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Activity of angiotensin-converting enzyme (ACE) in reproductive tissues of the stallion and effects of angiotensin II on sperm motility.

A testis-specific isoform of angiotensin-converting enzyme (ACE) has been identified in a number of mammalian species. The purpose of this study was to characterize the activity of ACE in equine spermatozoa, seminal plasma, and testis. Activity of ACE was determined in seminal plasma, ejaculated and epididymal spermatozoa from mature stallions as well as from pre- and postpubertal testis. The effect of addition of angiotensin II on equine sperm motility was also evaluated. The activity of ACE in detergent extracted sperm plasma membrane was approximately 13-fold higher than that detected in seminal plasma (93.7 mU/mg versus 7.0 mU/mg protein, respectively). Activity of ACE in equine testis was significantly higher in postpubertal than in prepubertal males (3.0 mU/mg versus 0.4 mU/mg protein, respectively), and ACE activity was reduced (P<0.001) in a dose-dependent fashion by the addition of captopril. The effect of angiotensin II on sperm motility was evaluated by computer-assisted semen analysis in sperm incubated with angiotensin II (0, 1, 10, 100 nM) at 38.5 degrees C. There was no significant effect of angiotensin II on the percent motile sperm; however, there was a significant main effect of angiotensin II (P<0.01) on the kinematic parameters beat cross frequency (BCF), average path velocity (VAP), and curvilinear velocity (VCL), respectively. In addition, there were significant stallionxconcentration interactions for amplitude lateral movement (ALH), BCF, linearity (LIN), straightness (STR), and VCL. This study demonstrates that ACE activity is present in sperm membrane from ejaculated and epididymal spermatozoa and in postpubertal testis. Further studies are required to determine the role of this testis-specific enzyme.

Angiotensin II↗

The Na,K-ATPase alpha4 isoform from humans has distinct enzymatic properties and is important for sperm motility.

In the rat, the Na,K-ATPase alpha4 isoform exhibits unique enzymatic characteristics and is important for sperm motility. In this work, we studied expression, localization and function of alpha4 in human spermatozoa. We show two catalytically active Na,K-ATPase alpha polypeptides with different ouabain affinity and identified expression of alpha1, alpha4, beta1 and beta3 isoforms in the gametes. In addition, human sperm presented two Na,K-ATPases composed of alpha4, alpha4beta1 and alpha4beta3. Kinetic analysis of these isozymes produced in insect cells showed that, compared with human alpha1beta1, alpha4beta1 and alpha4beta3 exhibit higher Na(+) and lower K(+) affinity and higher sensitivity to ouabain. These particular enzymatic properties suggested a role for alpha4 in sperm function. Using computer-assisted sperm analysis (CASA), we found that ouabain inhibition of alpha4 significantly decreased percentage sperm motility. In contrast, ouabain did not affect linearity of forward progression, amplitude of lateral head displacement, beat cross frequency and sperm straight-line, curvilinear or average path velocities. This suggests a primary role of alpha4 in flagellar motility. Accordingly, we found alpha4 in the sperm tail, predominating in the mid-piece of the flagellum. Therefore, similar to the rat ortholog, human Na,K-ATPase alpha4 isoform has a distinct activity that is essential for sperm function.

Animals↗

Sperm motility under conditions of weightlessness.

The aim of this study was to determine the differences in motility of frozen and thawed bull spermatozoa under conditions of weightlessness compared with ground conditions. The tests were performed within a series of scientific and technologic experiments under microgravity using sounding rockets in the Technologische Experimente unter Schwerelosigkeit (TEXUS) program launched in Kiruna, North Sweden. Using a computerized sperm motility analyzer, significant differences were found in sperm motility under microgravity compared with sperm under gravitational conditions on earth. Computer analysis showed alterations in straight line and curvilinear velocity, as well as in linearity values. The amount of progressively motile spermatozoa, including all spermatozoa with a velocity > 20 microns/second, increased significantly from 24% +/- 9.5% in the reference test to 49% +/- 7.6% in the microgravity test. In conclusion, there is strong evidence that gravity influences sperm motility.

Animals↗

The diagnostic value of seminal a-glucosidase enzyme index for sperm motility and fertilizing capacity.

OBJECTIVE: To find out the diagnostic value of seminal a-glucosidase enzyme index in the ejaculated semen samples of proven-fertility and primary infertility patients, for the sperm motility and fertilizing capacity. METHODS: The study which lasted approximately 2 years was carried out in the Andrology Department of the Arab Fertility Center, Jeddah, Kingdom of Saudi Arabia. Eighty-five healthy male patients (age 25-38 years) were admitted into the study. They were divided into 2 main groups as proven-fertility group and primary infertility group. Each group was again subdivided into 4 classes based on the sperm concentrations. The sperm motility assessment was carried out quantitatively by Semi automated Computer assisted Semen analysis using Autosperm analyzer. Simultaneously, a-glucosidase enzyme index was determined in all the semen samples. RESULTS: The results were tabulated as per the standard sperm motility grades of the World Health Organisation, under the classified sperm concentrations of the study. The progressive motility grades were compared with the enzyme index of a-glucosidase in both the groups. The total progressive motility was found higher in the high-density (classes I and II) than the low-density (classes III and IV) semen samples. The enzyme index readings of the various classes of sperm concentrations were significant (p<0.05) and showed a stepwise fall from 89% +/-9% to 52%+/-3% in the proven fertility group. Although, a similar trend has been observed (48%+/-5% to 20%+/-2%) in the primary infertility group, the enzyme index class II sperm concentration (44%+/-6%) was not found significant with class 1 (48%+/-5%) sperm concentration, but the significance was achieved with the other 2 classes. The readings of enzyme index between the groups were found highly significant (p, 0.001). CONCLUSION: The enzyme index of a-glucosidase estimations in semen samples may indicate a positive correlation with progressive motility and fertilizing capacity of spermatozoa and may serve as diagnostic value in individuals showing normal levels of sperm concentration and serum androgen.

Adult↗

The influence of peritoneal fluid from patients with minimal stage or treated endometriosis on sperm motility parameters using computer-assisted semen analysis.

The aim of this study was to determine the influence of peritoneal fluid from patients with minimal stage or treated endometriosis on sperm motility parameters. Peritoneal fluid aspirated at diagnostic laparoscopy for unexplained infertility from women during the luteal phase of the menstrual cycle (days 20-23) was incubated for 5 h with fresh semen samples obtained from men of recently proven fertility. Spermatozoa were prepared by a swim-up technique from unprocessed semen. Using computer-assisted semen analysis (Hamilton-Thorn Research, MA, USA), sperm motility and motion parameters were observed at 0, 120, 180 and 300 min. Compared with spermatozoa incubated in Earle's balanced salt solution/human serum albumin, the percentage motility, percentage progressive motility and progressive velocity of spermatozoa incubated in peritoneal fluid from patients without visible endometriosis were significantly higher (P < 0.05). Maximal effect was observed at 3 h and maintained until 5 h. We conclude that in an in-vitro study, in contrast to peritoneal fluid from patients with minimal stage endometriosis, peritoneal fluid from patients with unexplained infertility and no visible endometriosis can improve sperm motility when compared with culture medium.

Adult↗

Adverse effects of hydrosalpinx fluid on sperm motility and survival.

The negative impact of hydrosalpinx on IVF outcome is well recognized but some reports indicate that tubal infertility with hydrosalpinx is a heterogeneous entity and may have different effects on the outcome. The embryotoxic effects of hydrosalpinx fluid (HF) have been documented in mouse but not human embryos. This study examined the effects of HF on sperm motility and survival after various periods of incubation. Fifteen infertile patients with hydrosalpinx shown on ultrasound monitoring during ovarian stimulation underwent aspiration of HF after egg collection. Electrolytes, glucose and pyruvate concentrations were within the physiological ranges found in normal human tubal fluid. Sperm motility and velocities remained unchanged after 5 h of incubation with various concentrations of HF but the percentage of motile spermatozoa was significantly reduced after 24 h of incubation. Both 50 and 100% HF were potentially cytotoxic (survival indices <85%). The detrimental effect seemed to be dependent on the concentrations of HF. Low osmolarity, low lactate concentrations or the protein content may be responsible for the loss of sperm motility. A human sperm survival test using HF may be useful in selecting appropriate treatment options for patients with hydrosalpinx undergoing IVF treatment or tubal surgery.

Body Fluids↗

Effects of pentoxifylline on sperm motility characteristics and motility longevity of postthaw cryopreserved semen using computer-assisted semen analysis (CASA).

Computer-assisted sperm movement analysis was used to study the effect of pentoxifylline on human sperm motility characteristics and motility longevity of postthaw cryopreserved semen. This study focused on the following issues: the changes in individual movement characteristics in response to pentoxifylline, the persistence of the response during drug treatment. Computerized analysis was started at 30 min, 3 hrs and 24 hrs, after addition of pentoxifylline. Data obtained showed that pentoxifylline significantly increased percentage of sperm motility, average path velocity (VAP), curvilinear velocity (VCL), straight line velocity (VSL), amplitude of lateral head displacement (ALH) at two of the three time periods (p < 0.05). But, it did not significantly increase linearity (LIN), straightness (STR), and beat cross frequency (BCF) at any time. After 24 hrs, all motility variables were significantly decreased in both groups. However, the 24-hr motility longevity in the treatment group was greater than that of the control group. The present analysis shows interest in the use of pentoxifylline as a sperm movement enhancer for postthaw cryopreserved semen. It shows a beneficial effect in the majority of sperm movement parameters and motility longevity which may increase pregnancy rates after insemination. However, whether this change leads to an increase in fertilizing ability requires further study.

Cryopreservation↗

Intrauterine insemination of isolated motile sperm.

Semen from infertile males was filtered through liquid albumin, and sperm retrieved from the most distal fraction were used for intrauterine insemination. Compared with the ejaculate, the isolated specimens contained fewer sperm but an increased percentage of motile sperm. The fractions were also free of the debris (white cells, agglutinated sperm, immature forms, and particulate matter) commonly seen in the ejaculates. Despite the improvement in motility none of the 19 women who were treated for a total of 67 cycles became pregnant. It is possible that even the best sperm from a poor specimen may have intrinsic defects that interfere with their ability to fertilize.

Female↗

Improved separation of motile sperm in asthenospermia and its application to artificial insemination homologous (AIH).

A method is described for the separation of motile sperm from semen by "migration' into a diluent, providing a large area of interface between the semen and the diluent and resulting in a good yield of motile sperm (58%), with a mean percentage of motility of 95%. The method provides a specimen free of seminal plasma and debris and suitable for intrauterine insemination in less than two hours. The motile fraction was used for AIH treatment of 20 couples with a mean of 4 years' infertility, where the female factor had been excluded and the male was asthenospermic. After a mean of six cycles, three pregnancies were achieved. Improvement of the percentage of motility alone did not, in this study, give a significant improvement in conception rates when compared with accepted cycles with the placebo.

Adult↗

Quantitative analysis of Caenorhabditis elegans sperm motility and how it is affected by mutants spe11 and unc54.

The sperm of Caenorhabditis elegans translocate in a fashion similar to sperm of Ascaris suum even though their pseudopods are longer, more plastic in shape, and form multiple expansions zones around their perimeter. Mutants in spe-11 form primary spermatocytes with a defective perinuclear region, but the resulting spermatozoa can still crawl and fertilize eggs. However, the resultant zygotes die due to the absence of sperm-supplied spe-11. Computer-assisted analysis of translocating spe-11 sperm reveals a novel defect in the dynamic morphology of their pseudopods. A similar analysis of the C. elegans mutant unc-54, which lacks the most abundant isoform of myosin II, reveals no defect in sperm motility, as expected, since C. elegans sperm have substituted the protein MSP for actin in the process of pseudopod expansion. These results reveal an unexpected defect in the dynamic morphology of pseudopods of spe-11 sperm. This defect, however, does not significantly affect crawling velocity, and it demonstrates how computer-assisted motion analysis systems can reveal subtle behavioral phenotypes in C. elegans mutant spermatozoa.

Animals↗

Identification and characterization of a sperm motility promoting glycoprotein from buffalo blood serum.

Early investigators reported the occurrence of unidentified protein factors in biological fluids that may regulate sperm motility essential for fertility potential. This study reports for the first time purification of a forward motility stimulating protein (FMSF-I), to apparent homogeneity, from a biological fluid (buffalo blood serum) and its characterization. FMSF-I is the major motility protein of buffalo serum: a rich source of the factor. FMSF showed high protein specificity and affinity for activating forward motility of goat cauda epididymal spermatozoa. The motility promoter at 0.5 microM level showed maximal activity when nearly 60%-70% of spermatozoa expressed forward motility. It is a 66 kDa monomeric acidic protein rich in aspartate, glutamate, and leucine with isoelectric point of 3.7. FMSF: a Mg2+ -dependent protein binds to concanavalin A-agarose and the glycoprotein nature of FMSF has been confirmed by PAS staining. The factor lost activity completely when treated with alpha-mannosidase showing that the sugar part of the protein is essential for its biological activity. FMSF has no species specificity for its motility-activating potential. Sperm surface has specific receptors of FMSF, which is strongly immunogenic. The factor is present in testis and epididymis although liver is its richest source. Motility promoting efficacy of FMSF is markedly higher than the well-known non-protein motility activators: theophylline and bicarbonate or their combination. FMSF is a physiological activator of sperm motility and as a slaughterhouse byproduct it has potentiality for solving some of the problems of animal breeding, conservation of endangered species, and human infertility: a global social problem.

Amino Acid Sequence↗

Straw-thawing method interacts with sire and extender to influence sperm motility and conception rates of dairy cows.

Influence of interactions of straw-thawing method with sire and extender type (milk or egg yolk-based) on postthaw sperm motility and conception rates of dairy cows was assessed. In experiment 1, sperm from 10 Holstein sires were frozen in egg yolk citrate and heated whole-milk extenders using a split-ejaculate technique. Straws were thawed in a 37 degrees C water bath with or without 3 min of exposure to a 37 degrees C warming plate while wrapped in a paper towel (air-thaw). Percentage of motile sperm was assessed after 1 min and 3 h of incubation at 37 degrees C. In experiment 2, sperm from 3 Holstein sires were frozen in egg-yolk citrate and a nonheated whole milk extender using a split-ejaculate technique. Straws were thawed and motility was assessed as in experiment 1. Conception rates (n = 475) were compared in a single herd of Holstein cows after thawing straws in 35 degrees C water for 45 s or after air-thawing by wiping straws with a paper towel upon removal from the storage vessel and placing directly into the insemination gun. In each experiment, interactions between thaw method and sire, extender type, or both, indicated that conditions may be created that facilitate sperm tolerance of air-thaw procedures. When a significant thaw method effect was detected in either experiment, air-thaw was consistently associated with the lesser measure of sperm motility, conception, or both. In conclusion, although some sire x extender combinations seem to be tolerant of air-thaw procedures, other combinations are more sensitive, resulting in reduced postthaw sperm survival, conception rates of cows, or both, in response to air-thaw.

Animals↗