells are surrounded by a prominent vesicular area. The function of the vesicular area remains still unknown but might be correlated with the capacitation process inside the female. Further phylogenetic and functional implications of the spermatozoa and sperm conjugation are discussed. PMID:22982877

Lipke, Elisabeth; Michalik, Peter



Metabolism of motile zebrafish sperm  

Microsoft Academic Search

As metabolism of motile fish sperm is not well understood, the current study examined the metabolism of saline-activated zebrafish (Danio rerio) sperm. Activation of sperm with inhibitors of oxidative phosphorylation (potassium cyanide, 2,4 dinitrophenol or carbonyl cyanide 3-cholorophenylhydrazone) negatively impacted sperm motility by 60–90s postactivation. Incubation of quiescent sperm with 2,4 dinitrophenol prior to activation resulted in a 67% decrease

R. L. Ingermann; C. L. F. Schultz; M. K. Kanuga; J. G. Wilson-Leedy



20 CFR 718.103 - Pulmonary function tests.  

Code of Federal Regulations, 2013 CFR

...2013-04-01 2013-04-01 false Pulmonary function tests. 718.103 Section 718...Development of Medical Evidence § 718.103 Pulmonary function tests. (a) Any report of pulmonary function tests submitted in...



20 CFR 718.103 - Pulmonary function tests.  

Code of Federal Regulations, 2010 CFR

...2009-04-01 2009-04-01 false Pulmonary function tests. 718.103 Section 718...Development of Medical Evidence § 718.103 Pulmonary function tests. (a) Any report of pulmonary function tests submitted in...



20 CFR 718.103 - Pulmonary function tests.  

Code of Federal Regulations, 2010 CFR

...2010-04-01 2010-04-01 false Pulmonary function tests. 718.103 Section 718...Development of Medical Evidence § 718.103 Pulmonary function tests. (a) Any report of pulmonary function tests submitted in...



Sperm plasma membrane damage prior to intracytoplasmic sperm injection: a necessary condition for sperm nucleus decondensation.  


In the present study we investigated the relevance of sperm immobilization prior to intracytoplasmic sperm injection (ICSI) in the fertilization process. Using supravital staining of the spermatozoa with eosin and studying sperm decondensation with 2 mM dithiothreitol (DTT) in conditions imitating sperm handling during ICSI, we demonstrated that immobilization of the spermatozoon by squeezing its tail between the glass pipette and the bottom of the dish damages the sperm plasma membrane. Polyvinylpyrrolidone (PVP), which is usually present in the drop with the spermatozoon to facilitate its handling, was found to impede the access of both eosin and DTT to the sperm nucleus. We conclude that (i) sperm immobilization prior to ICSI damages the sperm plasma membrane, that (ii) this damage is sufficient for thiol-reducing agents to gain access to the sperm nucleus, and finally that (iii) PVP possibly interferes with sperm nucleus decondensation. PMID:8747054

Dozortsev, D; Rybouchkin, A; De Sutter, P; Dhont, M



Pulmonary Function Testing in Idiopathic Interstitial Pneumonias  

Microsoft Academic Search

Diffuse parenchymal lung diseases are a group of disorders that involve the space between the epithelial and endothelial basement membranes and are generally segregated into four major catego- ries. These include the idiopathic interstitial pneumonias, which are further categorized into seven clinical\\/radiologic\\/pathologic subsets.Thesedisordersgenerallyshareacommonpatternofphysi- ologic abnormality characterized by a restrictive ventilatory defect and reduced diffusing capacity (DLCO). Pulmonary function testing is

Fernando J. Martinez; Kevin Flaherty



Sperm dimorphism in Chilopoda: comparison of Scolopendromorpha and Geophilomorpha  

Microsoft Academic Search

Micro- and macrospermatozoa were found in the male genital duct and female spermatheca of Scolopendra cingulata (Scolopendromorpha), whereas sperm dimorphism was not encountered in Himantarium gabrielis (Geophilomorpha). On the basis of ultrastructural analysis, the micro-and macrosperm of S. cingulata and the single sperm type of H. gabrielis appear functional. They are very long, filiform cells with all the morphological parts




Immune Activation Reduces Sperm Quality in the Great Tit  

Microsoft Academic Search

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

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



Sperm proteins in teleostean and chondrostean (sturgeon) fishes.  


Sperm proteins in the seminal plasma and spermatozoa of teleostean and chondrostean have evolved adaptations due to the changes in the reproductive environment. Analysis of the composition and functions of these proteins provides new insights into sperm motility and fertilising abilities, thereby creating possibilities for improving artificial reproduction and germplasm resource conservation technologies (e.g. cryopreservation). Seminal plasma proteins are involved in the protection of spermatozoa during storage in the reproductive system, whereas all spermatozoa proteins contribute to the swimming and fertilising abilities of sperm. Compared to mammalian species, little data are available on fish sperm proteins and their functions. We review here the current state of the art in this field and focus on relevant subjects that require attention. Future research should concentrate on protein functions and their mode of action in fish species, especially on the role of spermatozoa surface proteins during fertilisation and on a description of sturgeon sperm proteins. PMID:18810648

Li, Ping; Hulak, Martin; Linhart, Otomar



Evaluation of Oxidative DNA Damage in Human Sperm and Its Association with Male Infertility  

Microsoft Academic Search

Recently, there is increasing evidence suggesting that oxidative sperm DNA damage is closely associated with impaired sperm function and male infertility. 8-Hydroxydeoxyguanosine (8- OHdG) is considered to be a precise and sensitive biomarker of oxidative DNA damage. The present study was thus designed to evaluate the extent of oxidative DNA damage in sperm and its as- sociation with male infertility



Transgenic zebrafish produced by retroviral infection of in vitro-cultured sperm  

Microsoft Academic Search

Transgenic modification of sperm before fertilization has distinct advantages over conventional transgenic methods. The primary advantage is that the mosaicism inherent in those other techniques is avoided. A culture system using primary cultures of zebrafish male germ cells, in which the differentiation from spermatogonia to functional sperm can occur in vitro, provides the opportunity for genetic modification of sperm in

Kayoko Kurita; Shawn M. Burgess; Noriyoshi Sakai



A Sperm Cytoskeletal Protein That Signals Oocyte Meiotic Maturation and Ovulation  

Microsoft Academic Search

Caenorhabditis elegans oocytes, like those of most animals, arrest during meiotic prophase. Sperm promote the resumption of meiosis (maturation) and contraction of smooth muscle-like gonadal sheath cells, which are required for ovulation. We show that the major sperm cytoskeletal protein (MSP) is a bipartite signal for oocyte maturation and sheath contraction. MSP also functions in sperm locomotion, playing a role

Michael A. Miller; Viet Q. Nguyen; Min-Ho Lee; Mary Kosinski; Tim Schedl; Richard M. Caprioli; David Greenstein



Cryopreservation of sperm from Murray cod, Maccullochella peelii peelii  

Microsoft Academic Search

Freshwater fish sperm are inactive in the male reproductive tract and seminal plasma. They are activated under hypotonic conditions, with a brief period of motility at the time of fertilization. The aims were to find a dilution medium that would maintain sperm inactivity, to assess the cytotoxicity of a range of cryoprotectants, and test the effectiveness of selected diluents for

Jonathan Daly; David Galloway; William Bravington; Michael Holland; Brett Ingram



The sperm nuclear matrix is required for paternal DNA replication  

Microsoft Academic Search

The mammalian sperm nucleus provides an excellent model for studying the relationship between the formation of nuclear structure and the initiation of DNA replication. We previously demonstrated that mammalian sperm nuclei contain a nuclear matrix that organizes the DNA into loop domains in a manner similar to that of somatic cells. In this study, we tested the minimal components of

Jeffrey A. Shaman; Yasuhiro Yamauchi; W. Steven Ward



The nuclear DNA longevity in cryopreserved boar spermatozoa assessed using the Sperm-Sus-Halomax.  


The aim of this experimental study was to evaluate the dynamics of nuclear DNA fragmentation in frozen-thawed (FT) boar spermatozoa incubated over time. Using the Sperm Chromatin Dispersion test (Sperm-Sus-Halomax), this study focused special attention on resolving the hypothesis that the original halo shapes around the sperm head could show dynamic changes over the postthawing incubation time. Twenty FT sperm samples from five boars (four per boar) were incubated at 37 °C during 168 hours and sperm motility (assessed using computer-assisted sperm analysis), viability (evaluated using the LIVE/DEAD Sperm Viability Kit), and nuclear DNA fragmentation were analyzed at 0, 0.5, 2, 4, 6, 24, 48, 72, and 168 hours. The percentages of motile and viable spermatozoa progressively decreased during incubation, with no motile and viable spermatozoa less than 10% in all boars at 24 hours of incubation. Four different halo shapes around the sperm head were considered in the Sperm Chromatin Dispersion test: normal, small, large scattered (typical fragmented nuclear DNA), and absent halo, all of them coexisting at the same time in the boar FT semen samples. Sperm with a large scattered halo did not change during postthaw, consistently showing percentages less than 5% over time in all boars. In contrast, the other three sperm populations showed a dynamic evolution over incubation time, characterized by a gradual reduction of sperm with normal halo, proportional to the increment in the sperm showing a small halo, followed by a switch between the sperm with a small halo and sperm with no halo. These results suggest that three of these four sperm populations, those showing small, large scattered, and absent halo, represent spermatozoa with different degrees of nuclear DNA damage, which should be taken into consideration to indicate the percentage of sperm with fragmented nuclear DNA in boar FT semen samples. PMID:23602080

Alkmin, Diego V; Martinez-Alborcia, Maria J; Parrilla, Inmaculada; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi



Potassium channels regulate hypo-osmotic shock-induced motility of common carp ( Cyprinus carpio) sperm  

Microsoft Academic Search

The osmolality and composition of fish seminal plasma usually suppress sperm motility in the testis and sperm duct. Change in the osmolality of the environment at spawning activates flagellar motion of the sperm tail. The effect of inhibitors of anion and cation channels on active motion was investigated by a microscopic motility test. While sodium channel inhibitors (amiloride and tetrodotoxin)

Zoltán Krasznai; Teréz Márián; László Balkay; Rezsó Gáspár; Lajos Trón



Sperm Whale Population Analysis.  

National Technical Information Service (NTIS)

Techniques which have been used to analyze whale populations and the resulting understanding of the sperm whale population of the North Pacific have been based on population estimates which are biased. Where variability is important to the understanding o...

C. W. Fowler R. J. Ryel L. J. Nelson



Tuning sperm chemotaxis.  


Sperm chemotaxis is a long-term puzzle and most of our knowledge comes from studying marine animals that are external fertilizers. Sperm are attracted by diffusible chemical factors (chemoattractants) released from the egg which redirect their swimming paths towards their source. This redirection is driven by increases in flagellar curvature that correlate with transient flagellar Ca(2+) increases. Recent experimental and modelling results provide insights into the signal flow underlying the translation of an external chemical gradient into an intracellular molecular and motor response. A fundamental element of sea-urchin sperm chemotaxis lies in the ability of these cells to suppress Ca(2+)-mediated increases in flagellar curvature while experiencing an increasing chemoattractant gradient. The article considers this new evidence and summarizes the known underlying cellular mechanisms and behavioural strategies that sperm use to locate and fertilize the oocyte. PMID:20863297

Guerrero, Adán; Wood, Christopher D; Nishigaki, Takuya; Carneiro, Jorge; Darszon, Alberto



Sperm Quality Control  

NSDL National Science Digital Library

New research from the University of Missouri-Columbia suggests how defective sperm are tagged for destruction. This 57 second audio recording from Science Update originally aired on Monday, February 11, 2008.

Science Update (AAAS;)



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

Microsoft Academic Search

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



Sperm studies in anesthesiologists  

SciTech Connect

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.

Wyrobek, A.J. (Lawrence Livermore National Lab., CA); Brodsky, J.; Gordon, l.; Moore, D.H., II; Watchmaker, G.; Cohen, E.N.



Metabolic rate limits the effect of sperm competition on Mammalian spermatogenesis.  


Sperm competition leads to increased sperm production in many taxa. This response may result from increases in testes size, changes in testicular architecture or changes in the kinetics of spermatogenesis, but the impact of each one of these processes on sperm production has not been studied in an integrated manner. Furthermore, such response may be limited in species with low mass-specific metabolic rate (MSMR), i.e., large-bodied species, because they cannot process energy and resources efficiently enough both at the organismic and cellular levels. Here we compare 99 mammalian species and show that higher levels of sperm competition correlated with a) higher proportions of seminiferous tubules, b) shorter seminiferous epithelium cycle lengths (SECL) which reduce the time required to produce sperm, and c) higher efficiencies of Sertoli cells (involved in sperm maturation). These responses to sperm competition, in turn, result in higher daily sperm production, more sperm stored in the epididymides, and more sperm in the ejaculate. However, the two processes that require processing resources at faster rates (SECL and efficiency of Sertoli cells) only respond to sperm competition in species with high MSMR. Thus, increases in sperm production with intense sperm competition occur via a complex network of mechanisms, but some are constrained by MSMR. PMID:24069461

Delbarco-Trillo, Javier; Tourmente, Maximiliano; Roldan, Eduardo R S



Quantification of mammalian sperm morphology by slit-scan flow cytometry  

SciTech Connect

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

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



Metabolic Rate Limits the Effect of Sperm Competition on Mammalian Spermatogenesis  

PubMed Central

Sperm competition leads to increased sperm production in many taxa. This response may result from increases in testes size, changes in testicular architecture or changes in the kinetics of spermatogenesis, but the impact of each one of these processes on sperm production has not been studied in an integrated manner. Furthermore, such response may be limited in species with low mass-specific metabolic rate (MSMR), i.e., large-bodied species, because they cannot process energy and resources efficiently enough both at the organismic and cellular levels. Here we compare 99 mammalian species and show that higher levels of sperm competition correlated with a) higher proportions of seminiferous tubules, b) shorter seminiferous epithelium cycle lengths (SECL) which reduce the time required to produce sperm, and c) higher efficiencies of Sertoli cells (involved in sperm maturation). These responses to sperm competition, in turn, result in higher daily sperm production, more sperm stored in the epididymides, and more sperm in the ejaculate. However, the two processes that require processing resources at faster rates (SECL and efficiency of Sertoli cells) only respond to sperm competition in species with high MSMR. Thus, increases in sperm production with intense sperm competition occur via a complex network of mechanisms, but some are constrained by MSMR.

delBarco-Trillo, Javier; Tourmente, Maximiliano; Roldan, Eduardo R. S.



Processes involved in assisted reproduction technologies significantly increase sperm DNA fragmentation and phosphatidylserine translocation.  


Sperm preparation techniques in assisted reproduction technologies (ART) are potential generators of exogenous stresses that cause additional DNA damage. DNA fragmentation tests, such as the sperm chromatin structure assay, involve freezing sperm samples in the absence of cryoprotectant. Thermal, oxidative stress (OS) and freezing are detrimental to sperm DNA fragmentation and phosphatidylserine (PS) translocation. The primary aim of this study was to subject mature sperm to environmental insults that normally occur during ART. We tested the hypotheses that OS, thermal stress and freeze-thawing caused sperm nuclear and membrane damage and that a positive correlation exists between PS translocation and DNA fragmentation. Sperm DNA integrity deteriorates in semen samples from men with advancing age and a sperm concentration of <15 m ml(-1) . The significant increase in sperm DNA fragmentation at 37 °C after merely 1 h is important clinically as semen liquefaction and short-term sperm storage in an ART cycle involve incubating samples at this temperature. Freezing without a cryoprotectant significantly increases the level of sperm nuclear damage, so it is important not to freeze neat semen prior to DNA fragmentation testing. This study highlights the importance of minimising the production of exogenous stresses during sperm preparation in ART. PMID:23230887

Balasuriya, A; Serhal, P; Doshi, A; Harper, J C



Recent Origins of Sperm Genes in Drosophila  

PubMed Central

Newly created genes often acquire testis-specific or enhanced expression but neither the mechanisms responsible for this specificity nor the functional consequences of these evolutionary processes are well understood. Genomic analyses of the Drosophila melanogaster sperm proteome has identified 2 recently evolved gene families on the melanogaster lineage and 4 genes created by retrotransposition during the evolution of the melanogaster group that encode novel sperm components. The expanded Mst35B (protamine) and tektin gene families are the result of tandem duplication events with all family members displaying testis-specific expression. The Mst35B family encodes rapidly evolving protamines that display a robust signature of positive selection within the DNA-binding high-mobility group box consistent with functional diversification in genome repackaging during sperm nuclear remodeling. The Mst35B paralogs also reside in a significant regional cluster of testis-overexpressed genes. Tektins, known components of the axoneme, are encoded by 3 nearly identical X-linked genes, a finding consistent with very recent gene family expansion. In addition to localized duplication events, the evolution of the sperm proteome has also been driven by recent retrotransposition events resulting in Cdlc2, CG13340, Vha36, and CG4706. Cdlc2, CG13340, and Vha36 all display high levels of overexpression in the testis, and Cdlc2 and CG13340 reside within testis-overexpressed gene clusters. Thus, gene creation is a dynamic force in the evolution of sperm composition and possibly function, which further suggests that acquisition of molecular functionality in sperm may be an influential pathway in the fixation of new genes.

Freeman, Zoe N.; Parker, Elizabeth R.; Heath, Benjamin D.; Karr, Timothy L.



Quantitative and functional changes in peripheral natural killer cells in women with reproductive failure after artificial insemination with donor sperm  

Microsoft Academic Search

Considerable evidence indicates that quantitative and\\/or functional abnormalities of natural killer (NK) cells are associated with reproductive failure. The objective of this study was to determine if peripheral NK percentage and activity are altered in women with reproductive failure after artificial insemination by donor (AID), and which parameters can best discriminate women with AID failure and normal controls. A case–control

Yuechao Lu; Bin Zeng; Yang Zhang; Wenpei Xiang; Lian Hu; Aihua Liao



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

PubMed Central

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, environmental, 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. Human sperm studies have complex requirements and, before sampling, careful consideration should be given to exposure details, group size and makeup, as well as animal and human data that indicate spermatogenic effects. Several study designs are possible and should include questionnaires covering medical and reproductive histories as well as known confounding factors. Animal sperm tests, such as the mouse morphology test, may be used to identify the toxic components of a complex mixture. Animal tests may also help assess the chemical effects on fertility and reproductive outcome in cases when human data are incomplete. Further efforts are needed in these areas to develop improved human sperm tests sensitive to induced spermatogenic damage, to develop improved animal models of induced spermatogenic damage, to understand the relationships among sperm changes, fertility, and reproductive outcome, and to develop sperm tests with express mutational end points.

Wyrobek, A J



Mitochondrial hydrogen peroxide and defective cholesterol efflux prevent in vitro fertilization by cryopreserved inbred mouse sperm.  


Recent advances in the cryopreservation of mouse sperm have resulted in dramatically improved in vitro fertilization (IVF) rates, but the biological mechanisms underlying the techniques remain unclear. Two different classes of compounds have been widely utilized to improve the IVF rates of cryopreserved mouse sperm: antioxidants and cyclodextrins. To determine how cryopreservation reduces mouse sperm IVF and how antioxidants and cyclodextrins mitigate this effect, we examined sperm function and oxidative damage after cryopreservation, with and without treatments, in mouse strains important for biomedical research. Our investigation revealed mouse strain-specific effects on IVF by modulation of oxidative stress and cholesterol efflux of cryopreserved sperm. Antioxidants improved IVF rates of C57Bl6/J cryopreserved mouse sperm by reducing hydrogen peroxide produced by sperm mitochondria and ameliorating peroxidative damage to the sperm acrosome. Enhancing cholesterol efflux with cyclodextrin restored capacitation-dependent sperm function and IVF after cryopreservation of C57Bl/6J, C57Bl/6N, and 129X1 mouse sperm. Our results highlight two accessible pathways for continued development of IVF techniques for mouse sperm and provide novel endpoints prognostic of IVF success. These insights may improve sperm cryopreservation methods of other mouse strains and species. PMID:23740947

Gray, Jeffrey E; Starmer, Joshua; Lin, Vivian S; Dickinson, Bryan C; Magnuson, Terry



A procedure for Poitou jackass sperm cryopreservation.  


We have tried to establish sperm banking for the endangered Poitou donkeys. No successful cryopreservation technique had been described for spermatozoa of this species; our preliminary work indicated that a particular medium and procedure may be effective for cryopreservation of Poitou jackass spermatozoa as evaluated by sperm motility, membrane integrity and pregnancy rate after AI with frozen-thawed semen. We found that glutamine at 80 mM and 10% (v/v) quail egg yolk in a basal medium containing 4% (v/v) glycerol (T2-94 medium) improved the post-thaw total and progressive motility and velocity assessed with the automated analyzer ATS-M. The T2-94 medium also preserved the sperm nuclear, acrosom, and plasma membrane integrity as assessed with the acridine orange method, fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA) lectin procedure, and hypo-osmotic swelling test, respectively. Semen frozen-thawed in T2-94 medium as used to artificially inseminate. 13 Poitou jennies from the beginning of estrus to ovulation during 4 cycles at a rate of one AI per day. Heigh pregnancies and 3 foals were obtained, but only when the glycerol was removed from sperm before AI. We conclude that the cryopreservation of Poitou jackass semen for sperm banking may succeed by using the T2-94 medium and removing the glycerol post-thaw, but before AI. PMID:10734453

Trimeche, A; Renard, P; Tainturier, D



Sperm counts and sperm sex ratio in male infertility patients.  


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

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



Sperm counts and sperm sex ratio in male infertility patients  

PubMed Central

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.

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



Healthy Sperm: Improving Your Fertility  


... Maintain a healthy weight. Some research suggests that obesity negatively affects sperm quality, reducing both sperm count ... America. 2011;38:359. The effects of workplace hazards on male reproductive health. Centers for Disease Control and Prevention. http://www. ...


[Effects of low doses of desmopressin (DDAPV) on gonadal and adrenal development, and on the testicular function and sperm motility].  


The present study proved that desmopressin (DDAVP) (1 microgram DDAVP/12 h/5 días) does not affect ovary, testis and adrenal development in immature Wistar rats (17 days old), because the DDAVP does not modify the weight of the aforementioned organs as compared with the control group. Nevertheless, the male adults Wistar rats (80 days old) showed lower serum testosterone concentrations than the control group, after injection of 4 micrograms/day (2 micrograms/12 h) or 8 micrograms/day (4 micrograms/12 h) of DDAVP during a 5 days period time. Moreover, paradoxical significant lower concentrations of serum testosterone were found in 4 micrograms DDAVP/day-treated rats than in 8 micrograms DDAVP/day-treated ones. The former also showed a decreased number of spermatozoa as compared with the latter and with the control group. The percentage of mobile spermatozoa was lower in rats treated with both concentrations of DDAVP as compared with the control group. Therefore, desmopressin does not delay gonadal and adrenal growth in immature rats, but, at low doses, it affects the testicular function and the mobility of the spermatozoa in male adult rats. PMID:8043737

García-Pascual, I J; Sánchez-Yagüe, J; Rodríguez Hernández, M C; Paniagua Gómez-Alvárez, R



Quantitative and functional changes in peripheral natural killer cells in women with reproductive failure after artificial insemination with donor sperm.  


Considerable evidence indicates that quantitative and/or functional abnormalities of natural killer (NK) cells are associated with reproductive failure. The objective of this study was to determine if peripheral NK percentage and activity are altered in women with reproductive failure after artificial insemination by donor (AID), and which parameters can best discriminate women with AID failure and normal controls. A case-control study of 20 fertile controls, 25 patients undergoing more than four treatment cycles without success (Group I), 25 experiencing biochemical pregnancy loss (Group II), and 25 undergoing embryo growth arrest or miscarriage (Group III) was performed. The peripheral NK percentage and cytotoxicity was evaluated by flow cytometry. Our results showed that a significantly increased percentage of NK cell cytotoxicity (NKCC) was detected in all study groups compared with the controls. The percentage of CD56(+) NK cells was elevated in Groups II and III compared with the controls. The percentage of CD56(+)CD16(+) NK cells in the study groups was higher than that in the controls. By using receiver operating characteristic curve and logistic regression analysis, we found that NKCC combined with CD56(+)CD16(+) NK percentage had good predictive power. The overall predicted percentage correct was 0.89. These results suggest that enhanced NKCC and an elevated cytotoxic NK percentage may be associated with AID failure and can be biomarkers for predicting AID-induced failure. PMID:21868103

Lu, Yuechao; Zeng, Bin; Zhang, Yang; Xiang, Wenpei; Hu, Lian; Liao, Aihua



Functional testing of the ATLAS SCT barrels  

NASA Astrophysics Data System (ADS)

The ATLAS SCT (semiconductor tracker) comprises 2112 barrel modules mounted on four concentric barrels of length 1.6 m and up to 1 m diameter, and 1976 endcap modules supported by a series of nine wheels at each end of the barrel region, giving a total silicon area of 60m. The assembly of modules onto each of the four barrel structures has recently been completed. In addition to functional tests made during the assembly process, each completed barrel was operated in its entirety. In the case of the largest barrel, with an active silicon area of approximately 10m, this corresponds to more than one million instrumented channels. This paper documents the electrical performance of the four individual SCT barrels. An overview of the readout chain is also given.

Phillips, Peter W.



Surgical recovery of sperm in non-obstructive azoospermia  

PubMed Central

The development of intracytoplasmic sperm injection (ICSI) opened a new era in the field of assisted reproduction and revolutionized the assisted reproductive technology protocols for couples with male factor infertility. Fertilisation and pregnancies can be achieved with spermatozoa recovered not only from the ejaculate but also from the seminiferous tubules. The most common methods for retrieving testicular sperm in non-obstructive azoospermia (NOA) are testicular sperm aspiration (TESA: needle/fine needle aspiration) and open testicular biopsy (testicular sperm extraction: TESE). The optimal technique for sperm extraction should be minimally invasive and avoid destruction of testicular function, without compromising the chance to retrieve adequate numbers of spermatozoa to perform ICSI. Microdissection TESE (micro-TESE), performed with an operative microscope, is widely considered to be the best method for sperm retrieval in NOA, as larger and opaque tubules, presumably with active spermatogenesis, can be directly identified, resulting in higher spermatozoa retrieval rates with minimal tissue loss and low postoperative complications. Micro-TESE, in combination with ICSI, is applicable in all cases of NOA, including Klinefelter syndrome (KS). The outcomes of surgical sperm retrieval, primarily in NOA patients with elevated serum follicle-stimulating hormone (FSH) (NOA including KS patients), are reviewed along with the phenotypic features. The predictive factors for surgical sperm retrieval and outcomes of treatment were analysed. Finally, the short- and long-term complications in micro-TESE in both 46XY males with NOA and KS patients are considered.

Ishikawa, Tomomoto



Oviducal sperm storage in poultry  

Technology Transfer Automated Retrieval System (TEKTRAN)

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


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


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

Favret, Karen P; Lynn, John W



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


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

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



Production of Transgenic Pigs Mediated by Pseudotyped Lentivirus and Sperm  

PubMed Central

Sperm-mediated gene transfer can be a very efficient method to produce transgenic pigs, however, the results from different laboratories had not been widely repeated. Genomic integration of transgene by injection of pseudotyped lentivirus to the perivitelline space has been proved to be a reliable route to generate transgenic animals. To test whether transgene in the lentivirus can be delivered by sperm, we studied incubation of pseudotyped lentiviruses and sperm before insemination. After incubation with pig spermatozoa, 62±3 lentiviral particles were detected per 100 sperm cells using quantitative real-time RT-PCR. The association of lentivirus with sperm was further confirmed by electron microscopy. The sperm incubated with lentiviral particles were artificially inseminated into pigs. Of the 59 piglets born from inseminated 5 sows, 6 piglets (10.17%) carried the transgene based on the PCR identification. Foreign gene and EGFP was successfully detected in ear tissue biopsies from two PCR-positive pigs, revealed via in situ hybridization and immunohistochemistry. Offspring of one PCR-positive boar with normal sows showed PCR-positive. Two PCR-positive founders and offsprings of PCR-positive boar were further identified by Southern-blot analysis, out of which the two founders and two offsprings were positive in Southern blotting, strongly indicating integration of foreign gene into genome. The results indicate that incubation of sperm with pseudotyped lentiviruses can incorporated with sperm-mediated gene transfer to produce transgenic pigs with improved efficiency.

Zhang, Yongliang; Xi, Qianyun; Ding, Jinghua; Cai, Weiguang; Meng, Fanmin; Zhou, Junyun; Li, Hongyi; Jiang, Qingyan; Shu, Gang; Wang, Songbo; Zhu, Xiaotong; Gao, Ping; Wu, Zhenfang



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


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

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



Remarkable longevity of dilute sperm in a free-spawning colonial ascidian.  


Many benthic marine invertebrates reproduce by releasing sperm into the sea (free-spawning), but the amount of time that sperm are viable after spawning may have different consequences for fertilization, depending on the type of free-spawner. In egg-broadcasting marine organisms, gamete age is usually assumed to be irrelevant because of the low probability of contact between dilute sperm and egg. However, direct dilution effects might be reduced in egg-brooding free-spawners that filter dilute sperm out of the water column, and sperm longevity may play a role in facilitating fertilization in these taxa. We investigated the effects of time, temperature, and mixing on the viability of naturally released sperm of the colonial ascidian Botryllus schlosseri. Our data indicate that B. schlosseri sperm have a functional life span that is considerably longer than those of the sperm of many other marine invertebrate taxa (half-life of approximately 16 to 26 h), are able to fertilize eggs at extremely low external sperm concentrations (ca. 10(1) sperm ml(-1)), and have a longevity that varies with temperature. It is possible that such prolonged sperm longevity may be achieved by reductions in motility, reactivation of quiescent sperm by chemical cues, or intermittent swimming. PMID:15198940

Johnson, Sheri L; Yund, Philip O



Testing a new version of the size-advantage hypothesis for sex change: sperm competition and size-skew effects in the bucktooth parrotfish, Sparisoma radians  

Microsoft Academic Search

A variety of field studies suggest that sex change in animals may be more complicated than originally depicted by the size-advantage hypothesis. A modification of the size-advantage hypothesis, the expected reproductive success threshold model, proposes that sperm competition and size-fecundity skew can strongly affect reproductive pay-offs. Size-fecundity skew occurs if a large female's fecundity is markedly higher than the aggregate

Roldan C. Muñoz; Robert R. Warner



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

SciTech Connect

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

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



Use of amides as cryoprotectants in extenders for frozen sperm of tambaqui, Colossoma macropomum.  


Amides were tested as cryoprotectants in comparison with glycerol and DMSO (more traditional cryoprotectants) for recovery of Colossoma macropomum (tambaqui fish) sperm. Milt was extended in Beltsville Thawing Solution, then frozen with the addition of 2%, 5%, 8%, or 11% of: (1) dimethylacetamide (DMA), (2) dimethylformamide (DMF), (3) methylformamide (MF), or with 5% glycerol or 10% dimethylsulfoxide. Fertilization rates were greatest (P<0.001) with amides; 8% DMF (91.6±1.3%), 5% DMF (88.9±1.6%), and 8% MF (83.0±1.6%), which did not significantly differ among themselves, when compared with glycerol (51.6±2.4%) and DMSO (61.9±3.1%). The best hatching rates (P<0.001) also occurred for 5% or 8% DMF and 8% MF (79.1±3.1, 87.6±1.5, and 74.8±3.0, respectively) and were also similar (P>0.05). For such treatments, both fertilization and hatching rates were similar (P>0.05) to those with fresh sperm (91.7±1.4 and 87.4±1.4, respectively). The best sperm motility across extenders (at least 55.7%) was with 5%, 8%, and 11% DMF (P<0.001). Those same treatments, along with 11% MF, provided the longest (P<0.001) period of motility (at least 1 min). The greatest sperm integrity (more than 54%) was with 5% and 11% MF and with DMA and DMF at all tested concentrations (P<0.001). The greatest (P<0.001) sperm viability (at least 31%) was for 5%, 8%, and 11% DMA, and with 8% and 11% MF, and also for DMF at all tested concentrations. Sperm DNA integrity was best (more than 50%) for 2%, 5%, and 8% MF and for DMA and DMF at all concentrations (P<0.001), whereas 2% DMA, 11% MF, 11% DMF, and the three amides at both 5% and 8% yielded the highest mitochondrial functionality (at least 44%; P<0.001); thus, 8% MF and both 5% and 8% DMF were the cryoprotectants with the best postthaw quality for C. macropomum sperm. PMID:22578629

Varela Junior, A S; Corcini, C D; Gheller, S M M; Jardim, R D; Lucia, T; Streit, D P; Figueiredo, M R C



Transgenic zebrafish produced by retroviral infection of in vitro-cultured sperm.  


Transgenic modification of sperm before fertilization has distinct advantages over conventional transgenic methods. The primary advantage is that the mosaicism inherent in those other techniques is avoided. A culture system using primary cultures of zebrafish male germ cells, in which the differentiation from spermatogonia to functional sperm can occur in vitro, provides the opportunity for genetic modification of sperm in vitro. Here, we report the production of transgenic zebrafish from cultured sperm. The sperm were differentiated from premeiotic germ cells infected with a pseudotyped retrovirus in vitro. The collected sperm were used to perform successful in vitro fertilizations, and transgenic embryos were identified. The transgenic fish transmitted the proviral integration to the next generation in a Mendelian fashion. We report the generation of a transgenic animal by cultured sperm and open the door to many exciting possibilities for the rapid generation of transgenic lines in model organisms such as zebrafish or other animal systems that are otherwise intractable to transgenesis. PMID:14745028

Kurita, Kayoko; Burgess, Shawn M; Sakai, Noriyoshi



OAIS Functional Model Conformance Test: A Proposed Measurement  

ERIC Educational Resources Information Center

|Purpose: The purpose of this paper is to develop a test for data centres, repositories and archives to determine OAIS functional model conformance. The test developed was carried out among the World Data Centre (WDC) member data centres. The method used to develop the OAIS functional model conformance test is discussed, along with the test

Laughton, Paul



Effects of sulfasalazine on sperm acrosome reaction and gene expression in the male reproductive organs of rats.  


Sulfasalazine (SASP) has been reported to depress the fertility in men and experimental male animals, but the fundamental mechanisms of infertility caused by SASP are still unknown. This study was designed to investigate the mechanisms of infertility in rats treated with SASP at a dose of 600 mg/kg for 28 days, including monitoring of sperm motility using computer associated sperm analysis system and acrosome reaction by FITC-concanavalin A lectin staining. The sperm motility and acrosome reaction, which are important for fertilization, were significantly reduced by SASP. Furthermore, to investigate the molecular mechanisms of infertility induced by SASP, mRNA expression analysis in the testes was performed using cDNA microarray as a first screening. It was revealed that CD59, which is located on the acrosomal membrane and is known to be important for the reproductive function of sperm, was affected in the testes; this was also confirmed by real-time PCR analysis, but the spermatogenesis-related genes examined in this study were not affected. Therefore, we focused on CD59 and two other acrosome membrane related-genes: MCP and DAF. CD59, MCP, and DAF in the epididymides of SASP-treated rats were significantly decreased as assessed by real-time RT-PCR analysis and additionally, the expression of CD59 protein was found to be decreased by Western blotting. These results allowed us to hypothesize that the suppression of epididymal acrosomal membrane proteins synthesis with their consequent reduced incorporation to the sperm membrane leads to a depressed sperm motility and acrosome reaction, and thereby leads to infertility in SASP treated male rats. PMID:15625186

Fukushima, Tamio; Kato, Masashi; Adachi, Tetsuya; Hamada, Yoshimasa; Horimoto, Masao; Komiyama, Masatoshi; Mori, Chisato; Horii, Ikuo



The monopulsed nature of sperm whale clicks  

NASA Astrophysics Data System (ADS)

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

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



The importance of redox regulated pathways in sperm cell biology.  


Redox regulated events are fundamental to our understanding of many cellular pathways and pathological processes. On the one hand, production of reactive oxygen species by mammalian spermatozoa has been associated with a loss of cell function and DNA integrity as a consequence of oxidative stress. These cells are exquisitely sensitive to such damage as a consequence of their relative lack of cytosolic antioxidant enzymes and relative abundance of polyunsaturated fatty acids. Given this susceptibility, it is surprising to discover that spermatozoa are intensely redox active cells and professional generators of reactive oxygen species. The latter are physiologically important to the spermatozoa in regulating every aspect of sperm function examined, including their movement characteristics, capacitation, sperm-zona interaction, the acrosome reaction and sperm-oocyte fusion. The molecular basis of this redox drive is still poorly understood in terms of the source of the reactive oxygen species and the mechanisms by which these reactive metabolites enhance sperm function. Recent advances include the discovery of NOX5 in the male germ line and elucidation of the role of reactive oxygen species in controlling a unique signal transduction cascade associated with sperm capacitation. Given the central importance of redox chemistry in the control of sperm function further research in this area may uncover valuable targets for contraceptive intervention. PMID:15109744

Baker, Mark A; Aitken, R John



Sperm storage mediated by cryptic female choice for nuptial gifts.  


Polyandrous females are expected to discriminate among males through postcopulatory cryptic mate choice. Yet, there is surprisingly little unequivocal evidence for female-mediated cryptic sperm choice. In species in which nuptial gifts facilitate mating, females may gain indirect benefits through preferential storage of sperm from gift-giving males if the gift signals male quality. We tested this hypothesis in the spider Pisaura mirabilis by quantifying the number of sperm stored in response to copulation with males with or without a nuptial gift, while experimentally controlling copulation duration. We further assessed the effect of gift presence and copulation duration on egg-hatching success in matings with uninterrupted copulations with gift-giving males. We show that females mated to gift-giving males stored more sperm and experienced 17% higher egg-hatching success, compared with those mated to no-gift males, despite matched copulation durations. Uninterrupted copulations resulted in both increased sperm storage and egg-hatching success. Our study confirms the prediction that the nuptial gift as a male signal is under positive sexual selection by females through cryptic sperm storage. In addition, the gift facilitates longer copulations and increased sperm transfer providing two different types of advantage to gift-giving in males. PMID:24153384

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



Cryopreservation of mutton snapper ( Lutjanus analis) sperm.  


This study aimed to develop a protocol of semen cryopreservation of the mutton snapper Lutjanus analis. The interaction between three extenders ( pH 6.1; 7.8 and 8.2) , two concentrations of dimethyl sulfoxide ( DMSO, 5 and 10%) and three cooling rates ( -90; -60 and -30°C.min-1) on the sperm motility rate and motility time were analyzed by a factorial experiment. A sample of 30 fishes ( 1,261 ± 449 g) collected in the nature was kept in floating net cages. The semen was frozen by using cryogenic straws, in nitrogen vapour and transferred, later, to liquid nitrogen. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The highest sperm motility rate and motility time ( P < 0.05) was achieved by combining extender C ( pH 8.2) with DMSO ( 10%) and cooling rate of -60°C.min-1 ( P < 0.05) . The use of cryopreserved sperm presented fertilization rates higher than 59% validating the present protocol for mutton snapper. PMID:23969847

Sanches, Eduardo G; Oliveira, Idili R; Serralheiro, Pedro C Da Silva; Cerqueira, Vinicius R



Detection and tracking of low contrast human sperm tail  

Microsoft Academic Search

Tracking sperm tail movement provides important information for clinical sperm research. It is also a crucial step for sperm immobilization in Intracytoplasmic Sperm Injection (ICSI). However, the low visibility of the sperm tail under optical microscopy, coupled with the sperm fast motility, render sperm tail identification and tracking challenging tasks to execute. This paper presents two approaches for sperm tail

Clement Leung; Zhe Lu; Navid Esfandiari; Robert F. Casper; Yu Sun



Short-term preservation of fowl sperm in buffered potassium chloride.  


Previous research demonstrated that sperm motility is dependent upon mitochondrial calcium cycling. Thus, sperm are inactivated when extracellular calcium ions are chelated. Mitochondrial calcium cycling, however, is driven by extracellular sodium ions. The hypothesis that sperm inactivation is subject to 2 variables was tested in the present work. Sperm motility was evaluated with computer-assisted sperm motion analysis in the first experiment. Sperm became immotile within minutes when suspended in buffered isotonic potassium chloride containing calcium ions. This outcome set the stage for the second experiment in which sperm were inactivated by centrifugation through 12% (wt/vol) Accudenz prepared with potassium chloride and tetrasodium 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Sperm mobility was the end point in the second and subsequent experiments. Potassium chloride was a suitable medium in regard to sperm inactivation with BAPTA followed by storage for 2 h at 10°C. Consequently, sperm so inactivated were reactivated after 1, 2, 3, 4, and 5 h of storage in the third experiment. Whereas pre- and postwash sperm mobility were equivalent, sperm mobility declined with time (P < 0.001) without exogenous energy in the storage medium. Therefore, the effect of 5 mM glucose was tested in the fourth experiment. In this case, recovery of sperm mobility was independent of time when sperm were stored at 10°C for 5 h (P > 0.05). Potassium chloride was replaced with potassium glutamate in the last experiment. Whereas reactivation was once again independent of time when sperm were stored with glucose (P > 0.05), greater variability was observed among observations in comparison with the potassium chloride-based medium. In summary, sperm motility was inactivated when calcium was chelated and extracellular sodium was replaced with potassium. Sperm reactivation was most consistent when chloride was the predominant extracellular anion. Collectively, these experiments demonstrate that short-term sperm storage can be achieved with simple media that promote sperm glycolysis and minimize energy demands imposed by the active transport of calcium and sodium ions. PMID:23571344

Froman, D P



Cryopreservation of Fish Sperm  

NASA Astrophysics Data System (ADS)

Present status of research activities in cryopreservation of fish gamete in aquaculture field was introduced. More than 59 fish species have been reported in the research histories and nearly half of them were studied during recent 10 years. This means that the research activities are increasing, though commercial profit have not obtained yet. Fish species of which sperm can successfully cryopreserved is still limited comparing to numerous species in telost. One of the major obstacle for improvement of the technique is existence of wide specie specific variance in the freezing tolerance of fish sperm. The varianc can possibly be explaind thorugh the informations obtained by the studies in comparative spermatology, which is recently activated field in fish biology.

Kurokura, Hisashi


Sperm proteins in teleostean and chondrostean (sturgeon) fishes  

Microsoft Academic Search

Sperm proteins in the seminal plasma and spermatozoa of teleostean and chondrostean have evolved adaptations due to the changes\\u000a in the reproductive environment. Analysis of the composition and functions of these proteins provides new insights into sperm\\u000a motility and fertilising abilities, thereby creating possibilities for improving artificial reproduction and germplasm resource\\u000a conservation technologies (e.g. cryopreservation). Seminal plasma proteins are involved

Ping Li; Martin Hulak; Otomar Linhart



The effect of human papillomavirus infection on sperm cell motility  

Microsoft Academic Search

Objective: To investigate the presence of human papillomavirus (HPV) in human sperm cells and to evaluate potential effects of HPV on the sperm functions.Design: A descriptive clinical study.Patient(s): Specimens of semen were collected from 24 randomly selected patients who attended the fertility clinics at Chang Gung Memorial Hospital.Main Outcome Measure(s): The presence of HPV DNA and RNA were examined by

Ying Ming Lai; Jo Fang Lee; Hong Yuan Huang; Yung Kuei Soong; Feng-Ping Yang; Chia C. Pao



Anorectal functional testing: review of collective experience  

Microsoft Academic Search

Anorectal manometry includes a number of specific tests that are helpful in the diagnostic assessment of patients with fecal incontinence and constipation; their purpose is to delineate the pathophysiological mechanism for these symptoms. Some of these tests may also provide helpful information in the assessment of patients with rectal pain or diarrhea, but their sensitivity and specificity are less well

Fernando Azpiroz; Paul Enck; William E. Whitehead



Intracytoplasmic Sperm Injection (ICSI)  

NSDL National Science Digital Library

Video microscopy demonstrating how sperm from an infertile male can be injected into a female egg. This video featured on the HHMI DVD, The Meaning of Sex: Genes and Gender, available free from HHMI. Also, this video is one minute and 15 seconds in length, and available in Quicktime (7 MB) and Windows Media (12 MB). All sex determination videos are located at:

Howard Hughes Medical Institute (HHMI;)



Sperm Chromatin and Environmental Factors  

Microsoft Academic Search

\\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

Aleksander Giwercman


Trajectory variance and autocorrelations within single-sperm tracks as population-level descriptors of sperm track complexity, predictability, and energy-generating ability.  


The objectives of the present study were to develop an alternative theoretical approach to the analysis of sperm motility and to develop motility parameters that would complement those more commonly used in current computer-assisted semen analysis procedures. We have defined a set of parameters and have tested them using boar spermatozoa undergoing bicarbonate-induced motility activation. The new parameters were calculated for a series of (x,y) coordinates of sperm head positions recorded at each move along the trajectory. The parameters were: mean velocity (MV), immobility ratio, fractal dimension (FD), the variance of the steplengths (VAR), and 2 autocorrelation function coefficients of the step-length time series for lags 1 and 2 (C(1) and C(2)). MV measures the average speed along the trajectory, and VAR is a measure of displacement variability that can be related to the specific mean (per step) kinetic energy of the spermatozoon. All of the parameters except MV and FD were affected by the sampling frequency (25 vs 50 Hz); inappropriately high sampling frequency in relation to magnification resulted in step-lengths between successive frames that were below the resolution threshold of the imaging system. The autocorrelation functions were especially informative; discrimination between sperm subpopulations was obvious within simple histogram formats, and complex statistical analyses were not needed for their identification. PMID:21474791

Abaigar, Teresa; Barbero, Javier; Holt, William V



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


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

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



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

SciTech Connect

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

Clark, A.G. [Pennsylvania State Univ., University Park, PA (United States); Prout, T.; Harshman, L.G. [Univ. of California, Davis, CA (United States)] [and others



Structure and evolution of the sea star egg receptor for sperm bindin.  


Selection on coevolving sperm- and egg-recognition molecules is a potent engine of population divergence leading to reproductive isolation and speciation. The study of receptor-ligand pairs can reveal co-evolution of male- and female-expressed genes or differences between their evolution in response to selective factors such as sperm competition and sexual conflict. Phylogeographical studies of these patterns have been limited by targeted gene methods that favour short protein-coding sequences amplifiable by PCR. Here, I use high-throughput transcriptomic methods to characterize the structure and divergence of full-length coding sequences for the gene encoding the protein component of a large complex egg surface glycopeptide receptor for the sperm acrosomal protein bindin from the sea star Patiria miniata. I used a simple but effective method for resolving nucleotide polymorphisms into haplotypes for phylogeny-based analyses of selection. The protein domain organization of sea star egg bindin receptor (EBR1) was similar to sea urchins and included a pair of protein-recognition domains plus a series of tandem repeat domains of two types. Two populations separated by a well-characterized phylogeographical break included lineages of EBR1 alleles under positive selection at several codons (similar to selection on sperm bindin in the same populations). However, these populations shared the same alleles that were under selection for amino acid differences at multiple codons (unlike the pattern of selection for population divergence in sperm bindin). The significance of positively selected EBR1 domains and alleles could be tested in functional analyses of fertilization rates associated with EBR1 (and bindin) polymorphisms. PMID:23432510

Hart, Michael W



Sperm chromatin structure assay (SCSA): a tool in diagnosis and treatment of infertility  

PubMed Central

Diagnosis of male infertility has mainly been based on the World Health Organization (WHO) manual-based semen parameter's concentration, motility and morphology. It has, however, become apparent that none of these parameters are reliable markers for evaluation of the fertility potential of a couple. A search for better markers has led to an increased focus on sperm chromatin integrity testing in fertility work-up and assisted reproductive techniques. During the last couple of decades, numerous sperm DNA integrity tests have been developed. These are claimed to be characterized by a lower intraindividual variation, less intralaboratory and interlaboratory variation and thus less subjective than the conventional sperm analysis. However, not all the sperm chromatin integrity tests have yet been shown to be of clinical value. So far, the test that has been found to have the most stable clinical threshold values in relation to fertility is the sperm chromatin structure assay (SCSA), a flow cytometric test that measures the susceptibility of sperm DNA to acid-induced DNA denaturation in situ. Sperm DNA fragmentation as measured by SCSA has shown to be an independent predictor of successful pregnancy in first pregnancy planners as well as in couples undergoing intrauterine insemination, and can be used as a tool in investigation, counseling and treatment of involuntary childlessness. More conflicting data exist regarding the role of sperm DNA fragmentation in relation to fertilization, pre-embryo development and pregnancy outcome in in vitro fertilization and intracytoplasmic sperm injection (ICSI).

Bungum, Mona; Bungum, Leif; Giwercman, Aleksander



Light, polarizing, and transmission electron microscopy: three methods for the evaluation of sperm quality.  


Semen from 33 patients were evaluated by light microscopy (LM) obtaining sperm concentration, percent motility, percentage of sperm with normal morphology (PAP staining), and percentage of dead sperm (Eosin Y stained). The samples were observed by polarizing microscopy (PM), that evaluates sperm morphology and the viability by birefringence of organelles, and it provides a PM index (percentage of birefringent, viable, motile sperm) and a percentage of dead, non-birefringent sperm. Sperm were processed for transmission electron microscopy (TEM) and TEM data were elaborated with a mathematical formula able to provide a fertility index (FI, number of sperm free of structural defects) and percentages of sperm immaturity and necrosis (dead sperm). To test the reliability of these techniques, the values of normal acrosome, nucleus, midpiece, and tail and the presence of cytoplasmic residues obtained with the three methods were compared. With the exception of cytoplasmic residues (P?=?0.40), significant differences in the evaluation of each organelle were observed and TEM analysis resulted as the most stringent screening. In addition, relationships among relevant sperm variables were investigated. Motility showed positive correlations with the percentage of normal tail, midpiece, and PM index (P?sperm death (non-birefringent sperm: P?sperm: P?sperm morphology: FI with PM index (P?sperm (PAP staining) (P?Sperm immaturity showed positive correlations (P?sperm quality. PM appears to offer several advantages 'midway' between LM and TEM and it should be considered in sperm analysis. PMID:23043672

Collodel, Giulia; Iacoponi, Francesca; Mazzi, Lucia; Terzuoli, Gaia; Pascarelli, Nicola Antonio; Moretti, Elena



Sperm ubiquitination and DNA fragmentation in men with occupational exposure and varicocele.  


Assessment of sperm ubiquitination and DNA fragmentation as sperm functional markers are proposed to complement routine semen analysis. This study focuses on the evaluation of these markers in infertile men with varicocele or exposed to occupational background. The results were compared with normozoospermic men. Semen parameters in both groups were lower than those in the control group. Ubiquitination median, as a marker for functionality of the ubiquitin-proteasome system, was also lower in both groups. The ubiquitination median showed a significant positive correlation with motility in both groups, while it showed only a negative correlation with sperm morphology in the varicocele group. DNA fragmentation showed a significant correlation with semen parameters, in total varicocele and also total exposure groups. In conclusion, significant difference of sperm ubiquitination between normal and study groups further validates that sperm ubiquitination as a potential molecular marker for sperm evaluation in addition to routine semen analysis in clinical laboratories. PMID:23594355

Hosseinpour, E; Shahverdi, A; Parivar, K; Sedighi Gilani, M A; Nasr-Esfahani, M H; Salman Yazdi, R; Sharbatoghli, M; Tavalaee, M; Chehrazi, M



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

NASA Astrophysics Data System (ADS)

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

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



Portable system for fast lung function test  

Microsoft Academic Search

The analysis of gas-concentration changes at mouth during normal breathing is nowadays a routine exam to infer lung functionality and several commercial instruments are available to carry out this kind of measurement. Unfortunately, most of these measuring systems are very specific, designed to be used in the hospital and costly. This paper describes a complete and versatile system which is

M. Parvis; C. Gulotta; R. Torchio



Testing the bipolarity of the Jungian functions.  


In this article, I offer an alternative explanation for recent studies that have questioned the assumed bipolarity of the Jungian functions measured by the Myers-Briggs Type Indicator (MBTI) due to the failure to observe negative correlations between the hypothesized opposite functions when a Likert format is used in place of the MBTI's forced-choice format (Cowan, 1989; Girelli & Stake, 1993; Loomis & Singer, 1980). It was suggested that the acquiescence response set might be partially responsible. A new instrument was constructed using agree-disagree items. Negative correlations between the hypothesized opposite functions were not observed. To deal with the acquiescence response set, participants were ranked on the number of statements endorsed, and the scores for the middle third of participants were analyzed again For these participants, correlations of -.67 and -.69 between each pair of theoretically opposite functions were found. I conclude thai this largely refutes criticisms of the bipolarity assumption, but there is still reason to develop a measure not relying on the assumption. PMID:16367681

Murray, W S



PH-20 but not acrosin is involved in sperm penetration of the macaque zona pellucida.  


In this study, we investigated the functions of PH-20 and acrosin during the interaction of macaque sperm with the zona pellucida. Both of these sperm enzymes have been reported to be present on the inner acrosomal membrane of acrosome reacted sperm, and have been suggested to play a role during secondary sperm-zona binding in other species. Anti-macaque PH-20 IgG, anti-pig acrosin IgG and soybean trypsin inhibitor (SBTI) were used as probes for immunolocalization of the two proteins at the ultrastructural level, and as reagents for blocking sperm penetration of the macaque zona pellucida in vitro. As a control, we performed similar studies with antibodies to CD-46, which is also located on the inner acrosomal membrane, but has no known function in sperm-zona pellucida interaction. After labeling with anti-acrosin IgG, gold label was not present on the sperm surface before the acrosome reaction, but was detected over the entire head of sperm that were induced to acrosome react with calcium ionophore A23187. In contrast, when sperm were induced to acrosome react by binding to intact zona pellucida, acrosin was present in the acrosomal shroud but not on the inner acrosomal membrane. Similar results were obtained when SBTI was used as a probe for enzyme localization. PH-20 and CD-46 were demonstrated on the inner acrosomal membrane of sperm induced to acrosome react by ionophore treatment and by zona binding. Neither anti-acrosin IgG nor anti-CD-46 IgG affected sperm penetration of the zona at concentrations up to 300 microg/ml, but zona penetration was blocked completely when anti-PH-20 IgG (100 microg/ml) was present during sperm-oocyte interaction. Ultrastructural observations of oocytes incubated with anti-PH-20 IgG showed that acrosomal shrouds were present on the zona surface but no sperm had begun to penetrate into the zona substance. We conclude that anti-PH-20 IgG prevented sperm penetration of the macaque zona pellucida by interference with secondary sperm-zona binding, rather than primary sperm-zona binding or the zona-induced acrosome reaction. Acrosin was not detected on the inner acrosomal membrane of sperm that are induced to acrosome react after zona binding, and acrosin does not appear to be critical for sperm penetration of the macaque zona pellucida. PMID:10369396

Yudin, A I; Vandevoort, C A; Li, M W; Overstreet, J W



Usng Subjective Percentiles and Test Data for Estimating Fragility Functions.  

National Technical Information Service (NTIS)

Fragility functions are cumulative distribution functions (cdfs) of strengths at failure. They are needed for reliability analyses of systems such as power generation and transmission systems. Subjective opinions supplement sparse test data for estimating...

L. L. George R. W. Mensing



Sperm-based contraceptive vaccines: current status, merits and development.  


At the present growth rate, the world population is estimated to reach a phenomenal 8.9 billion people by the year 2050, posing a great risk of overpopulation. Therefore, new strategies of contraception are required. A novel contraceptive strategy that is receiving considerable attention is that of immunocontraception. Spermatozoa have proteins that are unique, cell specific, immunogenic and accessible to antibodies. The targeting of antibodies to gamete-specific antigens implicated in sperm function, sperm-egg binding and fertilisation could block sperm binding and thus fertilisation. The present review highlights the current status, relative merits and future directions for various sperm-based candidate antigens with regard to the development of a contraceptive vaccine. PMID:16153335

Suri, Anil



Sperm antimotility properties of a seed extract of Abrus precatorius.  


This study examined the inhibitory effects of a methanol extract of Abrus precatorius seeds on the motility of washed human spermatozoa. The extract caused a concentration-related impairment of percentage sperm motility; with the EC50 concentration being 2.29 mg/ml. This effect on motility was essentially irreversible. With the highest concentration tested (20.0 mg/ml), the onset of the antimotility action was almost immediate. In addition, this concentration impaired the functional integrity of the plasma membrane (hypoosmotic swelling test) and viability (nigrosin-eosin stain) of spermatozoa. In contrast, with a lower concentration (5.0 mg/ml), such effects were not evident. It is concluded that at the lower concentrations the antimotility action may result from a rise in intracellular calcium (not via influx) and/or a decline in cAMP content and/or enhanced generation of a reactive oxygen species. PMID:1943179

Ratnasooriya, W D; Amarasekera, A S; Perera, N S; Premakumara, G A


Extraordinarily long sperm in the socially monogamous cichlid fish Pelvicachromis taeniatus  

Microsoft Academic Search

The main function of the spermatozoon is the transfer of the male haploid genome during fertilisation. In animals in general\\u000a 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

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



The effects of cryopreservation on the morphometric dimensions of caprine sperm heads  

Microsoft Academic Search

Cryopreserved semen has been utilised in the artificial insemination of livestock species for over 40 years, even though the detrimental effects of cryopreservation on sperm function and fertility are well documented. In the present study, computer-automated sperm-head morphometry was used to determine if goat sperm-head morphometry was affected by freezing and thawing. A microscope slide was prepared from single semen

C. G. Gravance; C. White; K. R. Robertson; Z. J. Champion; P. J. Casey



A Comparison of Statistical Significance Tests for Selecting Equating Functions  

ERIC Educational Resources Information Center

|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…

Moses, Tim




Microsoft Academic Search

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

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


A new test for gastric function  

Microsoft Academic Search

Summary  Hydrochloride of 2-Bencil 4-5 imidazoline (Priscol): a new chemical compound whose stimulant action on gastric secretion has\\u000a been tested as well by oral as by parenteral administration on a group of more than 70 gastrointestinal patients. The results\\u000a obtained and analyzed comparatively with other classes of chemical substances, also stimulants of gastric secretion, such\\u000a as alcohol or histamine, show that

Juan Nasio



Cooling and freezing of epididymal sperm in the common hippopotamus ( Hippopotamus amphibius)  

Microsoft Academic Search

Knowledge concerning reproduction in common hippopotamus is scarce and in particular very little is known about male reproductive physiology and sperm cryopreservation. Testes were obtained from nine castrated bulls and sperm extracted from the epididymides of eight of these individuals. Mean ± SEM values of reproductive parameters were: testicular weight (including epididymis and tunicas)—275.9 ± 54.1 g, total sperm motility—88.1

J. Saragusty; C. Walzer; T. Petit; G. Stalder; I. Horowitz; R. Hermes



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


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

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



Quantification of mammalian sperm morphology by slit-scan flow cytometry  

SciTech Connect

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

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



Osmotic tolerance limits and effects of cryoprotectants on the motility, plasma membrane integrity and acrosomal integrity of rat sperm.  


Osmotic stress is an important factor that can result in cell damage during cryopreservation. The objectives of this study were to determine: (1) isosmotic sperm cell volume; (2) osmotically inactive volume; (3) osmotic tolerance limits of rat sperm; and (4) the effects of addition and removal of glycerol (Gly), ethylene glycol (EG), propylene glycol (PG) or dimethyl sulfoxide (Me(2)SO) on rat sperm function. Sperm from Fischer 344 and Sprague-Dawley rats were used in this study. An electronic particle counter was used to measure the cell volume of rat sperm. Computer-assisted sperm motility analysis and flow-cytometric analysis were used to assess sperm motility, plasma membrane and acrosomal integrity. The isosmotic sperm cell volumes of the two strains were 37.0+/-0.1 and 36.2+/-0.2 microm(3), respectively. Rat sperm behaved as linear osmometers from 260 to 450 mOsm, and the osmotically inactive sperm volumes of the two strains were 79.8+/-1.5% and 81.4+/-2.2%, respectively. Rat sperm have very limited osmotic tolerances. The sperm motility and the sperm plasma membranes of both strains were sensitive to anisosmotic treatments, but the acrosomes of both strains were more sensitive to hyposmotic than hyperosmotic conditions. The one-step addition and removal of Me(2)SO showed the most deleterious effect on rat sperm motility, plasma membrane integrity, and acrosomal integrity among the four cryoprotectants. These data characterizing rat sperm osmotic behavior, osmotic and cryoprotectant tolerance will be used to design cryopreservation protocols for rat sperm. PMID:17084388

Si, Wei; Benson, James D; Men, Hongsheng; Critser, John K



Association of actin with sperm centrioles: isolation of centriolar complexes and immunofluorescent localization of actin  

PubMed Central

The centrioles of cnidarian sperm associate with striated specializations (pericentriolar processes) during spermiogenesis. Three functions have been proposed for the role of these structures: (a) an anchoring mechanism for the sperm flagellum, (b) a signal-transmitting mechanism for communication between sperm head and tal, and (c) a contractile mechanism involved in motor function of the sperm flagellum. To investigate these proposed functions, we developed a technique for the isolation and purification of Hydractinia sperm distal centriles with attached pericentriolar processes. SDS polyacrylamide electrophoretic profiles of whole sperm and pericentriolar process proteins revealed a prominent protein that comigrates with rabbit and penaeid shrimp muscle actin. To label and localize actin in hydroid spem, we produced in rabbits a highly specific antiserum to invertebrate actin that cross-reacts with both invertebrate and vertebrate muscle and nonmuscle actin. Immunofluorescent double antibody labeling of hydroid sperm with antiactin has demonstrated the presence of actin in the pericentriolar process region of the sperm. In earlier reports, it has been proposed that pericentriolar processes, if contractile, could alter the mid- piece asymmetry of hydroid sperm, facilitating the directional motility that these cells demonstrate in respone to egg-released chemoattractants. The present results support this hypothesis.



Sperm donation in Israel.  


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

Mor-Yosef, S; Schenker, J G



Testing the functional significance of tail streamers  

PubMed Central

Studies of the evolution of elaborate ornaments have concentrated on their role in increasing attractiveness to mates. The classic examples of such sexually selected structures are the elongated tails of some bird species. Elongated tails can be divided into three categories: graduated tails, pin tails and streamers. There seems to be little debate about whether graduated and pin tails are ornaments; i.e. costly signals used in mate choice. However, in the case of streamers there is considerable discussion about their function. It has been suggested that tail streamers could be (i) entirely naturally selected, (ii) entirely sexually selected, (iii) partly naturally and partly sexually selected. The prime example of a species with tail streamers is the swallow (Hirundo rustica) in which both sexes have tail streamers. In this paper we discuss the aerodynamic consequences of different types of manipulation of the streamer and/or outer tail feather. We make qualitative predictions about the aerodynamic performance of swallows with manipulated tail streamers; these predictions differ depending on whether streamers have a naturally or sexually selected function. We demonstrate that these hypotheses can only be separated if tail streamers are shortened and changes in aerodynamic performance measured during turning flight.

Evans, M. R.; Thomas, A. L. R.



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

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.

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



Large-male advantages associated with costs of sperm production in Drosophila hydei, a species with giant sperm.  


Males of the fruit fly Drosophila hydei were found to produce 23.47 +/- 0.46-mm-long spermatozoa, the longest ever described. No relationship was found between male body size and sperm length. We predicted that if these giant gametes are costly for males to produce, then correlations should exist between male body size, rates of sperm production, and fitness attributes associated with the production of sperm. Smaller males were found to make a greater relative investment in testicular tissue growth, even though they have shorter and thinner testes. Smaller males were also found to (i) be maturing fewer sperm bundles within the testes at any point in time than larger males, (ii) require a longer period of time post-eclosion to become reproductively mature, (iii) mate with fewer females, (iv) transfer fewer sperm per copulation, and (v) produce fewer progeny. The significance of these findings for body size-related fitness and the question of sperm size evolution are discussed. PMID:7937755

Pitnick, S; Markow, T A



Methods for Cryopreservation of Guinea Fowl Sperm  

PubMed Central

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.

Varadi, Eva; Vegi, Barbara; Liptoi, Krisztina; Barna, Judit



An update on contraindications for lung function testing  

Microsoft Academic Search

Guidelines on contraindications for lung function tests have been based on expert opinion from >30 years ago. High-risk contraindications to lung function testing are associated with cardiovascular complications such as myocardial infarct, pulmonary embolism or ascending aortic aneurysm. Slightly less risky but still serious contraindications are predominantly centred on recovery from major thoracic, abdominal or head surgery. Less serious surgical

Brendan G Cooper



Ethical issues in lung function testing in children  

Microsoft Academic Search

Most lung function tests are non-invasive and they are widely performed in children and adults for clinical reasons. The prevalence of respiratory disorders is such that there is a considerable amount of research in the area. For both clinical and research applications, professionals involved in lung function testing in children need to be aware of ethical issues pertaining to such

C. S. Beardsmore



Paediatric lung function testing : determinants and reference values  

Microsoft Academic Search

Paediatric lung function tests provide objective measures in diagnosis and follow-up of lung diseases, and give insight in the pathophysiology of lung growth and development. In the first part of the thesis, paediatric lung function tests were used to investigate determinants of lung growth and development. The implementation of smoke free legislation resulted in less pregnant women being exposed to

M. Koopman



Sperm mRNA transcripts are indicators of sub-chronic low dose testicular injury in the Fischer 344 rat.  


Current human reproductive risk assessment methods rely on semen and serum hormone analyses, which are not easily comparable to the histopathological endpoints and mating studies used in animal testing. Because of these limitations, there is a need to develop universal evaluations that reliably reflect male reproductive function. We hypothesized that toxicant-induced testicular injury can be detected in sperm using mRNA transcripts as indicators of insult. To test this, we exposed adult male Fischer 344 rats to low doses of model testicular toxicants and classically characterized the testicular injury while simultaneously evaluating sperm mRNA transcripts from the same animals. Overall, this study aimed to: 1) identify sperm transcripts altered after exposure to the model testicular toxicant, 2,5-hexanedione (HD) using microarrays; 2) expand on the HD-induced transcript changes in a comprehensive time course experiment using qRT-PCR arrays; and 3) test these injury indicators after exposure to another model testicular toxicant, carbendazim (CBZ). Microarray analysis of HD-treated adult Fischer 344 rats identified 128 altered sperm mRNA transcripts when compared to control using linear models of microarray analysis (q<0.05). All transcript alterations disappeared after 3 months of post-exposure recovery. In the time course experiment, time-dependent alterations were observed for 12 candidate transcripts selected from the microarray data based upon fold change and biological relevance, and 8 of these transcripts remained significantly altered after the 3-month recovery period (p<0.05). In the last experiment, 8 candidate transcripts changed after exposure to CBZ (p<0.05). The two testicular toxicants produced distinct molecular signatures with only 4 overlapping transcripts between them, each occurring in opposite directions. Overall, these results suggest that sperm mRNA transcripts are indicators of low dose toxicant-induced testicular injury in the rat. PMID:22952946

Pacheco, Sara E; Anderson, Linnea M; Sandrof, Moses A; Vantangoli, Marguerite M; Hall, Susan J; Boekelheide, Kim



Delineating the roles of males and females in sperm competition.  


Disentangling the relative roles of males, females and their interactive effects on competitive fertilization success remains a challenge in sperm competition. In this study, we apply a novel experimental framework to an ideally suited externally fertilizing model system in order to delineate these roles. We focus on the chinook salmon, Oncorhynchus tshawytscha, a species in which ovarian fluid (OF) has been implicated as a potential arbiter of cryptic female choice for genetically compatible mates. We evaluated this predicted sexually selected function of OF using a series of factorial competitive fertilization trials. Our design involved a series of 10 factorial crosses, each involving two 'focal' rival males whose sperm competed against those from a single 'standardized' (non-focal) rival for a genetically uniform set of eggs in the presence of OF from two focal females. This design enabled us to attribute variation in competitive fertilization success among focal males, females (OF) and their interacting effects, while controlling for variation attributable to differences in the sperm competitive ability of rival males, and male-by-female genotypic interactions. Using this experimental framework, we found that variation in sperm competitiveness could be attributed exclusively to differences in the sperm competitive ability of focal males, a conclusion supported by subsequent analyses revealing that variation in sperm swimming velocity predicts paternity success. Together, these findings provide evidence that variation in paternity success can be attributed to intrinsic differences in the sperm competitive ability of rival males, and reveal that sperm swimming velocity is a key target of sexual selection. PMID:24153386

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



Quantitative ultrastructural analysis of barley sperm  

Microsoft Academic Search

Summary Complete serial ultrathin sections of seven sperm pairs, computer-assisted measurements of cell, nuclear and organelle surface areas and volumes, and three-dimensional imagery were used to demonstrate that a process of cytoplasm and organelle elimination occurs during sperm maturation in barley. The number of mitochondria per sperm cell is reduced by 50%; sperm cell surface area and volume are reduced

H. L. Mogensen; Maxine Losoff Rusche



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


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

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



Identification and validation of mouse sperm proteins correlated with epididymal maturation  

PubMed Central

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.

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



Cryopreservation of sperm of red abalone (Haliotis rufescens)  

USGS Publications Warehouse

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

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



Lung function testing in adults with preferential nasal breathing  

Microsoft Academic Search

Three adult patients with asthma with preferential nasal breathing were found to have a typical pattern of lung function test results with substantial between test variation. This condition can be identified as a cause of unsatisfactory performance in respiratory tests by observing the patient's reaction after the nostrils have been occluded.

C F Stanford; B Martin; D P Nicholls; D Watson



Function Testing for Chemical Brain Damage: A Review  

Microsoft Academic Search

Testing of neurobehavioral functions for evaluation of the effects of chemicals on the human brain from community (i.e., environmental) exposures is logical and may be a preferred initial step. Sensitivity is improved (1) by adjusting individual tests for influential factors, found by regression modeling and by retaining significant coefficients; and (2) by the calculation of predicted values for each test

Kaye H. Kilburn



Intrasubject variability of pulmonary function testing in healthy children  

Microsoft Academic Search

The intrasubject variability of repeat pulmonary function testing was examined in 20 healthy children aged 10 to 16 years. The children were tested a maximum of 11 times over a period of two months. The tests examined were spirometry, maximum expiratory flow-volume curves, body plethysmograph determination of lung volumes, and single breath nitrogen washout. The time of day or the

A A Hutchison; A Erben; L A McLennan; L I Landau; P D Phelan



The ecological validity of tests of executive function  

Microsoft Academic Search

Ninety-two mixed etiology neurological patients and 216 control participants were assessed on a range of neuropsychological tests, including 10 neuropsychological measures of executive function derived from 6 different tests. People who knew the patients well (relatives or carers) completed a questionnaire about the patient's dysexecutive problems in everyday life, and this paper reports the extent to which the tests predicted




Functional and IDDQ testing on a static RAM  

Microsoft Academic Search

A layout defect analysis was performed on an 8K×8 static RAM. Simulations were done to determine the electrical behavior and detection possibilities of the defects. The authors introduce a test pattern for functional testing of SRAMs. It was found, by testing more than 7000 devices, that this 6N march pattern produces the same results as a 13N pattern in over

R. Meershoek; B. Verhelst; R. McInerney; L. Thijssen



Functional Search-based Testing from State Machines  

Microsoft Academic Search

The application of metaheuristic search techniques in test data generation has been extensively investigated in recent years. Most studies, however, have concentrated on the application of such techniques in structural testing. The use of search-based techniques in functional testing is less frequent, the main cause being the implicit nature of the specification. This paper investigates the use of search-based techniques

Raluca Lefticaru; Florentin Ipate



Receptor mediated amelioration of the detrimental effects of sperm agglutinating factor on sperm parameters.  


Sperm agglutinating factor (SAF) isolated from Staphylococcus aureus immobilizes spermatozoa by agglutination and causes sperm death. This interaction of SAF with spermatozoa is receptor mediated and this receptor has been isolated and purified from human spermatozoa. In this study we attempt to study whether the receptor could ameliorate the detrimental effects of SAF on sperm parameters. Receptor was evaluated against SAF mediated compromised sperm parameters such as Mg(2+) dependent ATPase activity, acrosome status and apoptosis, in vitro using fluorescent microscopy and flow cytometry as well as in vivo by studying the impact on fertility in mice. Incubation of SAF (80 ?g) with spermatozoa resulted in reduced Mg(2+) dependent ATPase activity and premature acrosomal loss whereas a higher concentration (100 ?g), induced apoptosis. However, in the presence of receptor a dose dependent blockage of SAF induced inhibition of Mg(2+) dependent ATPase activity was observed. At higher concentrations 100 and 125 ?g, receptor could inhibit both the premature acrosomal loss and apoptosis. In vivo studies showed that receptor (50 ?g) could alleviate SAF induced infertility in female Balb/c mice following a single intravaginal application before mating. The work highlights the efficacy of the receptor as a corrective measure against negative influence of SAF on functional parameters of spermatozoa as well as fertility and presents receptor as a potential therapeutic intervention against SAF induced infertility. PMID:23657873

Kaur, S; Prabha, V



Comparative sperm ultrastructure in Nemertea.  


Although the monophyly of Nemertea is strongly supported by unique morphological characters and results of molecular phylogenetic studies, their ingroup relationships are largely unresolved. To contribute solving this problem we studied sperm ultrastructure of 12 nemertean species that belong to different subtaxa representing the commonly recognized major monophyletic groups. The study yielded a set of 26 characters with an unexpected variation among species of the same genus (Tubulanus and Procephalothrix species), whereas other species varied in metric values or only one character state (Ramphogordius). In some species, the sperm nucleus has grooves (Zygonemertes virescens, Amphiporus imparispinosus) that may be twisted and give a spiral shape to the sperm head (Paranemertes peregrina, Emplectonema gracile). To make the characters from sperm ultrastructure accessible for further phylogenetic analyses, they were coded in a character matrix. Published data for eight species turned out to be sufficiently detailed to be included. Comparative evaluation of available information on the sperm ultrastructure suggests that subtaxa of Heteronemertea and Hoplonemertea are supported as monophyletic by sperm morphology. However, the data do not provide information on the existing contradictions regarding the internal relationships of "Palaeonemertea." Nevertheless, our study provides evidence that sperm ultrastructure yields numerous potentially informative characters that will be included in upcoming phylogenetic analyses. PMID:20544873

von Döhren, J; Beckers, P; Vogeler, R; Bartolomaeus, T



The influence of anti-zona and anti-sperm antibodies on sperm--egg interactions.  


Anti-zona antibodies are effective inhibitors of fertilization in vitro and, regardless of whether passive or active immunization techniques are used, in vivo. Antibodies raised against unfractionated zona pellucida antigens are chiefly directed against a group of carbohydrate-rich components localized on the outer surface of the zona. The interaction of anti-zona antibodies with these sites induces the formation of a surface precipitate which occludes the sperm binding sites by a process of steric hindrance, and stabilizes the zona structure against digestion by the proteolytic enzymes of the sperm head. Active immunization studies indicate that the long-term induction of infertility without adverse side effects is feasible in both laboratory rodents and primates when the zona pellucida is used as a target. Anti-sperm antibodies also exhibit a capacity for inhibiting fertilization in vivo and in vitro. To determine the most appropriate detection method to screen patients for anti-sperm antibodies several homologous and heterologous antisera were analysed by 5 different agglutination and immobilization techniques and then compared for their ability to inhibit the fertilizing capacity of human spermatozoa using the zona-free hamster egg penetration test. The results obtained with the Franklin--Dukes tube--slide test exhibited the closest correlation with the anti-fertility activity of a given antiserum; this activity could be amplified by the addition of complement to the medium. It is concluded that antibodies directed against the sperm head are responsible for limiting the fertilizing capacity of human spermatozoa in vitro and that it is these antibodies of which attention should be focused to unravel the role that immunological factors play in the aetiology of infertility in vivo. PMID:7019431

Aitken, R J; Rudak, E A; Richardson, D W; Dor, J; Djahanbahkch, O; Templeton, A A



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

PubMed Central

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

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



No association between sperm competition and sperm length variation across dung flies (Scathophagidae).  


Sperm length is extremely variable across species, but a general explanation for this variation is lacking. However, when the risk of sperm competition is high, sperm length is predicted to be less variable within species, and there is some evidence for this in birds and social insects. Here, we examined intraspecific variation in sperm length, both within and between males, and its potential associations with sperm competition risk and variation in female reproductive tract morphology across dung flies. We used two measures of variation in sperm size, and testis size was employed as our index of sperm competition risk. We found no evidence of associations between sperm length variation and sperm competition or female reproductive tract variation. These results suggest that variation in sperm competition risk may not always be associated with variation in sperm morphology, and the cause(s) of sperm length variation in dung flies remains unclear. PMID:24016061

Sharma, M D; Minder, A M; Hosken, D J



Shorter sperm confer higher competitive fertilization success.  


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

García-González, Francisco; Simmons, Leigh W



Containment purge and vent valve test program final report. [Function and leak testing  

Microsoft Academic Search

The test program investigated valve functionality and leak integrity. Three nuclear designed butterfly valves typical of those used in domestic nuclear power plant containment purge and vent applications were tested. For a comparison of response, two valves of the same size with differing internal designs were tested. For extrapolation insights, a larger sized valve was also tested. The valve experiments

R. Jr. Steele; J. C. Watkins



Association of Seminal Plasma Motility Inhibitors\\/Semenogelins with Sperm in Asthenozoospermia-Infertile Men  

Microsoft Academic Search

Seminal plasma motility inhibitors (SPMIs) are proteinase-resistant fragments of semenogelin I and II (Sgs), which are the major proteins of semen coagulum. SPMIs inhibit the motility of spermatozoa, and Sgs are thought to be natural regulators of human sperm function. The mechanism underlying sperm motility regulation and its association with defective motility in infertile men remain unclear. The purpose of

K. Terai; K. Yoshida; M. Yoshiike; M. Fujime; T. Iwamoto



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

Microsoft Academic Search

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

Maya D. Markova; Ralitsa S. Zhivkova



The giant bite of a new raptorial sperm whale from the Miocene epoch of Peru  

Microsoft Academic Search

The modern giant sperm whale Physeter macrocephalus, one of the largest known predators, preys upon cephalopods at great depths. Lacking a functional upper dentition, it relies on suction for catching its prey; in contrast, several smaller Miocene sperm whales (Physeteroidea) have been interpreted as raptorial (versus suction) feeders, analogous to the modern killer whale Orcinus orca. Whereas very large physeteroid

Olivier Lambert; Giovanni Bianucci; Klaas Post; Christian de Muizon; Rodolfo Salas-Gismondi; Mario Urbina; Jelle Reumer



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

Microsoft Academic Search

Summary Populations of sturgeon across the globe are threatened due to unregulated harvest and habitat loss, and the status varies among species across North America. Ready access to viable and functional sperm would contribute to recovery pro- grammes for these species. In this study, we examined the motility, viability (cell membrane integrity) of cryopreserved sperm from three North American acipenseriform

Á. Horváth; W. R. Wayman; J. C. Dean; B. Urbányi; T. R. Tiersch; S. D. Mims; D. Johnson; J. A. Jenkins



Proteomic characterization of sperm radial spokes identifies a novel spoke protein with an ubiquitin domain  

Microsoft Academic Search

Radial spokes are T-shaped protein complexes important for the regulation of axonemal dyneins in eukaryotic cilia and flagella. Using a functional proteomics approach, we identified six spoke proteins in sperm flagella of the ascidian Ciona intestinalis. Many of the domain\\/motif structures in spoke proteins are commonly found in flagella of both Ciona sperm and Chlamydomonas, but interestingly they often distribute

Yuhkoh Satouh; Kazuo Inaba



Testicular structure, spermatogenesis and sperm cryopreservation in the African clariid catfish Heterobranchus longifilis (Valenciennes, I 840)  

Microsoft Academic Search

The morphological and physiological characteris- tics of the testes and the sperm of the catfish Heterobranchus longifilis (Val.) are presented. The effect of cryopreservation on -the fertilizing capacity of the sperm was also evaluated. Testicular struc- ture and spermatogenesis are described using histological techniques. The coexistence in the lobules of spermatozoa and all the spermatogenic stages indicates that this species

Côte d'Ivoire


The sperm surface localization of the TRP-3/SPE-41 Ca2+ -permeable channel depends on SPE-38 function in Caenorhabditis elegans.  


Despite undergoing normal development and acquiring normal morphology and motility, mutations in spe-38 or trp-3/spe-41 cause identical phenotypes in Caenorhabditis elegans-mutant sperm fail to fertilize oocytes despite direct contact. SPE-38 is a novel, four-pass transmembrane protein and TRP-3/SPE-41 is a Ca(2+)-permeable channel. Localization of both of these proteins is confined to the membranous organelles (MOs) in undifferentiated spermatids. In mature spermatozoa, SPE-38 is localized to the pseudopod and TRP-3/SPE-41 is localized to the whole plasma membrane. Here we show that the dynamic redistribution of TRP-3/SPE-41 from MOs to the plasma membrane is dependent on SPE-38. In spe-38 mutant spermatozoa, TRP-3/SPE-41 is trapped within the MOs and fails to reach the cell surface despite MO fusion with the plasma membrane. Split-ubiquitin yeast-two-hybrid analyses revealed that the cell surface localization of TRP-3/SPE-41 is likely regulated by SPE-38 through a direct protein-protein interaction mechanism. We have identified sequences that influence the physical interaction between SPE-38 and TRP-3/SPE-41, and show that these sequences in SPE-38 are required for fertility in transgenic animals. Despite the mislocalization of TRP-3/SPE-41 in spe-38 mutant spermatozoa, ionomycin or thapsigargin induced influx of Ca(2+) remains unperturbed. This work reveals a new paradigm for the regulated surface localization of a Ca(2+)-permeable channel. PMID:22425620

Singaravelu, Gunasekaran; Chatterjee, Indrani; Rahimi, Sina; Druzhinina, Marina K; Kang, Lijun; Xu, X Z Shawn; Singson, Andrew



Sexual selection drives weak positive selection in protamine genes and high promoter divergence, enhancing sperm competitiveness  

PubMed Central

Phenotypic adaptations may be the result of changes in gene structure or gene regulation, but little is known about the evolution of gene expression. In addition, it is unclear whether the same selective forces may operate at both levels simultaneously. Reproductive proteins evolve rapidly, but the underlying selective forces promoting such rapid changes are still a matter of debate. In particular, the role of sexual selection in driving positive selection among reproductive proteins remains controversial, whereas its potential influence on changes in promoter regions has not been explored. Protamines are responsible for maintaining DNA in a compacted form in chromosomes in sperm and the available evidence suggests that they evolve rapidly. Because protamines condense DNA within the sperm nucleus, they influence sperm head shape. Here, we examine the influence of sperm competition upon protamine 1 and protamine 2 genes and their promoters, by comparing closely related species of Mus that differ in relative testes size, a reliable indicator of levels of sperm competition. We find evidence of positive selection in the protamine 2 gene in the species with the highest inferred levels of sperm competition. In addition, sperm competition levels across all species are strongly associated with high divergence in protamine 2 promoters that, in turn, are associated with sperm swimming speed. We suggest that changes in protamine 2 promoters are likely to enhance sperm swimming speed by making sperm heads more hydrodynamic. Such phenotypic changes are adaptive because sperm swimming speed may be a major determinant of fertilization success under sperm competition. Thus, when species have diverged recently, few changes in gene-coding sequences are found, while high divergence in promoters seems to be associated with the intensity of sexual selection.

Martin-Coello, Juan; Dopazo, Hernan; Arbiza, Leonardo; Ausio, Juan; Roldan, Eduardo R.S.; Gomendio, Montserrat



A Role for Acp29AB, a Predicted Seminal Fluid Lectin, in Female Sperm Storage in Drosophila melanogaster  

PubMed Central

Females of many animal species store sperm for taxon-specific periods of time, ranging from a few hours to years. Female sperm storage has important reproductive and evolutionary consequences, yet relatively little is known of its molecular basis. Here, we report the isolation of a loss-of-function mutation of the Drosophila melanogaster Acp29AB gene, which encodes a seminal fluid protein that is transferred from males to females during mating. Using this mutant, we show that Acp29AB is required for the normal maintenance of sperm in storage. Consistent with this role, Acp29AB localizes to female sperm storage organs following mating, although it does not appear to associate tightly with sperm. Acp29AB is a predicted lectin, suggesting that sugar–protein interactions may be important for D. melanogaster sperm storage, much as they are in many mammals. Previous association studies have found an effect of Acp29AB genotype on a male's sperm competitive ability; our findings suggest that effects on sperm storage may underlie these differences in sperm competition. Moreover, Acp29AB's effects on sperm storage and sperm competition may explain previously documented evidence for positive selection on the Acp29AB locus.

Wong, Alex; Albright, Shannon N.; Giebel, Jonathan D.; Ram, K. Ravi; Ji, Shuqing; Fiumera, Anthony C.; Wolfner, Mariana F.



Sperm speed is associated with sex bias of siblings in a human population  

PubMed Central

Recent studies investigating possible causes of male subfertility have largely focused on how lifestyle or environmental factors impact on the process of spermatogenesis. Markedly, fewer studies have investigated those risk factors that result in reduced sperm quality, such as poor sperm motility. The speed at which sperm swim is a major predictor of fertility and is extremely variable in human populations. It has been hypothesized that offspring sex may be adaptively manipulated to maximize the offspring's reproductive fitness (e.g., parents with genes for good male fertility traits, such as high sperm speed, would produce primarily sons and fewer daughters because the offspring will inherit advantageous male fertility genes). Conversely, parents with poor male fertility genes would produce primarily daughters. We tested whether there was an association between how fast a man's sperm swam and the sex bias of his siblings in a sample of men attending clinic for fertility investigations with their partner and with a wide range of semen characteristics, including sperm speed. We found that the sex bias of a man's siblings is associated with his sperm speed; men with female-biased siblings had significantly slower sperm (judged using computer-assisted sperm analysis (CASA)) than men from male-biased sibships. This observation suggests family composition is an important factor that needs to be considered in future epidemiological and clinical studies of human fertility.

Mossman, Jim A; Slate, Jon; Birkhead, Tim R; Moore, Harry D; Pacey, Allan A



Ionic regulation of sea urchin sperm motility, metabolism and fertilizing capacity.  

PubMed Central

In order to pursue the significance of the ionic regulation of sea urchin sperm behaviour, alterations in the cation composition of sea water were tested for their effects on sperm fertilizing capacity. Nearly all changes which resulted in lowered sperm intracellular pH, including lowered sea-water pH, inclusion of the divalent ion chelator EGTA, addition of dithiothreitol, or removal of sea-water Na+, enhanced sperm viability for periods of up to a week. These conditions caused decreased cell motility and elevated ATP concentrations, and prevented the acrosome reaction. Conversely, changes which increased the intracellular pH, decreased sperm ATP concentrations, or induced the acrosome reaction, reduced sperm viability. A single medium, high sea-water K+ concentrations (greater than 100 mM), provided an exception to these general trends. At elevated K+ concentrations sperm were quiescent but became completely infertile. These data show that sperm fertilizing capacity is generally extended by maintenance of the sperm in an inactive state, and the results suggest that decreased cellular energy levels contribute to decreased fertility.

Christen, R; Schackmann, R W; Shapiro, B M



A Semi-Automated Functional Test Data Analysis Tool  

SciTech Connect

The growing interest in commissioning is creating a demand that will increasingly be met by mechanical contractors and less experienced commissioning agents. They will need tools to help them perform commissioning effectively and efficiently. The widespread availability of standardized procedures, accessible in the field, will allow commissioning to be specified with greater certainty as to what will be delivered, enhancing the acceptance and credibility of commissioning. In response, a functional test data analysis tool is being developed to analyze the data collected during functional tests for air-handling units. The functional test data analysis tool is designed to analyze test data, assess performance of the unit under test and identify the likely causes of the failure. The tool has a convenient user interface to facilitate manual entry of measurements made during a test. A graphical display shows the measured performance versus the expected performance, highlighting significant differences that indicate the unit is not able to pass the test. The tool is described as semiautomated because the measured data need to be entered manually, instead of being passed from the building control system automatically. However, the data analysis and visualization are fully automated. The tool is designed to be used by commissioning providers conducting functional tests as part of either new building commissioning or retro-commissioning, as well as building owners and operators interested in conducting routine tests periodically to check the performance of their HVAC systems.

Xu, Peng; Haves, Philip; Kim, Moosung



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

PubMed Central

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

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



Test Review: Delis-Kaplan Executive Function System  

Microsoft Academic Search

The Delis-Kaplan Executive Function System (D-KEFS; Delis, Kaplan, & Kramer, 2001) represents the first set of executive tests co-normed on a large and representative national sample designed exclusively for the assessment of executive functions including flexibility of thinking, inhibition, problem solving, planning, impulse control, concept formation, abstract thinking, and creativity. The D-KEFS, composed of nine stand alone tests that can

Susan Homack; Donghyung Lee; Cynthia A. Riccio



Evolutionary Optimization of Dynamic Multi-objective Test Functions  

Microsoft Academic Search

Multi-objective as well as dynamic characteristics appear in many real-world problems. In order to use multi-objective evolutionary optimization algorithms (MOEA) efficiently, a systematic analysis of the algorithms' behavior in dynamic environments by means of dynamic test functions is necessary. These functions can be classified into problems with changing Pareto sets and\\/or Pareto fronts with different dynamic criteria. Thus, a test

Jorn Mehnen; Tobias Wagner; Gunter Rudolph


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

SciTech Connect

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

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



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

PubMed Central

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

Purchase, Craig F; Moreau, Darek T R



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


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

Purchase, Craig F; Moreau, Darek T R



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


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

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



Sperm trajectories form chiral ribbons.  


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. PMID:23588811

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



Sperm Trajectories Form Chiral Ribbons  

NASA Astrophysics Data System (ADS)

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.

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



Dephosphorylation of sperm guanylate cyclase during sea urchin fertilization  

SciTech Connect

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.

Ward, G.E.



Relationship between Testicular Volume and Conventional or Nonconventional Sperm Parameters  

PubMed Central

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.

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



Functional capacity evaluation: Ecological validity of three static endurance tests.  


Functional Capacity Evaluation's (FCE's) are designed to measure the functional capacity of injured workers. Static endurance tests are integrated aspects of FCE's. Little is known about the validity of the tests. In this study, three static endurance tests (overhead work, crouching and kneeling) of the Isernhagen Work Systems FCE are studied for ecological validity. By manipulating the environment in an experiment using three different conditions (normal, loud noise, high production requirement), the ecological validity of the tests was investigated. Results: the different conditions did not seem to influence the holding times, the perceived exertion and the productivity of the subjects. The results are discussed and it is concluded that the three static endurance tests meet conditions of ecological validity. In order to be able to state that the tests of the IWS FCE are ecologically valid, more research is needed to enable a generalization. PMID:12441452

Reneman, M.F.; Joling, C.I.; Soer, E.L.; Göeken, L.N.H.



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


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

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



Fault Modeling and Functional Test Methods for Digital Microfluidic Biochips  

Microsoft Academic Search

Dependability is an important attribute for microfluidic biochips that are used for safety-critical applications, such as point-of-care health assessment, air-quality monitoring, and food-safety testing. Therefore, these devices must be adequately tested after manufacture and during bioassay operations. Known techniques for biochip testing are all function oblivious (i.e., while they can detect and locate defect sites on a microfluidic array, they

Tao Xu; Krishnendu Chakrabarty



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

SciTech Connect

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

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



The test of functional health literacy in adults  

Microsoft Academic Search

OBJECTIVE: To develop a valid, reliable instrument to measure the functional health literacy of patients.\\u000a \\u000a \\u000a DESIGN: The Test of Functional Health Literacy in Adults (TOFHLA) was developed using actual hospital materials. The TOFHLA consists\\u000a of a 50-item reading comprehension and 17-item numerical ability test, taking up to 22 minutes to administer. The TOFHLA,\\u000a the Wide Range Achievement Test-Revised (WRAT-R), and

Ruth M. Parker; David W. Baker; Mark V. Williams; Joanne R. Nurss



State of the Art in Platelet Function Testing  

PubMed Central

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.

E. Kehrel, Beate; F. Brodde, Martin



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


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

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



A Powerful Test for Comparing Multiple Regression Functions  

PubMed Central

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).

Maity, Arnab



Test Items and Standards Related to Aerobic Functioning on the Brockport Physical Fitness Test  

Microsoft Academic Search

This article describes the procedures and rationale for the selection of test items and criterion-referenced standards associated with the aerobic functioning component of the Brockport Physical Fitness Test. Validity and reliability information is provided for the 1-mile run\\/walk, the PACER (16-m and 20-m), and the Target Aerobic Movement Test. The relevance of these test items and standards for youngsters withmental

Francis X. Short; Joseph P. Winnick



Epididymis cholesterol homeostasis and sperm fertilizing ability  

PubMed Central

Cholesterol, being the starting point of steroid hormone synthesis, is a long known modulator of both female and male reproductive physiology especially at the level of the gonads and the impact cholesterol has on gametogenesis. Less is known about the effects cholesterol homeostasis may have on postgonadic reproductive functions. Lately, several data have been reported showing how imbalanced cholesterol levels may particularly affect the post-testicular events of sperm maturation that lead to fully fertile male gametes. This review will focus on that aspect and essentially centers on how cholesterol is important for the physiology of the mammalian epididymis and spermatozoa.

Saez, Fabrice; Ouvrier, Aurelia; Drevet, Joel R



Dioxin-Induced Changes in Epididymal Sperm Count and Spermatogenesis  

PubMed Central

Background A single in utero exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on gestation day 15 decreased epididymal sperm count in adult rats and thus was used to establish a tolerable daily intake for TCDD. However, several laboratories have been unable to replicate these findings. Moreover, conflicting reports of TCDD effects on daily sperm production suggest that spermatogenesis may not be as sensitive to the adverse effects of TCDD as previously thought. Data sources We performed a PubMed search using relevant search terms linking dioxin exposure with adverse effects on reproduction and spermatogenesis. Data synthesis Developmental exposure to TCDD is consistently linked with decreased cauda epididymal sperm counts in animal studies, although at higher dose levels than those used in some earlier studies. However, the evidence linking in utero TCDD exposure and spermatogenesis is not convincing. Conclusions Animal studies provide clear evidence of an adverse effect of in utero TCDD exposure on epididymal sperm count but do not support the conclusion that spermatogenesis is adversely affected. The mechanisms underlying decreased epididymal sperm count are unknown; however, we postulate that epididymal function is the key target for the adverse effects of TCDD.

Foster, Warren G.; Maharaj-Briceno, Serena; Cyr, Daniel G.



Quantitative analysis of radiation-induced changes in sperm morphology  

SciTech Connect

When developing spermatogenic cells are exposed to radiation, chemical carcinogens or mutagens, the transformation in the morphology of the mature sperm can be used to determine the severity of the exposure. In this study five groups of mice with three mice per group received testicular doses of X irradiation at dosage levels ranging from 0 rad to 120 rad. A random sample of 100 mature sperm per mouse was analyzed five weeks later for the quantitative morphologic transformation as a function of dosage level. The cells were stained with gallocyanin chrome alum (GCA) so that only the DNA in the sperm head was visible. The ACUity quantitative microscopy system at Lawrence Livermore National Laboratory was used to scan the sperm at a sampling density of 16 points per linear micrometer and with 256 brightness levels per point. The contour of each cell was extracted using conventional thresholding techniques on the high-contrast images. For each contour a variety of shape features was then computed to characterize the morphology of that cell. Using the control group and the distribution of their shape features to establish the variability of a normal sperm population, the 95% limits on normal morphology were established. Using only four shape features, a doubling dose of approximately 39 rad was determined. That is, at 39 rad exposure the percentage of abnormal cells was twice that occurring in the control population. This compared to a doubling dose of approximately 70 rad obtained from a concurrent visual procedure.

Young, I.T.; Gledhill, B.L.; Lake, S.; Wyrobek, A.J.



Sea urchin sperm antigens mediating the acrosome reaction  

SciTech Connect

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

Trimmer, J.S.



Isolation and Proteomic Characterization of the Mouse Sperm Acrosomal Matrix*  

PubMed Central

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

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



Systematic characterization of sperm-specific membrane proteins in swine.  


To establish a systematic strategy for characterizing fertilization proteins of sperm cells, we prepared alloantisera by immunizing gilts with salt-washed membranes from boar spermatozoa. The antisera recognized a unique subset of sperm membrane proteins that migrated with M(r) 7500-66,000 in SDS-PAGE under nonreducing conditions. The antisera did not recognize proteins of erythrocyte membranes, and tissue absorption experiments further confirmed that the alloantigens were sperm-specific proteins. Each of these sperm-specific membrane proteins (SSMPs) possessed one or more disulfide bonds that were essential for its interaction with alloantibody. Enzymatic deglycosylation revealed that most of the SSMPs were glycoproteins, and their alloantigenicity was not dependent on the presence of N-linked oligosaccharides. The presence of disulfide bonds and glycosylation indicated that the SSMPs identified each comprise at least one extracellular domain. Two-dimensional electrophoresis resolved at least 14 distinct SSMPs, 13 of which possessed acidic pIs (range 4.2-4.8). By indirect immunofluorescence, the SSMPs localized to the cell surface overlying all major regions of the sperm cell. We conclude that the repertoire of immunodominant SSMPs in the pig is relatively small, which makes feasible the systematic elucidation of their functions in fertilization. PMID:11090456

Haden, N P; Hickox, J R; Whisnant, C S; Hardy, D M



Identification of calcium-binding proteins associated with the human sperm plasma membrane  

PubMed Central

Background The precise composition of the human sperm plasma membrane, the molecular interactions that define domain specific functions, and the regulation of membrane associated proteins during the capacitation process, still remain to be fully understood. Here, we investigated the repertoire of calcium-regulated proteins associated with the human sperm plasma membrane. Methods Surface specific radioiodination was combined with two-dimensional gel electrophoresis, a 45Ca-overlay assay, computer assisted image analysis and mass spectrometry to identify calcium-binding proteins exposed on the human sperm surface. Results Nine acidic 45Ca-binding sperm proteins were excised from stained preparative 2D gels and identified by mass spectrometry. Five of the calcium binding proteins; HSPA2 (HSP70-1), HSPA5 (Bip), HYOU1 (ORP150), serum amyloid P-component (SAP) and protein kinase C substrate 80K-H (80K-H) were found to be accessible to Iodo-Bead catalyzed 125I-labelling on the surface of intact human sperm. Agglutination and immunofluorescence analysis confirmed that SAP is situated on the plasma membrane of intact, motile sperm as well as permeabilized cells. Western blot analysis showed increased phosphorylation of human sperm 80K-H protein following in vitro capacitation. This is the first demonstration of the 80K-H protein in a mammalian sperm. Conclusion The presence of SAP on the surface of mature sperm implies that SAP has a physiological role in reproduction, which is thought to be in the removal of spermatozoa from the female genital tract via phagocytosis. Since 80K-H is a Ca2+-sensor recently implicated in the regulation of both inositol 1,4,5-trisphosphate receptor and transient receptor potential (TRP) cation channel activities, its detection in sperm represents the first direct signaling link between PKC and store-operated calcium channels identified in human sperm.



No evidence of sperm conjugate formation in an Australian mouse bearing sperm with three hooks  

PubMed Central

Sperm conjugation occurs when two or more sperm physically unite for motility or transport through the female reproductive tract. In many muroid rodent species, sperm conjugates have been shown to form by a single, conspicuous apical hook located on the sperm head. These sperm “trains” have been reported to be highly variable in size and, despite all the heads pointing in roughly the same direction, exhibit a relatively disordered arrangement. In some species, sperm “trains” have been shown to enhance sperm swimming speed, and thus have been suggested to be advantageous in sperm competition. Here, we assessed the behavior of sperm in the sandy inland mouse (Pseudomys hermannsburgensis), a muroid rodent that bears sperm with three apical hooks. First, we accrued genetic evidence of multiple paternity within “wild” litters to unequivocally show that sperm competition does occur in this species. Following this we utilized both in vitro and in vivo methodologies to determine whether sandy inland mouse sperm conjugate to form motile trains. Our observations of in vitro preparations of active sperm revealed that sandy inland mouse sperm exhibit rapid, progressive motility as individual cells only. Similarly, histological sections of the reproductive tracts of mated females revealed no in vivo evidence of sperm conjugate formation. We conclude that the unique, three-hooked morphology of the sandy inland mouse sperm does not facilitate the formation of motile conjugates, and discuss our findings in relation to the different hypotheses for the evolution of the muroid rodent hook/s.

Firman, Renee C; Bentley, Blair; Bowman, Faye; Marchant, Fernando Garcia-Solis; Parthenay, Jahmila; Sawyer, Jessica; Stewart, Tom; O'Shea, James E



No evidence of sperm conjugate formation in an Australian mouse bearing sperm with three hooks.  


Sperm conjugation occurs when two or more sperm physically unite for motility or transport through the female reproductive tract. In many muroid rodent species, sperm conjugates have been shown to form by a single, conspicuous apical hook located on the sperm head. These sperm "trains" have been reported to be highly variable in size and, despite all the heads pointing in roughly the same direction, exhibit a relatively disordered arrangement. In some species, sperm "trains" have been shown to enhance sperm swimming speed, and thus have been suggested to be advantageous in sperm competition. Here, we assessed the behavior of sperm in the sandy inland mouse (Pseudomys hermannsburgensis), a muroid rodent that bears sperm with three apical hooks. First, we accrued genetic evidence of multiple paternity within "wild" litters to unequivocally show that sperm competition does occur in this species. Following this we utilized both in vitro and in vivo methodologies to determine whether sandy inland mouse sperm conjugate to form motile trains. Our observations of in vitro preparations of active sperm revealed that sandy inland mouse sperm exhibit rapid, progressive motility as individual cells only. Similarly, histological sections of the reproductive tracts of mated females revealed no in vivo evidence of sperm conjugate formation. We conclude that the unique, three-hooked morphology of the sandy inland mouse sperm does not facilitate the formation of motile conjugates, and discuss our findings in relation to the different hypotheses for the evolution of the muroid rodent hook/s. PMID:23919134

Firman, Renée C; Bentley, Blair; Bowman, Faye; Marchant, Fernando García-Solís; Parthenay, Jahmila; Sawyer, Jessica; Stewart, Tom; O'Shea, James E



Computer-assisted sperm analysis (CASA) as a tool for monitoring sperm quality in fish  

Microsoft Academic Search

The use of motility as a measure of sperm quality in fish is reviewed. Computer assisted sperm analysis (CASA) provides a simple and rapid quantitative assessment of the quality of fish sperm and may predict its ability to fertilise eggs. It has been used to: monitor the effects of heavy metal pollutants, such as mercury and tributyltin, on sperm quality;

D. E Kime; K. J. W Van Look; B. G McAllister; G Huyskens; E Rurangwa; F Ollevier



Sperm precedence and sperm movement under different copulation intervals in the silkworm, Bombyx mori  

Microsoft Academic Search

Sperm precedence and sperm movement were investigated under different copulation intervals in the silkworm, Bombyx mori. The mass of sperm began to enter the spermatheca 1 h after copulation ended, and the number of sperm gradually increased in the spermatheca and attained a plateau when the spermatheca was filled at about 2.5 h after the end of copulation. Thereafter, the

Nobuhiko Suzuki; Takashi Okuda; Hitoshi Shinbo



Sperm competition games played by dimorphic male beetles  

PubMed Central

Reproductive strategies often consist of two alternative tactics whereby males either compete for and guard females, or sneak copulations. By their nature, alternative tactics expose males to differing risks of sperm competition; sneaks will always be subject to sperm competition but guards will be subject to sperm competition with low probability, dependent on the number of sneaks. Recent game-theoretical models predict that males in the sneak role should have the greater gametic expenditure but that the disparity in expenditure should decrease with increasing numbers of sneaks. Male dung beetles in the genus Onthophagus can be separated into two morphs: major males have horns and guard females whereas minor males are hornless and sneak copulations. Here we compare testis size and ejaculate characteristics between these alternative morphs. We find that in O. binodis 30% of males are sneaks, and sneaks have larger testes, ejaculate volumes, and longer sperm than guards. In O. taurus 60% of males are sneaks and there are no differences in gametic traits. Our data thus provide empirical support for game-theoretical models of sperm competition.

Simmons, L. W.; Tomkins, J. L.; Hunt, J.



Establishment of a transport system for mouse epididymal sperm at refrigerated temperatures.  


The exchange of genetically engineered mouse strains between research facilities requires transporting fresh mouse sperm under refrigerated temperatures. Although sperm generally maintains fertility for 48 h at cold temperatures, in vitro fertilization rates of C57BL/6 mouse sperm are low after 48-h cold storage. Furthermore, 48 h is often not sufficient for the specimens to reach their destinations. To increase the availability of this technology, we aimed to extend the cold storage period while maintaining sperm fertility. In this study, we determined the optimal medium for sperm preservation and evaluated the effect of reduced glutathione in the fertilization medium on sperm fertility after cold storage. We found that higher fertility levels were maintained after 72-h cold storage in the preservation medium Lifor compared with storage in paraffin oil, M2 medium, or CPS-1 medium. In addition, 1.0 mM glutathione enhanced sperm fertility. After transporting sperm from Asahikawa Medical University to our laboratory, embryos were efficiently produced from the cold-stored sperm. After transfer, these embryos developed normally into live pups. Finally, we tested the transport system using genetically engineered mouse strains and obtained similar high fertilization rates with all specimens. In summary, we demonstrated that cold storage of sperm in Lifor maintains fertility, and glutathione supplementation increased the in vitro fertilization rates of sperm after up to 96 h of cold storage. This improved protocol provides a simple alternative to transporting live animals or cryopreserved samples for the exchange of genetically engineered mouse strains among research facilities. PMID:22722060

Takeo, Toru; Tsutsumi, Aki; Omaru, Taichi; Fukumoto, Kiyoko; Haruguchi, Yukie; Kondo, Tomoko; Nakamuta, Yuko; Takeshita, Yumi; Matsunaga, Hiroko; Tsuchiyama, Shuuji; Sakoh, Kazuhito; Nakao, Satohiro; Yoshimoto, Hidetaka; Shimizu, Norihiko; Nakagata, Naomi



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


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

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



Motile sperm organelle morphology examination (MSOME): intervariation study of normal sperm and sperm with large nuclear vacuoles  

Microsoft Academic Search

BACKGROUND: Although the motile sperm organelle morphology examination (MSOME) was developed only as a selection criterion, its application as a method for classifying sperm morphology may represent an improvement in evaluation of semen quality, with potential clinical repercussions. The present study aimed to evaluate individual variations in the motile sperm organelle morphology examination (MSOME) analysis after a time interval. METHODS:

João Batista A Oliveira; Claudia G Petersen; Fabiana C Massaro; Ricardo LR Baruffi; Ana L Mauri; Liliane FI Silva; Juliana Ricci; José G Franco



Microsurgical Vasovasostomy versus Microsurgical Epididymal Sperm Aspiration/Testicular Extraction of Sperm Combined with Intracytoplasmic Sperm Injection  

Microsoft Academic Search

Purpose: Vasovasostomy (VVS) represents the standard therapy of choice for the treatment of obstructive azoospermia following vasectomy. However, recently, intracytoplasmic sperm injection (ICSI) has been suggested by some to represent the solution for all cases of malefactor infertility regardless of its etiology based on its success rates. Therefore, we compared VVS to microsurgical epididymal sperm aspiration (MESA)/testicular extraction of sperm

Axel Heidenreich; Petra Altmann; Udo H. Engelmann



Estrogens and falling sperm counts.  


Extravagant claims have been made repeatedly in recent years that human sperm counts are falling and that global exposure to environmental estrogens are responsible. The basis for these two distinct claims is reviewed. The claims of falling human sperm output, reviving an old debate, are prompted by a paper by Carlsen et al. (1992). This meta-analysis, however, is marred by numerous flaws that invalidate its claims. Major defects include severe heterogeneity of component studies, rendering them unsuitable for aggregation, and defective data analysis based on arithmetic mean rather than median, which showed no significant changes over time. This debate is likely to remain unresolved until valid, representative population-based studies of human sperm output can be achieved. None have been reported, or seem feasible in the near future, and so alternative strategies, based on surrogate variables for human male fertility not requiring sperm counts, need to be developed and validated. The plausible hypothesis that prenatal estrogen exposure might influence development of the human testis through effects on Sertoli cell replication and sperm carrying capacity has, however, been conclusively refuted by studies of boys born to women exposed to high doses of oral diethylstilbestrol during pregnancy. Neither fertility nor sperm output were adversely influenced by massive maternal estrogen exposure during pregnancy, although minor urogenital malformations did occur. The still wider claims of deteriorating male reproductive health, notably changes in prevalence or incidence of hypospadias or cryptorchidism, also lack convincing population-based evidence, although cancer registry data indicate a gradual increase in testis cancer in some countries. In summary, the available evidence does not support claims of falling sperm counts or any general deterioration in male reproductive health. Population-based studies of valid surrogate variables for male fertility not requiring semen analysis are needed. If population-based evidence regarding male fertility or sperm output could be generated, it is highly unlikely that prenatal estrogen exposure could be a valid explanation of any deterioration as massive maternal exposure to oral estrogen has negligible effects on male fertility or sperm output. PMID:11800170

Handelsman, D J



The Importance of Functional Tests in Personalized Medicine  

PubMed Central

Cardiovascular disease is the most prevalent disease mainly in the Western society and becoming the leading cause of death worldwide. Standard methods by which healthcare providers screen for cardiovascular disease have only minimally reduced the burden of disease while exponentially increasing costs. As such, more specific and individualized methods for functionally assessing cardiovascular threats are needed to identify properly those at greatest risk, and appropriately treat these patients so as to avoid a fate such as heart attack, stroke, or death. Currently, endothelial function testing—in both the coronary and peripheral circulation—is well established as being associated with the disease process and future cardiovascular events. Improving such testing can lead to a reduction in the risk of future events. Combining this functional assessment of vascular fitness with other, more personalized, testing methods should serve to identify those at the greatest risk of cardiovascular disease earlier and subsequently reduce the affliction of such diseases worldwide.

Widmer, R. Jay; Lerman, Amir



Nuclear microscopy of sperm cell elemental structure.  

National Technical Information Service (NTIS)

Theories suggest there is a link between protamine concentrations in individual sperm and male fertility. Previously, biochemical analyses have used pooled samples containing millions of sperm to determine protamine concentrations. These methods have not ...

G. S. Bench R. Balhorn A. M. Friz S. P. H. Freeman



Supplementation of the dilution medium after thawing with reduced glutathione improves function and the in vitro fertilizing ability of frozen-thawed bull spermatozoa  

Microsoft Academic Search

In this study, we evaluated the effects of glutathione\\u000a (L-gamma-glutamyl-L-cysteinylglycine; GSH) supplementation of the\\u000a thawing extender on bull semen parameters to compensate for the\\u000a decrease in GSH content observed during sperm freezing. To address\\u000a these questions fully, we used a set of functional sperm tests. These\\u000a included tests of sperm motility assayed by computer-assisted semen\\u000a analysis, membrane lipid packing disorder,

Joaquin Gadea; David Gumbao; Sebastian Canovas; Francisco Alberto Garcia-Vazquez; Luis Alberto Grullon; Juan Carlos Gardon



The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis  

PubMed Central

Objective The aim of the present study was to examine the relationship among male age, strict morphology, and sperm chromatin structure and condensation. Methods Sperm samples from a total of 100 men underwent semen analysis, and sperm chromatin structure and condensation were assessed with toluidine blue (TB) and aniline blue (AB) tests. Results Prevalence of strict morphology of less than 4%, and abnormal sperm chromatin structure and condensation did not show any statistically significant differences according to male age (p=0.605, p=0.235, and p=0.080). No significant correlation was demonstrated among age of male partners, strict morphology, and abnormal sperm chromatin structure using TB and AB tests. Howeve