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PubMed1989

Measurement of boar sperm motility by the trans-membrane migration method.

The conventional microscopic methods for evaluating sperm motility of domestic animals are mostly inadequate due to their subjectivity and lack of precision. Recently, a trans-membrane migration method, originally developed for the examination of human sperm motility, has substantially overcome these problems. This study investigated the applicability of the method to boar sperm motility measurement. The apparatus used was simple and consisted only of syringe plungers, poriferous membranes, and modified multi-well culture plates. It measured the proportion of sperm in the semen that moved across the membrane after incubation at 37 degrees C for 3 hr. The sperm motility as measured by this method correlated well with that measured by direct microscopic examinations. The measurement was more reliable using an 8-microns instead of a 5-microns pore-size membrane. The method was found to work equally well for the sperm motility measurement of the semen with a sperm concentration between 1.5 x 10(8)/ml and 6.0 x 10(8)/ml. The results indicate that this method is a simple, objective, quantitative, and reproducible design for the measurement of boar sperm motility.

Animals↗

PubMed1997

Sperm interaction with fallopian tube apical membrane enhances sperm motility and delays capacitation.

OBJECTIVE: To understand the effect of sperm contact with the apical plasma membrane of tubal epithelial cells on sperm motility, velocity, and capacitation. DESIGN: Prospective, controlled in vitro study. SETTING: University medical center. PATIENT(S): Women of reproductive age undergoing hysterectomy for benign gynecologic indications and normozoospermic donors of proved fertility. MAIN OUTCOME MEASURE(S): Sperm motility as measured manually, velocity as measured by computer-assisted sperm motility analysis, and capacitation status as measured by the chlortetracycline fluorescence assay. RESULT(S): Sperm incubated with apical membrane vesicles had a significantly higher motility at 12 (87.4% +/- 3.4% versus 69.2% +/- 4.8% [mean +/- SEM]), 24 (85.2% +/- 3.1% versus 60.5% +/- 7.2%) and 48 hours (78.9% +/- 5.3% versus 42.4% +/- 11.3%) compared with control (sperm incubated with human tubal fluid media in the absence of apical membrane vesicles) (n = 4). Progressive velocity was significantly higher at 12 (78.2 +/- 1.4 versus 61.7 +/- 16.1 microns/s) and 24 (66.2 +/- 3.9 versus 34.4 +/- 9.8 microns/s) hours (n = 4). Incubation with apical membrane vesicles significantly slowed the transition from uncapacitated to capacitated chlortetracycline fluorescence pattern (n = 5). CONCLUSION(S): Contact with the apical plasma membrane of tubal epithelial cells enhances sperm motility and delays sperm capacitation in vitro.

Acrosome↗

PubMed2003

Requirement of Ca2+ on activation of sperm motility in euryhaline tilapia Oreochromis mossambicus.

Euryhaline tilapia Oreochromis mossambicus acclimates to the external spawning environment by modulating its mechanism for regulating sperm motility. Adaptation of sperm was performed by acclimating fish in various environments. In this paper, regulatory mechanisms of freshwater-acclimated tilapia were studied in detail. Tilapia sperm motility was vigorous in hypotonic conditions and decreased with increasing osmolality. Sperm motility was reduced in hypotonic conditions when extracellular Ca(2+) was chelated; however, extracellular Ca(2+) was not a major factor for motility activation since sperm were motile even when extracellular Ca(2+) levels were nominally depleted by EGTA. The Ca(2+) indicator, fluo 3, showed that intracellular [Ca(2+)] increased on motility activation independently of extracellular [Ca(2+)], accompanied by swelling of the sperm neck region called the sleeve structure. Intracellular [Ca(2+)] was not increased under hypertonic conditions, in which sperm were immotile, even on addition of extracellular Ca(2+). It is possible that Ca(2+) is stored in the neck region. Demembranated sperm were reactivated in the presence of Ca(2+), but cAMP failed to reactivate the motility. Furthermore, we detected phosphorylation and dephosphorylation of three proteins at serine and threonine residues on motility activation. It is likely that hypotonic shock causes an increase in intracellular [Ca(2+)] that activates motility activation via phosphorylation of some flagellar proteins.

Acclimatization↗

PubMed1990

The inhibitory effect of anticonvulsants on human sperm motility: measured with a trans-membrane migration method.

The in-vitro effects of four anticonvulsant drugs (phenytoin, phenobarbitone, carbamazepine and valproate) on human sperm motility were studied with a trans-membrane migration method. Sperm motility was measured either immediately after semen had been mixed with the drug or after a 2-hour pre-incubation at 37 degrees C. When the drug effect was evaluated after the semen-drug mixture had been pre-incubated for 2 hours, the concentration of phenytoin, carbamazepine and valproate that inhibited sperm motility to 50% of control (EC50) was 1.59, 4.23, and 5.00 mM, respectively. If sperm motility was immediately measured without preincubation, the EC50 was increased to 8.47 and 100.00 mM for carbamazepine and valproate, respectively, whereas phenytoin could not inhibit sperm motility to less than 50% of the control at the tested concentrations. Both with and without pre-incubation, phenobarbitone, even up to 12.92 mM, could not inhibit sperm motility to less than 50% of the control. The result was compatible with previous findings that drugs with a membrane stabilizing effect may inhibit human sperm motility. This study provided further information regarding the application of human sperm motility as a cellular model for future pharmacological research.

Adult↗

PubMed1989

Regulation of mammalian sperm motility.

The physiological regulation of sperm motility has become more amendable to investigation since the demonstration that cAMP and calcium play a role in modulating the functioning of the flagellar axoneme. Although the external triggering mechanisms that initiate motility and capacitation are still unknown, evidence supports a modification of the calcium balance by gated Ca2+ channels, accompanied by shifts in the internal pH. Ca2+ and pH may in turn act indirectly through cAMP and cAMP-dependent kinase (kinase(a] to control the phosphorylation state of functional proteins in the flagellar axoneme. The role of calcium is of central importance, but it is clear that several separate Ca2+-dependent mechanisms are involved. Ca2+ controls the curvature of the sperm flagellum and, so, can change the motility of the sperm from progressive swimming to tumbling. Under the appropriate conditions, calcium appears to have the capacity to deactivate motility by activating phosphodiesterase and phosphatase. The deactivating effect of Ca2+ may be offset under some circumstances by coactivation of adenyl cyclase, so phosphorylation of the axoneme and the motility are maintained. The specific factors determining the predominant calcium effect are not yet known, but internal pH of the sperm may play a major role.

Animals↗

PubMed1990

Sperm motility after passage through the hen's egg yolk medium.

This study aimed to observe the percentage of motility of human spermatozoa after passage through 20 per cent hen's egg yolk medium. Twenty-one samples of freshly ejaculated human semen having a sperm density over 20 million per ml and a percentage of motile sperm over 50 per cent were used in the study. The percentage of sperm motility was estimated under the microscope by conventional method. The sperm motility was evaluated within 30 minutes after ejaculation and every one hour thereafter for 8 hours. After the first observation, the semen was divided into two parts, one part kept as a control and the other used for the experiment. The latter was washed in Tyrode's solution and overlaid on 20 per cent egg yolk column and allowed to migrate toward the bottom of the column for two and a half hours. The spermatozoa at the bottom of the column were brought out and washed in Tyrode's solution. The percentage of the sperm motility was observed every hour for 5 hours. After passing through the 20 per cent egg yolk medium, percentage of sperm motility was observed to increase from 66.2 to 87.2 while that of the control declined to 49.3. Five hours after passing through the egg yolk medium the percentage of motile sperm was still high, an average of 80 as compared to 17.4 for the control. Results of this study indicate that hen's egg yolk has a sperm motility stimulating property. Whether hen's egg yolk can be used as a sperm motility stimulant in the treatment of such conditions as asthenospermia and oligospermia is subjected for further study.

Animals↗

PubMedDate not supplied

Effects of Chinese herbal medicine on sperm motility and fluorescence spectra parameters.

Hochu-ekki-to is a Chinese herbal medicine that has been used for unexplained male infertility with some favorable clinical results. However, the detailed mechanisms of the effect of this medicine on male infertility have not been disclosed. This study investigated the effects of Hochu-ekki-to on sperm motility of human semen. Sixteen semen samples were obtained from 14 normal volunteers. Semen samples were incubated with 1, 10, and 100 micrograms/mL of Hochu-ekki-to solution in 5% CO2, 37 degrees C for 1-6 h, and the motility was compared with that of the control groups (the untreated semen and the semen incubated with phosphate-buffered saline). The motility of sperm in 10 and 100 micrograms/mL of Hochu-ekki-to solution for 1-6 h was significantly higher than that of the control groups. One microgram/milliliter of Hochu-ekki-to solution also maintained higher sperm motility after 4-6 h incubation compared with the control groups. Previous reports showed that sperm motility correlated significantly with fluorescence intensity peaks from both spermatozoa and semen plasma. However, the fluorescence intensity at 622 nm excited at 488 nm from spermatozoa fractions in each group was not significantly different. It would appear that the enhanced sperm motility induced by Hochu-ekki-to is mediated by improvement of the circumstances of semen rather than that of spermatozoa-related factors.

Adult↗

PubMed2003

Characteristics of sperm motility induced on the egg-jelly in the internal fertilization of the newt, Cynops pyrrhogaster.

Most urodeles undergo internal fertilization and sperm are directly inseminated onto the surface of egg-jelly. Feature of sperm motility induced on the egg-jelly was examined in the newt, Cynops pyrrhogaster. When sperm were directly inseminated onto an egg-jelly, sperm motility was immediately induced on its surface. The egg-jelly of C. pyrrhogaster was composed of six sublayers that were added by turns in oviduct. When the eggs with various sets of the sublayers were obtained and sperm were inseminated onto the egg-jelly, the immediate activity for the initiation of sperm motility was observed only on the outermost sublayer. Similarly, the immediate initiation of sperm motility was induced in the sperm suspended in the extract of the egg-jelly (JE). The initiation of sperm motility was affected by the external pH, and the motility was activated in the moving sperm. A K(+)-channel antagonist, charybdotoxin (CTX), or a Ca(2+)-channel antagonist, gallopamil inhibited the initiation of sperm motility in a dose dependent manner. These results demonstrated the feature of the mechanism regulating sperm motility under stable surroundings in the internal fertilization of amphibians.

Animals↗

PubMedDate not supplied

Effect of leukaemia inhibitory factor on long-term sperm motility and survival.

Leukaemia inhibitory factor (LIF) is expressed at high constitutive levels in the human Fallopian tubal epithelium. In this study, the effect of human recombinant LIF on sperm motility and survival in vitro was investigated. Human spermatozoa were incubated in sperm washing medium that contained various concentrations of LIF at 37 degrees C and under 5% of CO(2) in air for up to 48 h. Sperm motion characteristics were measured using a sperm motility analyser. Sperm survival was determined by the hypo-osmotic swelling test. The effect of LIF on sperm motility was concentration-dependent and maximal effect was observed at a concentration of 5 ng/ml. Sperm motility was significantly higher after 24 h exposure to LIF compared with control (P < 0.001). Sperm survival was also prolonged in a concentration-dependent manner. LIF significantly enhanced sperm survival at higher concentrations (10 ng/ml) and the result was significant after 48 h exposure (P < 0.05). LIF increased long-term sperm motility and survival in vitro.

Dose-Response Relationship, Drug↗

PubMed2003

Sperm mitochondria of patients with normal sperm motility and with asthenozoospermia: morphological and functional study.

Studies were performed on ejaculated human spermatozoa (32 subjects with normal sperm motility and 25 subjects with low sperm motility). Morphology of sperm midpiece was evaluated in light, fluorescent and transmission or scanning electron microscope. Changes in mitochondrial membrane potential (delta(psi)m) and mass of mitochondria were analysed by flow cytometry using mitochondrial specific probes JC-1 and Mito Tracker Green FM. Moreover, oxidoreductive capability of sperm mitochondria was assessed using cytochemical reaction for NADH-dependent dehydrogenases. In flow cytometry analysis of JC-1-stained spermatozoa, two asthenozoospermic subpopulations were distinguished: patients with a high percentage (76 +/- 11%, 13 subjects) and patients with a low percentage (29 +/- 14%,12 subjects) of spermatozoa with functional-polarized mitochondria with high delta(psi)m. Our microscopic investigations of spermatozoa of seven asthenozoospermic patients reveal that the deformed and unusually thickened sperm midpieces (50-70% of cells), occasionally with persistent cytoplasmic droplet, contain supernumerary mitochondria with normal substructure, full oxidoreductive capability and high delta(psi)m. The midpiece deformations cause nonprogressive movement or immotility. They can also appear in smaller number of spermatozoa (5-35% of cells) in patients with normal sperm motility. Moreover, in three cases of asthenozoospermia midpiece malformations were accompanied by abnormal morphology of outer dense fibers and axoneme. The cytochemical, fluorescence and SEM studies showed the absence of midpieces in many (60-80%) spermatozoa in some other cases of asthenozoospermia. The morphological observations corresponded with flow cytometry analysis of Mito Tracker Green FM-stained spermatozoa. Our results suggest that in some cases of asthenozoospermia the sperm mitochondria can be functionally active and display high delta(psi)m in large number of cells. The results may suggest that asthenozoospermia does not necessarily result from energetic disturbances of sperm mitochondria. The low sperm motility may be associated with deformations of the mitochondrial sheath containing functional mitochondria. The combination of fluorescence microscopy and flow cytometry with electron microscopic investigations is a sensitive, precise and comprehensive examination which helps discover sperm abnormalities responsible for asthenozoospermia.

Adult↗

PubMed1995

Nitric oxide inhibition of human sperm motility.

OBJECTIVE: To determine the effect of nitric oxide (NO) on sperm motility in vitro. DESIGN: Normal human sperm separated by centrifugation through a discontinuous Percoll gradient and subsequent swim-up were incubated for up to 24 hours with NO donors, with and without the known NO quencher hemoglobin, as well as with agents that raise intracellular cyclic 3',5'-guanosine monophosphate (cGMP). Sperm respiration was determined by a tetrazolium-formazan spectrophotometric assay. SETTING: Andrology laboratory. MAIN OUTCOME MEASURES: Absolute sperm motility and respiration. RESULTS: Sperm incubated with the NO donors 1 mM nitroprusside, 100 to 125 microM 3-morpholinosydnonimine, and 25 to 125 microM pure nitric oxide gas dissolved in buffer were inhibited in motility in a dose-dependent fashion. The inhibition could be reversed by the NO quencher hemoglobin. Agents that raise cellular cGMP (dibutyryl cGMP or 8-bromo-cGMP) did not inhibit motility. Nitric oxide inhibited sperm respiration, as measured by the tetrazolium-formazan assay. CONCLUSIONS: Nitric oxide reduces sperm motility, possibly by a mechanism involving inhibition of cellular respiration independent of an elevation of intracellular cGMP. Nitric oxide elaborated in the female or male genital tract in vivo could adversely influence sperm function and fertility.

Cyclic GMP↗

PubMed2005

Concerted action between Ca2+ and hyperosmolality initiates sperm motility in amphioxus Branchiostoma belcheri tsingtauense.

This study investigated the effects of different environmental conditions on the initiation and maintenance of sperm motility in amphioxus Branchiostoma belcheri tsingtauense. The findings were that: (1) hyperosmolality in the absence of Ca(2+) ions did not initiate amphioxus sperm motility; (2) addition of Ca(2+) into EGTA-containing Ca(2+)-free artificial sea water (ASW), in which no sperm were motile, restored sperm motility; (3) Ca(2+) failed to induce sperm motility under conditions of hypoosmolality; (4) K(+) channel blockers quinine and 4-aminopiridine did not suppress the initiation of sperm motility; and (5) changes in pH did not cause sperm motility in a solution isotonic to seawater without Ca(2+). In conclusion, we inferred that a concerted action between Ca(2+) and hyperosmolality was essential to initiate motility of amphioxus sperm, whereas K(+) and pH were indispensable to maintain motility.

4-Aminopyridine↗

PubMedDate not supplied

Effect of cytokines on sperm motility and ionophore-stimulated acrosome reaction.

The effects of soluble products from leukocytes (WBC) on sperm motility and calcium ionophore-stimulated acrosome reaction (AR) were examined. Supernatants of mononuclear WBC, isolated from peripheral blood and stimulated by the lectins Phytolacca americana (pokeweed mitogen) or concanavalin A, caused a weak but significant inhibition of progressive sperm motility. The recombinant cytokine interferon-gamma (IFN-gamma) in high concentrations inhibited motility of sperm from 5 of 8 donors tested. The recombinant cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) did not show any effect on sperm motility. Reactive oxygen species (ROS) produced by stimulation of polymorphonuclear WBC with a phorbol ester (PMA) tended to inhibit sperm motility. Neither supernatants from lectin-stimulated mononuclear WBC, IFN-gamma, TNF-alpha, IL-8, nor ROS showed any significant effect on the ionophore-challenged AR.

Acrosome↗

PubMed1985

Acquisition of sperm motility and its maintenance during storage in the lizard, Lacerta vivipara.

Lizard spermatozoa, which are non-motile in the testis, develop the ability to swim as they pass along the excurrent duct. The addition of caffeine, a phosphodiesterase inhibitor, induced forward motility in spermatozoa from the caput epididymidis and increased the velocity of spermatozoa from the distal part of the epididymis. Caffeine had no effect on the motility of testicular spermatozoa. This suggests that sperm motility in this species is cyclic AMP-dependent but this factor alone is not sufficient to induce testicular sperm motility. In samples from the distal region of the epididymis, sperm motility was maximal in April just after the breeding season and then decreased significantly during the following months. A parallel can be drawn between these data and the levels of testosterone in the plasma. In the lizard, as in mammals, the epididymis may play an important role in the maturation of spermatozoa.

Animals↗

PubMed1998

Human sperm NADH and NADPH diaphorase cytochemistry: correlation with sperm motility.

OBJECTIVES: We have examined the correlation between the retention of residual sperm cytoplasm and sperm motility in semen from men presenting for infertility evaluation. METHODS: Semen samples (n = 12) were obtained from nonazoospermic men presenting for infertility evaluation at our institution. Samples were fractionated into high-, intermediate-, and low-density subpopulations by Percoll gradients in order to examine the correlation between the retention of residual sperm cytoplasm and sperm motility. Residual sperm cytoplasm retention was detected by cytochemical staining of sperm for nicotinamide adenine dinucleotide (NADH)- or nicotinamide adenine dinucleotide phosphate (NADPH)-dependent diaphorase activity. RESULTS: The different sperm subpopulations (low, intermediate, and high density) had significantly different percentages of sperm with droplet retention (analysis of variance, P < 0.05). Using either NADH or NADPH diaphorase staining as a marker of the cytoplasmic space, a significant negative correlation was observed between the percentage of sperm with residual cytoplasmic droplets and the percentage of motile sperm (r = -0.58 and -0.61, respectively, P < 0.05). CONCLUSIONS: Assessment of residual sperm cytoplasm retention is a simple diagnostic test. Although this test is of unproven value in the management of infertile men, this and other studies suggest that it may provide useful data on sperm function.

Dihydrolipoamide Dehydrogenase↗

PubMedDate not supplied

Effects of antiepileptic drugs on sperm motility of normal controls and epileptic patients with long-term therapy.

The in vitro effects of four antiepileptic drugs (AEDs) on human sperm motility were studied with a transmembrane migration method. Sperm motility of epileptic patients receiving chronic AED therapy was also investigated. Sperm motility was measured immediately after semen had been mixed with AED and after a 2-h preincubation at 37 degrees C. Both in vitro and in vivo studies demonstrated that AEDs inhibited sperm motility. When the drug effect was evaluated after the semen-AED mixture had been preincubated for 2 h, sperm motility was inhibited to 50% of control at concentrations of 1.59, 4.23, and 5.00 mM for phenytoin, carbamazepine, and valproate, respectively. Both with and without preincubation, phenobarbital, even up to 12.92 mM, did not inhibit the motility to less than 50% of the control. In the in vivo study, poor sperm motility was noted in epileptic patients with long-term AED therapy despite serum levels within the therapeutic range. Shorter duration of activity of spermatozoa was also observed in these patients. Interference with sperm membrane function by AEDs may be the underlying mechanism.

Adult↗

PubMed2000

Cryopreservation of sperm in common carp Cyprinus carpio: sperm motility and hatching success of embryos.

In this study, fish sperm cryopreservation methods were elaborated upon for ex situ conservation of nine strains of Bohemian common carp. Common carp sperm were diluted in Kurokura medium and chilled to 4 degrees C and dimethyl sulfoxide was added. Cryotubes of sperm with media were then cooled from +4 to -9 degrees C at a rate of 4 degrees C min(-1) and then from -9 to -80 degrees C at a rate of 11 degrees C min(-1), held for 6 min at -80 degrees C, and finally transferred into liquid N(2). The spermatozoa were thawed in a water bath at 35 degrees C for 110 s and checked for fertilization yield, hatching yield of embryos, and larval malformations. Fresh and frozen/thawed sperm were evaluated for the percentage and for the velocity of motile sperm from video frames using image analysis. The percentage and velocity of sperm motility at 15 s after activation of frozen/thawed sperm was significantly lower than that of fresh sperm (nine males). ANOVA showed a significant influence of fresh vs frozen/thawed sperm on fertilization rate (P < 0.0001), but differences in hatching rate and in larval malformation (0-6.8%) were not significant, and different males had a significant influence on fertilization and hatching rate (P < 0.003 and P < 0.007, respectively). Multiple range analysis (LSD) showed significant differences between fresh and frozen/thawed sperm regarding fertilization rate (68 +/- 11 and 56 +/- 10%, respectively) and insignificant differences between fresh and frozen/thawed sperm on the hatching rate (50 +/- 18 and 52 +/- 9%, respectively). The percentage and velocity of fresh sperm motility were correlated, respectively, with the fertilization yield of frozen/thawed sperm at the levels r = 0.51 and r = 0.54.

Animals↗

PubMed1999

Effect of panax notoginseng extracts on inferior sperm motility in vitro.

The purpose of this study was to investigate the effects of Panax notoginseng extracts on inferior sperm motility in vitro. Semen samples were collected from 23 patients with sperm motility between 20% and 40%. The sperm count was over 20 x 10(6)/ml in accordance with the World Health Organization standard. 1.0 mg/ml and 2.0 mg/ml of Panax notoginseng extracts including aqueous extract, n-butanol extract, and polysaccharide fraction on sperm motility and progression were evaluated by computer assisted semen analysis. The results demonstrated that sperm motility as well as progression on inferior sperm motility were enhanced at 1 hour and 2 hours after incubation with all three types of extracts.

Adult↗