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PubMed1995

[Effect of caffeine and pentoxifylline on human sperm motility after one week storage at 3 degrees C].

The effect of caffeine (C) and pentoxifylline (P) on the sperm motility was examined after a 3 degrees C storage in a TEST yolk buffer for one week. Thus stored 24 semen samples from infertile patients were incubated with or without C or P at 37 degrees C. The motility was analysed for up to 240 minutes. The poststorage addition of C (1 mM) and P (1 mM) significantly increased the serum motility rate (p < 0.01). The stimulant effect of C and P was rather transient lasting no more than 2 hours. The prestorage addition of C and P in a preservant TEST yolk buffer was less effective than the poststorage administration. According to the time-related profiles of the sperm motility, 24 cases were divided into three groups. In group A (11 cases), the sperm motility rate increased from 30 to 90 minutes even without C or P. This group was responder of C and P. In group B (8 cases), the motility rate increased only in response to C and P and, in group C (5 cases), no such responses were observed. The prestorage semen qualities in terms of the sperm concentration, the motility rate, the velocity, the linearity, and the amplitude of the lateral head displacement were compared in these three groups. There was a significant difference between these 3 groups (p < 0.05) on the parameters in terms of the sperm concentration, motility rate and velocity. It was suggested that these prestorage parameters predict the sperm activity in response to C and P after a one week 3 degrees C storage.

Adult↗

PubMed1993

Earlier testing after vasectomy, based on the absence of motile sperm.

OBJECTIVE: To review the results of postvasectomy testing when clearance was based on the absence of motile sperm and to compare them with regimens based on complete azoospermia. DESIGN: A review of 2,260 seminal assay results from 3,178 consecutive vasectomies performed during a 17-year period. SETTING: An outpatient vasectomy service in a private group practice in suburban Sydney. RESULTS: Clearance was given sooner and with less testing than with other reported regimens, without loss of reliability. CONCLUSIONS: Testing can be done 4 weeks after vasectomy, regardless of the number of postvasectomy ejaculations. If specimens are examined within 12 hours of collection, clearance may safely be given if motile sperm are absent. Repeat tests are essential if any motile sperm remain but are not needed if only nonmotile sperm are found.

Humans↗

PubMedDate not supplied

Reproducibility of sperm motility measurements in asthenozoospermic and normozoospermic individuals using the Hamilton-Thorn motility analyzer.

Computer-assisted semen analysis provides a useful, quantitative method for analyzing sperm movement parameters. However, few experimental data have been published on the measurement characteristics of CASA. This study measured the reproducibility of measurements of sperm motility using a Hamilton-Thorn motility analyzer. Twenty normozoospermic and 20 asthenozoospermic patients provided semen samples. Ten samples from each group were analyzed as unprocessed semen; the remainder were analyzed after washing. An aliquot from each sample was analyzed by CASA 10 times over an 18-min period. The mean coefficient of variation for all sperm motility parameters was less than 10%, except for the percentage of motile cells in the asthenozoospermic unprocessed samples, which was 17.1%. The mean coefficient of variation for percentage motility was reduced to 9.6% in the washed asthenospermic group. Other motility parameters were unaltered by washing in the asthenospermic samples. There were no substantial differences between the washed and unwashed normospermic measurements. Normospermic samples showed better reproducibility in measurement than asthenozoospermic samples. Washing reduced the difference, probably because of the removal of debris in the asthenozoospermic samples. The results indicate that serial measurements of sperm motility in both unprocessed and washed semen samples can be reliably made using the Hamilton-Thorn motility analyzer, and that reproducibility can be improved in asthenospermic samples by washing.

Humans↗

PubMed1997

Role of extracellular and intracellular pH in carp sperm motility and modifications by hyperosmosis of regulation of the Na+/H+ exchanger.

Recently developed flow cytometric methods (Trón et al.: Mol Immunol 27:1307-1311, 1990) to measure the intracellular pH (pHi) and intracellular potassium concentration in mammalian cells by using the fluorescent pH-indicator dye 2',7bis-(2-carboxyethyl)-5 (and-6)-carboxyfluorescein (BCECF) were adopted for measuring these parameters in carp sperm. The intracellular potassium concentration of the carp sperm was 62.4 +/- 5.3 mM. This is very similar to the potassium concentration of the seminal plasma (87 +/- 16 mM), and it suggests a depolarized state of the sperm cell in the semen. An average pHi value of 7.06 +/- 0.11 was obtained by measuring sperm samples taken from ten animals. Changes in the ionic composition of the environment did not alter pHi. Sperm motility was initiated by transferring the cells to an environment of 110 mOsm osmolality. This hypoosmotic shock induced fast changes in the membrane structure that could be reversed by restoring physiologic osmolality. Activation was accompanied by a fast alkalinization of the sperm cells. This pH change was amiloride sensitive, suggesting the involvement of the Na+/H+ exchanger in the activation process. Alkalinization of acid-loaded sperm cells depended on the osmolality of the environment. Equilibrium pHi of these cells in hyperosmotic buffers was substantially lower relative to cells in an isoosmotic environment. Effects of the extracellular and intracellular pH on carp sperm motility were also examined. Extracellular pH below 5.5 abolished sperm motility completely. Alkaline extracellular pH did not alter the duration of sperm motility even at extreme values (pHe = 9.6). Duration of the flagellar motion did not depend on the pHi between values of 6.5 and 8.5; however, it was significantly reduced both below and above this range. No motility was observed below pHi = 6.0 or above pHi = 9.5 with a 10 min incubation time at these pH values prior to activation. Effects of the extracellular and intracellular pH on sperm motility were partially reversible.

Animals↗

PubMed1999

Enhancement or initiation of testicular sperm motility by in vitro culture of testicular tissue.

OBJECTIVE: To describe different techniques of testicular tissue culture and their effect on sperm motility, mainly in cases of totally immotile spermatozoa, and to compare the effect of in vitro culture with that of motility stimulants. DESIGN: Prospective study. SETTING: University teaching hospital. PATIENT(S): Ten patients undergoing testicular biopsy for diagnostic purposes or for intracytoplasmic sperm injection. INTERVENTION(S): Dissected testicular biopsy samples and tissue blocks were cultured at 37 degrees C for up to 96 hours. Immediately after dissection, immotile testicular spermatozoa were incubated for 30 minutes in pentoxifylline and 2-deoxyadenosine. MAIN OUTCOME MEASURE(S): Sperm motility and vitality. RESULT(S): Overall, dissected samples showed improved sperm motility, which peaked within 48 hours of culture. Unlike motility, vitality declined linearly, from 56.3%+/-19% at initial assessment to 18.8%+/-11% at 96 hours. Five samples had initially immotile spermatozoa, of which four acquired motility at 48 hours. In vitro culture showed results comparable with those of incubation with pentoxifylline and 2-deoxyadenosine. Culture of tissue blocks did not improve motility or vitality compared with dissected tissue. CONCLUSION(S): The motility of testicular spermatozoa was enhanced or initiated after in vitro culture. Testicular biopsy culture may be an alternative to the use of motility stimulants to obtain motile spermatozoa for intracytoplasmic sperm injection, particularly when oocytes are not immediately available.

Adult↗

PubMed2000

A role for phosphorylation of glycogen synthase kinase-3alpha in bovine sperm motility regulation.

The long-term goal of our work is to understand biochemical mechanisms underlying sperm motility and fertility. In a recent study we showed that tyrosine phosphorylation of a 55-kDa protein varied in direct proportion to motility. Tyrosine phosphorylation of the protein was low in immotile compared to motile epididymal sperm. Inhibition or stimulation of motility by high calcium levels or cAMP, respectively, results in a corresponding decrease or increase in tyrosine phosphorylation of the 55-kDa protein. Here we report purification and identification of this motility-associated protein. Soluble extracts from bovine caudal epididymal sperm were subjected to DEAE-cellulose, Affi-Gel blue, and cellulose phosphate chromatography. Tyrosine phosphate immunoreactive fractions contained glycogen synthase kinase-3 (GSK-3) activity, suggesting a possible correspondence between these proteins. This suggestion was verified by Western blot analyses following one-dimensional and two-dimensional gel electrophoresis of the purified protein using monoclonal and affinity-purified polyclonal antibodies against the catalytic amino-terminus and carboxy-terminus regions of GSK-3. Further confirmation of the identity of these proteins came from Western blot analysis using antibodies specific to the tyrosine phosphorylated GSK-3. Using this antibody, we also showed that GSK-3 tyrosine phosphorylation was high in motile compared to immotile sperm. Immunocytochemistry revealed that GSK-3 is present in the flagellum and the anterior portion of the sperm head. These data suggest that GSK-3, regulated by phosphorylation, could be a key element underlying motility initiation in the epididymis and regulation of mature sperm function.

Amino Acid Sequence↗

PubMed1996

Purification and characterization of the active precursor of a human sperm motility inhibitor secreted by the seminal vesicles: identity with semenogelin.

Human seminal plasma contains a sperm motility inhibitor that originates from seminal vesicles as a precursor form. This precursor is degraded into smaller peptides by prostatic proteases shortly after ejaculation. The seminal plasma sperm motility inhibitor (SPMI) precursor was purified by a combination of cation-exchange chromatography on S-Sepharose followed by C4 reverse-phase high-performance liquid chromatography directly from seminal vesicle fluid or washed seminal coagulum. The purification procedure yielded a protein of apparent homogeneity, with a molecular mass of 52 kDa by SDS-PAGE. It migrated as a 105-kDa protein by molecular sieving under denaturing conditions. The purified SPMI precursor was digested by the prostatic protease prostate-specific antigen (PSA), causing a 76 +/- 4% drop in biological activity and transformation into low molecular mass SPMI polypeptides (5-20 kDa) similar to those observed in liquefied semen. The N-terminal amino acid sequences of three degradation peptides were obtained by Edman degradation and found to correspond to residues 45-50, 85-90, and 137-143 of semenogelin, a protein characterized as the major structural component of semen coagulum. The amino acid composition of SPMI precursor was found to be almost identical to that of semenogelin. Moreover, the mass of the precursor was estimated at 49,620 daltons by electrospray-ionization mass spectrometry, a value in close agreement with the expected mass of semenogelin according to its cDNA sequence. The SPMI precursor was found to inhibit sperm motility in a dose-dependent manner, with complete immobilization at 500 U/ml of SPMI. The motility of completely immobilized spermatozoa was partially recovered after washing of the cells. The results suggest that SPMI precursor is the major component of the seminal vesicle secretions and seminal coagulum. It can be degraded by PSA in a manner reminiscent of its processing in whole semen. Taken together these results indicate that the SPMI precursor is semenogelin and that intact semenogelin can immobilize spermatozoa.

Amino Acid Sequence↗

PubMed1977

The effects of prostaglandins on sperm motility.

Inflammatory exudates around intrauterine devices were found to produce large concentrations of prostaglandin F2alpha. To evaluate the effect of this substance, 100 semen specimens were incubated in concentrations 100 times greater than that found in normal semen. Motility indices were calculated, and significant inhibition of sperm motility was found. The effect of prostaglandin E on sperm motility is also reviewed. The physiology of prostaglandins and their mechanism of action via the cyclic nucleotide system are discussed.

Humans↗

PubMed1994

Effect of peritoneal fluid supplemented with exogenous progesterone on sperm motility in vitro.

Since progesterone has been claimed to induce acrosomal reaction and hyperactivated motility of human spermatozoa, the present study was undertaken to determine if its presence at concentrations similar to those of peri-ovulatory follicular fluid could influence the effect of peritoneal fluid on sperm motility in vitro. To this end, 11 sperm samples were incubated at 37 degrees C with five peritoneal fluids with/without exogenous progesterone, and sperm motility was assessed using a computer-assisted analyser at time (t) = 0, 2.5, 5 and 24 h. Overall there was no observable constant trend for enhancement or inhibition of sperm motility. Progesterone generally induced a negative effect on those sperm samples with high velocities in the native peritoneal fluids and a positive effect on those sperm samples demonstrating low motility in the native peritoneal fluids. The incorporation of progesterone into the incubation medium seemed to result in a 'tuning' of sperm velocity to around 30-50 micron/s. However, a given sperm sample reacted differently when incubated with various peritoneal fluids and, reciprocally, different semen samples incubated with the same peritoneal fluid showed very variable motility patterns. The greater variability of the effects exerted by progesterone on sperm motility could arise from the fact that each sperm sample may contain subpopulations of gametes with different sensitivity to progesterone.

Adult↗

PubMed1989

Intracellular pH regulates bovine sperm motility and protein phosphorylation.

Bovine sperm in neat caudal epididymal fluid become motile in response to either pH elevation or dilution of the fluid. Buffers containing permeant weak acids at physiologic concentrations are able to mimic these effects of caudal fluid. These observations lead to the hypothesis that a pH-dependent epididymal fluid quiescence factor regulates bovine sperm motility by modulating sperm intracellular pH (pHi). Here we report that sperm pHi, measured with the fluorescent pH probe carboxyfluorescein, increases by approximately 0.4 units in response to either of these motility-initiating manipulations. At least 26 discrete phosphoprotein bands are distinguishable by sodium dodecylsulfate-polyacrylamide gel electrophoresis after incubation of intact caudal sperm with 32PO4. A prominent phosphoprotein, with Mr approximately 255,000 (pp255) and a relatively high specific radioactivity, is reversibly dephosphorylated in response to elevations in pHi that initiate sperm motility. Unlike most of the sperm phosphoproteins, the extraction of pp255 requires reducing agents. This phosphoprotein cosediments with the sperm heads but not the tail, midpiece, soluble, or plasma membrane fractions. No other pHi-dependent phosphorylation changes are apparent in gels of whole sperm extracts. However, subcellular fractionation allows the detection of increased phosphorylation of two plasma membrane phosphoproteins (Mr approximately 105,000 and 97,000) and decreased phosphorylation of another plasma membrane phosphoprotein (Mr approximately 120,000) in response to increasing pHi. This is the first report describing changes in endogenous phosphoproteins from intact motile and nonmotile bovine sperm that are regulated by pHi.

Animals↗

PubMed1976

[The effect of kallikrein on sperm motility (author's transl)].

The effect of kallikrein (240 KE/week i.m. or 600 KE/day orally) on the sperm count, sperm motility and morphology was investigated in 27 patients with pathological ejaculate findings. Kallikrein was administered over a period of 9 to 12 weeks. A significant increase in sperm motility was observed in patients with asthenospermia. No influence was noted on the ejaculate findings in patients with oligospermia. Interference between the kallikrein-kinin system and sperm motility is postulated.

Adult↗

PubMed1980

Effect of in vitro selenium supplementation on bovine sperm motility.

The effects of selenium (Se) on motility of bovine spermatozoa were evaluated in two experiments. In Experiment 1, increasing doses of Se alone or with vitamin E were found to increase motility in Se concentrations up to 1 mug/ml. However, Se alone had a greater effect than did Se with vitamin E. The sperm motility response to Se concentrations from background to 1 mug/ml on motility best fit a quadratic equation with a maximum at about .65 mug Se/ml. In Experiment 2, the effect of Se addition before and after freezing was evaluated. This experiment demonstrated that treatment of semen with Se before freezing significantly increased the post-thaw motility as compared with portions of the same ejaculate which were not treated. Treatment after thawing produced motilities which were intermediate and not significantly different from either pre-freezing supplementation or no supplementation at all. These data indicate that some semen exhibits a greater percentage of motile sperm when Se is added prior to freezing, however, no information is yet available on the fertility of Se-treated semen.

Journal Article↗

PubMed1997

Studies on the relationship between urokinase plasminogen activator (uPA) and human sperm motility.

To clarify the role of urokinase plasminogen activator (uPA) in the mechanisms of regulating sperm motility and the ability of fertilizing, we investigated the quantities and activities of uPA in human seminal plasma and on the membrane of spermatozoa. Semens were harvested from 22 infertile patients with asthenospermia and 20 healthy fertile men according to WHO standards. To quantify the membrane-bound uPA in the samples, polyclonal antibodies against human urokinase were employed by means of a sandwich ELISA. The uPA activities in seminal plasma and on the surface of spermatozoa were determined using A-garose-Fibrine-Plate method and the experiment of immunological identification with polyclonal antibodies against urokinase. In lysates of spermatozoa, significantly lower levels of uPA (23.1 +/- 7.35 mu/10(6) cells) and uPA activity (5.13 +/- 3.85 mu/10(6) cells) were found in patient group as compared to healthy fertile men exhibiting normospermia (29.89 +/- 9.40 mu/10(6) cells and 10.17 +/- 6.18 mu/10(6) cells). In seminal plasma, uPA activity in patient group (2134 +/- 1581.3 IU/L) was also found significantly lower than that of normal group (3365 +/- 1859.5 IU/L). Positive correlations were observed between sperm motility and uPA quantities (r = 0.48, P < 0.005), as well as with uPA activities (r = 0.45, P < 0.005). Thus, it is inferred that membrane associated uPA on human spermatozoa may be related directly to sperm motility and fertility.

Adult↗

PubMedDate not supplied

Stimulation of ejaculated domestic cat sperm motility with caffeine, pentoxifylline, and 2'-deoxyadenosine.

Sperm motility patterns of cryopreserved domestic cat ejaculates treated at 37 degrees C with 1 mM caffeine, pentoxifylline, or 2'-deoxyadenosine were analyzed using computer-assisted semen analysis (CASA). The percent motility (MOT), curvilinear velocity (VLC), straight-line velocity (VSL), linearity (LIN), and amplitude of lateral head displacement (ALH) were measured for each group following 15 min of treatment. Motility indices were examined during a 6-h treatment period to determine the effect of each chemical on sperm longevity. Caffeine, pentoxifylline, and 2'-deoxyadenosine each increased (p > .05) the MOT and VCL of the ejaculates compared to the controls. The longevity of the treated and control samples were not significantly different throughout the incubation period. These results, similar to previous findings with cryopreserved epididymal cat sperm, demonstrate that motility stimulants can significantly elevate the MOT and VCL of cryopreserved ejaculated cat sperm without having deleterious effects on longevity.

Animals↗

PubMed1992

Evaluation of Astroglide, a new vaginal lubricant: effects of length of exposure and concentration on sperm motility.

Traditional vaginal lubricants have been shown to adversely affect sperm motility. Astroglide, a new vaginal lubricant, and K-Y Jelly were tested at varying concentrations to assess their suitability for infertility patients requiring a lubricant. The vaginal lubricants tested impaired sperm motility in a concentration-dependent but not time-dependent manner. We conclude that all traditional vaginal lubricants should be avoided in patients desiring conception. Future studies should attempt to mimic in vivo conditions and focus on concentration-dependent effects.

Cellulose↗

PubMed2002

Reactive oxygen species-mediated loss of bovine sperm motility in egg yolk Tris extender: protection by pyruvate, metal chelators and bovine liver or oviductal fluid catalase.

Improvement of bovine semen cryopreservation requires a better understanding of the properties of the currently used extenders. At present, about half of the spermatozoa die or become immotile following cryopreservation. The implication of an oxidative stress during or following the process of cryopreservation has been suspected to alter sperm functions. However, insufficient information is available on the effect of oxidative stress on sperm functions in their surrounding environment, the extender, such as the one based on egg yolk, Tris and glycerol. In this study, we investigated the effects of hydrogen peroxide (H2O2) and superoxide anion (O2*-) on bovine sperm motility in a widely used egg yolk Tris glycerol (EYTG) extender in comparison to a reference medium, the Tyrode's albumen lactate pyruvate (TALP). Bovine sperm were incubated for 6 h with or without concentrations of H2O2 ranging from 12.5 microM to 1.25 mM and with the hypoxanthine/xanthine oxidase system (X/XOD) that generates O2*-. Sperm motility was established by computer assisted semen analysis (CASA) in four similar experiments using the same frozen pool of semen. We have found that sperm motility was reduced significantly by H2O2 concentrations 20-fold lower in EYTG than in TALP medium. The differential resistance of the two media was explained by pyruvate present in TALP that acts as an antioxidant and metals ions, chelated by ethylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DETAPAC), found in egg yolk that might react with H2O2. Addition of only 5 U/ml of bovine liver catalase or oviductal fluid catalase (OFC) were sufficient to overcome the loss of sperm motility caused by 100 microM H2O2 in both EYTG and TALP. However, OFC was the most effective of the two catalases in EYTG. In addition to maintain sperm motility, catalase (5 U/ml) and pyruvate (5 mM) increased the intracellular sperm ATP level in comparison to sperm incubated alone for 6 h at 38.5 degrees C in EYTG. Moreover, EDTA, pyruvate and catalase prevented sperm ATP loss in presence of 100 mM of H2O2 in EYTG. These results indicated that EYTG has a very limited capacity to neutralize H2O2, and the addition of low amounts of catalase and millimolar concentrations of pyruvate greatly improved the antioxidant properties of a commonly used extender.

Animals↗

PubMedDate not supplied

Swim-down separation of progressively motile sperm from poor quality human semen by the modified funnel procedure.

The simultaneous swimming up and down from identical semen specimens suggested that the separation efficiency of progressively motile sperm by the swim-down method was superior to that of the swim-up method. The swimming down was performed in a small plastic funnel, the bottom of which was plugged with a 2.5 ml disposable syringe. In this funnel, 2.0 ml of 70% Percoll was poured and then the sperm resuspension was overlaid. After 1 h, progressively motile sperm penetrating into the lower part of Percoll layer (1.6 ml) was collected in the syringe. The technical strategies to yield higher density of progressively motile sperm were found to be centrifugal concentration of the sperm from whole ejaculate prior to the swimming down, and subsequent re-centrifugation of the separated sperm after the swimming down. Oligo-asthenozoospermic semen (22 +/- 4.4 x 10(6) ml1, 5.5 +/- 4.4% motility, n = 8) was processed by the present method, yielding sperm qualities of 32 +/- 19 x 10(6) ml1, 74 +/- 14% in the final preparations. Overall improvement in fertility index (sperm density ml1 x motility % x 10(8)) reached 27-folds on average.

Cell Separation↗

PubMed2006

An attempt to improve standardization of sperm motility class assessment using the superimposed image analysis system (SIAS) software.

AIM: An attempt was made to improve the standardization of sperm motility assessment. METHODS: A computerized system based on image superimposition producing final image with a motion effect was employed. Numerical definition of straight-line velocity (VSL), curvilinear velocity (VCL) and linearity (LIN) related to these motility classes were made, at different frame/rates (20 frames/s, 30 frames/s and 60 frames/s). RESULTS: At least 4 motility class categories were found to be necessary (instead of 3, WHO) for a valid assessment. According to the cut-offs suggested by the ROC curves, motility classes were defined as follows: Class 1 (straight progressive motility): VSL = or > 23 microm/s; LIN = or > 0.58 at 20 frames/s; LIN = or > 0.50 at 30 frames/s; LIN = or > 0.44 at 60 frames/s. Class 2 (straight slow motility): VSL > 10 microm/s and < 23 microm/s; LIN = or > 0.58 at 20 frames/s; LIN = or > 0.51 at 30 frames/s; LIN = or > 0.43 at 60 frames/s. Class 3 (progressive non straight motility): VSL > 10 microm/s; LIN < 0.58 at 20 frames/s; LIN < 0.51 at 30 frames/s; LIN<0.43 at 60 frames/s. Class 4 (non progressive motility): VSL <10 microm/s. A frame rate of 20 frames/s was found to be sufficient to distinguish sperm motility classes in standard semen analysis. CONCLUSIONS: The numerical definition of sperm motility classes may contribute towards standardization in the objective evaluation of sperm kinematics.

Cytological Techniques↗