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Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro.

AIM: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. METHODS: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 deg. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. RESULTS: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. CONCLUSION: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.

Adult↗

[Effects of the gene transfer into sperm mediated by liposomes on sperm motility and fertilization in vitro].

To clarify the problems of gene transfer in the fertility clinic, we investigated the safety and effectiveness of the gene transfer of mouse sperm using the lipofection method. We used the pCAGGS-lacZ plasmid connected with cytomegarovirus enhancer/chicken beta-actin promoter as an adventitious gene. After incubation of the DNA-liposome complex and mouse sperm of epididymis in the medium, we investigated the sperm motility after gene transfer, in vitro fertilization rate of the gene transferred sperm, and gene expression of the fertilized ovum and fetus. Sperm motility was lowered significantly with the increase in DNA density. The fertilization rate for in vitro fertilization was lowered significantly with the increase in DNA density. Gene expression was observed in some of the fertilized ova. In the fetus 12.5 days after embryo transfer, gene transfer was confirmed by the PCR method, but we could not find gene expression by X-gal staining. We proved that the sperm with a gene transferred by the lipofection method could fertilize the ovum and transfer adventitious gene to ova. We speculate that sperm motility and fertility rate were lowered because the introduction of DNA and the DNA-liposome complex destroyed parts of the cell. Development of a sperm fractionation method for DNA transfer and establishment of a method for effective and safe DNA transfer are awaited.

Animals↗

Seminal prolactin and its relationship to sperm motility in men.

Semen assessment and levels of serum and seminal plasma prolactin (PRL) were determined in 81 men. In subjects with both normal sperm concentrations and normal sperm motility, the levels of serum and seminal plasma PRL were 14.7 +/- 1.48 and 10.0 +/- 1.05 ng/ml (mean +/- standard error of the mean, SEM, P less than 0.05), respectively. This difference is not observed either in subjects with polyzoospermia with normal sperm motility, or in those with oligozoospermia. Serum PRL was higher in azoospermia and also in subjects with lower levels of seminal citric acid. Seminal plasma PRL was directly related to sperm motility (r = 0.70, P less than 0.01) and inversely related to sperm concentration (r = -0.42 P less than 0.05). Seminal PRL was increased in subjects with higher levels of seminal citric acid and decreased in subjects with lower levels of corrected seminal fructose. Serum and seminal plasma PRL did not change significantly in subjects with different concentrations of serum testosterone.

Adult↗

[Correlation of total antioxidant capacity in seminal plasma with sperm motility of infertile men].

OBJECTIVE: To evaluate the correlation of total antioxidant capacity (TAC) in seminal plasma with sperm motility in infertile men, and explore the clinical significance of TAC in seminal plasma in male fertility. METHODS: One hundred and thirteen infertile men with normal sperm density were included in the experiment group and 28 fertile men in the normal control. The seminal parameter analysis was performed by computer-assisted semen analysis (CASA) system. Seminal plasma TAC was measured with spectroscopic analysis. RESULTS: Seminal plasma TAC was (14.37 +/- 8.45) U in the infertile men with normal sperm density and (19.82 +/- 6.33) U in the fertile control. Compared with the fertile men, seminal plasma TAC in the experiment group was significantly lower (P < 0.01). There was significant correlation between seminal plasma TAC and sperm motility, grade a sperm (r = 0.208, P < 0.05), grade (a + b) sperm (r = 0.231, P < 0.05), straightness (STR) (r = 0.200, P < 0.05), linearity (LIN) (r = 0.208, P < 0.05), curvilinear velocity (VCL) (r = 0.189, P < 0.05), straight line velocity (VSL) (r = 0.210, P < 0.05), average path velocity (VAP) (r = 0.215, P < 0.05), and beat cross frequency (BCF) (r = -0.248, P < 0.01). There was no significant correlation among the average motion degree (MAD), the amplitude of lateral head displacement (ALH) and wobbly (WOB). CONCLUSION: TAC in seminal plasma is closely related to male fertility, appropriate TAC provides a favourable environment for sperm swimming. The decreased level of TAC in seminal plasma may be one of the causes of male infertility. The analysis of TAC in seminal plasma may afford valuable evidence in exploring the mechanism of male infertility and in clinical medication.

Adult↗

Fluorescence supravital stain of human sperm: correlation with sperm motility measured by a transmembrane migration method.

We developed a supravital stain of human sperm with fluorescent dyes. Either Hoechst 33,258 or fluorescein isothiocyanate could be used, the former stained sperm head while the later stained the whole sperm. Sperm vitality assessed with any of these two fluorescent dyes correlated well with that determined by eosin-nigrosin counterstain. When sperm vitality was compared with sperm motility measured with a transmembrane migration method, we found that many vital sperm were immotile because sperm vitality was higher than sperm motility in tested samples.

Bisbenzimidazole↗

Pathophysiology of sperm motility.

Mammalian spermatozoa acquire the ability to swim during their transit from the testis to the oviduct under the control of several external and intracellular factors. These factors play also a pivotal role in regulating acquisition of hyperactivated motility and during the process of chemotaxis. This review summarizes the involvement of such factors in acquisition and maintenance of sperm motility, hyperactivation and chemotaxis, focusing in particular on the molecular bases of asthenozoospermia, a pathology of seminal plasma characterized by reduced sperm motility, which is one of the main causes of male infertility. Current in vitro treatments of asthenozoospermia are shown, together with the most recent findings on pharmacological and physiological molecules capable of stimulating sperm motility. The structure, function and mechanism of sperm flagellum responsible for the development of active motility are also analyzed in details.

Allergy and Immunology↗

Longitudinal study of semen quality of unexposed workers. Sperm motility characteristics.

As part of a longitudinal study of human semen characteristics of unexposed workers, sperm motility measurements were made using computer-assisted sperm analysis. Motility analyses were conducted on monthly samples from 46 men for 9 months. Measurements of curvilinear velocity, straight-line velocity (VSL), linearity, amplitude of lateral head displacement (ALH), and beat-cross frequency were collected in eight microscope fields for each semen sample. The variability within a sample, between samples from the same individual (between monthly samples), and between individuals were calculated using a nested analysis of variance. For all sperm motility measurements, at least 90% of the variation was observed between cells within a semen sample. For all variables, the component of variation between subjects was the smallest percentage (ranging from 1.3% for ALH to 4.0% for VSL). When sample means were used in the nested analysis of variation, at least 75% of the variation was observed between samples from the same individual. These results will be useful in power calculations for future studies.

Adult↗

Stabilization of adenylate energy charge and its relation to human sperm motility.

The adenylate energy charge of human ejaculated spermatozoa was studied when the sperm motility was perturbed by varying pH, prolonged incubation, and caffeine. Between pH 8 and 9, which was optimal for the sperm motility, the energy charge was in the physiological range of 0.8 to 0.9. Above pH 9, the mobility, ATP content, and adenine nucleotide pool declined rapidly but the energy charge was maintained slightly below 0.8. Below pH 8, the motility also dropped drastically, but the ATP, nucleotide pool, and energy charge fell only slightly. Prolonged incubations of the spermatozoa decreased the motility, ATP, and nucleotide pool. However, the energy charge would remain above 0.6. Caffeine stimulation of the motility caused a rapid fall of ATP and the reduction of the physiological energy charge by 0.2 unit, unless glucose was added. Imidazole which reduced the caffeine-stimulated motility did not alter the physiological energy charge of the spermatozoa. The study showed that the spermatozoa could maintain the energy charge above 0.6 under stress.

Adenine Nucleotides↗

Inhibitory action of halothane on rat masculine sexual behavior and sperm motility.

Adult male rats were exposed to inhale halothane in the following regime: 15 ppm/4 h/5 days/week/9 weeks. Sexual behavior observations and sperm motility test were made before halothane exposure (0 days) and at 15, 30, 45 and 60 days of exposure. Fifteen days after halothane exposure, this anesthetic inhibited the proportion of animals displaying ejaculation. In those animals ejaculating, halothane produced an inhibition of masculine sexual behavior reflected as an increase in the intromission latency, number of mounts and postejaculatory interval. At 30 days after exposure, only an increase in the intromission latency was observed. At 45 and 60 days, the inhibitory effect of halothane on sexual behavior disappeared. Similarly, at 15 and 30 days, but not at 45 or 60 days of halothane exposure, a reduced sperm motility was observed. Such transient effects of halothane suggest the development of tolerance to the inhibitory actions of this anesthetic on sexual behavior and sperm motility. These halothane effects are in line with an inhibition of masculine sexual behavior after stimulation of the GABAergic system.

Administration, Inhalation↗

Seminal immunoreactive relaxin in domestic animals and its relationship to sperm motility as a possible index for predicting the fertilizing ability of sires.

Although immunoassayable relaxin has been detected in human and boar seminal plasma, there is no evidence suggesting the existence of immunoreactive relaxin in the seminal plasma of other domestic animals. The first objective of this study was to determine whether immunoreactive relaxin was present in the seminal plasma of bulls, rams and he-goats. In addition, the correlation of immunoreactive relaxin with sperm motility as an index for predicting the fertilizing ability of bull sires was investigated. Semen with normal sperm motility was collected from bulls, rams and he-goats, and the relaxin immunoreactivity of the semen samples was measured using a time-resolved fluoroimmunoassay (TR-FIA) for porcine relaxin that we developed. The presence of relaxin immunoreactivity was demonstrated in seminal plasma from bulls, rams and he-goats. The level of immunoreactive relaxin in seminal plasma was the highest in bulls followed by humans, rams, boars and he-goats in that order, when relaxin levels in boar and human semen having normal sperm motility were also assayed under the same conditions. When the correlation between the seminal plasma level of immunoreactive relaxin and sperm motility was examined in bull semen samples as an index for predicting fertilizing ability, it was found that the relaxin level was significantly correlated with the percentage of spermatozoa showing the most intensive motility (r = 0.64, p < 0.05). These results indicate that immunoreactive relaxin is widely found in the seminal plasma of domestic animals and that measuring the relaxin concentration of seminal plasma may be useful to identify subfertile sires or predict the fertility potential of individual sires.

Animals↗

A possible role of heat shock proteins in human sperm motility.

The testicular spermatogenic stem and seminiferous tubular cells selectively synthesize heat shock proteins (Hsps) during heat stress. Hsps, synthesized from testicular cells and leukocytes, are identical in molecular masses as well as chemical properties. In this study, we induced the Hsps from leukocytes and investigated their in vitro effects on human sperm motility. Semen samples were divided into two parts, washed and unwashed. The whole blood was heated in 43 degrees C for 15 minutes for induction of Hsps. A trans-membrane migration method was used to examine the effect of heated blood plasma on human sperm motility. The main heat-induced proteins of leukocytes were detected by 2-D electrophoresis and Coomassie blue stain. Leukocytes treated by heat produced a large amount of Hsp72 and Hsp80, while only a small amount was observed in that of non-heated leukocytes. The heated blood plasma inhibited motility of washed sperm in a manner that was dose-dependent. In the presence of seminal plasma fluid, however, the inhibitory effects of heated plasma on human sperm motility could not be observed. It was concluded that the heat-induced substance(s) from leukocytes, which being highly possible the Hsps, interfered the mobility of wash human sperm and the inhibition might be antagonized by seminal plasma.

Heat-Shock Proteins↗

Effects of cotinine on sperm motility, membrane function, and fertilizing capacity in vitro.

We evaluated the effect of cotinine on sperm fertilizing capacity in vitro. Human spermatozoa were washed and re-suspended in medium containing albumin and various concentrations of cotinine (0, 100, 200, 400, or 800 ng/ml). After an 8-h incubation period, sperm motility, hypoosmotic swelling test (HOST) outcome, and the percentage of hyperactivated spermatozoa were assayed. Aliquots of spermatozoa were then processed for the zona-free hamster oocyte sperm penetration assay (SPA) or hamster ooplasmic injections. Spermatozoa exposed to concentrations of cotinine equal to 400 or 800 ng/ml demonstrated significantly smaller outcomes for all of the above with the exception of after hamster ooplasmic injections, where high cotinine concentrations did not affect sperm viability or sperm capacity to undergo decondensation and activate hamster oocytes. It appears that cotinine concentrations of 400 or 800 ng/ml exert a detrimental effect on sperm motility, membrane function, and the ability to undergo capacitation. In addition, the current findings suggest that smokers with a high seminal plasma cotinine concentration who participate in assisted reproduction programs may be treated with intracytoplasmic sperm injections (ICSI) rather than conventional in vitro fertilization (IVF) trials.

Animals↗

Treatment of severe male-factor infertility with high concentrations of motile sperm by microinsemination in embryo cryopreservation straws.

A microinsemination technique was evaluated for treating our program's most severe cases of male-factor infertility. Oocytes were inseminated with high concentrations of motile sperm (1 to 9 x 10(6)/ml) in 10 to 150 microliters within embryo cryopreservation straws. Fertilization was obtained in 20 of 29 (69%) couples treated by this technique. In the 15 patients in which only embryos generated from the straw technique were transferred, 7 clinical pregnancies resulted (46.7% per transfer). The implantation rate for couples receiving embryos from the straw technique only (12/58; 20.7%) compared favorably to that observed for other cases treated during this same time period with regular insemination techniques (111/766; 14.5%). Clinical pregnancy rates per transfer for IVF-ET, TET, and PROST were 33.0% (1/3), 0% (0/2), and 60.0% (6/10), respectively. The percentage of polyploidic embryos was significantly lower (P less than 0.0001) for male-factor patients treated by the straw technique with high sperm concentrations than for non-male-factor patients treated during this same time period with standard sperm concentrations. Normal births have resulted from straw inseminations with 3.4 x 10(6) and ongoing pregnancies with 5.0 x 10(6) motile sperm/ml. The results of this study suggest that some cases of male-factor infertility can be successfully treated by insemination with high concentrations of motile sperm in embryo cryopreservation straws. A technique of centrifuging sperm in straws was also developed to concentrate the entire fraction of washed sperm into 10 microliters. Further development of this technique may allow treatment of more severe cases of oligo/asthenospermia by microinsemination with high concentrations of motile sperm than is presently possible with standard washing techniques.

Adult↗

Factors affecting sperm motility. III. Influence of visible light and other electromagnetic radiations on human sperm velocity and survival.

Specimens of semen from fertile and infertile patients were exposed to different electromagnetic radiations, including visible light, ultraviolet (UV) light, x-rays, and high-frequency radio waves. Sperm motility was analyzed before, during, and after irradiation by the multiple exposure photography (MEP) method. No significant difference was found between controls and specimens exposed to various doses of visible and UV light and x-rays either immediately or several hours after exposure. In contrast to spermatozoa of other species that were reported to be adversely affected by visible and UV light and x-rays, human spermatozoa seem to be highly resistant to similar doses of these radiations. A deleterious influence was observed when high-frequency radio waves were applied to human spermatozoa. This may be attributed to an intracellular diathermic effect. The informative value of this study in relation to routine semen analyses and experimental studies in the physiology and comparative anatomy of spermatozoa is discussed.

Cell Survival↗

Cytogenetic studies in motile sperm from normal men.

Most studies on human sperm chromosomes from normal men involve the heterologous fertilization of zona free hamster eggs by unselected human sperm. In this work, we have performed cytogenetic studies of highly motile sperm, selected by a swim-up method. A total of 505 motile human sperm complements from three normal donors was analysed. The total frequency of sperm with chromosomal abnormalities (10.9%; 6.9% structural aberrations and 4.0% aneuploidy) and the sex ratio (50.4% X:49.6% Y) were similar to those obtained from whole semen samples. Our results indicate that the selection of motile sperm does not imply chromosomal selection.

Adult↗

Purification and characterization of a sperm motility inhibitor in human seminal plasma.

A sperm motility inhibitor from human seminal plasma was purified and characterized. The purification procedure includes dialysis, ion exchange chromatography on SP-Sephadex C-25 and adsorption chromatography on hydroxylapatite. With this procedure, the seminal plasma motility inhibitor was purified 290-fold with a 24% recovery in inhibitory activity. Its molecular weight has been estimated at 18,000 to 22,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, but at 13,000 to 15,000 according to molecular sieving under native conditions. The mobility inhibitor has an isoelectric point pH 9.1. It is stable over a wide range of pH (5 to 10) and at temperatures up to 60 C. The observation that the seminal plasma factor inhibited purified bull dynein ATPase in a concentration-dependent manner may suggest that it blocks the motility of demembranated spermatozoa by interfering with dynein arm function.

Adenosine Triphosphatases↗

The effects of beta-adrenergic drugs on the human sperm motility in vitro. II. The effects of isoprenaline and fenoterol.

Thirty semen samples from men with asthenozoospermia were divided into three groups. 0.025 mM of Isoprenaline and 0.65 x 10(-2) mM Fenoterol were added to the first group, a double dose to the second one and five times higher dose to the last one. The lowest concentration used in our experiment results from the applied intravenous therapeutic doses for people. The sperm motility was evaluated under phase-contrast microscope in the 5, 30, 60, 120 and 240 minute of the experiment. The lowest doses of both drugs were most active in relation to the sperm motility in 120 minute after their administration, but the effect appeared to be much higher for Isoprenaline. Isoprenaline resulted in the higher increase of the sperm motility after the addition of the double dose in 240 Minute of the experiment. The effects of the applied B-adrenergic drugs on the sperm motility were discussed.

Adult↗

Enhancement of sperm motility by bradykinin and kinin analogs.

Washed human spermatozoa were incubated with different concentrations of bradykinin, des-arg9-bradykinin, and Ile-Ser-bradykinin (T-kinin). Bradykinin significantly increased the total motility of human spermatozoa at a very low concentration (10(-9) M corresponding to about 1 ng/ml). Enhancement of sperm motility was dependent on the bradykinin concentration, and the effect showed saturation at concentrations above 10(-8) M bradykinin (i.e., sperm motility does not increase further with increasing bradykinin concentration). In the concentration range between 10(-11) to 10(-6) M, des-arg9-bradykinin and T-kinin showed no significant change in motility of washed human spermatozoa. The low effective concentration and the saturation of the effect at higher concentrations of kinin makes it probable that the action of bradykinin is mediated by a specific bradykinin receptor. The kinin receptor of human spermatozoa should be of subtype B2 because the specific B1-agonist, des-arg9-bradykinin, showed no influence on sperm motility.

Bradykinin↗