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Cinematographic analysis of bovine epididymal sperm motility: epididymal maturation and forward motility protein.
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Standardization and quality control of sperm concentration and sperm motility counts in semen analysis.
The paper reports a study of standardization and quality control of sperm concentration counts and visual motility assessments in human semen analyses performed for infertility investigations and from internal quality control procedures. Sperm concentration determinations were performed in Improved Neubauer haemocytometers on volumetric dilutions made using a positive displacement pipettor for sampling the liquefied semen. In addition to a standard 1 + 19 dilution a second dilution of either 1 + 9, 1 + 19 or 1 + 49 was made according to whether the estimated sperm concentration was less than 20, 20-100 or greater than 100 X 10(6)/ml respectively. The duplicate determinations of sperm concentration were highly significantly correlated (P much less than 0.001) with less than 5% variability. Parallel visual sperm motility assessments were made by two pairs of technicians and showed highly significant correlations (P much less than 0.001) between technicians in the determination of the percentages of motile and progressive spermatozoa as well as the subjective rating of sperm progressivity. When these values were incorporated into a calculated motility index which gave added weight to the progressive spermatozoa and to their quality of progression the correlations between technicians remained highly significant (P much less than 0.001) with average differences of the order of 1.0%. Therefore, provided that sufficient attention is paid to technician training, regular standardization checks and the use of only proven reliable procedures, quantitatively accurate values for sperm concentration and motility can be obtained in routine semen analyses.
Expression of aquaporins in the efferent ductules, sperm counts, and sperm motility in estrogen receptor-alpha deficient mice fed lab chow versus casein.
Estrogens play an important role in the male reproductive tract, and this is especially so for the efferent ductules, where alpha-estrogen receptors (ERalpha) have been localized. Mice deficient in ERalpha (alphaERKO mice) are infertile, and the effect appears to be due in part to retention of water at the level of the efferent ductules. In the present study, we examined the consequences of ERalpha deletion on the distribution of certain aquaporins (AQPs), water protein channels, in the efferent ductules and on sperm numbers and motility. In addition, the effects of feeding mice a regular lab chow diet, which contains phytoestrogens, known to affect male reproductive tract functions, and a casein diet, which lacks phytoestrogens, were also assessed. Light microscope immunolocalizations of AQP-1 and AQP-9 revealed dramatic reduction and patchier staining in alphaERKO mice with distal areas of the efferent ductules being more affected than proximal areas. No other changes in immunolocalizations were noted as a consequence of diet. Computer-assisted sperm analyses demonstrated a 62% reduction in cauda epididymal sperm/ml in alphaERKO mice fed lab chow, whereas 87% fewer sperm/ml were observed in alphaERKO mice fed casein, suggesting an enhanced role for sperm production and concentration in a diet containing phytoestrogens. All sperm motility parameters were altered to some degree in alphaERKO mice fed lab chow. Alterations in sperm motility parameters were also detected, but were less dramatic in alphaERKO mice fed casein. These data suggest that the decrease in AQP expression in the efferent ductules of alphaERKO mice contributes in part to water retention in this tissue, eventually leading to backflow of water into the testis, with subsequent decreases in sperm concentration and motility. The data also suggest that phytoestrogens, which are present in regular lab chow, can influence the male reproductive tract with and without the presence of ERalpha, promoting efferent ductule and epididymal functions when ERalpha is expressed, but inhibiting these same functions when ERalpha is missing. Taken together the data underscore the importance of estrogens and ERalpha in maintaining sperm maturation and preventing male infertility.
Tales from the tail: what do we really know about sperm motility?
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Objective evaluation of sperm motility and values of velocity.
Differences between the sperm velocity values of normozoospermic men and of those with proved fertility, above all in younger age groups, are quoted. There is a reference to the possibility of the diagnostic and prognostic importance of following the sperm velocity in various time intervals, at a minimum rate within 1 and 5 hours after ejaculation.
Sialic acid concentration and sperm motility.
Sialic acid (SA) forms an integral part of the sperm membrane, and the masking properties of SA are well documented. Masking of estrogen and melatonin receptors on the sperm midpiece by SA has been demonstrated. In this paper we report on the correlation of sperm bound SA concentration and motility. We found a negative correlation between the spermbound SA concentration and both the quantitative and qualitative motility. The spermbound SA correlated positively although not significantly with the spermatozoal ATP concentration. SA may mask the receptors on the sperm membrane, affecting sperm metabolism. This will inhibit the breakdown of ATP to its lower nucleotides resulting in poor motility. This may be another reason to believe that SA may play a key role in sperm motility.
Protein kinase A-anchoring inhibitor peptides arrest mammalian sperm motility.
Cyclic AMP-dependent protein kinase (PKA) is anchored at specific subcellular sites through the interaction of the regulatory subunit (R) with protein kinase A-anchoring proteins (AKAPs) via an amphipathic helix binding motif. Synthetic peptides containing this amphipathic helix domain competitively disrupt PKA binding to AKAPs and cause a loss of PKA modulation of cellular responses. In this report we use S-Ht31, a cell-permeant anchoring inhibitor peptide, to study the role of PKA anchoring in sperm. Our analysis of three species of mammalian sperm detected three isoforms of PKA (RIIalpha, RIIbeta, and RIbeta) and one 110-kDa AKAP. The addition of S-Ht31 to bovine caudal epididymal sperm inhibits motility in a time- and concentration-dependent manner. A control peptide, S-Ht31-P, identical to S-Ht31 except for a proline for isoleucine substitution to prevent amphipathic helix formation, had no effect on motility. The inhibition of motility by S-Ht31 is reversible but only if calcium is present in the suspension buffer, suggesting a role for PKA anchoring in regulating cellular calcium homeostasis. Surprisingly, inhibition of PKA catalytic activity had little effect on basal motility or motility stimulated by agents previously thought to work via PKA activation. These data suggest that the interaction of the regulatory subunit of PKA with sperm AKAPs, independent of PKA catalytic activity, is a key regulator of sperm motility and that disruption of this interaction using cell-permeable anchoring inhibitor peptides may form the basis of a sperm-targeted contraceptive.
Regulation of sperm motility at the axonemal level.
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Electroejaculation does not impair sperm motility in the beagle dog: a comparative study of electroejaculation and collection by artificial vagina.
Sperm samples obtained by electroejaculation (EEJ) in men with anejaculatory infertility have a markedly lower quality than those obtained by normal ejaculation. An electrical effect of the EEJ procedure has been implicated by some investigators as a direct cause of low sperm quality. To determine whether the EEJ procedure causes direct sperm damage, we compared ejaculates obtained from dogs by EEJ and by artificial vagina (AV). In seven adult beagle dogs, semen was collected weekly, alternating between the two procedures. The average (antegrade) sample from AV had a volume of 2.6 ml., sperm concentration of 150.1 x 10(6) per ml., total sperm count of 381.7 x 10(6) and motility of of 26.3%. The average antegrade sample from EEJ had a volume of 1.8 ml., a concentration of 129.6 x 10(6) per ml., a total sperm count of 166.8 x 10(6) and a motility of 30.1%. Of these differences only the total sperm counts and the total motile sperm counts were statistically significant (p < 0.05). There were no significant differences between the antegrade motilities, total sperm counts (antegrade plus retrograde-381.7 versus 243.4 x 10(6), for AV and EEJ, respectively) or the total motile sperm counts from the two procedures (103.9 versus 78.0 x 10(6)). There were no differences in the average curvilinear velocity (VCL) measured by computer-assisted sperm motion analysis (56.9 mu. per second for AV, 47.4 mu. per second for antegrade EEJ specimens and 41.7 mu. per second for retrograde EEJ specimens). Since routine semen parameters between artificial vagina and electroejaculation did not differ in dogs, we conclude that the electroejaculation procedure is not responsible for the gross semen abnormalities reported in electroejaculation of anejaculatory men.
Contributions of extracellular and intracellular Ca2+ to regulation of sperm motility: Release of intracellular stores can hyperactivate CatSper1 and CatSper2 null sperm.
In order to fertilize, mammalian sperm must hyperactivate. Hyperactivation is triggered by increased flagellar Ca(2+), which switches flagellar beating from a symmetrical to an asymmetrical pattern by increasing bending to one side. Thimerosal, which releases Ca(2+) from internal stores, induced hyperactivation in mouse sperm within seconds, even when extracellular Ca(2+) was buffered with BAPTA to approximately 30 nM. In sperm from CatSper1 or CatSper2 null mice, which lack functional flagellar alkaline-activated calcium currents, 50 microM thimerosal raised the flagellar bend amplitudes from abnormally low levels to normal pre-hyperactivated levels and, in 20-40% of sperm, induced hyperactivation. Addition of 1 mM Ni(2+) diminished the response. This suggests that intracellular Ca(2+) is abnormally low in the null sperm flagella. When intracellular Ca(2+) was reduced by BAPTA-AM in wild-type sperm, they exhibited flagellar beat patterns more closely resembling those of null sperm. Altogether, these results indicate that extracellular Ca(2+) is required to supplement store-released Ca(2+) to produce maximal and sustained hyperactivation and that CatSper1 and CatSper2 are key elements of the major Ca(2+) entry pathways that support not only hyperactivated motility but possibly also normal pre-hyperactivated motility.
Vaginal lubricants: effect of glycerin and egg white on sperm motility and progression in vitro.
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Inhibition of sperm motility by the volatile anesthetic halothane.
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In-vitro culture of hamster epididymal epithelium and induction of sperm motility.
Epithelium from the proximal corpus region of the epididymis of adult hamsters was cultured in modified RPMI 1640 medium supplemented with growth factors and androgens at 37 degrees C in 5% CO2 in air. Prepared plaques of epithelium formed spheres of tissue with epithelial cells outermost. At the light and electron microscope level, these epithelial balls displayed morphology consistent with continued secretory and absorptive function. After 3-5 days, cultured cells either plated out over the bottom of plastic wells or formed vesicles which expanded as their interior became fluid filled. Spermatozoa recovered from the caput epididymidis were co-cultured with epithelium. After 8 and 24 h, a proportion of spermatozoa (30%) exhibited slow but persistent flagellum beats with slow progressive motility. Spermatozoa in control incubations were immotile. This change in motility pattern would suggest that some sperm maturation processes had occurred in vitro.
Sperm motility in fertile men and males in infertile units: in vitro test.
An in vitro test evaluating spermatozooan motility was used to compare percentage of forward progressive spermatozooa at initial observation, within 1 h of obtaining samples, with forward progression after 8 h in Petri dishes (humid chambers). A constant motility profile was found in the sperm of fertile men after 8 h, whereas a different profile was found in 94% of the infertile male groups.
Ultrastructural and rheological properties of bovine vaginal fluid and its relation to sperm motility and fertilization: a review.
The tubular genital organs of the mammalian female reproductive tract provide different luminal microenvironments that can act as an aid or a barrier to gamete transport and the first steps of embryonic development. The mechanism of sperm transport through the genital tract has not been fully established but, first of all, sperm cells encounter and need to overcome mucous-like secretions to reach the site where fertilization occurs. One of these secretions, vaginal fluid, is a complex biological product, which is mainly derived from cervical mucus. In this review, we focus on recent studies that describe the relationships between the ultrastructural and rheological properties of bovine vaginal fluid and sperm motion. In the light of this, we propose bovine vaginal fluid as a morphofunctional model for the study of sperm transport in mammals.
Protective role of superoxide dismutase in human sperm motility: superoxide dismutase activity and lipid peroxide in human seminal plasma and spermatozoa.
The levels of superoxide dismutase (SOD), a highly specific scavenging enzyme for superoxide anion radicals (O2-), and lipid peroxide produced by oxygen free radicals were measured in human seminal plasma and spermatozoa. Seminal plasma contained 366.8 +/- 20.9 U/ml (mean +/- SE) of SOD activity. SOD activity in human spermatozoa showed a significant correlation to the number of motile spermatozoa, while the activity in seminal plasma did not relate to the sperm concentration or motility. The lipid peroxide concentration in seminal plasma was 6.22 +/- 0.46 nmol/ml and had no significant relationship to sperm concentration or motility. The malondialdehyde (MDA) concentration in spermatozoa was significantly related to the number of immotile spermatozoa. A decrease in the motility of spermatozoa incubated in medium without seminal plasma was observed after 120 min, while the MDA concentration of the spermatozoa increased. Addition of exogenous SOD (400 U/ml) to the sperm suspension significantly decreased this loss of motility and the increase of the MDA concentration. These data suggest a significant role for SOD in sperm motility. It seems that lipid peroxidation of human spermatozoa may cause loss of motility and that SOD may inhibit this lipid peroxidation. These results suggest that SOD may have a possible clinical application in the use of spermatozoa for in-vitro fertilization (IVF) or artificial insemination.