scispace - formally typeset
Search or ask a question

Showing papers on "Sperm published in 1996"


Journal ArticleDOI
TL;DR: It is suggested that estrogen action is required for fertility in male mice and that the mutation of the ER in ERKO males leads to reduced mating frequency, low sperm numbers, and defective sperm function.
Abstract: The reproductive system of male mice homozygous for a mutation in the estrogen receptor (ER) gene (ER knock-out; ERKO) appears normal at the anatomical level. However, these males are infertile, indicating an essential role for ER-mediated processes in the regulation of male reproduction. Adult ERKO male mice have significantly fewer epididymal sperm than heterozygous or wild-type males. Although spermatogenesis is occurring in some seminiferous tubules of 3- to 5-month-old ERKO males, other tubules either have a dilated lumen and a disorganized seminiferous epithelium with few spermatogenic cells or lack a lumen and contain mainly Sertoli cells. There are no obvious differences in seminiferous tubules at 10 days of age between wild-type and ERKO mice, but the lumen in ERKO males is dilated in all seminiferous tubules by 20 days. However, spermatogenesis progresses and similar numbers of sperm are present in the cauda epididymis of ERKO and wild-type males until 10 weeks of age. Disruption of spermatogenesis and degeneration of the seminiferous tubules become apparent after 10 weeks in the caudal pole of the testis and progresses in a wave to the cranial pole by 6 months. However, the seminal vesicles, coagulating glands, prostate, and epididymis do not appear to be altered morphologically in ERKO mice. Serum testosterone levels are somewhat elevated, but LH and FSH levels are not significantly different from those in wild-type males. Sperm from 8- to 16-week-old mice have reduced motility and are ineffective at fertilizing eggs in vitro. In addition, ERKO males housed overnight with hormone-primed wild-type females produce significantly fewer copulatory plugs than do heterozygous or wild-type males. These results suggest that estrogen action is required for fertility in male mice and that the mutation of the ER in ERKO males leads to reduced mating frequency, low sperm numbers, and defective sperm function.

835 citations


Journal ArticleDOI
TL;DR: Treatment of asthenospermic patients with oral Vitamin E significantly decreased the MDA concentration in spermatozoa and improved sperm motility, and nine of the 52 treated patients successfully impregnated their spouses.
Abstract: Asthenospermia is the main factor of male infertility among patients consulting the Asir Infertility Center in Abha, Saudi Arabia. Lipid peroxidation occurring in both the seminal plasma and spermatozoa was estimated by malondialdehyde (MDA) concentration. Spermatozoal MDA concentration was higher in men with decreased sperm motility. The MDA concentration in the seminal plasma exhibited no relationship with sperm concentration, sperm motility, the number of immotile spermatozoa, or even the absence of spermatozoa. The MDA concentration in sperm pellet suspensions of asthenospermic and oligoasthenospermic patients was almost twice that of the normospermic males. The MDA concentration in the sperm pellet suspension from normospermic or oligospermic patients was about 10% that in the seminal plasma. However, the MDA concentration in the sperm pellet suspension of asthenospermic or oligoasthenospermic patients was about 15% that in the seminal plasma. Treatment of asthenospermic patients with oral Vitamin E significantly decreased the MDA concentration in spermatozoa and improved sperm motility. Eleven out of the 52 treated patients (21%) impregnated their spouses; nine of the spouses successfully ended with normal term deliveries, whereas the other two aborted in the first trimester. No pregnancies were reported in the spouses of the placebo-treated patients.

577 citations


Journal ArticleDOI
TL;DR: The possibility that paternal smoking causes mutations in sperm that lead to cancer, birth defects, and genetic diseases in offspring is discussed.
Abstract: Our previous studies have shown that men with low ascorbate intake have markedly increased oxo8dG in the DNA of their sperm. Because cigarette smoke is high in oxidants and depletes plasma and tissue antioxidants, oxidative DNA damage in sperm and tocopherol and ascorbate levels in seminal plasma were determined in smokers and non-smokers. The level in sperm DNA of oxo8dG, an oxidative lesion of guanine, was 50% higher in smokers compared to nonsmokers (p = 0.005). The concentration of alpha-tocopherol in seminal plasma was decreased in smokers by 32% (p = 0.03). Smoking and low antioxidant levels increase oxidative damage to sperm DNA. We discuss the possibility that paternal smoking causes mutations in sperm that lead to cancer, birth defects, and genetic diseases in offspring.

470 citations


Journal ArticleDOI
24 Feb 1996-BMJ
TL;DR: Data confirm previously published data from other countries that semen quality is changing, declining by about 2.1% per year Research is urgently required to examine the function as well as the number of sperm and to assess whether these changes are affecting human health and male fertility.
Abstract: Objective: To determine whether the quality of semen has changed in a group of over 500 Scottish men born between 1951 and 1973. Design: Retrospective review of data on semen quality collected in a single laboratory over 11 years and according to World Health Organisation guidelines. Setting: Programme of gamete biology research funded by Medical Research Council. Subjects: 577 volunteer semen donors. Of these, 171 were born before 1959, 120 were born in 1960-4, 171 in 1965-9, and 115 in 1970-4. Main outcome measures: Conventional criteria of semen quality including semen volume (ml), sperm concentration (106/ml), overall motility (% motile), total number of sperm in the ejaculate (106), and total number of motile sperm in the ejaculate (106). Results: When the four birth cohort groups were compared a later year of birth was associated with a lower sperm concentration, a lower total number of sperm in the ejaculate, and a lower number of motile sperm in the ejaculate. The median sperm concentration fell from 98x106/ml among donors born before 1959 to 78x106/ml among donors born after 1970 (P=0.002). The total number of sperm in the ejaculate fell from 301x106 to 214x106 (P=0.0005), and the total number of motile sperm in the ejaculate fell from 169.7x106 to 129.0x106 (P=0.0065). Conclusion: This study provides direct evidence that semen quality is deteriorating, with a later year of birth being significantly associated with a reduced number of sperm in adult life. Key messages Key messages When men born in the 1970s were compared with men born in the 1950s, the total number of motile sperm in the ejaculate was reduced by almost 25% These data confirm previously published data from other countries that semen quality is changing, declining by about 2.1% per year Research is urgently required to examine the function as well as the number of sperm and to assess whether these changes are affecting human health and male fertility

454 citations


Journal ArticleDOI
TL;DR: Results suggest that divergent pathways might regulate tyrosine phosphorylation of p105 and p81 and sperm capacitation after cAMP-dependent phosphorylations of an intermediate protein, and suggest that protein phosphatase PP1 is involved in the two latter phenomena while PP2A mediates sperm capacitations.
Abstract: The involvement of cAMP in the process of sperm capacitation has been the subject of several studies. In addition, the importance of protein-tyrosine phosphorylation in this process has been investigated, although only a few studies have been reported in the human. Since agents regulating the intracellular concentrations of cAMP affect sperm capacitation rates, the role of cAMP on the expression of phosphotyrosine-containing proteins was investigated during human sperm capacitation. Fetal cord serum ultrafiltrate, a known capacitation inducer in human spermatozoa, caused an increase in the phosphotyrosine content of 105- and 81-kDa proteins (p105 and p81), the two major phosphotyrosine-containing proteins of human spermatozoa. Similar effects were observed when spermatozoa were incubated with phosphodiesterase inhibitors or cell-permeant cAMP analogs, suggesting that cAMP is involved in these two processes. Forskolin, an adenylyl cyclase activator, also caused an increase in both sperm capacitation rates and tyrosine phosphorylation of p105 and p81, while 12-O-tetradecanoyl phorbol 13-acetate stimulated both capacitation and tyrosine phosphorylation of p105 and p81 only when spermatozoa were incubated in the presence of bicarbonate, in agreement with its reported effects on cAMP production and hamster sperm capacitation. The inhibition of these phenomena by cAMP-dependent protein kinase inhibitors, and the stimulation by protein phosphatase inhibitors, suggest that Ser/Thr protein phosphorylation plays an important role in the regulation of both sperm capacitation and protein-tyrosine phosphorylation pathways. However, observations that both calyculin A and okadaic acid stimulated sperm capacitation, whereas only calyculin A increased p105 and p81 phosphotyrosine content and sperm velocity, suggest that protein phosphatase PP1 is involved in the two latter phenomena while PP2A mediates sperm capacitation. These results suggest that divergent pathways might regulate tyrosine phosphorylation of p105 and p81 and sperm capacitation after cAMP-dependent phosphorylation of an intermediate protein.

397 citations


Journal ArticleDOI
TL;DR: The data show no decline in sperm counts over a 25-year period in 1,283 men who banked sperm before vasectomy at three distinct geographical sites in the United States.

379 citations


Journal ArticleDOI
25 Jan 1996-Nature
TL;DR: A soluble sperm protein is identified that exhibits Ca2+ oscillation-inducing ('oscillogen') activity in eggs and may represent the physiological trigger for development in mammals.
Abstract: At fertilization in mammals, the sperm induces a characteristic series of Ca2+ oscillations in the egg which serve as the essential trigger for egg activation and early development of the embryo. It is not known how the sperm initiates this fundamental process, however, nor has any pathway linking sperm-egg membrane-receptor binding with intracellular Ca2+ release been demonstrated. Microinjection of sperm extracts into mammalian eggs elicits Ca2+ oscillations identical to those occurring at fertilization, which suggests that sperm may introduce a Ca2+ oscillation-inducing factor into the egg on gamete membrane fusion. Here we identify a soluble sperm protein that exhibits Ca2+ oscillation-inducing ('oscillogen') activity in eggs. Sperm oscillogen exists as an oligomer with a subunit of M(r) 33K and a specific intracellular localization at the equatorial segment of the sperm head. Cloning of the 33K oscillogen complementary DNA indicates similarity with a hexose phosphate isomerase found in prokaryotes. This sperm-derived oscillogen, termed oscillin, may represent the physiological trigger for development in mammals.

373 citations


Journal ArticleDOI
TL;DR: One of the factors involved in the etiology of defective sperm function is the incomplete extrusion of germ cell cytoplasm during spermiogenesis as a consequence of which the spermatozoa experience a loss of function associated with the induction of oxidative stress.
Abstract: A method has been developed for quantifying the residual cytoplasm present in the midpiece of human spermatozoa, based upon the imaging of NADH oxidoreductase activity. This procedure used NADH and nitroblue tetrazolium as electron donor and acceptor, respectively, and resulted in the discrete staining of the entire midpiece area, including the residual cytoplasm. Image analysis techniques were then used to generate binary images of the midpiece, from which objective measurements of this cellular domain could be undertaken. Such data were found to be highly correlated with biochemical markers of the cytoplasmic space, such as creatine kinase (CK) and glucose-6-phosphate dehydrogenase (G-6-PDH), in sperm populations depleted of detectable leukocyte contamination. Morphometric analysis of the sperm midpiece was also found to reflect semen quality in that it predicted the proportion of the ejaculate that would be recovered from the high-density region of Percoll gradients and was negatively correlated with the movement and morphology of the spermatozoa in semen. Variation in the retention of excess residual cytoplasm was also associated with differences in the functional competence of washed sperm preparations, both within and between ejaculates. Thus, within-ejaculate comparisons of high- and low-density sperm subpopulations revealed a relative disruption of sperm function in the low-density fraction. This disruption was associated with the presence of excess residual cytoplasm in the midpiece, high concentrations of cytoplasmic enzymes, and the enhanced-generation reactive oxygen species (ROS). Functional differences between individual high-density Percoll preparations were also negatively correlated with the area of the midpiece and the corresponding capacity of the spermatozoa to generate ROS. These findings suggest that one of the factors involved in the etiology of defective sperm function is the incomplete extrusion of germ cell cytoplasm during spermiogenesis as a consequence of which the spermatozoa experience a loss of function associated with the induction of oxidative stress.

353 citations


Journal ArticleDOI
TL;DR: It is postulate that poor chromatin packaging and/or damaged DNA may contribute to failure of sperm decondensation after ICSI and result in failure of fertilization.
Abstract: In this study we investigated whether morphology and chromatin anomalies in human spermatozoa can influence fertilization after intracytoplasmic sperm injection (ICSI). We examined unfertilized oocytes, using the fluorochrome Hoechst 33342, to determine whether a relationship exists between failure of fertilization and sperm chromatin quality. Sperm chromatin packaging quality was assessed using the chromomycin A3 (CMA3) fluorochrome, and the presence of DNA damage in spermatozoa, using in-situ nick translation, Normal males present sperm parameters with a normal morphology of > 20%, CMA3 fluorescence of < 30% and exhibit endogenous nicks in < 10% of their spermatozoa. When patients were separated according to these values no difference was observed in their fertilization rates after ICSI. When the unfertilized ICSI oocytes were examined, we found that patients with CMA3 fluorescence of <30% and nicks in < 10% of their spermatozoa had only 17.5 and 21.6% respectively of their unfertilized oocytes containing spermatozoa that remained condensed. In contrast, patients with higher CMA3 and nick values had a significantly higher number, 41.2 and 48.9%, of their unfertilized oocytes containing condensed spermatozoa. sperm morphology did not show any such pattern. The percentage of spermatozoa which had initiated decondensation in unfertilized oocytes was not influenced by morphology, CMA3 fluorescence or nicks. In light of these results we postulate that poor chromatin packaging and/or damaged DNA may contribute to failure of sperm decondensation after ICSI and result in failure of fertilization.

321 citations


Journal ArticleDOI
TL;DR: It is found that bicarbonate causes a rapid increase in the ability of live boar spermatozoa to bind merocyanine, apparently by perturbing enzymic control processes.
Abstract: Bicarbonate/CO2 is believed to be the key in vitro effector of sperm capacitation, a process which induces major changes in the sperm plasma membrane in preparation for fertilization. In a flow cytometric study, we examined the effect of bicarbonate on boar spermatozoa using merocyanine, an impermeant lipophilic probe which binds to plasma membranes with increasing affinity as their lipid components become more disordered. We found that bicarbonate causes a rapid increase in the ability of live boar spermatozoa to bind merocyanine. First detected about 100 sec after exposure to bicarbonate and largely complete by 300 sec, this increase appears to result from individual cells within the sperm population switching from a low merocyanine-binding state to a high binding state. The majority of live spermatozoa are capable of responding in this way, and do so in proportion to bicarbonate concentration, half-maximal response being induced by about 3 mM bicarbonate; however, overall population response varies greatly between ejaculates. Increased merocyanine stainability is observed over the whole surface area of the cell, and is reversible both with respect to temperature (it is only manifested above 30 degrees C) and with respect to presence of bicarbonate. A similar effect can be induced by phosphodiesterase inhibitors such as isobutylmethylxanthine, and enhanced by a permeant cyclic nucleotide analogue. We conclude that bicarbonate causes a major alteration in sperm plasma membrane lipid architecture, apparently by perturbing enzymic control processes. This novel action of bicarbonate may represent an initial permissive event in the capacitation sequence.

306 citations



Journal ArticleDOI
TL;DR: Three sperm competition game models devised mainly for externally fertilizing group-spawning species such as many fish predict that males should decrease their sperm expenditure as the estimated number of competitors present at a given spawning increases above two.
Abstract: A distinction is made between sperm competition risk (where there is typically a low probability of competition between two ejaculates) and sperm competition intensity (where typically two or more ejaculates compete). The relation between sperm competition intensity and sperm expenditure can be radically different across species from that within a species. Across species, the average ejaculate expenditure will increase with the average intensity of sperm competition. But within a species, the reverse trend is generally predicted for greater than two males competing for the same set of eggs. These effects are demonstrated with three sperm competition game models. They are devised mainly for externally fertilizing group-spawning species such as many fish, in which males group around a female and ejaculate when the female sheds her eggs. Fertilization is assumed to be instantaneous and each male gains a proportion of the eggs equal to his sperm number divided by the total sperm. In the first model, males cannot assess the number of competitors, and their ejaculate effort is shaped by the average number of males for the species or locally isolated deme. The proportion of reproductive effort expended on the ejaculate is predicted to increase as (N-1)/N, where N = the mean number of competing males present at a spawning. Thus if N is large, ejaculate expenditure dominates reproductive effort. In the second model, males can estimate whether there are more or less than average numbers of competitors present at a spawning, and in the third model, males can assess the number of competitors exactly. As in the first model, these models confirm that the mean ejaculate effort should increase with the mean number of competitors for the species. However, they predict that males should decrease their sperm expenditure as the estimated number of competitors present at a given spawning increases above two. These conclusions do not apply to sperm competition risk: there is thus no conflict with earlier models based on risk.

Journal ArticleDOI
TL;DR: Spermatozoa do not need to be structurally intact for intracytoplasmic injection, the plasma and acrosomal membranes and all tail components are not essential for normal embryo development, at least in the mouse, and the cryopreservation conditions required for maintenance of the genetic integrity of sperm nuclei are less stringent than those necessary for keeping plasma membrane-intact spermatozoA alive.
Abstract: MATERIALS AND METHODS To determine whether spermatozoa must be structurally intact before microsurgical injection into oocytes for normal fertilization, intact spermatozoa, as well as sperm heads separated from tails by sonication, were individually injected into oocytes. When whole spermatozoa were injected immediately after their immobilization, the majority of the oocytes were fertilized and developed normally. Sonication in the presence or absence of Triton X-100 decapitated more than 95% of spermatozoa. Although all decapitated spermatozoa were diagnosed as "dead" by live/dead sperm staining, separated sperm heads (nuclei) could participate in normal embryo development when injected into the oocytes. The ability of isolated sperm heads (nuclei) to participate in normal embryo development was maintained under cryopreservation conditions that were not suitable for the survival of plasma membrane-intact spermatozoa. These results indicate that 1) spermatozoa do not need to be structurally intact for intracytoplasmic injection, 2) the plasma and acrosomal membranes and all tail components are not essential for normal embryo development, at least in the mouse, and 3) the cryopreservation conditions required for maintenance of the genetic integrity of sperm nuclei are less stringent than those necessary for keeping plasma membrane-intact spermatozoa alive.

Journal ArticleDOI
TL;DR: The above data show that testicular biopsies may have an important therapeutic role in the management of infertility in azoospermic patients.
Abstract: A comprehensive study is presented of a series of 124 infertile men undergoing testicular sperm retrieval for intracytoplasmic sperm injection (ICSI). In this study we correlated the histological changes observed in the testicular tissue with the results of the wet preparation and the outcome after ICSI using testicular spermatozoa. In all patients with normal spermatogenesis and hypospermatogenesis spermatozoa were recovered from the wet preparation. The sperm recovery rate as 84% in patients with incomplete germ-cell-aplasia and maturation arrest, while in patients with complete germ-cell aplasia or maturation arrest this figure was 76%. In these patients more specimens were sampled and fewer spermatozoa were recovered. Since no spermatozoa were recovered in only 10 patients, ICSI with testicular sperm was performed in the remaining 114 couples (91.9%). The normal fertilization rate was 57. 8%. The fertilization rate was significantly lower in couples among whom the husband showed germ-cell aplasia and maturation arrest. Overall, 55.2% of normally fertilized oocytes developed into embryos showing <=50% of anucleate fragments. There were no major differences between the different histological categories in terms of embryonic development in vitro. The overall pregnancy rates per testicular sperm extraction (TESE) procedure, per ICSI procedure and per transfer were respectively 36.3, 39.5 and 43.7%. The overall implantation rate per embryo (sacs/embryos replaced) was 20.3%. A lower implantation rate was observed in couples among whom the husband had maturation arrest (not statistically significant). The above data show that testicular biopsies may have an important therapeutic role in the management of infertility in azoospermic patients.

Journal ArticleDOI
TL;DR: A specific composition of phospholipids and a significant concentration of polyunsaturated fatty acids (PUFA) have been shown to be present in sperm membranes and Seminal plasma has a highly specialized scavenger system that defends the sperm membrane against lipoperoxidation.
Abstract: This article is, in part, a review of present knowledge regarding the lipid metabolism of the sperm cell and the lipid composition of the sperm plasma membrane. It is also a summary of our research on this topic, reporting published and unpublished data. The article tries to cover both basic and clinical research. Sperm cells use lipid metabolic pathways for the production of part of their energy. The double leaflets of the membrane are not simply a passive, bilayer, lipidic film in which the receptors receive their molecular specific signals, but are a very specialized structure. Complete maturation of the lipids of the sperm cell membrane is reached after passage of the spermatozoon through the epididymis. A specific composition of phospholipids and a significant concentration of polyunsaturated fatty acids (PUFA) have been shown to be present in sperm membranes. Plasmalogen is a special kind of phospholipid found exclusively in spermatozoa and other cells with the capacity to react promptly to stimuli. In addition, we have found a high concentration of the n-3 PUFA family in the sperm membrane. Seminal plasma has a highly specialized scavenger system that defends the sperm membrane against lipoperoxidation. Various pathologies and systemic predisposition can lead to an antioxidant/pro-oxidant disequilibrium. Clinical trials with natural scavengers could be a useful research area in which to seek a treatment for these pathologies. Of the natural scavengers, glutathione has been shown to restore the physiological constitution of PUFA in the cell membrane under certain conditions.

Journal ArticleDOI
TL;DR: Results are discussed in terms of the costs of producing longer sperm, the correlated evolution of sperm length and body size, the relationship between breeding system and sperm production patterns, and the nature of differences between vertebrates and invertebrates in sperm production and the size of testes.
Abstract: Relationships among body mass, testis mass, sperm length, and the number of sperm produced were examined among 11 Drosophila species, after controlling for phylogenetic ef- fects. This is the first study to examine many of these relationships comparatively in an inverte- brate taxon; patterns observed among these variables were fundamentally different from those consistently reported in studies of vertebrates. In regression analyses, testis mass increased with body mass with an exponent greater than one, which indicates that males of larger-bodied Drosophila species make a proportionately greater energetic investment in testes than do males of smaller-bodied species. The positive allometry of testis mass is hypothesized to be a combined consequence of the unusual positive relationship between body mass and sperm length and the positive relationship between sperm length and testis mass. Interspecific variation in testis mass was found to be a function of variation in sperm length rather than variation in the number of sperm produced. Significant trade-offs were identified between sperm length and the number of sperm produced and transferred per copulation. Results are discussed in terms of the costs of producing longer sperm, the correlated evolution of sperm length and body size, the relation- ship between breeding system and sperm production patterns, and the nature of differences between vertebrates and invertebrates in sperm production and the size of testes.

Journal ArticleDOI
TL;DR: It is shown that a vacuolar (H+)–ATPase is expressed at high levels on the luminal plasma membrane of specialized cells in the epididymis, which closely resemble acid–secreting kidney intercalated cells and that similar cells are also present in the vas deferens.
Abstract: An acidic luminal pH (ref. 1–3) is involved in sperm maturation, and in maintaining sperm in an immotile state in the epididymis and vas deferens2,4–6. Neutralization by prostatic fluid is one of a complex series of events that triggers sperm motility2,7,8. Failure of the acidification mechanism might, therefore, result in poor sperm maturation, premature motility and infertility. We have shown that a vacuolar (H+)–ATPase is expressed at high levels on the luminal plasma membrane of specialized cells in the epididymis9, which closely resemble acid–secreting kidney intercalated cells10,11. We now show that similar cells are also present in the vas deferens, and that a bafilomycin–sensitive proton flux can be detected using a noninvasive proton–selective vibrating probe. Up to 80% of the net proton secretion in the vas deferens is inhibited by bafilomycin, consistent with a major role of a vacuolar–type (H+)–ATPase in this process. This acidification mechanism is a potential target for novel strategies aimed at modulating the acidification capacity of parts of the male reproductive tract and, therefore, in regulating male fertility.

Journal ArticleDOI
TL;DR: The findings suggest that capacitation is a series of positive destabilizing events that eventually lead to cell death, and that a succession of spermatozoa in the correct physiological state may be provided for the freshly ovulated egg.
Abstract: Capacitation, the process whereby spermatozoa are rendered capable of interacting with and fertilizing the egg, was discovered more than 40 years ago. However, our understanding of it is still far from satisfactory. Several factors conspire to obfuscate studies of capacitation mechanisms: the inherent functional heterogeneity of sperm populations, the range of functions used as parameters of capacitation (whence the endpoint of the process has become conceptually uncertain), and the several profound differences between model in vitro fertilization (IVF) systems and the situation in vivo in the female reproductive tract. Recent investigations in the author's laboratory have shown that bicarbonate/CO2, an essential component for successful IVF, causes rapid changes in lipid architecture of the sperm plasma membrane and slower changes in surface coating. These changes are accompanied by membrane destabilization and cell death. Evidence suggests that bicarbonate's actions are mediated through cyclic nucleotide signalling. Of particular note is the heterogeneity in rate of response to bicarbonate shown by individual cells in the sperm populations. Taken together with other observations, the findings suggest that capacitation is a series of positive destabilizing events that eventually lead to cell death. The 'capacitated' state would then be a window of destabilization within which spermatozoa can undergo a zona-induced acrosome reaction and display hyperactivated motility. Further along the destabilization pathway, spontaneous acrosome reactions would occur before total membrane degeneration. In vivo, capacitation would be a conflict between destabilization and sperm survival. Concentrations of bicarbonate are maintained low in the cauda epididymidis, where sperm survive for long periods, and one may speculate that hormonal control of local bicarbonate/CO2 in oviducal 'storage' sites in the female tract could allow 'safe' sequestering of live spermatozoa until around the time of ovulation; the environment may then change to produce a 'capacitating' effect, whence, due to the inherent functional heterogeneity of the sequestered population, small numbers of capacitated spermatozoa are released sequentially. In this way, a succession of spermatozoa in the correct physiological state may be provided for the freshly ovulated egg.

Journal ArticleDOI
TL;DR: In sperm, inhibition of T channels during gamete interaction inhibits zona pellucida-dependent Ca2+ elevations, as demonstrated by ion-selective fluorescent probes, and also inhibits acrosome reactions, which suggests a mechanism for the reported contraceptive effects of those compounds in human males.
Abstract: The sperm acrosome reaction is a Ca2+-dependent exocytotic event that is triggered by adhesion to the mammalian egg’s zona pellucida. Previous studies using ion-selective fluorescent probes suggested a role of voltage-sensitive Ca2+ channels in acrosome reactions. Here, whole-cell patch clamp techniques are used to demonstrate the expression of functional T-type Ca2+ channels during mouse spermatogenesis. The germ cell T current is inhibited by antagonists of T-type channels (pimozide and amiloride) as well as by antagonists whose major site of action is the somatic cell L-type Ca2+ channel (1,4-dihydropyridines, arylalkylamines, benzothiazapines), as has also been reported for certain somatic cell T currents. In sperm, inhibition of T channels during gamete interaction inhibits zona pellucida-dependent Ca2+ elevations, as demonstrated by ion-selective fluorescent probes, and also inhibits acrosome reactions. These studies directly link sperm T-type Ca2+ channels to fertilization. In addition, the kinetics of channel inhibition by 1,4-dihydropyridines suggests a mechanism for the reported contraceptive effects of those compounds in human males.

Journal ArticleDOI
TL;DR: It is concluded that in this population of healthy young men there has not been any decline in semen quality in the past 21 years.

Journal ArticleDOI
TL;DR: The observed deterioration of sperm characteristics is considered to reflect degeneration of sperm production among men aged between 20 and 40 years.
Abstract: We have retrospectively analysed the sperm characteristics of 416 consecutive healthy young men who presented themselves in the past 19 years as candidate sperm donors. Ejaculate volume increased slightly (P = 0.067), and average sperm concentration decreased (P = 0.035) by 12.4 x 10(6)/ml over the observation period, so that sperm count per ejaculate remained unchanged (P = 0.91). In contrast, sperm morphology (r = - 0.23, P < 0.0001), rapid progressive motility (r = - 0.42, P < 0.0001) and total motility (r = - 0.33, P < 0.0001) presented an important and time-related decrease. When a quadratic model was used rather than a linear one to analyse the data on rapid progressive motility, there appeared to have been no further decline since 1990. The average proportion of spermatozoa with normal morphology decreased from 39.2% in the period 1977-1980 to 26.6% in 1990-1995 (P < 0.0001), and the mean percentage of spermatozoa with rapid progressive motility decreased from 52.7 to 31.7% (P < 0.0001). The percentage of candidate donors with sperm characteristics below the 5th percentile cut-off value of a normal fertile population increased from 13 to 54% during the observation period (P < 0.0001). Since the technique of semen analysis has remained essentially unchanged in-so-far as has been practically possible, as has the method of recruitment of candidate sperm donors, the observed deterioration of sperm characteristics is considered to reflect degeneration of sperm production among men aged between 20 and 40 years.

Journal ArticleDOI
24 Feb 1996-BMJ
TL;DR: Sperm concentration has not changed with time in the Toulouse area, contrary to the Parisian results, which were recently confirmed in the Paris area of France.
Abstract: Objectives: To investigate whether sperm production has changed during the past 16 years in the Toulouse area of France. Design: Time series analysis of sperm donors9 specimens between 1977 and 1992. Setting: Sperm bank of university hospital in Toulouse, France. Subjects: 302 healthy fertile men candidate sperm donors more than 20 and up to 45 years old and without any infertile brothers. Main outcome measure: Spermatozoa concentration. Results: Donors9 mean age at time of donation was 34.05 (SD 5.13), but this increased significantly (P 0.05). Conclusion: Sperm concentration has not changed with time in the Toulouse area. Key messages Key messages This decline in sperm count was recently confirmed in the Paris area of France We studied sperm production of healthy fertile men in the Toulouse area of south west France The men were recruited according to the same selection criteria as in the Parisian study, but, contrary to the Parisian results, the sperm count of the semen samples had remained constant during the past 16 years These discrepant findings could be explained by different environmental conditions noted between the two areas

Journal ArticleDOI
TL;DR: One aspect of mouse sperm capacitation is the selective activation of one major pHi regulator, which attenuates alkaline shifts in pHi during capacitation as well as the ability of sperm to produce a secretory response to zona pellucida agonists.

Journal ArticleDOI
22 Feb 1996-Nature
TL;DR: It is shown that in the nematode Caenorhabditis elegans, mating with males reduces the lifespan of hermaphrodites by a mechanism independent of egg production or receipt of sperm.
Abstract: THEORIES of life-history evolution propose that trade-offs occur between fitness components, including longevity and maximal reproduction1–3. In Drosophila, female lifespan is shortened by increased egg production4, receipt of male accessory fluid5 and courting6. Male lifespan is also reduced by courting and/or mating7. Here we show that in the nematode Caenorhabditis elegans, mating with males reduces the lifespan of hermaphrodites by a mechanism independent of egg production or receipt of sperm. Conversely, males appear unaffected by mating. Thus, in C. elegans there is no apparent trade-off between longevity and increased egg or sperm production, but there is a substantial cost to hermaphrodites associated with copulation.

Journal ArticleDOI
TL;DR: It is demonstrated that the essential role of the epididymis in male fertility is highlighted and a highly specific function of the c-ros receptor tyrosine kinase during development of distinct epithelial cells is demonstrated.
Abstract: The c-ros gene was originally identified in mutant form as an oncogene. The proto-oncogene encodes a tyrosine kinase receptor that is expressed in a small number of epithelial cell types, including those of the epididymis. Targeted mutations of c-ros in the mouse reveal an essential role of the gene in male fertility. Male c-ros -/- animals do not reproduce, whereas the fertility of female animals is not affected. We demonstrate that c-ros is not required in a cell autonomous manner for male germ cell development or function. The gene, therefore, does not affect sperm generation or function in a direct manner. The primary defect in the mutant animals was located in the epididymis, showing that c-ros controls appropriate development of the epithelia, particularly regionalization and terminal differentiation. The epididymal defect does not interfere with production or storage of sperm but, rather, with sperm maturation and the ability of sperm to fertilize in vivo. Interestingly, sperm isolated from c-ros -/- animals can fertilize in vitro. Our results highlight the essential role of the epididymis in male fertility and demonstrate a highly specific function of the c-ros receptor tyrosine kinase during development of distinct epithelial cells.

Journal ArticleDOI
TL;DR: Computer assisted sperm analysis is shown for the first time to provide a rapid and quantitative measure of the effects of environmental pollutants on the quality of fish sperm, suggesting concentrations of cadmium which have been found to be present in fish gonads as a result of bioaccumulation from the water or diet may decrease the motility of sperm during maturation or storage in the testis.

Journal ArticleDOI
TL;DR: This article found that males of Acheta domesticus and G. supplicans increased the amount of sperm transferred as apparent competition increased and that male A. domesticus transferred more sperm when encountering larger females.
Abstract: There is evidence to suggest that males of various species can respond to the threat of sperm competition by varying the amount of sperm transferred during copulation. We tested this in two species of cricket, Acheta domesticus and Gryllodes supplicans (Orthoptera: Gryllidae) by varying the apparent threat of intermale competition experimentally. The results showed that males of both species increased the amount of sperm transferred as apparent competition increased and that male A. domesticus transferred more sperm when encountering larger females. The results also showed that male G. supplicans produced a larger spermatophylax when a larger ampulla was transferred, a relationship consistent with a sperm protection function.

Journal ArticleDOI
TL;DR: NOS is present in human spermatozoa and that eNOS and bNOS are abundant in normozoospermic samples, and nitric oxide (at endogenous concentrations) appears to be necessary for adequate sperm motility.
Abstract: The aim of this study was to investigate the presence of nitric oxide synthase (NOS) and the production of nitric oxide (NO) by human spermatozoa. Immunoreactivity was examined using a polyclonal antibody raised against porcine cerebellar nitric oxide synthase and monoclonal endothelial (elMOS) and brain (bNOS) antibodies. Using each antibody, NOS was observed localized in the head and midpiece regions of the spermatozoon. Immunofluorescence observed for eNOS and bNOS was more intense in normozoosperm ic samples. Sperm motility was assessed by computer-assi sted semen analysis (CASA) in the presence and absence of N G-nitro-L-arginine methyl ester (L-NAME; 10~5M), an NO synthesis inhibitor or tumour necrosis factor (TNF)-a (20 lU/ml), a superoxide inducer. In the presence of L-NAME, percentage progressive motility, average path velocity (VAP), straight line velocity (VSL) and curvilinear velocity (VCL) were significantly reduced after 30 min. Sperm viability was not decreased by TNFoc or L-NAME. The accumulation of nitrite (the stable end-product of the NOS/NO pathway) by spermatozoa was measured using the Griess reaction. After 8 h, nitrite concentrations were lower in asthenozoospermic compared to normozoosperm ic samples. In the presence of TNFcc, nitrite accumulation was significantly reduced in normozoosperm ic samples. We conclude that NOS is present in human spermatozoa and that eNOS and bNOS are abundant in normozoospermic samples. Nitric oxide (at endogenous concentrations) appears to be necessary for adequate sperm motility.

Journal ArticleDOI
TL;DR: The data suggest that the disassembly of the sperm tail during bovine fertilization occurs as a series of precisely orchestrated events involving the destruction and transformation of particular sperm structures into zygotic and embryonic components.
Abstract: Sperm incorporation and the conversion of the sperm-derived components into zygotic structures during in vitro fertilization of bovine oocytes was explored by combining ultrastructural studies with observations of the fertilizing sperm tagged with a mitochondrion-specific vital dye MitoTracker green FM. The zygotes fertilized by the MitoTracker-labeled sperm were fixed at various times after fertilization and then processed for immunocytochemistry to examine the distribution of DNA, microtubules, and sperm tail components, including the fibrous sheath and axonemal microtubules. We show here that the complete incorporation of the sperm, but not sperm-oocyte binding and oocyte activation, depends upon the integrity of oocyte microfilaments and is inhibited by the microfilament disrupter cytochalasin B. After sperm incorporation, the mitochondria are displaced from the sperm's connecting piece, and the sperm centriole is exposed to the egg cytoplasm. This event is followed by the formation of the microtubule-based sperm aster, which is responsible for the union of male and female pronuclei. Concomitantly, the major structure of the sperm principal piece, the fibrous sheath, disappears. After the first mitosis, the compact mitochondrial sheath can be seen in one of the blastomeres. An aggregate of the sperm mitochondria is observed at the entry of the second mitosis, although they remain in the vicinity of the nucleus and can later be seen at one pole of the metaphase spindle. The mitochondrial cluster is occasionally found in one of the blastomeres in the early-stage four-cell embryos, but it is no longer detected by the beginning of the third mitotic cycle. These data suggest that the disassembly of the sperm tail during bovine fertilization occurs as a series of precisely orchestrated events involving the destruction (fibrous sheath and mitochondrial sheath) and transformation (DNA, sperm centriole) of particular sperm structures into zygotic and embryonic components.

Journal ArticleDOI
TL;DR: Nonobstructive azoospermia caused by maturation arrest may be treated with testicular sperm extraction with ICSI apparently as successfully as Sertoli cell only.