[Sperm motility and evaluation of reproductive capacity].
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Urogenital infections are considered important factors in male infertility. In this in vitro study we have evaluated the impact of leucocytes in association with an artificial infection with Escherichia coli on the motility of human spermatozoa. Ejaculates and blood samples were obtained from healthy donors with normal semen parameters. Ejaculates were prepared by swim-up technique and five fractions were isolated for incubation. Leucocyte subtypes were separated from blood samples by gradient centrifugation. Purified sperm suspensions were adjusted to a concentration of 20 x 106 ml-1 and incubated with lymphocytes/ monocytes, polymorphonuclear granulocytes (PMN), and E. coli. Samples were incubated for up to 6 h at 37 degrees C. Motility analysis was performed using a computer-assisted sperm analyzer (CASA). Spermatozoa incubated with 3 x 106 PMN ml-1 revealed a significant (P=0.003) decrease in progressive motility after 2 h. This decrease remained weakly significant (P=0.024) after 4 and 6 h. Lymphocytes and monocytes had no effect on sperm motility. Spermatozoa incubated with granulocytes and E. coli demonstrated highly significant alterations in motility after 4 and 6 h of incubation (P < 0.001). The PMN indicate an effect on motility of spermatozoa under experimental conditions. However, the results suggest that bacteria are the primary agents that interfere with sperm motility.
This study was undertaken to evaluate the effects of gamma-aminobutyric acid (GABA) and GABAergic agonists and antagonists on sperm kinematic parameters and hyperactivation, evaluated by a computer-assisted semen analysis (CASA) system, and intracellular cAMP content in 22 normozoospermic semen samples. Because of the possible interaction of progesterone with the GABAA receptor, we also evaluated the effects of progesterone on these parameters. GABA increased beat cross frequency, curvilinear velocity (VCL), the percentage of spermatozoa moving with an average path velocity > 10 microns/s (active) and hyperactivation, and decreased linearity and straightness. Bicuculline, a GABAA receptor antagonist, antagonized the effects of GABA on all these parameters except the percentage of active spermatozoa. Muscimol, a GABAA receptor agonist, increased VCL, the percentage of active spermatozoa, and hyperactivation by about the same extent as GABA, suggesting the involvement of the GABAA receptor. However, the GABAB receptor also seems to mediate some of the effects of GABA, because baclofen, a selective agonist for this receptor, increased significantly the percentage of active spermatozoa and hyperactivation. The effect of baclofen on this latter parameter was, however, less pronounced than that obtained with GABA or muscimol. Progesterone had the same effects as GABA on sperm kinematic parameters and hyperactivation and the simultaneous presence of both compounds was not more effective than each single one. GABA and progesterone did not have any effect on intracellular cAMP content. In conclusion, GABA modulated sperm kinematic parameters and increased hyperactivation. These effects have the same magnitude of those produced by progesterone and seem to be mediated mainly by the GABAA receptor. We speculate that GABA may be a physiological regulator of sperm function.
The male reproductive organ of rainbow trout and chum salmon consists of a pair of testes and sperm ducts. Spermatozoa in the distal portion of the sperm ducts exhibit full motility in the K+-free medium. However, spermatozoa from the testis were almost immotile in this medium. This suggests that the spermatozoa acquire a capacity for movement during their passage from the testis along the sperm duct. In chum salmon migrating into a bay, the sperm duct was almost empty. However, after the fish have travelled upstream for 1 km to their spawning ground in the river, the spermatozoa have left the testis, moved into the sperm duct and are capable of becoming motile. Thus it is probable that the process of acquiring the ability to move occurs within a relatively short period in this simple reproductive organ. Additionally, testicular spermatozoa demembranated with Triton X-100 exhibited motility, although the motility was less than that of demembranated spermatozoa from the sperm duct, suggesting that the acquisition of motility may correspond with the development of some function of the plasma membrane.
Spermatozoa of rainbow trout and chum salmon, which have no potential for motility in the testis, acquire that potential in the sperm duct. This paper demonstrates that there is little difference between the levels of sodium, potassium, calcium, magnesium, chloride and osmolality of the seminal plasma in the testis and in the sperm duct. However, the bicarbonate concentration of the seminal plasma and the pH value of semen were higher in the sperm duct than in the testis. When immotile spermatozoa obtained from the testis were incubated in artificial seminal plasma with a high pH and containing HCO3-, spermatozoa became motile within 1 h. These results suggest that spermatozoa of salmonid fish acquire the potential for motility as a result of the increase in seminal bicarbonate concentration and pH that occurs as spermatozoa pass from the testis to the sperm duct.
To minimize overestimation of motility, it is recommended that fresh semen be diluted with seminal plasma prior to automated analysis. However, for glycerolated or cryopreserved semen this is impractical, and alternative methods are needed to minimize automated motility bias. In the present study, the proportion of motile spermatozoa was determined in fresh, diluted and cryopreserved semen (n = 25 ejaculates) using visual and automated methods. The effect of software settings on motility was investigated by assessing samples at a range of modified settings. At standard settings, automated motility was biased in fresh semen (+7.2%) after dilution with cryopreservative (-2.9%) and after cryopreservation (-7.8%) (P < 0.0001 versus visual). Automated motility was inversely related to the minimum number of frames for motility sampling (P < 0.0001), with mean estimates of 41.0, 46.1, 52.0 and 58.2% generated at settings of 8, 4, 2 and 1 frame(s) respectively (n = 15 fresh, diluted and cryopreserved samples). Based on an arbitrary ordinal scale, a method was developed whereby motility sampling was adjusted prior to analysis according to sperm density. Analysis of an independent set of semen samples with density-adjusted software settings reduced bias in automated estimates (n = 30) before and after freezing (P < 0.0001). In addition, bias was no longer related to sperm density. In conclusion, modification of software settings is an effective alternative to dilution to minimize bias in automated motility estimates in fresh, diluted and cryopreserved human semen.
Sampling conditions that affect the biological validity of computer-assisted analysis of ram sperm motion were examined using a continuous real-time computerized semen analysis system (Hobson Sperm Tracker). Search radius (SR, 10 settings) and minimum track point (MTP, 10 settings) were varied factorially to evaluate their effects on the inclusion of sperm subpopulations within derived datasets. Low SR (< 12 microns) or high MTP values (> 26 frames) precluded measurements of rapidly moving cells, whereas high SR (> 17 microns) and low MTP settings (< 22 frames) led to erroneous tracking and poor data quality. Suitable settings for these set-up parameters were derived and tested for their biological consistency with four methods of preparing ram semen for computerized analysis. The preparation techniques tested were: centrifugation through sucrose-based Ficoll and Percoll media, a swim-up technique and simple dilution in Tyrode's media. The "selective' Percoll and swim-up methods generated sperm populations with significantly higher linear velocities and a lower tendency to deviate from linear trajectories than from either the Ficoll method or dilution technique. Deleterious effects of centrifugation were evident, particularly on sperm survival in vitro over several hours. It is concluded that computer-assisted semen analysis provides useful information about the behaviour of ram spermatozoa in vitro, but the measurement conditions must be defined carefully.
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The effect of six metabolically active compounds, namely caffeine, kallikrein, cyclic adenosine 3':5'-monophosphate, prostaglandin E, triiodothyronine, and insulin on spermatozoal motility was investigated. Hypokinetic and normokinetic semen specimens from fertile and infertile patients were incubated with various concentrations of these compounds. Spermatozoal velocity and percentage of motility were determined periodically with the objective multiple exposure photography (MEP) method before and during the following 4 to 8 hours. No significant differences was found between experimental and control specimens as to these parameters under the influence of the tested compounds, with the exception of caffeine. Caffeine immediately increased the percentage of motile spermatozoa by 30% to 50% of initial values in approximately two-thirds of cases. No influence on sperm velocity was detected during this period. By using the combined MEP and supravital staining techniques it was found that nonmotile live spermatozoa were activated by caffeine without any effect on the percentage of dead sperm. Some practical implications of these findings, the difference between our results and those reported by others, and the mechanism of action of these compounds are reviewed and discussed.
In the human male, Leydig cell function of the testis declines slowly but significantly with age. A similar decline in the spermatogenic activity, however, is not well proven by consistent data. Cross-section studies quoted in the literature mostly describe only minor changes in different sperm parameters with age. In our database we identified 253 men who had attended our department since 1989 for subsequent semen analyses at intervals of more than 1 year. The semen analyses were performed according to the guidelines of the WHO with the exception that motility parameters were measured by CASA. The mean interval between the two analyses was 967 days with a range of 341-2,731 days. None of the parameters evaluated showed a significant increase or decrease with time. The mean seminal volume was identical in the two analyses. Mean sperm count increased during the observation period by 12. 10(6) spermatozoa/year. In 177 patients an increase and in 73 patients a decrease occurred. Also a corresponding increase in total sperm count was observed. Sperm motility increased in 123 patients, and it decreased in 123 patients. Only slight differences in the increase or decrease of the two parameters between the groups of patients with and without a history of genital diseases and/or varicocele were found. The results of our study do not support the suggestion that the sperm parameters decrease with age.
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The effect of serum from patients with severe chronic renal failure on the motility of ejaculated spermatozoa was objectively evaluated by the multiple exposure photography method. Uremic serum was not found to have a deleterious effect on the motility of ejaculated spermatozoa. On the contrary it caused a significant increase of spermatozoal velocity and a slighter increase in the percentage motility. These effects cannot be only attributed to the lowering of seminal viscosity and to the reduction of sperm concentration, but to various substances contained in blood serum.
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