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PubMed2006

Cholesterol-loaded cyclodextrins added to fresh bull ejaculates improve sperm cryosurvival.

Damage occurring to spermatozoa during cryopreservation results in a loss of motile cells and cells that are functionally normal, compared with fresh sperm samples. Treating bull sperm with cholesterol-loaded cyclodextrins (CLC) before cryopreservation results in increased sperm cryosurvival. However, in previous studies, CLC were always added to sperm samples that had been highly diluted. The aim of this study was to develop a procedure for adding CLC to whole bull ejaculates that would optimize sperm cryosurvival. Adding 2 or 4 mg of CLC/120 x 10(6) sperm to sperm samples ranging in concentration from 120 to 2,000 x 10(6) sperm/mL resulted in greater (17 to 28 percentage points; P < 05) numbers of live cells compared with control samples (no CLC treatment), regardless of the sperm concentration, except for samples at 120 x 10(6) sperm/mL treated with 4 mg of CLC. Incubating sperm with CLC at 23 or 37 degrees C before cryopreservation resulted in similar sperm cryosurvival. The cooling rate used to cryopreserve CLC-treated cells did not affect sperm cryosurvival. Finally, adding CLC to undiluted ejaculates (2 mg of CLC/120 x 10(6) sperm) resulted in greater percentages of live sperm compared with the control samples (62 vs. 45%; P < 0.05), although the percentages of motile sperm were similar for both CLC-treated and control samples (58%). In conclusion, bull sperm cryo-survival can be improved if spermatozoa are treated with CLC before freezing. In addition, CLC can be added to fresh ejaculates at either 23 or 37 degrees C. This technique is simple, practical, and can be easily integrated into current cryopreservation protocols.

Animals↗
PubMed1983

Effects of in vitro storage on the ATP-phosphohydrolase activity and ultrastructure of ejaculated bull sperm.

Extended bull ejaculates were stored in vitro at 4 +/- 1 degree C in the dark. The pH of the ejaculates did not change significantly during storage. The percentage of sperm with intact plasmalemmae was 70% on day 2 and negligible by day 36. On day 2, acrosomes were absent in 3% of the sperm and partial in about 20%; acrosomal damage increased with storage, and by day 36 acrosomes were absent in 12% of the sperm and partially degraded in 72%. The Mg++-ATPase activity decreased from 2.6 to 1.9 microM Pi liberated per 2 X 10(7) sperm per 30 min incubation at 37 degrees between days 2 and 9, but remained at about 65% of its initial value between days 9 and 36. Thin-sectioned sperm sampled from ejaculates at days 2, 9, and 36 of storage showed a gradual but steady deterioration, which began and was most drastic in the head. Flagellar deterioration was negligible on day 9, but by day 36 subtle evidence of deterioration appeared. The persistence of Mg++-ATPase at 65% of initial levels in sperm stored 36 days in vitro is probably attributable to the stability of the movement-related flagellar ATPases.

Adenosine Triphosphatases↗
PubMedDate not supplied

Analysis of ejaculates from patients with cystic fibrosis.

Ejaculates from two men with cystic fibrosis (CF) were examined. Both had azoospermia. A considerable decrease in volume and fructose content was noted. The absolute amounts of calcium, magnesium, and zinc per ejaculate showed normal values in one of the patients but were two to three times increased the the other compared to mean values of a control group. Thus the concentrations of these cations were increased at least fivefold in both patients. The amount of Mg(2+)-and Ca(+)-dependent ATPase was comparable to that of controls, but values were higher than in men with oligospermia. Both the divalent cations and the Mg2+- and Ca2+-dependent ATPase curve profiles of split ejaculate fractions were atypical. Secretory granules and vesicles were plentiful in the seminal plasma of both patients while amorphous substance was practically absent. The present findings agree with a less affected function of the prostate gland and a dysfunction of the seminal vesicles in these patients.

Adenosine Triphosphatases↗
PubMed1985

Volume, sperm concentration, and fertilizing capacity of turkey ejaculates obtained from successive cloacal strokes during semen collection.

Ejaculate fractions (EF), obtained as the ejaculate from each of four successive cloacal strokes (EF1, EF2, EF3, and EF4, respectively), were collected manually by abdominal massage from Large White breeder turkeys. Semen volume and sperm concentration were the greatest for EF 1 and decreased with each successive fraction. Ejaculate Fraction 1 and EF (1 + 2) represented 50 and 79% of the total sperm collected, respectively. Approximately 6, 9, 10, and 11 hens could each be inseminated with 200 X 10(6) sperm from semen collected from each male using EF1, EF (1 + 2), EF (1 + 2 + 3), and EF (1 + 2 + 3 + 4), respectively. There were no differences in duration of fertility, percentage fertility, or percentage hatchability of eggs from hens inseminated with semen from EF1, EF3, and pooled control semen samples.

Animals↗
PubMed1990

Participation of the GABA-benzodiazepine system in the inhibition of defensive burying produced by ejaculation.

We recently reported a reduction in defensive burying behaviour after ejaculation. The present study analyzed whether this reduction was mediated via an increase in GABAergic transmission. Two experimental series were included: in the first series the reduction in burying behaviour was blocked by Ro 15-1788 (5 and 10 mg/kg), bicuculline (2.5 mg/kg) and picrotoxin (0.5 mg/kg). In the second series the effect of ejaculation on defensive burying was potentiated by diazepam (0.5 mg/kg) or muscimol (0.5 mg/kg). With the exception of 10 mg/kg of Ro 15-1788 these treatments did not affect the basal burying behaviour levels. The results support the hypothesis that the GABA-benzodiazepine system participates in the reduction in burying behaviour found after ejaculation, and are discussed on the basis of neurochemical and pharmacological data indicating an increase in GABAergic transmission during the postejaculatory interval.

Journal Article↗
PubMed2003

Evaluation of deoxyribonuclease activity in seminal plasma of ejaculated chicken semen.

AIM: To confirm the stability of exogenous genes in the generation of transgenic chickens using ejaculated chicken sperm, the deoxyribonuclease (DNase) activity was evaluated in the seminal plasma of ejaculated semen and the stability of DNA was examined by adding lipofection reagents. METHODS: A PCR fragment (249 bp) of pEGFPN-1 vector was used as the DNA substrate and was incubated with the seminal plasma at 40 degree C for 30 min. Then, the whole reaction solution was subjected to agarose gel electrophoresis and the DNA size was evaluated under UV light. RESULTS: The DNA substrate was completely diminished after incubation with seminal plasma. However, the substrate was intact after incubation with heat-treated seminal plasma or incubation with seminal plasma in the presence of 0.5 mmol/L approximately 5 mmol/L EDTA. The substrate was stabilized in the seminal plasma by the addition of commercially available lipofection reagents. CONCLUSION: The DNase activity is present in the seminal plasma of ejaculated chicken semen. However, DNA is stable in the liposomal-DNA complex.

Animals↗
PubMed2006

[Correlation of epididymal protease inhibitor Eppin and Semenogelin on human ejaculated spermatozoa].

OBJECTIVE: To evaluate the correlation of epididymal protease inhibitor(Eppin) and Semenogelin(Sg) on human ejaculated spermatozoa. METHODS: The experimental approaches include: (1) Immunoprecipitation of Eppin with anti-Eppin from semen; (2) Colocalization of Eppin and Sg by immunofluorescence; (3) Immunoprecipitation of rEppin and rSg;(4) Far-Western blotting of rEppin and rSg;(5) Competition of saturated 125I-rSg binding to rEppin with unlabeled Sg, and direct binding of 125I-rSg to rEppin on a blot; (6) Autoradiography of 125I-rSg with rEppin. RESULTS: Eppin-Sg complex present on the surface of human ejaculated spermatozoa, Cys-239 is the only cystein for rEppin binding rSg. Reduction and carboxymethylation of Cys-239 blocks binding of 125I-rEppin to rSg. CONCLUSION: Our study demonstrates that Eppin and Sg bind to each other on human ejaculated spermatozoa. A disulfide linkage occurs between Sg and Eppin, indicating the specificity of binding.

Humans↗
PubMed1989

[Diagnostic value of the hypo-osmotic swelling test in pathologic ejaculation findings].

304 ejaculates with oligozoospermia from 227 partners of women suffering from sterility have been investigated by hypoosmotic swelling test, the detection of the motility rate and sperm motile efficiency. In a comparison between selected parts of the group and 104 semen samples of 15 fertile donors from the AID-program is was possible to show, that the swelling test has a better discrimination between fertile and infertile ejaculates than the other two methods. In spite of that the motility rate and the swelling test had a positive correlation (r = 0.73) in the whole group. Because of the high standard deviation of the sperm motile efficiency the diagnostic value of it is doubtful. In borderline cases of pathologic ejaculate factor the hypoosmotic swelling test is useful as a selection criterium for IVF and related techniques.

Female↗
PubMed1989

[Fertilizing capacity of the ejaculate of nutria (Myocastor coypus) after the removal of the seminal vesicles as evaluated by the penetration test and natural mating].

The fertility of male coypu sperm following seminal vesicle extirpation was investigated using the penetration test into the egg of Syrian golden hamster (Mesocricetus auratus). Ejaculates were obtained from five males by means of electro-ejaculation under halothane narcosis. The results of the zona-free hamster eggs (ZFHE) penetration test showed that the ejaculates of all the surgically treated coypu males were fertile and that ZFHE value fluctuated from 54 to 76.6%. The results obtained in experiments with natural mating revealed that the extirpation of male coypu seminal vesicles did not affect their fertility. In total 47 foetuses were found post mortem in ten coypu females covered by surgically treated males, which on average represented 4.7 foetuses per female.

Animals↗
PubMedDate not supplied

Josamycin concentration in human ejaculate and its influence on sperm motility--a contribution to antibiotic therapy in andrological patients.

The concentration of josamycin was determined in the split ejaculate of 5 volunteers after oral administration for several days. One aim of this investigation was to examine the penetration of the macrolide antibiotic into the prostate and the seminal vesicles. 2.23 +/- 1.8 micrograms/ml josamycin was found in fraction I of the ejaculate, consisting mostly of prostatic secretion, and 1.56 +/- 1.37 micrograms/ml josamycin in fraction II comprising mainly secretions from the seminal vesicles. The concentrations of josamycin found in both fractions of the ejaculate are clearly comparable with serum levels of the antibiotic. Josamycin thus attains concentrations in the prostate and seminal vesicles which are effective against Mycoplasma and Chlamydia, pathogens of increasing importance in infections of the urogenital tract. In vitro studies on samples from 30 andrological patients showed that josamycin (0.5 micrograms/ml) did not impair, but even increased the motility of spermatozoa (p less than or equal to 0.01). On the basis of these results josamycin is recommended for the treatment of andrological patients. In particular, the specific antibacterial spectrum also indicates the use of this antibiotic for treatment of the partner when children are desired. The usual precautionary measures for pregnancy must then be adhered to.

Administration, Oral↗
PubMed1985

[Intratesticular administration of testosterone in the therapy of disorders of the quality of ejaculates in boars].

The clinical effect of the bilateral intratesticular administration of a microcrystalline water suspension of testosterone was studied in ten boars of Duroc breed exhibiting semen quality disorders. The ejaculate quality disorders included separate or combined deviations of the values of sperm concentration (hypozoospermia to oligozoospermia), sperm activity (asthenozoospermia), or the occurrence of abnormal spermatozoa (teratozoospermia) from the spermiological standard. After intratesticular infiltration, all the criteria of spermiogram were completely restored in four cases and the boars could be returned to insemination use. A partial improvement of the spermiogram (the number of spermiological criteria with positive responses and the development of their values) without fully meeting all the requirements of the spermiological standard was recorded in four breeding boars. No positive response of ejaculate quality characteristics was obtained in two boars. As to the evaluated criteria of the spermiogram, sperm activity was the most frequent positively influenced parameter in the cases of idiopathic or combined asthenozoospermia, and the increased percentage of abnormal spermatozoa was the least frequent positively influenced parameter in the cases of idiopathic or combined teratozoospermia. It is supposed that the easily practicable methods of intratesticular infiltration with a microcrystalline testosterone suspension could expand the therapeutic possibilities available to veterinary andrologists in the treatment of ejaculate quality disorders in boars.

Animals↗
PubMed1984

Antibody-coated bacteria in ejaculate in bacterial prostatitis.

In patients with bacteriologically proved prostatitis (Stamey test) ejaculate was examined for antibody-coated bacteria (ACB). Forty-four of the 68 patients (65%) with bacteriologically proved prostatitis had ACB in the ejaculate, but only 7 of 89 patients (8%) with prostatodynia. Concomitantly, coeruloplasmin and complement (C3) concentrations in the ejaculate were examined by radial immunodiffusion (RID). The ACB test had much higher specificity (92%) than RID, and the sensitivity of ACB was 65 per cent.

Adult↗
PubMed1978

A comparison of the properties of the two portion split ejaculate in normal and in oligospermic subjects.

The whole ejaculate and a two-portion split ejaculate from each of fifteen normal and seventeen oligospermic men were analysed from sperm density, total sperm population, sperm motility and morphology and seminal plasma fructose. These parameters did not differ significantly between the two fractions. In either group of subjects, although a higher motility was noted in the "normal" samples when compared with the corresponding "Oligo-spermic" samples, the usefulness of the split ejaculate for the purposes of artificial insemination is questioned.

Fructose↗
PubMedDate not supplied

[Karyological analysis of the count of immature germ cells in the ejaculate].

A quantitative karyological analysis of immature male sex cell (IMSC) composition in ejaculate of males with normal spermogram (a relative standard), oligo-, astheno-, terato- or azoospermia has been made. The specimens were obtained from 100 patients. Because of IMSC absence, a part of the 150 specimens lacked informative value. The proportion of IMSC from ejaculate of fertile patients with normospermia was less than 4%. The cells were mainly spermatids, 26% of which remained unseparated in meiotic division. In oligozoospermia the number of ejaculate IMSC increased (up to 26.4%) as well as that of spermatocytes at the stage of meiotic pachytene and diplotene. The same trend persisted in asthenozoospermic males. In azoospermia the percent of IMSC averaged 52.4% varying greatly from individual to individual. Characteristic for this condition was also increased number of unseparated spermatids. The advantages and prospects of the method are discussed.

Humans↗
PubMedDate not supplied

[The protein spectrum of ejaculates with different fertilities].

Immunochemical assay for protein composition was made in 22 fertile and 110 subfertile human sperm samples. Changes in the levels of albumin, prostatic acid phosphatase, sperm-specific tripsin inhibitor, prostate-specific antigen and alpha-2-microglobulin of fertility in spermoplasm were studied in different disturbances of ejaculate fecundating ability. The latter was determined by 14 morphofunctional and physicochemical criteria of sperm fertility. With an increase of the ejaculate volume above 5 ml protein homeostasis showed abnormality. The authors consider functional value of the study proteins in ejaculate, pattern of spermatozoid motor activity, maintenance of normal spermoplasmic pH; discuss practical opportunities of the data obtained for increasing fertility of the sperm used in artificial fertilization, diagnosis and treatment of male infertility.

Adult↗
PubMed1987

Analysis of the flagellar bending waves of ejaculated ram sperm.

The variability of flagellar movement, illustrated by the highly heterogeneous nature of the ejaculated sperm population of the ram, was analyzed by the use of a stroboscopic technique and an adapted microphotographic 24 X 36 camera system. The multiple-moving-exposures (MME) records give very distinct successive sequences of the flagellar beats and are particularly suitable for the analysis of bend development and propagation along the tail. With this technique, the parameters of the flagellar bending waves of ejaculated ram sperm have been determined. Most of the sperm have planar flagellar beatings; few are rolling under the conditions of observation. The trajectories of the gametes are mostly linear; nevertheless, some have circular paths. The analysis of bending has been focused on two examples for which the difference in the progressiveness ratio was maximum. The circular pathways for ram spermatozoa are linked to an asymmetry between principal and reverse bend probably induced by differences in wave propagation evidenced along the flagellum. A typical sperm flagellar movement may be related either to the conditions of the observations or to some differences in the maturation process of the sperm.

Animals↗
PubMedDate not supplied

Immunofluorescence reveals ubiquitination of retained distal cytoplasmic droplets on ejaculated porcine spermatozoa.

The purpose of the present study was twofold: 1) to determine if antibodies raised against ubiquitin would recognize antigens associated with the porcine cytoplasmic droplet (CD), and 2) to determine if the same antibody would identify ubiquitinated substrates on the surface of morphologically abnormal boar spermatozoa. Permeabilization with the detergent Triton X-100 (0.05%) showed virtually all CDs to be ubiquitin positive. Distal droplets (DDs) retained in situ on boar spermatozoa were readily labeled following Triton permeabilization, whereas DDs present on nonpermeabilized cells were not. Negative control preparations lacked the ubiquitin staining on the DD. The use of microtubes for fixation and incubation provided clearer images as well as better sperm cell distribution and density than an initial slide-mounted technique. Immunoblotting indicated that larger amounts of ubiquitinated proteins were present in extracts from sperm cells from an ejaculate with an abnormally high percentage of retained DDs (52% DDs) compared to a morphologically normal sample (6% DDs). The primary antibody recognized both mono-ubiquitin of bovine origin (8.5 kd) and human ubiquitin conjugate (35 kd), as demonstrated by Western blot. Preabsorption of the anti-ubiquitin antibody with purified bovine ubiquitin was successful in preventing diaminobenzidine staining of sperm extract from the high DD ejaculate. The presence of antigens recognized by anti-ubiquitin antibodies in the boar sperm CD, coupled with the possibility that superfluous ubiquitin species are detrimental to embryonic development by targeting critical paternally contributed zygotic organelles, raises concerns that retained DDs may be more detrimental to fertility than previously suspected.

Animals↗