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PubMed1989

[A simple microphotographic method for the evaluation of sperm motility].

A simple objective method for examination of sperm quality, both for practical and research purposes, is described. The results obtained show that the evaluation of samples must be performed by 15 minutes after the removal of the spermatozoa from the culture system and by 5 minutes after the placement on the microscope slide, in order to avoid the influence of external conditions on sperm motility. From the comparison of parameters examined (percentage of sperm motility, mean velocity, lateral head displacement, number of waves per second, distribution in individual speed intervals) it follows that media used for sperm incubation (B2 and F 10) do not affect negatively sperm motility and may be exchanged with each other if necessary. The results contribute to the improvement of diagnostic and treatment of andrological disturbances. The study will be used in our programme of treatment of infertile couples.

Humans↗

PubMed1993

Rises of intracellular Ca2+ and pH mediate the initiation of sperm motility by hyperosmolality in marine teleosts.

Spermatozoa of marine teleosts, puffers and flounder, were completely quiescent when they were washed to remove electrolytic components of the seminal plasma and then diluted in nonelectrolyte solutions isotonic to the seminal plasma. Sperm motility was initiated upon dilution in hypertonic nonelectrolyte solutions. These observations suggest that sperm motility is suppressed by seminal osmolality and motility is triggered solely by the increase in external osmolality which occurs at natural spawning in hypertonic seawater. Extracellular Ca2+ had no influence on the osmolality-dependent initiation of sperm motility. However, sperm motility was initiated even in isotonic solution when Ca2+ was introduced into the sperm cells by Ca2+ ionophore. Intracellular Ca2+ increased at the osmolality-dependent initiation of sperm motility under Ca(2+)-free conditions. These results suggest that the release of Ca2+ from intracellular storage in response to the increase in external osmolality has a key role in the initiation of sperm motility. A transient increase in intracellular pH was also observed at the hyperosmolality-dependent initiation of sperm motility. Furthermore, initiation of sperm motility was induced even in isotonic solutions when intracellular pH increased by the treatment with ammonium salts. These results suggest that an increase in intracellular pH, as well as the rise in intracellular Ca2+, has an important role in the initiation of sperm motility in marine teleosts.

Acetates↗

PubMed1993

Use of a sealed minichamber for direct observation and evaluation of the in vitro effect of cigarette smoke on sperm motility.

OBJECTIVE: To determine the in vitro effect of cigarette smoke on sperm motility and survival in an attempt to find a possible association with clinical studies that had pointed to the existence of such an effect in vivo. DESIGN: Laboratory experiments in which the effect of cigarette smoke on human sperm placed in a sealed minichamber was directly observed and determined by motion analysis. PATIENTS: High-quality fresh semen specimens were obtained from healthy donors and used for the experimental studies. INTERVENTIONS: Various doses of filtered and nonfiltered cigarette smoke, collected by an improvised manual smoking machine, were streamed at a rate of 2 mL/s into sealed chambers containing samples of washed spermatozoa. Evaluation of the changes in sperm motility was performed in real time by direct microscopical observation and repeated determinations by the multiple exposure photography technique until their complete immobilization. RESULTS: Streaming of 100 mL of cigarette smoke induced a dramatic drop in sperm motility, which caused sperm immobilization in about 15 minutes. This effect showed a clear dose-response relationship either with the amounts streamed or with the time during which sperm were exposed to cigarette smoke. The harmful effect was almost the same whether sperm were exposed to filtered or nonfiltered cigarette smoke. The role of carbon monoxide and hydrogen cyanide on these findings was studied separately, and it was shown that the inhibitory effect was not due to the presence of these two important components of cigarette smoke. CONCLUSIONS: The clear evidence of the in vitro harmful effect of crude cigarette smoke on sperm motility points to the possibility that in heavy smokers certain toxic agents may reach mature spermatozoa or their precursors via the blood stream and induce their harmful effect on sperm motility in vivo.

Carbon Monoxide↗

PubMed1979

Elimination of errors induced during a routine human sperm motility analysis.

Influence of some variables on the accuracy of sperm motility evaluation was studied by analyzing normal seminal specimens with the aid of themultiple exposure photography (MEP) method. Results of this study demonstrated the existence of variations in sperm motility between view areas of the same sample or between different samples of the same specimen. Variations were found even when the same view area was analyzed intermittently during a short period of time. Motility was practically unaffected when a regular sized drop was kept within the preparation for about 20 min. The light from the microscope neither stimulated nor depressed sperm motility when they were illuminated continuously for that period of time. Percent of motility and sperm velocity was directly related to the thickness of the examined drop when the latter ranged between zero and ten micrometers. According to these findings it is suggested that several view areas from various drops of the same specimen should be analyzed with each view area being inspected several times before motility of that field is assessed. Motility should be evaluated from drops of standard and constant thickness; otherwise major errors in sperm motility assessment are to be expected.

Diagnostic Errors↗

PubMed1982

Studies on the effects of sugars on washed human sperm motility.

The motility of ejaculated human sperm which was washed thoroughly by Ficoll gradient centrifugation to remove seminal plasma was investigated in the presence of various concentrations of sugars. Washed human sperm showed high motility in an artificial medium containing glucose and fructose, which were major sugar components of semen. However, mannitol and 3-O-methyl-D-glucose had no effect on sperm motility. Dose-response curves of sperm motility in the additions of glucose or fructose were bell-shaped. The maximum sperm motility was found in the medium with 1 mM glucose or 15 mM fructose. When, however, both 1 mM glucose and 15 mM fructose were added to the medium, sperm motility was higher than that with an individual sugar. Although phlorizin decreased glucose activated sperm motility, it has no effect in case fructose was the activator. The results suggest that the effect of glucose on human sperm motility is due to a different mechanism from that of fructose.

Carbohydrates↗

PubMed1997

Effect of motile sperm count after swim-up on outcome of intrauterine insemination.

OBJECTIVE: To analyze the prognostic value of motile sperm count after swim-up in IUI with husband's sperm in a large group of subfertile couples. DESIGN: Retrospective study. SETTING: University hospital. PATIENT(S): Nine hundred two couples undergoing 3,037 treatment cycles. INTERVENTION(S): Intrauterine insemination with husband's sperm after swim-up was performed after mild ovarian stimulation with clomiphene citrate and hCG under hormonal and ultrasonographic control of follicle development. MAIN OUTCOME MEASURE(S): Pregnancy rate (PR) in correlation to motile sperm count after swim-up. RESULT(S): A nonlinear increase in PR per treatment cycle was observed with increasing numbers of motile sperm used for IUI. Insemination with < 0.8 x 10(6) motile sperm after swim-up resulted in a PR of < 1% per treatment cycle. When the motile sperm count was above this level, the PR per cycle reached a plateau of 6.9% to 10.2%, with a minor tendency for increase with higher sperm numbers. CONCLUSION(S): Strict analysis of motile sperm count after swim-up is a useful prognostic factor for PRs after IUI. There is a good chance for conception if > or = 0.8 x 10(6) motile sperm are available after appropriate selection methods. Intrauterine insemination performed with considerably higher numbers of motile sperm does not lead to a significant increase in PRs.

Adolescent↗

PubMed1995

The life span of sperm motility and pattern in cumulus coculture.

OBJECTIVE: To study the effect of cumulus cell coculture on human sperm motility, life span, and patterns. DESIGN: Controlled prospective study of 50 normal semen samples. Sperm motility and forward grade was observed in droplets of Ham's F-10 media with cumulus cell coculture and compared with sperm in droplets of Ham's F-10 only. SETTING: The Egyptian IVF-ET Center. MAIN OUTCOME MEASURES: Sperm motility, life span, and forward progressive pattern. RESULTS: The time course of loss of sperm motility in cumulus coculture was significantly slower than without cumulus. Also, the percentage of motile sperm with linear progressive motility of grade 4,3 was significantly higher in cumulus coculture. CONCLUSION: Cumulus cell coculture has a positive effect on prolongation of sperm motility life span and its forward progression.

Cell Communication↗

PubMed1987

Importance of sperm motility after capacitation in interpreting the hamster ovum sperm penetration assay.

Routine semen analysis and a zona-free hamster ovum sperm penetration assay (SPA) were carried out for 220 men from a group of infertile couples. The grade of sperm motility was determined after a preincubation period. For both the normal and abnormal semen samples, the number of positive SPAs was significantly higher when the postcapacitation motility was normal. This difference was observed also when the routine semen characteristics of density, motility, and morphologic features were considered separately regarding their relationship with the SPA. Motility after preincubation showed the most significant positive correlation with the SPA result. Therefore, the SPA score should be limited to semen samples showing adequate postincubation motility.

Animals↗

PubMed2006

Ram sperm motility after intermittent scrotal insulation evaluated by manual and computer-assisted methods.

AIM: To study whether additional measurements of motility characteristics of spermatozoa by computer assisted semen analysis (CASA) were more sensitive indicators of reduced semen quality than estimates of percentages of motile, rapid or progressive cells. METHODS: Intermittent scrotal insulation was applied to 6 rams for 16 h per day for 21 days or to 2 of these for 12 h per day for 28 days in the following year. Semen was collected and evaluated by CASA immediately and either frozen or stored at 30 degrees Celsius or 5 degrees Celsius before re-evaluation. RESULTS: Intermittent scrotal insulation caused falls in the percentage of motile, progressive and rapid sperm, as did freezing-thawing and storage at 30 degrees Celsius or 5 degrees Celsius. Motility characteristics (amplitude of lateral head displacement, mean path velocity, mean progressive velocity and curvilinear velocity), as determined by CASA fell only when the percentage of motile sperm was already reduced. Freezing and thawing or liquid storage of the semen from insulated rams caused a greater fall in the percentage of motile and rapid sperm than control semen, but only affected the motility characteristics when the percentage of motile sperm was already reduced. CONCLUSION: Intermittent scrotal insulation affected not only the motility of the freshly collected sperm, but also their ability to withstand the additional stress of storage. The additional data on motility characteristics obtained by CASA appeared to be no more a sensitive indicator than the percentage of motile cells of reductions in semen quality.

Animals↗

PubMed2005

Molecular aspects of declining sperm motility in older men.

OBJECTIVE: To investigate the molecular mechanism for declining sperm motility in older men. DESIGN: Retrospective and prospective study. SETTING: University-based andrology unit. PATIENT(S): Semen was collected from 2,111 patients and sperm donors. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Age, sperm concentration, ejaculate volume, sperm motility (including velocity average path, velocity straight line, velocity curvilinear, linearity, and lateral head displacement), normal sperm morphology, percentage of abnormally blue-stained flagella, and total and free T, FSH, and LH concentrations were analyzed. In addition, the zinc concentration in the seminal fluid, flagella, and the whole spermatozoa was measured by means of atomic absorption spectrometry. RESULT(S): Patient age correlated negatively with T concentration and sperm motility, including velocity, but positively with the percentage of abnormally stained flagella and the flagellar zinc. Whereas the percentage of abnormally stained flagella correlated negatively with motility and sperm velocity, there was a positive relationship with the flagellar zinc content. Flagellar zinc content was negatively correlated with motility. Testosterone showed a positive relationship with motility and sperm concentration. CONCLUSION(S): Because the epididymis is functionally T-dependent, our data suggest that the removal of zinc from the outer dense fibers during epididymal sperm maturation is affected in aging men, which in turn will result in decreased sperm motility.

Adolescent↗

PubMed2006

[Vas-to-Epididymis antidromic injection of 30% ethanol reduces sperm motility in rats].

OBJECTIVE: To investigate the reduction of sperm motility in rats induced by vas-to-epididymis antidromic injection of 30% ethanol and its mechanism. METHODS: Forty male adult Sprague-Dawley rats were randomized into 3 groups: bilateral vas injection (n = 15) , sham operation control (n = 15) and normal (n = 10). An aliquot of 0.5 ml of 30% ethanol was injected from vas to epididymis bilaterally. After 1 month, all the rats'vasa and epididymides were ablated for studies of the sperm motility, construction changes of the vas and contents of IL-6, IFN-gamma and carnitine of the epididymis. RESULTS: There was markedly significant difference in sperm motility in the injection group (P < 0.01). The number of sperms in the bilateral vas injection group was 31, while in the sham operation control and normal groups was 64 and 68, respectively. The contents of IL-6 and IFN-gamma increased, and the carnitine reduced significantly (P < 0.05). However, no significant differences were noted between the control and the normal groups (P > 0.05). The contents of IL-6, IFN-gamma and carnitine in the bilateral vas injection group were 772.7 pg/ml, 350.7 pg/ml and 491.1 mol/L. But the same indexes in the sham operation and normal groups were 308.5 pg/ml, 172. 2 pg/ml and 664. 6 mol/L and 287. 8 pg/ml, 163. 8 pg/ml and 605.5 mol/L. CONCLUSION: The antidromic injection of ethanol from vas to epididymis can not only interfere the environment for sperm maturation but also activate the immunologic cells that secrete many cytokines (CK) in the genital system. All the factors can induce the reduction of sperm motility.

Animals↗

PubMed2001

Fertilization of sea urchin eggs and sperm motility are negatively impacted under low hypergravitational forces significant to space flight.

Sperm and other flagellates swim faster in microgravity (microG) than in 1 G, raising the question of whether fertilization is altered under conditions of space travel. Such alterations have implications for reproduction of plant and animal food and for long-term space habitation by man. We previously demonstrated that microG accelerates protein phosphorylation during initiation of sperm motility but delays the sperm response to the egg chemotactic factor, speract. Thus sperm are sensitive to changes in gravitational force. New experiments using the NiZeMi centrifugal microscope examined whether low hypergravity (hyperG) causes effects opposite to microG on sperm motility, signal transduction, and fertilization. Sperm % motility and straight-line velocity were significantly inhibited by as little as 1.3 G. The phosphorylation states of FP130, an axonemal phosphoprotein, and FP160, a cAMP-dependent salt-extractable flagellar protein, both coupled to motility activation, showed a more rapid decline in hyperG. Most critically, hyperG caused an approximately 50% reduction in both the rate of sperm-egg binding and fertilization. The similar extent of inhibition of both fertilization parameters in hyperG suggests that the primary effect is on sperm rather than eggs. These results not only support our earlier microG data demonstrating that sperm are sensitive to small changes in gravitational forces but more importantly now show that this sensitivity affects the ability of sperm to fertilize eggs. Thus, more detailed studies on the impact of space flight on development should include studies of sperm function and fertilization.

Animals↗

PubMed1985

Effects of pH, lactate, and viscoelastic drag on sperm motility: a species comparison.

Little or no motility is observed when sperm from 5 mammalian species are incubated in vitro in their cauda epididymal fluid (CEF). We examined the effects of pH, lactate, and viscoelastic drag on sperm motility to determine whether these factors are responsible for this inhibition of motility. The pHs of CEF from bull, dog, rat, guinea pig, and hamster were 5.8, 6.2, 6.9, 6.9, and 7.2, respectively. The lactate concentration of epididymal semen collected from anesthetized animals ranged from 0.6 to 0.9, but increased almost 10-fold in samples from rats or dogs when measured 2 h postmortem. Increasing the pH of CEF to 7.0 resulted in the initiation of full motility for bull and dog sperm. Suspensions of sperm in buffer at various pHs (from 4.0 to 7.6) produced a sigmoidal motility curve for all species. All species, including bull and dog, showed almost full motility in buffer at a pH equal to the pH of their own CEF. Motility of bull and dog sperm showed greater inhibition with decreasing pH when suspended in CEF instead of buffer. The addition of 15 mM lactate, which has been shown to lower sperm intracellular pH, shifted the motility versus pH curves of all species toward higher pH. In bull and dog the addition of lactate produced a motility profile that was indistinguishable from that in their own CEF. The viscoelastic drag of the CEF of only two species, rat and hamster, was sufficiently high to inhibit sperm motility. We conclude that the low pH of the CEF from bulls and dogs plus the presence of lactate is sufficient to cause inhibition of motility.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid-Base Equilibrium↗

PubMed1998

Bull sperm motility and membrane integrity in media varying in osmolality.

Studies were designed to evaluate bull sperm motility and membrane integrity following exposure to Tyrode's solution varying in osmolality from 100 to 1537 mOsm. Congo red and bisbenzimide (HOECHST 33258) stains were used to distinguish between sperm with intact versus disrupted plasma membranes. Sperm motility was subjectively evaluated. No significant differences were found between motile and unstained sperm (sperm with intact membranes) in solutions with nearly physiologic osmolalities between 200 and 300 mOsm. However, in 100- and 150-mOsm solutions, the percentage of motile sperm (5 and 19%), respectively) was lower than the percentage of sperm unstained with Congo red (18 and 35%). With HOECHST 33258 stain, the corresponding values were 7 and 14% versus 26 and 31%. The percentage of motile sperm declined greatly in the 500-mOsm medium, but the proportion of unstained sperm was affected little until the osmotic pressure exceeded 732 mOsm. Partial recovery of motility occurred when sperm were returned to the isotonic medium. This study indicated that the sperm plasma membrane was more resistant to osmotic damage than were the mechanisms responsible for sperm motility. Lack of motility in hypertonic media was not an absolute indicator of cell death, and unstained sperm overestimated sperm viability, which are factors to consider when sperm are being evaluated, especially relative to cryopreservation.

Animals↗

PubMed1990

Time-related decline in sperm motility patterns in men with cytotoxic antibodies.

Sperm motility patterns in semen from 10 fertile nonautoimmune men (fertile control group) and 33 infertile men with various titers of cytotoxic sperm antibodies in their seminal plasma (group 1: titers less than or equal to 16, n = 6; group 2: titers 64 to 512, n = 12; group 3: titers greater than or equal to 1024, n = 15) were evaluated every 2 hours for 12 hours and finally at 24 hours. A significant decline in sperm swimming speed and linearity was observed at 6 hours in semen from 27 infertile men with sperm antibodies. Beginning at 8 hours, semen from sperm antibody-positive men in group 2 showed a significant decline in percentage motile sperm (p less than 0.001) compared to the fertile controls. The percentage motility in semen of donors in groups 1 and 3 was significantly lower than that in semen of fertile donors at 10 hours (p less than 0.05), 12 hours (p less than 0.01), and 24 hours (p less than 0.001). The mean velocity in groups 2 and 3 was significantly less than that in fertile controls at 10 and 12 hours (p less than 0.05). The linearity of sperm motility started to decline 4 hours after semen samples were obtained from sperm antibody-positive groups 2 and 3 in contrast to sperm antibody-negative fertile or infertile groups (p less than 0.05). It is concluded that the presence of cytotoxic sperm antibodies in the seminal plasma adversely affects sperm linearity as early as 4 hours after semen collection.(ABSTRACT TRUNCATED AT 250 WORDS)

Autoantibodies↗

PubMed1983

Caffeine and sperm motility.

Six different doses of caffeine were used to study its effect on human spermatozoal motility. The recently developed method for assessment of sperm motility by videomicrography was used to determine the percentage of sperm motility, swimming speed (velocity), percentage of progressive sperm, and progressive swimming speed. In doses of 3 and 6 mM/ml, caffeine significantly increased the percentage of motile sperm, but it had no influence on sperm velocity. In doses greater than 6 mM/ml, caffeine exhibited a slight insignificant stimulation that reversed later into an inhibitory effect on all parameters of sperm motility in a dose of 60 mM/ml. Reaching a dose of 120 mM/ml, caffeine caused complete immobilization of human spermatozoa.

Caffeine↗

PubMed2004

CatSper gene expression in postnatal development of mouse testis and in subfertile men with deficient sperm motility.

BACKGROUND: The search for Ca2+ channels residing in sperm has led to the recent cloning and characterization of a novel gene, named CatSper, which codes for a unique Ca2+ channel expressed exclusively in the testis. It plays an essential role in sperm motility, penetration into the oocyte, and ultimately in male fertility. In this study, we assessed the temporal profile of CatSper gene expression during mouse testis development and performed a semi-quantitative evaluation of expression levels in a group of subfertile men which lack sperm motility. METHODS: A small piece of testicular tissue obtained by either multi-site testicular biopsy or orchidectomy was used for semi-quantitative RT-PCR of CatSper and beta2-microglobulin (beta2m, as an internal control) genes. RESULTS: Our results reveal that: (i) the expression of mouse CatSper is developmentally regulated with a direct correlation between CatSper expression and mouse sexual maturation. CatSper gene expression is first detected at 3 weeks of age and coincides with the appearance of round spermatids in the developing mouse testis. (ii) There is a significant reduction in the level of CatSper gene expression (up to 3.5-fold difference) among patients which lack sperm motility as compared with patients whose infertility cannot be ascribed to a deficiency in motility (used as a control). CONCLUSIONS: The data obtained in this study support a potential role for CatSper in sperm motility and fertility in mouse and human. CatSper is therefore implicated as a potential target to explore the molecular mechanisms of male infertility.

Adult↗

PubMed1998

Characterization of recombinant precursor proteins of the human seminal plasma sperm motility inhibitor synthesized in insect cells.

Human seminal plasma sperm motility inhibitor (SPMI) proteins which are exclusively secreted from seminal vesicles, inhibit sperm motility. It is secreted as biologically active 52 kDa and a mixture of 71 and 76 kDa precursor forms, which are identical to semenogelin-I and II (Sg-I and Sg-II), respectively. To understand the molecular mechanism underlying the inhibition of sperm motility by SPMI proteins, we expressed human Sg-I and Sg-II genes in insect cells using a baculovirus system. The baculoviruses expressing full-size Sg-I and Sg-II proteins that were N-terminally-tagged with a hexahistidine were selected, and were infected with Sf 21 cells. The Sg-I and Sg-II proteins were purified from infected cells by column chromatography using Ni-NTA resin 48 h after infection. The full-size Sg-I and Sg-II proteins were obtained in soluble forms. However, they tended to aggregate to form a gel, as expected from naturally occurring semenogelin. Both the purified recombinant Sg-I and Sg-II proteins showed strong SPMI activities with a complete inhibition of sperm motility at 60 units/mg, equivalent to the natural proteins. This production system that permits the generation of purified Sg-I and Sg-II proteins, as well as mutant derivatives, will be helpful for further study on male infertility.

Animals↗