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At least 163 records · Page 9Linked to original sources

PubMed2005

Intracytoplasmic sperm injection: a novel selection method for sperm with normal frequency of chromosomal aneuploidies.

OBJECTIVE: To test a newly invented intracytoplasmic sperm injection (ICSI) sperm selection method based on sperm hyaluronic acid (HA) binding. DESIGN: Comparison of chromosomal disomy and diploidy frequencies in sperm arising from semen and in HA-bound sperm. SETTING: Academic andrology laboratory. PATIENT(S): Men presenting for semen analysis. INTERVENTION(S): Washed sperm fractions of 32 semen samples were applied to Petri dishes or glass slides coated with immobilized HA. The unbound sperm were rinsed gently, and the HA-bound sperm were removed with an ICSI pipette. The control sperm population was the unselected sperm. Both HA-selected and unselected sperm were treated with fluorescence in situ hybridization with centromeric probes for the X, Y, and 17 chromosomes. MAIN OUTCOME MEASURE(S): Chromosomal disomy and diploidy frequencies. RESULT(S): In the HA-bound sperm (495-2,079 per man, 41,670 in all) compared with unselected sperm (4,770 per man, 162,210 in all), the chromosomal disomy frequencies were reduced to 0.16% from 0.52%, diploidy to 0.09% from 0.51%, and sex chromosome disomy to 0.05% from 0.27% (a 5.4-fold reduction vs. 4-fold respective increase in ICSI offspring). CONCLUSION(S): The HA sperm selection method for ICSI, which is based on a relationship between sperm receptors for zona pellucida and HA, will likely reduce the potential genetic complications and adverse public health effects of ICSI.

Andrology↗

PubMed1998

Coculture of human sperm with bovine oviduct epithelial cells decreases sperm chromatin structural changes seen during culture in media alone.

OBJECTIVE: To compare sperm chromatin structural changes seen in media only culture or in coculture with bovine oviduct epithelial cells. DESIGN: Three freshly ejaculated and three cryopreserved sperm samples in media culture or in oviduct epithelial cell coculture. Sperm in each treatment were evaluated by the sperm chromatin structure assay during a 72-hour time course. SETTING: An academic research laboratory. PATIENT(S): Normospermic donors with children. INTERVENTION(S): Semen collection through masturbation after 48 hours of abstinence. MAIN OUTCOME MEASURE(S): The sperm chromatin structure assay using flow cytometry to detect the susceptibility of sperm in either treatment to denaturation of DNA in situ. RESULT(S): The sperm chromatin structure assay data differed for sperm type (fresh or cryopreserved), over time, and between treatments within 6 hours of culture. In oviduct epithelial cell coculture, fresh sperm chromatin structure assay values for fresh sperm were stable, whereas in control medium higher chromatin degeneration levels were seen by 10 hours. For cryopreserved sperm, chromatin degeneration had increased by 1 hour postthaw in both treatments, although levels were higher in the control treatment thereafter. CONCLUSION(S): Sperm chromatin structural changes occur over time in culture. Such changes were observed within 2 hours for cryopreserved sperm. Coculture of sperm with oviduct epithelial cells results in a stabilizing effect for sperm against chromatin changes.

Animals↗

PubMed2005

Beta-defensin 126 on the cell surface protects sperm from immunorecognition and binding of anti-sperm antibodies.

Beta-defensin 126 (DEFB126), formerly known as epididymal secretory protein 13.2 (ESP13.2), coats the entire primate sperm surface until completion of capacitation, and it is a candidate for providing immune protection in the female reproductive tract. To further examine the potential role of DEFB126 as a means of protection from immune recognition, cynomolgus macaque sperm were exposed to a number of treatments that are known to alter sperm surface coats, including capacitation. We used a novel in vivo assay to determine immune recognition: aldehyde-fixed whole sperm injections into rabbits. Following booster injections, immunoblot analyses of whole sperm prepared in various manners was conducted. On Days 60 and 80 post-initial immunization, the antisera showed a remarkably strong reaction to a single 34-36 kDa protein, which was shown to be DEFB126. Sera from rabbits that were immunized with sperm washed more rigorously using Percoll gradients showed an increase in the number and intensity of proteins recognized on whole sperm Western blots, although DEFB126 was still the major immune response. When capacitated sperm, from which most DEFB126 had been released, were used as the immunogen, there was a dramatic increase in the immune recognition to a variety of protein bands. Sperm treated with neuraminidase to remove sialic acid on DEFB126 before fixation were shown to still possess DEFB126, but lacked the sialic acid component of the glycoprotein. These sperm were as immunogenic as capacitated sperm even though the desialylated DEFB126 still covered the entire cell surface. These sperm lost their highly negative charge (the isoelectric point of DEFB126 shifted from pI 3.0 to pI 6.4). Experiments using different sperm plasma membrane protein-specific Igs showed that recognition did not occur when DEFB126 was present, but following capacitation these Igs readily recognized the exposed sperm membrane. Our data suggest that DEFB126 protects the entire primate sperm surface from immune recognition and that the sialic acid moieties are responsible for the cloaking characteristic of this unique glycoprotein.

Animals↗

PubMedDate not supplied

Fractions of Percoll eliminated sperm: increased sperm yield in percoll wash.

Sperm preparation by Percoll is one of the most widely used techniques, but it has the disadvantage of producing a low final yield of motile sperm. This study compared the quality of Percoll-eliminated sperm with that of Percoll recovered sperm to determine if the profile of the Percoll preparation could be improved. Semen specimens were fractionated by discontinuous Percoll gradient column. This resulted in 4 sperm fractions: sperm retained in the semen (SRS), sperm clumped at the interface of 40 and 80% Percoll (SCI), sperm scattered in the column (SSC), and sperm-forming pellet, called Percoll-recovered sperm (PRS). The sperm fractions were evaluated for count, motility, viability, vitality, and morphology. The SRS exhibited the lowest motility, viability, and vitality (40, 68, and 64%, respectively), which were significantly different from those of SCI, SSC, and PRS. The motility, viability, and vitality differences between SSC and PRS did not reach a statistically significant level. Good motility (grades a and b) was found in SSC (73%) when compared with that of PRS (82%). The lower part of SSC and PRS exhibited identical sperm morphology. The Percoll sperm recovery was 18%, but if SSC is combined with PRS, the total recovery becomes 40%. These data suggest that the SSC fraction, particularly the lower part, which maintains the physical contact with the PRS, can be pooled together to increase the final sperm yield without compromising the quality of the sperm to be used for insemination.

Cell Survival↗

PubMed2005

Discrepancy between sperm acrosin activity and sperm morphology: significance for fertilization in vitro.

BACKGROUND: In routine semen analysis, discrepancies may occur between sperm acrosin activity test results and sperm acrosomal morphology. METHODS: Discrepant test results of sperm acrosin activity (spectrophotometric assay) vs. sperm morphology (strict criteria) in the initial diagnostic investigation of 107 infertile couples were evaluated with respect to fertilization rate (% oocytes with 2 pronuclei) further obtained in IVF treatment. RESULTS: Acrosin activity positively correlated with sperm morphology (% normal forms) (r=0.537) and fertilization rate (r=0.526). ROC curves for the prediction of > or =50% fertilization rate were comparable for acrosin activity and sperm morphology, with optimal cutoff values at 25 microIU/10(6) sperm and 10%, respectively. In multiple regression analysis, sperm acrosin activity (P=0.002) predicted fertilization rate independently of sperm morphology (P<0.001) and sperm vitality (eosin-nigrosin stain) (P=0.03). Acrosin activities > or =25 microIU/10(6) sperm were observed in 36% of severe teratozoospermic samples (< or =4% normal spermatozoa) associated with low fertilization rate. Twenty percent of the morphologically normal ejaculates showed a low acrosin activity (<25 microIU/10(6) sperm) and low hypoosmotic swelling test (HOST) scores (31.4+/-7.6%) and were associated with low fertilization rate. CONCLUSION: The sperm acrosin assay can help to predict sperm fertilizing capacity in IVF independently of sperm morphology.

Acrosin↗

PubMed2005

Sperm viability matters in insect sperm competition.

Experimental studies in insects have shown how sperm competition can be a potent selective force acting on an array of male reproductive traits . However, the role of sperm quality in determining paternity in insects has been neglected, despite the fact that sperm quality has been shown to influence the outcome of sperm competition in vertebrates . A recent comparative analysis found that males of polyandrous insect species show a higher proportion of live sperm in their stores . Here, we test the hypothesis that sperm viability influences paternity at the within-species level. We use the cricket Teleogryllus oceanicus to conduct sperm competition trials involving prescreened males that differ in the viability of their sperm. We find that paternity success is determined by the proportion of live sperm in a male's ejaculate. Furthermore, we were able to predict the paternity patterns observed on the basis of the males' relative representation of viable sperm in the female's sperm-storage organ. Our findings provide the first experimental evidence for the theory that sperm competition selects for higher sperm quality in insects. Between-male variation in sperm quality needs to be considered in theoretical and experimental studies of insect sperm competition.

Animals↗

PubMed1998

Successful sperm retrieval by percutaneous epididymal and testicular sperm aspiration.

PURPOSE: We evaluated the safety and efficacy of percutaneous sperm aspiration from the epididymis or testicle as a diagnostic technique to confirm sperm production and as a therapeutic technique to harvest sperm for use in intracytoplasmic sperm injection. MATERIALS AND METHODS: We present our experience with 69 sperm aspiration procedures in men considered to have nonreconstructible obstructive azoospermia. This short outpatient procedure was performed using a butterfly needle with the patient under intravenous sedation and local anesthesia. RESULTS: Of the 32 diagnostic aspirations 20 demonstrated mature and motile sperm, 9 maturation arrest and 3 germ cell aplasia. In 35 of 37 therapeutic sperm aspirations (95%) adequate samples of sperm after processing (mean of 5.4 million) were obtained. Of 24 epididymal aspirations 13 (54%) had sufficient residual sperm for cryopreservation of 1 to 5 vials (mean 2.5) containing an average of 170,000 sperm per vial. In the 32 intracytoplasmic sperm injection cycles 221 of 392 eggs (56.3%) fertilized (2PN) and 6 resulted in ongoing pregnancies (21.4% per transfer). There have been no acute or chronic complications in this patient population. Ten men underwent a second successful aspiration procedure for intracytoplasmic sperm injection and 3 underwent a third aspiration without added difficulty. CONCLUSIONS: Percutaneous epididymal or testis sperm aspiration is a minimally invasive sperm retrieval technique and appears to be an effective alternative to microsurgical epididymal sperm aspiration, which is more invasive, costly and technically difficult.

Adult↗

PubMed2003

Sperm length influences fertilization success during sperm competition in the snail Viviparus ater.

Sperm form and size is tremendously variable within and across species. However, a general explanation for this variation is lacking. It has been suggested that sperm size may influence sperm competition, and there is evidence for this in some taxa but not others. In addition to normal fertilizing sperm, a number of molluscs and insects produce nonfertile sperm that are also extremely morphologically variable, and distinct from fertilizing forms. There is evidence that nonfertile sperm play an indirect role in sperm competition by decreasing female remating propensity in Lepidopterans, but in most taxa the function of parasperm is unknown. We investigated the role of nonfertile (oligopyrene) sperm during sperm competition in the fresh water snail Viviparus ater. Previous studies found that the proportion of oligopyrene sperm increased with the risk of sperm competition, and hence it seems likely that these sperm influence fertilization success during competitive matings. In mating experiments in which females were sequentially housed with males, we examined a range of male characteristics which potentially influence fertilization success. We found that the size of oligopyrene sperm was the best predictor of fertilization success, with males having the longer sperm siring the highest proportion of offspring. Furthermore, we found a positive shell size and sperm concentration effect on paternity, and females with multiply sired families produced more offspring than females mating with only one male. This result suggests polyandry is beneficial for female snails.

Analysis of Variance↗

PubMed1992

Sperm size and sperm competition in birds.

In a sample of 20 species of North American passerine birds we found no relation between sperm size and mating system like that previously reported in mammals (Gomendio & Roldan (Proc. R. Soc. Lond. B 243, 181 (1991)). Instead, we found a positive correlation between sperm length and the length of female sperm storage tubules (SSTS) and a negative correlation between sperm length and the number of SSTS. Both of these correlations suggest that the more than fivefold variation in sperm size we found among species can be explained by sperm competition for access to storage sites (SSTS) in females. As longer sperm appear to be able to swim faster, selection should favour long sperm when SSTS are in short supply; sperm long enough to fill an SST might also prevent access to SSTS by the sperm of other males. Conversely, selection should favour shorter sperm when there is an advantage to sperm layering within an SST promoting a last-male mating advantage. Although we conclude that sperm competition influences sperm size in birds, little is known about the interactions between sperm and SSTS. It seems clear, however, that detailed study of this interaction will provide a new dimension to the study of avian mating systems.

Animals↗

PubMed2003

Characterization of a novel ZP3-independent sperm-binding ligand that facilitates sperm adhesion to the egg coat.

During mammalian fertilization, sperm adhere to the extracellular coat of the egg, or zona pellucida, in a species-specific manner. In mouse, evidence suggests that sperm recognize and bind to specific oligosaccharide ligands within the zona pellucida glycoprotein, ZP3, via beta1,4-galactosyltransferase I (GalT I), a lectin-like receptor on the sperm surface. Although in vitro experiments using isolated gametes lend support to this model, recent in vivo studies of genetically altered mice question whether ZP3 and/or GalT I are solely responsible for sperm-egg binding. In this regard, sperm from GalT I-null mice bind poorly to ZP3 and fail to undergo a zona-induced acrosome reaction; however, they still bind to the ovulated egg coat in vitro. In this report, we characterize a novel ZP3- and GalT I-independent mechanism for sperm adhesion to the egg coat. Results show that the ovulated zona pellucida contains at least two distinct ligands for sperm binding: a ZP3-independent ligand that is peripherally associated with the egg coat and facilitates gamete adhesion; and a ZP3-dependent ligand that is present in the insoluble zona matrix and is recognized by sperm GalT I to facilitate acrosomal exocytosis. The ZP3-independent ligand is not a result of contamination by egg cortical granules, nor is it the mouse homolog of oviduct-specific glycoprotein. It behaves as a 250 kDa, WGA-reactive glycoprotein with a basic isoelectric point, distinguishing it from the acidic glycoproteins that form the insoluble matrix of the egg coat. When eluted from isoelectric focusing gels, the acidic matrix glycoproteins possess sperm-binding activity for wild-type sperm, but not for GalT I-null sperm, whereas the basic glycoprotein retains sperm-binding activity for both wild-type and GalT I-null sperm. Thus, GalT I-null sperm are able to resolve gamete recognition into at least two distinct binding events, leading to the characterization of a novel, peripherally associated, sperm-binding ligand on the ovulated zona pellucida.

Acetylglucosaminidase↗

PubMed2003

Human sperm DNA integrity: correlation with sperm cytoplasmic droplets.

OBJECTIVES: To examine the retention of sperm cytoplasmic droplets (CD) and DNA denaturation (DD) in semen from fertile and infertile men. METHODS: Semen samples were obtained from consecutive nonazoospermic men presenting for infertility evaluation (n = 101) and fertile men presenting for vasectomy (n = 13). The standard semen parameters (sperm concentration, motility, and morphology), sperm DD, and sperm CD were monitored. Sperm DD was evaluated by flow cytometry analysis of acridine orange-treated spermatozoa and expressed as the percentage of spermatozoa demonstrating denatured DNA. RESULTS: The mean (+/-SE) percentages of spermatozoa with CD and DD were significantly higher in infertile than in fertile men (sperm CD 15.7% +/- 0.8% versus 4.8% +/- 0.7% and sperm DD 22.0% +/- 1.5% versus 10.8% +/- 1.8%, respectively). Sperm CD and DD were positively correlated (r = 0.59). Also, sperm CD and DD values correlated inversely with the standard semen parameters. CONCLUSIONS: Our data demonstrate that the retention of sperm CD correlates positively with sperm DD and that significantly higher sperm DD and CD are found in infertile than in fertile men. These data suggest that the enhanced susceptibility of sperm DNA to denaturation is associated with the abnormal disposal of residual sperm cytoplasm in the testis and/or epididymis.

DNA↗

PubMed2000

Sperm binding and penetration of the zona pellucida in vitro but not sperm-egg fusion in an Australian marsupial, the brushtail possum (Trichosurus vulpecula).

Sperm capacitation and in vitro fertilisation (IVF) have been achieved in most eutherian mammals and American marsupials under relatively simple culture conditions. In contrast sperm capacitation in Australian marsupials has not been achieved in vitro and attempts at IVF have previously been characterised by a complete lack of sperm-zona pellucida (ZP) binding. Recently, co-culture of sperm with oviduct epithelial cell monolayers or with oviductal explant conditioned media has been shown to prolong the viability and motility of brushtail possum spermatozoa, as well as to induce capacitation-associated changes such as transformation of sperm to the T-shape orientation. In this study we report that these in vitro produced T-shaped sperm, and in vivo derived T-shaped sperm flushed from the oviduct of artificially inseminated possums as a control, are able to bind to and penetrate the ZP of approximately 25% of eggs recovered from PMSG/LH-superovulated possums in vitro. Development of ZP receptivity and penetrability towards sperm was also identified as a major factor affecting the outcome of IVF. Neither in vivo nor in vitro derived T-shaped sperm were able to bind to or penetrate the ZP if eggs were obtained from animals that were treated with pLH less than 76 h after PMSG. Thus this study provides preliminary evidence for the necessity of sperm-oviduct epithelial cell interactions for capacitation in Australian species and lends further support to the suggestion that the T-shape head orientation is indicative of sperm capacitation. Despite the occurrence of sperm-ZP binding and penetration, sperm-egg membrane fusion and egg activation were not observed. Although the factor(s) responsible for the lack of sperm-egg membrane fusion in the possum have not been identified it is possible that egg capacity for membrane fusion develops independently of zona receptivity and is defective in these eggs, or alternatively that membrane fusion requires strictly defined ionic conditions which are not provided by the IVF media used in this study.

Animals↗

PubMed1997

A role for the disintegrin domain of cyritestin, a sperm surface protein belonging to the ADAM family, in mouse sperm-egg plasma membrane adhesion and fusion.

Sperm-egg plasma membrane fusion is preceded by sperm adhesion to the egg plasma membrane. Cell-cell adhesion frequently involves multiple adhesion molecules on the adhering cells. One sperm surface protein with a role in sperm-egg plasma membrane adhesion is fertilin, a transmembrane heterodimer (alpha and beta subunits). Fertilin alpha and beta are the first identified members of a new family of membrane proteins that each has the following domains: pro-, metalloprotease, disintegrin, cysteine-rich, EGF-like, transmembrane, and cytoplasmic domain. This protein family has been named ADAM because all members contain a disintegrin and metalloprotease domain. Previous studies indicate that the disintegrin domain of fertilin beta functions in sperm-egg adhesion leading to fusion. Full length cDNA clones have been isolated for five ADAMs expressed in mouse testis: fertilin alpha, fertilin beta, cyritestin, ADAM 4, and ADAM 5. The presence of the disintegrin domain, a known integrin ligand, suggests that like fertilin beta, other testis ADAMs could be involved in sperm adhesion to the egg membrane. We tested peptide mimetics from the predicted binding sites in the disintegrin domains of the five testis-expressed ADAMs in a sperm-egg plasma membrane adhesion and fusion assay. The active site peptide from cyritestin strongly inhibited (80-90%) sperm adhesion and fusion and was a more potent inhibitor than the fertilin beta active site peptide. Antibodies generated against the active site region of either cyritestin or fertilin beta also strongly inhibited (80-90%) both sperm-egg adhesion and fusion. Characterization of these two ADAM family members showed that they are both processed during sperm maturation and present on mature sperm. Indirect immunofluorescence on live, acrosome-reacted sperm using antibodies against either cyritestin or fertilin beta showed staining of the equatorial region, a region of the sperm membrane that participates in the early steps of membrane fusion. Collectively, these data indicate that a second ADAM family member, cyritestin, functions with fertilin beta in sperm-egg plasma membrane adhesion leading to fusion.

ADAM Proteins↗

PubMedDate not supplied

Influence of sperm processing on the fertilizing capacity and recovery of motile sperm from thawed human semen.

Artificial insemination programs now rely almost exclusively on frozen semen preparations as their source of sperm. Unfortunately, several reports indicate that conception rates using frozen-thawed semen are inferior to freshly ejaculated specimens. The present study was designed to investigate the fertilizing capacity of sperm and the recovery of motile sperm from thawed semen following different sperm processing methods. Washed resuspended pellets contained sperm with at least the same fertilizing potential as sperm from swim-up techniques. However, the recovery of motile sperm from the pellets was more than 3 times greater than from the swim-up techniques. Percentage motility, progressive velocity, and amplitude of lateral head displacement were generally higher in the sperm from swim-up techniques than in the sperm from the pellets, despite equivalent fertilizing potentials. It can be argued from these results that washed resuspended sperm preparations provide a significantly greater number of motile sperm without a loss in fertility when compared to sperm from swim-up techniques. Accordingly, this study raises questions about the use of the sperm swim-up as a procedure for processing thawed semen for use in intrauterine insemination in which maximal numbers of motile sperm are required. It also demonstrates the need to identify new methods for processing frozen-thawed specimens for assisted reproductive procedures.

Animals↗

PubMed2004

Spermatozoal traits and sperm competition in Atlantic salmon: relative sperm velocity is the primary determinant of fertilization success.

Sperm competition occurs when sperm from more than one male compete for fertilizations. This form of post-copulatory sexual selection is recognized as a significant and widespread force in the evolution of male reproductive biology and as a key determinant of differential male reproductive success. Despite its importance, however, detailed mechanisms of sperm competition at the gamete level remain poorly understood. Here, we use natural variation in spermatozoal traits among wild Atlantic salmon (Salmo salar), a species naturally adapted to sperm competition, to examine how the relative influences of sperm (i) number, (ii) velocity, (iii) longevity, and (iv) total length determine sperm competition success. Atlantic salmon fertilize externally, and we were therefore able to conduct controlled in vitro fertilization competitions while concurrently measuring spermatozoal traits within the aqueous micro-environment to which salmon gametes are naturally adapted. Microsatellite DNA fingerprinting revealed that a male's relative sperm velocity was the primary determinant of sperm competition success. There was no significant relationship between fertilization success and either relative sperm number or total length; sperm longevity showed an inverse relationship with competition success. These relationships were consistent for two experimental repeats of the in vitro fertilization competitions. Our results therefore show, under the natural microenvironment for salmon gametes, that relative sperm velocity is a key spermatozoal component for sperm competition success. Atlantic salmon sperm can be considered to enter a competition analogous to a race in which the fastest sperm have the highest probability of success.

Animals↗

PubMed1996

Cholesterol and phospholipid levels of washed and percoll gradient centrifuged mouse sperm: presence of lipids possessing inhibitory effects on sperm motility.

Levels of DNA, cholesterol, and phospholipids of mouse caudal epididymal and vas deferens sperm that were processed through simple washing and Percoll gradient centrifugation were measured. The DNA and cholesterol contents of washed sperm and Percoll gradient centrifuged (PGC) sperm (DNA = 3.6 +/- 0.3 pg/sperm and 3.4 +/- 0.3 pg/sperm, respectively; cholesterol = 0.219 +/- 0.057 nmole/microgram DNA and 0.224 +/- 0.030 nmole/microgram DNA, respectively, for washed and PGC sperm) were not significantly different from each other; however, the phospholipid level of PGC sperm was only one half of that of washed sperm (0.315 +/- 0.071 nmole/microgram DNA versus 0.720 +/- 0.075 nmole/microgram DNA, respectively). The presence of 0.3% bovine serum albumin (BSA) in the culture medium used in sperm washing did not change the cholesterol and phospholipid contents of washed sperm. Similarly, the cholesterol and phospholipid levels of washed sperm and PGC sperm that were further incubated in BSA-containing medium for 30 min remained the same. Interestingly, substantial amounts of lipids, as determined by the cholesterol and phospholipid levels, were released into the supernatants of the sperm washes, and sperm needed to be washed at least twice to ensure their stable levels of cholesterol and phospholipids. The lipid mixture in the first sperm wash supernatant was shown to have inhibitory effects on PGC sperm motility.

Animals↗

PubMed1994

Flow cytometric analysis of mouse sperm using monoclonal anti-sperm antibody A-1.

We have previously prepared an anti-mouse sperm monoclonal antibody (A-1) which inhibited sperm penetration into the egg zona pellucida. By indirect immunofluorescence (IIF), the A-1 antibody was shown to recognize an antigen localized in the acrosomal area of sperm. This antibody bound negligibly to fresh sperm, while binding to methanol-fixed sperm was almost complete. After methanol fixation, no sperm that penetrated into the zona were immunoreactive for this antibody. In the present study we examined the localization and fate of A-1 antigen during the acrosome reaction by IIF and flow cytometry (FCM). Cauda epididymal sperm were treated with either calcium ionophore A23187 or zona solution, immunostained indirectly, and subjected to FCM. Treatment with A23187 reduced the percentage of immunoreactive sperm to 59% from the 80% obtained in the untreated sperm. The treatment also reduced the average fluorescence intensity per fluorescence-positive spermatocyte to 65 channels, while this intensity was 89 channels in the untreated sperm. A similar result was obtained from treatment with zona solution. The proportion of sperm that was immunoreactive with A-1 antibody was reduced to 55% by incubation in zona-containing media from the 80% obtained in zona-free media. On the other hand, neither A23187 nor the zona solution affected the immunoreactivity or the fluorescence intensity of caput epididymal sperm, while the A-1 antigen was present in both the immature sperm from the caput epididymis of adult mice and in the mature sperm from the cauda epididymis of the same mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrosome↗

PubMed1988

Human sperm injection into the perivitelline space (SI-PVS) of hamster oocytes: effect of sperm pretreatment by calcium-ionophore A23187 and freezing-thawing on the penetration rate and polyspermy.

Calcium-ionophore A23187 and freezing-thawing were used as sperm treatments before human sperm injection into the perivitelline space (SI-PVS) of hamster oocytes. The penetration rate (PR) was higher when SI-PVS was performed with calcium-ionophore-treated (28%) or frozen-thawed (51%) sperm than with untreated sperm (8%). Optimal PR occurred when five calcium-ionophore-treated (57%) or frozen-thawed (71%) sperm were injected under the zona pellucida. When the sperm:egg ratio was 1:1, PR was higher for calcium-ionophore-treated (18.5%) or frozen-thawed (27.8%) sperm than for untreated sperm (0.0%). Calcium-ionophore sperm treatment had no effect on the polyspermic oocyte rate (POR) or the mean number of swollen sperm nuclei per penetrated oocyte (Pd) or per injected sperm (SR). This may result from premature oocyte activation induced by Ca-ionophore. However, POR was higher with frozen-thawed (74%) than with untreated (50%) or Ca-ionophore-treated (50%) sperm. Whatever the sperm treatment, there was a trend toward a lower SR as the number of injected sperm increased. Cytoplasmic regulation of polyspermy in the hamster oocyte is discussed.

Animals↗