Showing papers on "Sperm published in 2007"
TL;DR: Comparison of protamine gene and amino-acid sequences suggests that the family evolved from specialized histones through protamine-like proteins to the true protamines.
Abstract: The protamines are a diverse family of small arginine-rich proteins that are synthesized in the late-stage spermatids of many animals and plants and bind to DNA, condensing the spermatid genome into a genetically inactive state. Vertebrates have from one to 15 protamine genes per haploid genome, which are clustered together on the same chromosome. Comparison of protamine gene and amino-acid sequences suggests that the family evolved from specialized histones through protamine-like proteins to the true protamines. Structural elements present in all true protamines are a series of arginine-rich DNA-anchoring domains (often containing a mixture of arginine and lysine residues in non-mammalian protamines) and multiple phosphorylation sites. The two protamines found in mammals, P1 and P2, are the most widely studied. P1 packages sperm DNA in all mammals, whereas protamine P2 is present only in the sperm of primates, many rodents and a subset of other placental mammals. P2, but not P1, is synthesized as a precursor that undergoes proteolytic processing after binding to DNA and also binds a zinc atom, the function of which is not known. P1 and P2 are phosphorylated soon after their synthesis, but after binding to DNA most of the phosphate groups are removed and cysteine residues are oxidized, forming disulfide bridges that link the protamines together. Both P1 and P2 have been shown to be required for normal sperm function in primates and many rodents.
612 citations
TL;DR: Direct protein interactions among CatSpers, the sperm specificity of these proteins, and loss of ICatSper in each of the four CatSper−/− mice indicate that CatSper are highly specialized flagellar proteins.
Abstract: Mammalian spermatozoa become motile at ejaculation, but before they can fertilize the egg, they must acquire more thrust to penetrate the cumulus and zona pellucida. The forceful asymmetric motion of hyperactivated spermatozoa requires Ca2+ entry into the sperm tail by an alkalinization-activated voltage-sensitive Ca2+-selective current (ICatSper). Hyperactivation requires CatSper1 and CatSper2 putative ion channel genes, but the function of two other related genes (CatSper3 and CatSper4) is not known. Here we show that targeted disruption of murine CatSper3 or CatSper4 also abrogated ICatSper, sperm cell hyperactivated motility and male fertility but did not affect spermatogenesis or initial motility. Direct protein interactions among CatSpers, the sperm specificity of these proteins, and loss of ICatSper in each of the four CatSper−/− mice indicate that CatSpers are highly specialized flagellar proteins.
479 citations
TL;DR: Diabetes is associated with increased sperm nuclear and mtDNA damage that may impair the reproductive capability of men attending fertility clinics and this work aims to address this concern.
Abstract: BACKGROUND: Diabetes mellitus (DM) is increasing in men of reproductive age. Despite this, the prevalence of diabetes in men attending fertility clinics is largely unknown. Furthermore, studies examining the effects of DM on sperm fertility potential have been limited to conventional semen analysis. METHODS: Conventional semen analysis (semen volume, sperm count, motility and morphology) was performed for 27 diabetic (mean age 34 ± 2 years) and 29 non-diabetic subjects (control group, men undergoing routine infertility investigations, mean age 33 ± 1 years). Nuclear DNA (nDNA) fragmentation was assessed using the alkaline Comet assay and mitochondrial DNA (mtDNA) deletions by Long-PCR. RESULTS: Other than a small, but significant, reduction in semen volume in diabetic men (2.6 versus 3.3 ml; P < 0.05), conventional semen parameters did not differ significantly from control subjects. Diabetic subjects had significantly higher mean nDNA fragmentation (53 versus 32%; P < 0.0001) and median number of mtDNA deletions (4 versus 3; P < 0.05) compared with control subjects. CONCLUSIONS: Diabetes is associated with increased sperm nuclear and mtDNA damage that may impair the reproductive capability of these men.
440 citations
TL;DR: In this article, the role of nutritional and biochemical factors in reproduction and sub-fertility is discussed and a literature search is performed using MEDLINE, Science Direct and bibliographies of published work with both positive and negative results.
Abstract: Current treatments of subfertile couples are usually empiric, as the true cause of subfertility often remains unknown. Therefore, we outline the role of nutritional and biochemical factors in reproduction and subfertility. A literature search was performed using MEDLINE, Science Direct and bibliographies of published work with both positive and negative results. The studies showed that folate has a role in spermatogenesis. In female reproduction, folate is also important for oocyte quality and maturation, implantation, placentation, fetal growth and organ development. Zinc has also been implicated in testicular development, sperm maturation and testosterone synthesis. In females, zinc plays a role in sexual development, ovulation and the menstrual cycle. Both folate and zinc have antioxidant properties that counteract reactive oxygen species (ROS). Thiols, such as glutathione, balance the levels of ROS produced by spermatozoa and influence DNA compaction and the stability and motility of spermatozoa. Oocyte maturation, ovulation, luteolysis and follicle atresia are also affected by ROS. After fertilization, glutathione is important for sperm nucleus decondensation and pronucleus formation. Folate, zinc, ROS and thiols affect apoptosis, which is important for sperm release, regulation of follicle atresia, degeneration of the corpus luteum and endometrial shedding. Therefore, the concentrations of these nutrients may have substantial effects on reproduction. In conclusion, nutritional and biochemical factors affect biological processes in male and female reproduction. Further research should identify pathways that may lead to improvements in care and treatment of subfertility.
430 citations
TL;DR: This study has written and validated a free CASA software primarily for analysis of fish sperm, and improved upon the traditional velocity straight line (VSL) algorithm, eliminating inaccurate characterization of highly curved fish sperm paths.
392 citations
TL;DR: The data suggest that sperm from infertile patients, especially those with oligospermia, may carry a higher risk of transmitting incorrect primary imprints to their offspring, highlighting the need for more research into ART.
Abstract: Recent studies suggest that assisted reproductive technologies (ART), which involve the isolation, handling and culture of gametes and early embryos, are associated with an increased incidence of rare imprinting disorders. Major epigenetic events take place during this time and the process of ART may expose the epigenome to external influences, preventing the proper establishment and maintenance of genomic imprints. However, the risks of ART cannot be simply evaluated because the patients who receive ART may differ both demographically and genetically from the general population at reproductive age. In this study, we examined the DNA methylation status of seven imprinted genes using a combined bisulphite-PCR restriction analysis and sequencing technique on sperm DNA obtained from 97 infertile men. We found an abnormal paternal methylation imprint in 14 patients (14.4%) and abnormal maternal imprint in 20 patients (20.6%). The majority of these doubly defective samples were in men with moderate or severe oligospermia. These abnormalities were specific to imprinted loci as we found that global DNA methylation was normal in these samples. The outcome of ART with sperm shown to have an abnormal DNA methylation pattern was generally poor. However, one sample of sperm with both paternal and maternal methylation errors used in ICSI produced a child of normal appearance without any abnormalities in their imprinted methylation pattern. Our data suggest that sperm from infertile patients, especially those with oligospermia, may carry a higher risk of transmitting incorrect primary imprints to their offspring, highlighting the need for more research into ART.
392 citations
TL;DR: Catalase (CAT) activities were higher in the group that was applied 25mM taurine as an antioxidant, than in all of the other groups in this study, and the addition of antioxidants did not cause significant differences in levels of malondialdehyde (MDA), glutathione (GSH), and glutATHione peroxidase (G SH-Px), after thawing, when compared to groups with no additives.
308 citations
TL;DR: The opening of interchain disulphide bridges in protamines may explain this aspect, as antioxidant vitamins, especially vitamin C, are able to open the cystin net, thus interfering with paternal gene activity during preimplantation development.
Abstract: Reactive oxygen species (ROS) have a negative impact on sperm DNA, leading to the formation of oxidative products such as 8-oxo-7,8-dihydroxyguanosine. This compound causes fragmentation and, thus, has a mutagenic effect. Patient treatment with oral antioxidant vitamins is, therefore, standard practice for male infertility, in an attempt to decrease formation of ROS and improve fertility. In this study, the DNA fragmentation index and the degree of sperm decondensation were measured using the sperm chromatin structure assay before and after 90 days treatment with antioxidant vitamins associated with zinc and selenium. Antioxidant treatment led to a decrease in sperm DNA fragmentation (–19.1%, P < 0.0004), suggesting that at least part of the decay was linked to ROS. However, it also led to an unexpected negative effect: an increase in sperm decondensation with the same order of magnitude (+22.8%, P < 0.0009). The opening of interchain disulphide bridges in protamines may explain this aspect, as antioxidant vitamins, especially vitamin C, are able to open the cystin net, thus interfering with paternal gene activity during preimplantation development. This observation might explain the discrepancy observed concerning the role of these antioxidant treatments in improving male fertility.
290 citations
TL;DR: This is the first report of a broad epigenetic defect associated with abnormal semen parameters and the underlying mechanism for these epigenetic changes may be improper erasure of DNA methylation during epigenetic reprogramming of the male germ line.
Abstract: BackgroundMale-factor infertility is a common condition, and etiology is unknown for a high proportion of cases. Abnormal epigenetic programming of the germline is proposed as a possible mechanism compromising spermatogenesis of some men currently diagnosed with idiopathic infertility. During germ cell maturation and gametogenesis, cells of the germ line undergo extensive epigenetic reprogramming. This process involves widespread erasure of somatic-like patterns of DNA methylation followed by establishment of sex-specific patterns by de novo DNA methylation. Incomplete reprogramming of the male germ line could, in theory, result in both altered sperm DNA methylation and compromised spermatogenesis.Methodology/Principal FindingWe determined concentration, motility and morphology of sperm in semen samples collected by male members of couples attending an infertility clinic. Using MethyLight and Illumina assays we measured methylation of DNA isolated from purified sperm from the same samples. Methylation at numerous sequences was elevated in DNA from poor quality sperm.ConclusionsThis is the first report of a broad epigenetic defect associated with abnormal semen parameters. Our results suggest that the underlying mechanism for these epigenetic changes may be improper erasure of DNA methylation during epigenetic reprogramming of the male germ line.
274 citations
TL;DR: A number of laboratory tests have been developed to determine properties of sperm function but few have been adopted into routine clinical use and international collaborations should be initiated to develop clinically relevant molecular and functional tests.
Abstract: Traditionally, the diagnosis of male infertility has relied upon microscopic assessment and biochemical assays to determine human semen quality. The conventional parameters given most importance have been the concentration, motility, and morphology of sperm in the ejaculate. Most laboratories also include 'sperm suitability' tests where the subpopulations of sperm more likely to finish the marathon journey to the oocyte are separated by density centrifugation. These tests are essential to provide the fundamental information on which clinicians base their initial diagnosis. However, none of these parameters addresses sperm function and their clinical value in predicting fertility is questionable. The advent of intracytoplasmic sperm injection (ICSI) has further reduced the significance and perceived need for sperm quality tests since ICSI requires only one sperm, not subject to classic, or indeed any, tests for the procedure to be successful. Over the past decade, a number of laboratory tests have been developed to determine properties of sperm function. These include quantitative sperm motion parameters, capacitation, basal and induced acrosome reactions, sperm-zona pellucida interactions and nuclear and mitochondrial sperm DNA but few have been adopted into routine clinical use. International collaborations should be initiated to develop clinically relevant molecular and functional tests with agreed protocols and clinical thresholds as a matter of urgency.
274 citations
TL;DR: In a cell with few organelles and minimal cytoplasmic volume, internal Ca2+ concentration, [Ca2+]i, regulates almost all these activities; how does such a simple cell achieve this?
Abstract: Thanks to a worrying decrease in male fertility, understanding how sperm 'work' is a matter both of interest and great importance. Sperm of all animals detect various environmental cues. The 'behavioural' and physiological responses of sperm must be specific, appropriate and correctly timed. Strangely, in a cell with few organelles and minimal cytoplasmic volume, internal Ca(2+) concentration, [Ca(2+)](i), regulates almost all these activities. How does such a simple cell achieve this - and is it as simple as it seems?
TL;DR: It would be sensible to advise men to abstain from smoking to avoid decreased fecundity, and a positive dose-response relationship between smoking and testosterone, LH and the LH/free testosterone ratios is found.
Abstract: BACKGROUND: Previous studies suggest a deleterious effect of cigarette smoking on semen quality, but their results have not been consistent. We studied the association between current smoking and semen characteristics and hormonal levels in a large group of healthy men. METHODS: From 1987 to 2004, seven separate occupational or environmental semen quality studies were co-ordinated by our department. A total of 2562 men participated, each providing semen and blood sample and answering a questionnaire about lifestyle and factors related to health. Appropriate semen and smoking data were available for 2542 men. RESULTS: Adjusting for study, age and other covariates, we observed an inverse dose–response relation between smoking and semen volume, total sperm count and percentage motile sperm. Heavy smokers had a 19% lower sperm concentration than non-smokers. We found a positive dose–response relationship between smoking and testosterone, LH and the LH/free testosterone ratios. CONCLUSION: Current smoking in adult life moderately impairs the semen quality. It is well known that semen quality is associated to fecundity. Therefore, it would be sensible to advise men to abstain from smoking to avoid decreased fecundity.
TL;DR: Insight into the molecular basis of seminal fluid signaling in the female reproductive tract may inform new interventions and management practices to ensure maximal fertility and reduce embryo mortality in pigs and, potentially, other livestock species.
Abstract: Seminal fluid contains potent signaling agents that influence female reproductive physiology to improve the chances of conception and pregnancy success. Cytokines and prostaglandins synthesized in the male accessory glands are transferred to the female at insemination, where they bind to receptors on target cells in the cervix and uterus, activating changes in gene expression that lead to modifications in structure and function of the female tissues. The consequences are increased sperm survival and fertilization rates, conditioning of the female immune response to tolerate semen and the conceptus, and molecular and cellular changes in the endometrium that facilitate embryo development and implantation. Male-female tract signaling occurs in rodents, livestock animals, and all other mammals examined thus far, including humans. In mice, the key signaling moieties in seminal plasma are identified as members of the transforming growth factor-beta family. Recent studies indicate a similar signaling function for boar factors in the pig, whereby the sperm and plasma fractions of seminal fluid appear to synergize in activating an inflammatory response and downstream changes in the female tract after insemination. Seminal plasma elicits endometrial changes, with induction of proinflammatory cytokines and cyclooxygenase-2, causing recruitment of macrophages and dendritic cells. Sperm contribute by interacting with seminal plasma factors to modulate neutrophil influx into the luminal cavity. The cascade of changes in local leukocyte populations and cytokine synthesis persists throughout the preimplantation period. Exposure to seminal fluid alters the dynamics of preimplantation embryo development, with an increase in the number of fertilized oocytes attaining the viable blastocyst stage. There is also evidence that seminal factors influence the timing of ovulation, corpus luteum development, and progesterone synthesis. Insight into the molecular basis of seminal fluid signaling in the female reproductive tract may inform new interventions and management practices to ensure maximal fertility and reduce embryo mortality in pigs and, potentially, other livestock species.
TL;DR: The latest insights into the mechanisms underlying the process of making meiotic diploids and DH individuals are discussed, and the use of doubled haploids and clones in quantitative trait locus mapping and selective breeding is explored.
TL;DR: Sperm DNA fragmentation measured 2 to 5 months before the assisted reproduction procedure was a prognostic indicator of the fertilization, pregnancy, and miscarriage rates and the pregnancy outcome.
TL;DR: The DNA of STI pathogens was detected in semen from a high percentage of asymptomatic male infertility patients, and was associated with poor semen quality, and efforts to diagnose and treat subclinical genital-tract infections should be intensified.
TL;DR: In vivo changes in the intra-luminal milieu of the oviduct of pigs and cows are reviewed which relate to the modulation of sperm capacitation around spontaneous ovulation, thus maximizing the chances of normal fertilization.
TL;DR: It is concluded that sampling a concentration field of chemoattractant along circular and helical swimming paths is a robust strategy for chemotaxis that works reliably for a vast range of parameters.
Abstract: We develop a theoretical description of sperm chemotaxis. Sperm cells of many species are guided to the egg by chemoattractants, a process called chemotaxis. Motor proteins in the flagellum of the sperm generate a regular beat of the flagellum, which propels the sperm in a fluid. In the absence of a chemoattractant, sperm swim in circles in two dimensions and along helical paths in three dimensions. Chemoattractants stimulate a signaling system in the flagellum, which regulates the motors to control sperm swimming. Our theoretical description of sperm chemotaxis in two and three dimensions is based on a generic signaling module that regulates the curvature and torsion of the swimming path. In the presence of a chemoattractant, swimming paths are drifting circles in two dimensions and deformed helices in three dimensions. The swimming paths can be described by a dynamical system that exhibits different dynamic regimes, which correspond to different chemotactic behaviours. We conclude that sampling a concentration field of chemoattractant along circular and helical swimming paths is a robust strategy for chemotaxis that works reliably for a vast range of parameters.
TL;DR: Evaluation of seminal ROS levels and extent of sperm DNA damage especially in an infertile male may help develop new therapeutic strategies and improve success of assisted reproductive techniques (ART).
Abstract: Oxidative stress (OS) in the reproductive tract is now a real entity and concern due to the potential harmful effects of high levels of reactive oxygen species (ROS) on sperm number, motility, quality, and function including damage to sperm nuclear DNA. Evaluation of OS related damage to non-functional sperm is highly relevant as intracytoplasmic sperm injection (ICSI) technique, an effective therapy for severe male factor infertility, bypasses the majority of reproductive tract deficiencies. Despite the controversial findings in the existing literature, there is now enough evidence to show that sperm DNA damage is detrimental to reproductive outcomes. In addition, spermatozoa of infertile men are suggested to carry more DNA damage than do the spermatozoa from fertile men. Besides impairment of fertility such damage is likely to increase the transmission of genetic diseases during the assisted reproductive procedures. Standardization of protocols to assess reactive oxygen species and DNA damage is very important in introducing these tests in such clinical practice. Thus evaluation of seminal ROS levels and extent of sperm DNA damage especially in an infertile male may help develop new therapeutic strategies and improve success of assisted reproductive techniques (ART).
TL;DR: The observed significant synergistic effect of BPb and blood cadmium on increasing serum testosterone, and additive effect of a decrease in serum selenium on Increasing serumosterone, may have implications on the initiation and development of prostate cancer because testosterone augments the progress of prostatecancer in its early stages.
TL;DR: It is demonstrated that the decrease in DHT induced by 5ARIs is associated with mild decreases in semen parameters that appear reversible after discontinuation.
Abstract: Context: Dutasteride and finasteride are 5-reductase inhibitors (5ARIs) that dramatically reduce serum levels of dihydrotestosterone (DHT). Objective: Because androgens are essential for fertility, we sought to determine the impact of 5ARI administration on serum testosterone (T), DHT, and spermatogenesis. Design, Setting, Subjects, and Intervention: We conducted a randomized, double-blinded, placebo-controlled trial in 99 healthy men randomly assigned to receive dutasteride (D; 0.5 mg) (n 33), finasteride (F; 5 mg) (n 34), or placebo (n 32) once daily for 1 yr. Main Outcome Measures: Blood and semen samples were collected at baseline and 26 and 52 wk of treatment and 24 wk after treatment and were assessed for T, DHT, and semen parameters. Results: D and F significantly (P 0.001) suppressed serum DHT, compared with placebo (D, 94%; F, 73%) and transiently increased serum T. In both treatment groups, total sperm count, compared with baseline,wassignificantlydecreasedat26wk(D,28.6%;F,34.3%) but not at 52 wk (D, 24.9%; F, 16.2%) or the 24-wk follow-up (D, 23.3%; F, 6.2%). At 52 wk, semen volume was decreased (D, 29.7%; F, 14.5%, significantly for D) as was sperm concentration (D, 13.2%; F, 7.4%, neither significant). There was a significant reduction of 6 to 12% in sperm motility during treatment with both D and F and at follow-up. Neither treatment had any effect on sperm morphology. Conclusions: This study demonstrates that the decrease in DHT induced by 5ARIs is associated with mild decreases in semen parameters that appear reversible after discontinuation. (J Clin Endocrinol Metab 92: 1659–1665, 2007)
TL;DR: Investigation of the associations between male age and sperm DNA damage and the influence of several lifestyle factors in a healthy non-clinical group of 80 non-smokers with no known fertility problems indicates that older men have increased sperm DNADamage associated with alkali-labile sites or single-strand DNA breaks and independent of age, men with substantial daily caffeine consumption have increased DNA damage associated with double-stranded DNA breaks.
Abstract: The trend for men to have children at older ages raises concerns that advancing age may increase the production of genetically defective sperm, increasing the risks of transmitting germ-line mutations. We investigated the associations between male age and sperm DNA damage and the influence of several lifestyle factors in a healthy non-clinical group of 80 non-smokers (age: 22-80) with no known fertility problems using the sperm Comet analyses. The average percent of DNA that migrated out of the sperm nucleus under alkaline electrophoresis increased with age (0.18% per year, p=0.006); but there was no age association for damage measured under neutral conditions (p=0.7). Men who consumed >3 cups coffee per day had {approx}20% higher % tail DNA under neutral but not alkaline conditions compared to men who consumed no caffeine (p=0.005). Our findings indicate that (a) older men have increased sperm DNA damage associated with alkali-labile sites or single-strand DNA breaks, and (b) independent of age, men with substantial daily caffeine consumption have increased sperm DNA damage associated with double-strand DNA breaks. DNA damage in sperm can be converted to chromosomal aberrations and gene mutations after fertilization increasing the risks for developmental defects and genetic diseases among offspring.
TL;DR: In HA-selected spermatozoa the frequency of chromosomal disomy and diploidy is reduced 4- to 6-fold compared with semen sperm fractions, similar to the increase in numerical chromosomal aberrations in ICSI children.
Abstract: The testis-expressed chaperone protein, HspA2 (previously creatine kinase M isoform) was established as a measure of human sperm cellular maturity, function and fertility. The presence of HspA2 in the synaptonemal complex is likely to link low HspA2 expression and increased frequency of chromosomal aneuploidies in arrested-maturity spermatozoa. A relationship also exists between HspA2 expression in elongating spermatids and the associated spermatogenetic events, including plasma membrane remodelling and the formation of zona pellucida and hyaluronic acid (HA) binding sites. The HA receptor of mature spermatozoa, when coupled with HA-coated slides and/or Petri dishes, allows visual observation of sperm–HA binding, providing a basis for sperm maturity testing, a major improvement in semen evaluation, and selection of mature spermatozoa for intracytoplasmic sperm injection (ICSI). Thus, in HA-selected spermatozoa the frequency of chromosomal disomy and diploidy is reduced 4- to 6-fold compared with semen sperm fractions. This reduction is similar to the increase in numerical chromosomal aberrations in ICSI children. Combined studies of sperm shape and chromosome probes demonstrated that sperm morphology does not aid selection of haploid spermatozoa. The HA-mediated sperm selection is a novel and efficient technique that may alleviate potential problems related to ICSI fertilization with visually selected spermatozoa.
TL;DR: It is shown that both ZPBP proteins play an earlier structural role during spermiogenesis, and molecular phylogenetic analysis of ZPBPs from amphibians, birds, and mammals suggests that these paralogous genes coevolved to play cooperative roles during sPermiogenesis.
Abstract: Zona pellucida binding protein 1 (ZPBP1), a spermatid and spermatozoon protein that localizes to the acrosome, was originally identified in pigs and named for its binding to the oocyte zona pellucida. In an in silico search for germ cell-specific genes, Zpbp1 and its novel paralog, Zpbp2, were discovered and confirmed to be expressed only in the testes in both mice and humans. To study the in vivo functions of both ZPBP proteins, we disrupted Zpbp1 and Zpbp2 in mice. Males lacking ZPBP1 were sterile, with abnormal round-headed sperm morphology and no forward sperm motility. Ultrastructural studies demonstrated that absence of ZPBP1 prevents proper acrosome compaction, resulting in acrosome fragmentation and disruption of the Sertoli-spermatid junctions. Males null for ZPBP2 were subfertile, demonstrated aberrant acrosomal membrane invaginations, and produced dysmorphic sperm with reduced ability to penetrate zona pellucida. Molecular phylogenetic analysis of ZPBPs from amphibians, birds, and mammals suggests that these paralogous genes coevolved to play cooperative roles during spermiogenesis. Whereas ZPBP1 was discovered for an in vitro role in sperm-egg interactions, we have shown that both ZPBP proteins play an earlier structural role during spermiogenesis.
TL;DR: It is shown that alkalinization also has a dramatic effect on membrane potential, producing a rapid hyperpolarization, which is primarily mediated by a weakly outwardly rectifying K+ current (IKSper) originating from the principal piece of the sperm flagellum.
Abstract: Mature mammalian spermatozoa are quiescent in the male reproductive tract. Upon ejaculation and during their transit through the female reproductive tract, they undergo changes that enable them to fertilize the egg. During this process of capacitation, they acquire progressive motility, develop hyperactivated motility, and are readied for the acrosome reaction. All of these processes are regulated by intracellular pH. In the female reproductive tract, the spermatozoan cytoplasm alkalinizes, which in turn activates a Ca2+-selective current (ICatSper) required for hyperactivated motility. Here, we show that alkalinization also has a dramatic effect on membrane potential, producing a rapid hyperpolarization. This hyperpolarization is primarily mediated by a weakly outwardly rectifying K+ current (IKSper) originating from the principal piece of the sperm flagellum. Alkalinization activates the pHi-sensitive IKSper, setting the membrane potential to negative potentials where Ca2+ entry via ICatSper is maximized. IKSper is one of two dominant ion currents of capacitated sperm cells.
TL;DR: The data suggest that microvilli may participate in sperm-oocyte fusion, and it is found that oocyte CD9 co-immunoprecipitates with two Ig superfamily cis partners, EWI-2 and E WI-F, which could have a role in linking CD9 to the oocyte microvillar actin core.
TL;DR: CFTR in sperm may be involved in the transport of HCO3− important for sperm capacitation and that CFTR mutations with impaired CFTR function may lead to reduced sperm fertilizing capacity and male infertility other than CBAVD.
Abstract: Cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel, mutations of which cause cystic fibrosis, a disease characterized by defective Cl− and HCO3− transport. Although >95% of all CF male patients are infertile because of congenital bilateral absence of the vas deferens (CBAVD), the question whether CFTR mutations are involved in other forms of male infertility is under intense debates. Here we report that CFTR is detected in both human and mouse sperm. CFTR inhibitor or antibody significantly reduces the sperm capacitation, and the associated HCO3−-dependent events, including increases in intracellular pH, cAMP production and membrane hyperpolarization. The fertilizing capacity of the sperm obtained from heterozygous CFTR mutant mice is also significantly lower compared with that of the wild-type. These results suggest that CFTR in sperm may be involved in the transport of HCO3− important for sperm capacitation and that CFTR mutations with impaired CFTR function may lead to reduced sperm fertilizing capacity and male infertility other than CBAVD.
TL;DR: The origin and function of sperm borne RNA transferred into the oocyte is discussed along with their putative role in early embryogenesis, which still needs to be experimentally proven.
TL;DR: The results of this study provide a new approach to the cryopreservation of sperm from rams and related breeds, and thereby contribute to the improvement of these breeds for the world sheep industry.
Abstract: Uysal O., M. N. Bucak: Effects of Oxidized Glutathione, Bovine Serum Albumin, Cysteine and Lycopene on the Quality of Frozen-Thawed Ram Semen. Acta Vet. Brno 2007, 76: 383-390. Free radicals are known to be involved in lipid peroxidation as well as DNA and sperm membrane damages that may lead to decreased sperm motility or cell death. The balance between free radical production and their detoxifi cation may be an important factor in sperm survival and function before, during and after cryopreservation. The aim of this study was to determine the effects of the addition of the antioxidants of oxidized glutathione (GSSG), bovine serum albumin (BSA), cysteine and lycopene to freezing media on the post-thawing sperm characteristics, including motility, morphology, acrosome integrity, viability and membrane integrity. A total number of 42 ejaculates were collected using the artifi cial vagina from 4 Akkaraman rams and 10 replicates of the ejaculates were diluted with a Tris-based extender containing additives and no additives as control. GSSG (5 mM), BSA (20 mg/ml), cysteine (10 mM) and lycopene (800 μg) showed more positive effects than other concentrations of the supplements and controls in protecting sperm characteristics after the freezing-thawing process (P < 0.001). Many aspects of sperm protection, e.g. sperm motility, viability and membrane stabilisation of the sperm cells during relative cryopreservation, are the key factors in determining the preservation of sperm function. The results of this study provide a new approach to the cryopreservation of sperm from rams and related breeds, and thereby contribute to the improvement of these breeds for the world sheep industry. Antioxidants, ram semen, freezing, extender
TL;DR: The inventory of proteins present in the human sperm proteome includes a number of notable discoveries including the first description of a nicotinamide adenine dinucleotide phosphate oxidase, dual‐oxidase 2, finally laying to rest any doubts about the presence of such enzymes in spermatozoa.
Abstract: A comprehensive analysis of the proteins found in human spermatozoa is essential for understanding the events leading up to, and including, fertilization and development. Proteomics offers a platform for investigating this process, provided that the dynamic range is relatively low. In this report, spermatozoa from a number of human sperm ejaculates were isolated in a pure state using discontinuous Percoll gradient centrifugation. Triton X-100 soluble and insoluble proteins were recovered and separated by SDS-PAGE. The separation lanes were dissected into 96 fractions and analyzed individually by LC-MS(n) . A comprehensive protocol, involving LC-MS/MS analysis eventually down to the ninth most intense peak found in the MS-survey scan, was performed. Analysis of purified human sperm populations resulted in the identification of 1056 gene products, of which approximately 8% have not previously been characterized. The data were supported by the large number of proteins represented by expressed sequence tags in the testis. Bioinformatic analysis demonstrated that 437 of the gene products were involved in various metabolic pathways including glycolysis and oxidative phosphorylation. The inventory of proteins present in the human sperm proteome includes a number of notable discoveries including the first description of a nicotinamide adenine dinucleotide phosphate oxidase, dual-oxidase 2, finally laying to rest any doubts about the presence of such enzymes in spermatozoa. Furthermore, a number of different classes of receptor have also been detected in these cells and are potential regulators of sperm function. This list includes at least six seven-pass transmembrane receptors, six tyrosine kinase receptors, a tyrosine phosphatase receptor, glutamate-gated ion channel receptors, transient receptor potential cation channels, and a non-genomic progesterone receptor. This is the first published list of identified proteins in human spermatozoa using LC-MS/MS analysis.