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Levels of P34H, a sperm protein of epididymal origin, as a predictor of conventional in vitro fertilization outcome.

A prospective double-blind study was performed to determine if Western blot detection of P34H, a sperm protein of epididymal origin that is involved in the binding to the zona pellucida, is predictive of standard IVF outcome. Our results demonstrate that the proportion of positive P34H cases that produced embryos in vitro clearly differs from cases with undetectable levels of P34H (P<.001).

Adolescent↗

Intra-abdominal ovarian-type mucinous cystadenoma associated with fallopian tube-like structure and aberrant epididymal tissue in a male patient.

Ovarian-type mucinous tumors may occasionally occur in the retroperitoneum, pancreas, and liver exclusively or almost exclusively in women. In men, only few cases of such neoplasms arising within or around the testis have been reported. We describe a unique case of an ovarian-type mucinous cystadenoma occurring in the peritoneal cavity of a 65-year-old male patient with secondary adrenocortical insufficiency and hypogonadism. There was a typical fallopian tube enclosed in the capsule of the tumor. Contiguous to that, accessory ectopic epididymal tissue was found. We have interpreted this case as the result of a minor disorder of embryonic development involving structures of both müllerian and wolffian origin. The disproportion of sexual hormones might have had a role in inducing intra-abdominal müllerian remnants to give rise to the mucinous cystadenoma.

Aged↗

Immunisation of the male tammar wallaby (Macropus eugenii) with spermatozoa elicits epididymal antigen-specific antibody secretion and compromised fertilisation rate.

Immunocontraception has been proposed as an effective and humane means of controlling overabundant kangaroo populations in Australia. We have examined the feasibility of using a sperm-based vaccine for this purpose using a model macropod species, the tammar wallaby (Macropus eugenii). This study has demonstrated immunocontraception in a marsupial species following immunisation of males with homologous spermatozoa. Serum anti-sperm IgG titres were associated with a significant reduction in fertilisation rates following mating with superovulated female wallabies. Antigen-specific IgG penetrated the reproductive tract at the rete testis and bound spermatozoa in vivo. IgG was detected bound to the acrosome and midpiece regions of both epididymal and ejaculated spermatozoa. The absence of adverse testicular pathology and sperm movement effects suggests that contraception may have been achieved by antibody-mediated blocking of sperm surface antigens essential for fertilisation. This study demonstrates that a contraceptive vaccine targeting sperm antigens has potential for fertility control in male macropods.

Acrosome↗

Molecular diffusion in sperm plasma membranes during epididymal maturation.

Fluorescence recovery after photobleaching (FRAP) analysis has been used to measure lipid diffusion in different regions of the sperm plasma membrane. Our goal has been to understand how some membrane components are confined to specific surface domains, whilst others are freely diffusing and in some cases are able to migrate against large concentration gradients. Results with a variety of fluorescent lipid reporter probes (ODAF, NBD-PC, NBD-cholesterol) show that diffusion coefficients (D) are generally three to four times higher on the sperm acrosome than on the principal piece of the tail and increase significantly during epididymal maturation (ram, mouse, goat, dog and monkey sperm). Cholesterol diffusion is approximately 10 times faster on the sperm head than the tail and has a heterogenous distribution when detected with filipin. Lipid diffusion is very temperature sensitive but remarkably insensitive to changes in external pH and osmotic pressure. There was no evidence that the posterior ring or annulus functioned as diffusion barriers to lipids. On this basis it was possible to construct models of increasing complexity to describe the behaviour of a lipid molecule on the sperm surface, beginning with simple linear diffusion progressing to random diffusion and eventually to constrained diffusion.

Animals↗

Epithelial response to experimentally introduced intraluminal bacteria in the avian epididymal ducts.

The response of the epithelial cells of the various ducts of the avian epididymis, whose function is poorly understood, to intraluminal bacteria was evaluated by the injection of an avirulent strain of Salmonella gallinarium into the RT for 24 h. Ultrastructurally, bacteria and invading mononuclear cells were present in the lumina of the RT, proximal efferent ducts (PED) and distal efferent ducts. However, only the non-ciliated (Type I) cells of the PED epithelium ingested bacteria from the lumen. Fragments of bacteria also occurred in several intercellular spaces in the epithelium of the PED. Some mononuclear cells also contained fragments of bacteria. Neither cell death in the various epithelia nor mononuclear infiltration of the periductal tissue occurred. Therefore, in addition to the established function of absorbing most of the testicular fluid entering the epididymis, the Type I cells also appear capable of recognising and removing foreign particulate matter from the epididymal through-flow in the proximal part of the epididymis.

Animals↗

Post mortem time and season alter subpopulation characteristics of Iberian red deer epididymal sperm.

We have studied the effect of post mortem time and season on sperm subpopulation pattern and characteristics. We used epididymal samples from free-ranging Iberian red deers harvested during the hunting season. We studied samples at different moments of the year (rut, transition period and post-rut), and at different times post mortem (up to 4 days). Sperm were extracted from the cauda epididymis and their motility was evaluated by means of a CASA system. A principal component and clustering analysis were carried out to identify subpopulations. Post mortem time caused a significant decrease in motility quality, and a general deterioration in subpopulation characteristics. We found three subpopulations the first day, and the one indicating good sperm quality decreased with post mortem time until it disappeared on the fourth day. This may indicate considerable impairment of the samples after 72 h post mortem, which could compromise their use in AI programs. With regard to season, subpopulation pattern and characteristics were better in the transition and post-rut periods. Moreover, we found one subpopulation formed by mature spermatozoa, which increased from rut to post-rut. This might be a negative fact, because samples collected after the rut may undergo hypermaturation, which possibly impairs fertility. Our results are of interest for the management of wildlife germplasm banks based on post mortem sperm recovery.

Animals↗

Comparison between different markers for sperm quality in the cat: Diff-Quik as a simple optical technique to assess changes in the DNA of feline epididymal sperm.

The majority of wild felids, as well as some domestic cats, have low sperm concentration in their ejaculates, and a high proportion of abnormal spermatozoa. We have employed several possible semen quality markers to further characterize cat epididymal sperm. Methods included possible apoptotic reporters, such as the annexin V assay to monitor exposure of phosphatidylserine (PS) on the outer leaflet of the plasma membrane, as well as cell integrity; and the TUNEL assay to quantify DNA breaks. Sperm surface ubiquitination, another putative marker of sperm quality, was also monitored. The annexin V assay revealed a high percentage of sperm with PS exposure, and the TUNEL assay pointed to high levels (13+/-12%) of sperm with DNA breaks. Correlations were found between apoptotic markers (but not ubiquitination) and semen parameters. In parallel to this analysis, cat sperm morphology was evaluated using the Diff-Quik optical stain, which has been used in human reproduction laboratories. Several types of abnormalities could be characterized with this method. Remarkably, head staining abnormalities detected using the Diff-Quik staining method were strongly correlated with, and could accurately predict, sperm DNA defects detected in the same sample using the TUNEL assay. We therefore suggest that sperm morphology analysis using Diff-Quik could be used in field conditions to assess sperm status, due to the simplicity of the procedure and the equipment involved.

Animals↗

Birth of live Spanish ibex (Capra pyrenaica hispanica) derived from artificial insemination with epididymal spermatozoa retrieved after death.

As a consequence of increasing limitations to maintaining genetic variability in endangered wildlife species, methods of assisted reproduction widely used in domestic animals are being applied to nondomestic species. However, practical efforts have met limited success to date. The Spanish ibex (Capra pyrenaica hispanica) is a wild caprine originating exclusively in the mountains of Spain. This study was designed to evaluate the fertilizing capability of cryopreserved Spanish ibex epididymal spermatozoa recovered postmortem. For this purpose, we have previously evaluated the effect of time elapsed between death and sperm recovery on spermatic parameters, and the fertilization ability of frozen-thawed spermatozoa using heterologous in vivo fertilization by intrauterine insemination in domestic goat (Capra hircus). The time of death significantly affected most sperm quality parameters (motility, viability and intact acrosomes). The fertility obtained by heterologous artificial insemination was 18.7%, and only goats inseminated with spermatozoa recovered within 8h after death became pregnant. Our findings showed that heterologous in vivo fertilization is a useful method to evaluate the fertilizing capacity of sperm samples in rare or wild species. Sperm samples, with verified fertilization ability in the previous trial, were used to inseminate a total of six ibex females. Inseminations resulted in one pregnancy. The study demonstrated for the first time the feasibility of applying artificial insemination in Spanish ibex.

Animals↗

Epididymal tuberculosis: case report and review of the literature.

The incidence of tuberculosis (TB) is increasing worldwide, with more than 20% of cases exhibiting extrapulmonary manifestations. The genitourinary tract is the most common site of extrapulmonary TB. Epididymal TB, although rare, may be the initial location of infection and may cause infertility. The diagnosis depends on culture of the organism but has been facilitated by the development of urine polymerase chain reaction analysis. Treatment for TB remains the combination of three or four anti-TB drugs for 6 to 9 months. Genitourinary TB remains relatively rare in the United States and requires a high index of suspicion to make the diagnosis.

Epididymis↗

Expression of constitutively active Notch1 in male genital tracts results in ectopic growth and blockage of efferent ducts, epididymal hyperplasia and sterility.

The Notch signaling pathway is involved in a variety of developmental processes. Here, we characterize the phenotypes developing in the reproductive organs of male transgenic (Tg) mice constitutively expressing the activated mouse Notch1 intracellular domain (Notch1(intra)) under the regulatory control of the mouse mammary tumor virus (MMTV) long terminal repeat (LTR). Tg expression was detected in testis, vas deferens and epididymis by Northern blot analysis. In situ hybridization with a Notch1-specific probe lacked sensitivity to detect expression in normal-appearing cells, but demonstrated expression in hyperplastic epithelial cells of the vas deferens, epididymis and efferent ducts. Tg males from three independent founder lines were sterile. Histological analysis of reproductive organs of young Tg males (postnatal ages 8 and 21) showed no difference compared to those of non-Tg males. In contrast, in adult Tg mice from day 38 onwards, the efferent ducts, the vas deferens and most epididymal segments revealed bilateral epithelial cell hyperplasia with absence of fully differentiated epithelial cells. Electron microscopy confirmed the uniformly undifferentiated state of these cells. Immunohistochemistry with anti-PCNA antibody also revealed enhanced proliferation of Tg epididymis. In adult Tg testis, the different generations of germ cells of seminiferous tubules appeared normal, although some tubules were highly dilated and revealed an absence of early and/or late spermatids. The epithelial cells of the Tg tubuli recti and rete testis were not abnormal, but the rete testis was highly dilated and contained numerous spermatozoa, suggesting a downstream blockage. Consistent with a blockage of efferent ducts often seen at the rete testis/efferent duct interface, spermatozoa were absent in epididymis of all adult Tg mice and in all highly hyperplastic efferent duct tubules of these Tg mice. Such a blockage was visualized by injection of Evans blue dye into the rete testis lumen. Finally, the presence of ectopic hyperplastic efferent duct tubules was observed within the testicular parenchyma itself, outside their normal territory, suggesting that Notch1 signaling is involved in the establishment of these borders. This phenotype seems to represent a novel developmental defect in mammals. Together, these results show that constitutive Notch1 signaling significantly affects the development of male reproductive organs.

Animals↗

Pregnancy achieved by intracytoplasmic sperm injection using cryopreserved vasal-epididymal sperm from a man with spinal cord injury.

Anejaculation and poor semen quality are two major causes of infertility in men with spinal cord injury (SCI). The poor motility of retrieved sperm usually has low fertilization potential and is thought to be unfavorable for cryopreservation. This report describes a pregnancy after intracytoplasmic sperm injection (ICSI) with cryopreserved vasal-epididymal sperm from a man with SCI and anejaculation. An attempt was made to obtain sperm through electroejaculation, but no motile sperm were found in two trials. Therefore, the subject underwent vasal aspiration. The retrieved sperm had a concentration of 26 x 10(6)/mL and a motility of 3%. ICSI was considered to be the best choice for the couple, but the wife did not become pregnant in the first cycle of treatment. A successful pregnancy was achieved by ICSI in the second cycle using frozen-thawed sperm, supernumerary in the previous cycle, with a density of 5 x 10(6)/mL and 1% motility. A set of healthy twins, one boy and one girl, were delivered via cesarean section at 36 weeks of gestation. Complementary to other assisted reproductive techniques, ICSI may provide men with SCI a greater opportunity to father children. The supernumerary sperm, regardless of quality, should be cryopreserved to avoid the necessity and risk of repeated assisted ejaculations and aspirations of the genital tract.

Adult↗

Use of NAD(P)H and flavoprotein fluorescence signals to characterize the redox state of pyridine nucleotides in epididymal bull spermatozoa.

The redox behaviour of the NAD(P) system and flavoproteins was registered by simultaneous fluorescence measurements in epididymal bull spermatozoa. The flavoprotein fluorescence signal can nearly exclusively be attributed to an NAD-linked enzyme, alpha-lipoamide dehydrogenase (Em7.4 = -286 mV). A comparison of intact with digitonin-permeabilized spermatozoa revealed that about 50% of the total NAD(P)H fluorescence signal was of mitochondrial origin. Under equilibrium conditions, the midpoint potentials of the NAD(P)H fluorescence signal of both compartments were almost identical (-300 mV). When lactate was present as substrate, 1 mM caffeine increased respiration oxidizing the NAD(P)H system in both mitochondria and cytosol. This indicates a close relationship of the two NAD pools in spermatozoa.

Adenine Nucleotides↗

Effect of insulin on glucagon binding to isolated rat epididymal adipocytes.

Effect of insulin on glucagon binding to rat epididymal adipocytes was studied in vitro. [125I]iodoglucagon binding to isolated adipocytes was increased by preincubation of the cells with insulin. Maximal increase was observed with 7 X 10(-10) M insulin. In Scatchard analysis, [125I]iodoglucagon competition data generated one binding site with a single affinity for glucagon binding in the cells pretreated with buffer alone. Pretreatment of the cells with insulin increased the affinity without changes in the number of binding sites. [125I]iodoglucagon binding to isolated adipocytes was not affected by pretreatment of the cells with luteinizing hormone, follicle-stimulating hormone, growth hormone, or with prolactin. These results suggest that insulin stimulates glucagon binding to adipocytes.

Adipose Tissue↗

Decrease in bicarbonate transport activities during epididymal maturation of porcine sperm.

Kinetic studies of [14C]HCO3- and [35S]SO4(2-) transport show the existence of a common transport system for the two anions on the plasma membranes of porcine epididymal sperm. Km value for bicarbonate (8.3mM) is within the range of physiological concentration of bicarbonate. The anion transport activity is dependent on the temperature but independent of co-existence of Na+. Its pH dependency and sensitivity to the several anion transport inhibitors are similar to those observed in erythrocyte. Anion transport activity decreases during sperm maturation in epididymis.

Adenylyl Cyclases↗

Effects of calcium on brazilin-induced glucose transport in isolated rat epididymal adipocytes.

Brazilin increased [3H]2-deoxyglucose uptake in isolated rat epididymal adipocytes. The fact that calcium may be required for the stimulatory effects of insulin on glucose transport suggests that brazilin might also require calcium for its glucose transport-stimulating action. Changes in the concentration of extracellular calcium had no significant effect on brazilin-induced glucose transport. Nifedipine and verapamil decreased brazilin-induced glucose transport, and quin2-AM abolished the effect of brazilin on glucose transport. A23187, however, showed no effect on brazilin action. 45Ca2+ uptake into adipocytes was not influenced by brazilin treatment, and trifluoperazine significantly inhibited the effect of brazilin on glucose transport. These data suggest that calmodulin and the maintenance of the intracellular calcium concentration, rather than an increase in it, may be essential for the stimulatory action of brazilin on glucose transport.

Adipocytes↗

Effects of brazilin on GLUT4 recruitment in isolated rat epididymal adipocytes.

The effects of brazilin on glucose transport into isolated rat epididymal adipocytes were investigated. Brazilin increased [3H]2-deoxy-D-glucose uptake, which was characterized by an increase in Vmax with no effect on the Km value. Phenylarsine oxide, which inhibits the translocation of glucose transporters, decreased brazilin-stimulated glucose transport to the basal level. The inhibition of phosphatidylinositol 3-kinase (PI3-kinase) with wortmannin also blocked brazilin-stimulated glucose transport. Western blot analysis with an anti-GLUT4 antibody revealed that brazilin increased the translocation of GLUT4 from intracellular pools to the plasma membrane. Brazilin, in combination with phorbol ester, showed an additive effect on glucose transport. The stimulating effect of phorbol ester on glucose transport was inhibited by staurosporine, but the effect of brazilin remained unchanged. Protein kinase C activity was not influenced by brazilin treatment. The inhibition of protein synthesis showed no effect on brazilin-stimulated glucose transport, and GLUT4 content in the total membrane fraction was not altered as a result of treatment with brazilin for 4 hr. Metabolic labeling of GLUT4 with [35S]methionine showed that de novo synthesis of GLUT4 was not induced by brazilin. These data suggest that brazilin may increase glucose transport by recruitment of GLUT4 from intracellular pools to the plasma membrane of adipocytes via the activation of PI3-kinase. However, the effect of brazilin may not be mediated by GLUT4 synthesis and protein kinase C activation.

Adipocytes↗