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Analysis of surface proteins of rat spermatozoa during epididymal transit and identification of antigens common to spermatozoa, rete testis fluid and cauda epididymal plasma.

Spermatozoa from the testis and cauda epididymidis of the rat were surface labelled with radioactive iodide, extracted with detergent, and the radioactive proteins separated by two-dimensional polyacrylamide gel electrophoresis. In some instances spermatozoa were also surface labelled with tritiated borohydride in the presence of galactose oxidase. Soluble proteins in blood serum, rete testis fluid and cauda epididymal plasma were also iodinated and separated by gel electrophoresis. In addition, aliquants of the radioactive sperm extracts, blood serum and reproductive tract fluids were each immunoprecipitated with polyspecific antisera directed against either testicular sperm membranes, caudal sperm membranes, blood serum, rete testis fluid or cauda epididymal plasma before gel electrophoresis. From the patterns of radioactive proteins detected on the resultant gels, a two-dimensional map was created for each of the sperm extracts and for the various fluids. Proteins which were nonhomologous between testicular and caudal spermatozoa were identified, as well as proteins which were common to spermatozoa and reproductive tract fluids. Epididymal transit was characterized by the loss of certain proteins from the sperm surface, including three borohydride-labelled proteins of Mr 130 000, and by the addition of others, most notably a highly abundant protein of Mr 42 000. Several of the proteins lost from spermatozoa accumulated in the epididymal plasma whilst some of those added to the sperm surface could be identified as direct secretory products of the epididymis. Rete testis fluid contained blood proteins in addition to others presumed to be testis-specific, whilst the composition of cauda epididymal plasma was markedly different from blood serum or rete testis fluid.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

In vitro culture of brushtail possum (Trichosurus vulpecula) epididymal epithelium and induction of epididymal sperm maturation in co-culture.

A medium modified from eutherian systems was used to culture epididymal epithelial cells of the brushtail possum (Trichosurus vulpecula) for more than 2 months. Epididymal tubule fragments from the caput, corpus and cauda epididymides were used to generate cell monolayers. All three epididymal cell culture systems supported maturational changes in marsupial spermatozoa and enabled immature possum spermatozoa to differentiate from a T-shape to a streamlined shape, accompanied by the development of progressive motility after co-culture with 7-day-old cultured epididymal cell monolayers. This epididymal cell and sperm co-culture system for marsupial species may facilitate the identification of specific epithelial factors that affect sperm maturation, particularly in a species in which morphological maturation is readily visible.

Animals↗

Effect of alpha-chlorohydrin on epididymal sperm and epididymal plasma in swine.

Investigation of the contraceptive mechanism of alpha-chlorohydrin was done by analyses of epididymal plasma and certain epididymal sperm characteristics after oral administration of 0, 5, 10 or 30 mg/kg, day of the drug to boars for 15 days. Water resorption in caput epididymidis was slightly decreased in all treatment groups. Sodium, potassium, chloride, glycerylphosphorylcholine levels, and seminal antigens in epididymal plasma were not altered significantly by 10 or 30 mg/kg of the drug. The boars on 5 mg/kg exhibited significantly elevated sodium, potassium, or chloride values in various segments. Motility was significantly lower on corpus and proximal cauda epididymal spermatozoa from alpha-chlorohydrin treated boars. Only boars receiving 30 mg/kg exhibited impaired sperm motility in the distal cauda. The movement of the cytoplasmic droplets to the distal position was retarded in boars on the two highest dose levels. The results suggest that the contraceptive effects of alpha-chlorohydrin in the boar is probably not mediated via an impaired epididymal function.

Animals↗

Amiodarone-associated epididymitis: drug-related epididymitis in the absence of infection.

An atypical epididymitis syndrome developed in 6 of 56 men (11 per cent) treated with amiodarone, an effective new anti-arrhythmic agent. Of the 6 patients 5 (87 per cent) had bilateral epididymal enlargement and pain. The mean dosage in the patients with epididymitis was 700 mg. per day compared to 377 mg. per day in all patients (p less than 0.01). No infectious etiology was implicated in any patient. Temporary discontinuation or decrease in dosage is recommended for patients who suffer noninfectious epididymitis while on amiodarone therapy.

Adult↗

Thyroid-epididymal relationship. I. Influence of hypothyroidism on epididymal lipids.

The influence of thyroidectomy on the lipid composition of caput and cauda epididymides have been studied. The analysis was conducted in epididymal tissues free from fluids and sperm. A general tendency towards accumulation of epididymal lipids was observed in hypothyroid rats. Hypothyroidism also brought about a differential regional response, which may be age-related. The existence of a relationship between triacylglycerols, phospholipids and diacylglycerols has been suggested. Since immediate thyroxine replacement to thyroidectomised rats maintained epididymal lipids at control levels, it is concluded that hypothyroidism has a definite influence on the epididymal lipid composition.

Animals↗

Percutaneous epididymal sperm aspiration versus microsurgical epididymal sperm aspiration for irreparable obstructive azoospermia--experience with 100 cases.

PURPOSE: This study investigated the sperm retrieval success rates, fertilization rates, pregnancy rates, and complications of percutaneous epididymal sperm aspiration (PESA) and microsurgical epididymal sperm aspiration (MESA) in cases of irreparable obstructive azoospermia. METHODS: During a period of 36 months, 100 men with irreparable obstructive azoospermia underwent 109 cycles of sperm retrieval procedures and intracytoplasmic sperm injection (ICSI). We routinely performed PESA first in each retrieval cycle; MESA and/or testicular sperm extraction (TESE) were performed if PESA failed. The sperm retrieval success rates, mean fertilization rates, and pregnancy rates of PESA and MESA were evaluated. RESULTS: PESA was performed in all 109 retrieval cycles with a successful sperm retrieval rate of 61%. When PESA failed to retrieve a sufficient number of viable sperm, MESA was subsequently performed with a sperm retrieval rate of 93%. Three cases, which had failed retrieval with both the PESA and MESA procedures, received TESE successfully. The rates of fertilization and pregnancy were 56% and 39% in the 66 PESA-ICSI cycles, respectively, and 47% and 45% in the 40 MESA-ICSI cycles. No significant differences were found in fertilization rates or pregnancy rates among the various sperm retrieval methods and obstruction etiologies. The overall mean fertilization rate and pregnancy rate were 51% and 41%, respectively. CONCLUSION: Both PESA and MESA can be used successfully to obtain sufficient sperm for ICSI. PESA cannot replace MESA in some cases as some epididymal pathologies prevent its success. The results of this study indicate that PESA should be the treatment of choice for patients with ductal obstruction distal to the epididymis, owing to its higher initial success rate. In contrast, patients with irreparable epididymal obstruction might achieve better success rates with MESA.

Epididymis↗

Candida epididymitis: newly recognized opportunistic epididymal infection.

Epididymitis is common, yet it is rarely associated with fungal pathogens. We report a case of Candida albicans epididymitis in a diabetic which was ultimately treated by orchiectomy. Opportunistic infections of the genitourinary tract in immunosuppressed patients are becoming more prevalent; examples include fungal infections in patients with acquired immune deficiency syndrome or after organ transplant. The fact that opportunistic organisms can invade the epididymis and produce infection suggests that in cases of persistent epididymitis, which have failed to respond to conventional therapy, more aggressive diagnostic procedures should be considered. Needle aspiration with cultures for fungus and viral organisms should be performed. This is especially true in patients with preexisting chronic illness or an immune compromised state.

Aged↗

Potential contraceptive use of epididymal proteins: evidence for the participation of specific antibodies against rat epididymal protein DE in male and female fertility inhibition.

Previous studies in our laboratory indicated that immunization of male and female Wistar and Lewis rats with epididymal protein DE, resulted in the development of anti-DE antibodies in over 90% of the animals, with a significant and reversible reduction of fertility. In the present study, ELISA assays performed to analyze the evolution of the immune response indicated that antibody levels in the sera of immunized animals reached a maximum at 8 weeks after the initial injection and then gradually decreased, returning to control values by the end of the sixth month. Western blot experiments demonstrated that the immune sera specifically recognized DE in epididymal sperm extracts and epididymal cytosol, while no reaction was observed with different reproductive and essential organs. The immune sera were also capable of recognizing DE on the surface of both fresh and capacitated sperm as indicated by indirect immunofluorescence experiments. Finally, the exposure of sperm to immune sera prior to uterine insemination resulted in a significant (P < 0.05) reduction in the percentage of fertilized eggs compared to controls, with no effect on sperm motility and viability, nor on their ability to undergo capacitation. Together, these results support the participation of the raised antibodies as mediators of the antifertility effect and suggest a specific interference at the sperm-egg interaction level.

Animals↗

The clinical use of epididymal aspiration cultures in the management of selected patients with acute epididymitis.

Culture and sensitivity testing of specimens obtained by percutaneous and intraoperative needle aspiration of the epididymis have been of clinical importance in choosing proper antimicrobial therapy in selected patients with acute non-venereal epididymitis. We have found aspiration cultures to be valuable in patients 1) with indwelling urethral catheters, 2) who fail to respond to initial antimicrobial therapy, 3) who have epididymitis found on surgical exploration for torsion of the teticle and 4) with recurrent epididymitis in whom the etiologic agent is uncertain.

Acute Disease↗

Clusterin (SGP-2) in epididymal luminal fluid and its association with epididymal spermatozoa in androgen-deprived rats.

Clusterin is a heterodimeric glycoprotein synthesized and secreted by rat Sertoli cells and epididymal epithelium. The goal of this study was to determine the presence of clusterin in the luminal fluid of the cauda epididymides and its association with the membranes of developing spermatozoa in the presence and absence of androgen. We have previously demonstrated by two-dimensional (2-D) Western blot probing for clusterin that in epididymal fluid the amounts of clusterin were: caput greater than corpus greater than cauda. Luminal fluid from cauda epididymides was collected from control and orchiectomized rats (6 and 12 days) and orchiectomized animals that received testosterone implants. Equal volumes of fluid were analyzed by 2-D Western blot probing for clusterin. Following orchiectomy, there was an increase in clusterin in the luminal fluid after 6 days and maximal amount after 12 days compared with control cauda fluid. Orchiectomized animals which received testosterone treatment showed levels of clusterin comparable to that of controls. Serum clusterin was detected in fluid of orchiectomized animals with and without testosterone. Western blots of cauda sperm membrane extracts of control animals and orchiectomized animals treated with testosterone had a very low level of epididymal clusterin, whereas extracts collected from orchiectomized animals revealed high levels of clusterin. We suggest that, in the normal animal, clusterin is secreted into the lumen of the proximal epididymis where it binds to the sperm membrane. In the distal epididymis, clusterin dissociates from sperm and is processed (proteolysis/endocytosis). We hypothesize that, in the absence of androgen, the processing and regulation of clusterin is disrupted.

Animals↗

Phospholipid requirement of epididymal testosterone 5 alpha-reductase and phospholipid composition of epididymal microsomes.

We have investigated phospholipid requirement for testosterone 5 alpha-reductase solubilized from microsomal and nuclear fractions of rat epididymis. The 5 alpha-reductase from microsomal fraction was stimulated by phosphatidylcholine (PC) with long acyl-chain lengths, but inhibited by short chain PC. The nuclear enzyme activity was weakly activated by PC with various acyl-chain lengths tested. Synthetic phosphatidylserine (PS), such as dioleoylPS, most strongly stimulated the microsomal enzyme activity, but did not exhibit any activation of the nuclear enzyme activity. Endogenous phospholipids, such as PC, PS, and phosphatidylethanolamine (PE) separated from bovine epididymal microsomes were tested for their stimulatory effects on microsomal and nuclear enzymes. Among these endogenous phospholipids, PS most greatly stimulated the microsomal 5 alpha-reductase activity, whereas both PC and PE weakly activated the enzyme activity. On the other hand, endogenous PC and PS had no ability to support the nuclear enzyme activity. The fatty acid compositions of PC and PS from bovine epididymal microsomes were determined, in order to elucidate the relationship between 5 alpha-reductase activation by these phospholipids and the structure of their acyl chains. The relative content of fatty acids in PC, in a decreasing order, was palmitate > linoleate > oleate; that in PS was stearate > oleate > palmitate. Based on these observations, the roles of microsomal PS and PC in epididymal 5 alpha-reductase reaction will be discussed.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Evaluation of motility, freezing ability and embryonic development of murine epididymal sperm after coculture with epididymal epithelium.

Murine sperm from the caput, corpus and cauda epididymis were cocultured with epididymal epithelial cells of their own region or more distal regions, in the presence and absence of androgens (testosterone and dihydrotestosterone). Epithelial cell cultures were used 3 or 10 days after preparation in a complex tissue culture medium (Chang's) as plated tubules. The coculture studies involving spermatozoa and oocytes with epithelial cells were carried out in T6 medium. Motility of caput spermatozoa was maintained for 24 h in the presence of day 3 corpus and cauda epithelial cells and hormones but not under other conditions. Likewise, the motility of corpus spermatozoa was maintained for 24 h in the presence of day 3 cauda epithelial cells and hormones but not other conditions. Fertilization of zona-intact oocytes by epididymal spermatozoa was not affected by their coculture for 24 h with epithelial cells but fertilization rates for zone-free oocytes were increased for caput spermatozoa cocultured with more distal epithelial cells. Fertilization rates for both zona-intact and zone-free oocytes were increased for corpus spermatozoa cocultured with more distal cauda epithelial cells. The developmental capacity of embryos derived from caput spermatozoa was not significantly increased by coculture with epithelial cells but those derived from corpus spermatozoa cocultured with cauda epithelial cells were significantly increased. We conclude that the presence of more distal epithelial cells of the mouse epididymis maintains motility in culture, increases the ability of caput and corpus spermatozoa to fertilize zona-free oocytes and increases the developmental capacity of embryos formed from corpus spermatozoa. These observations demonstrate the function of epididymal regions in the maturation of murine spermatozoa for fertilization and embryo development.

Animals↗

Movement characteristics of bovine epididymal spermatozoa: effects of forward motility protein and epididymal maturation.

Movement characteristics of untreated bovine caudal epididymal spermatozoa were compared by high-speed cinemicrography with those of theophylline-activated caput epididymal spermatozoa with and without added forward motility protein (FMP). Comparison of individual movement characteristics clearly established the importance of FMP in converting the nonprogressive motility of theophylline-activated caput sperm into the progressive swimming of mature caudal sperm. Although the total or curvilinear distance traveled in 1 sec by theophylline-activated caput sperm was not changed by the addition of FMP, the linear progression was doubled and the percentage of progressively motile sperm was tripled by this protein. Untreated caudal sperm were 80% motile and theophylline-activated caput sperm were nearly 50% motile; the percentage of motile sperm that were progressive was the same for theophylline-activated caput sperm with FMP and for untreated caudal sperm. Caput sperm without FMP roll infrequently, if at all, but caput sperm with FMP and caudal sperm roll at 4.7 Hz. The beat frequency increases significantly with the addition of FMP and is even higher for caudal sperm. The hydrodynamic power output rises concomitantly with the beat frequency. Perhaps the most striking difference between caput sperm without FMP and those with it is in the swimming paths they follow. Caput sperm without FMP exhibit frequent reversals in direction, or yawing of the sperm heads as they loop back and cross over their tails in an apparently very flexible bending. Their average swimming paths are circles. Caput sperm with FMP and caudal sperm do not show this behavior, but swim in average paths which are linear. The minimum radius of curvature of the tail of caput sperm without FMP is much smaller than that for the other two cell types. These studies clarify the role of FMP in epididymal development of sperm motility.

Animals↗

The effect of epididymal and testicular fluids on the fertilising capacity of testicular and epididymal spermatozoa.

It is possible that the fertilising capacity of spermatozoa in the epididymis is influenced by the epididymal secretion. We have studied this problem by obtaining spermatozoa before entry into the epididymis and after passage through it, incubating both types of spermatozoa in fluids from the rete testis and cauda epididymidis and then checking their fertilising capacity. While spermatozoa from the rete testis were infertile, rete testis fluid did not decrease the fertilising capacity of epididymal sperm from the cauda epididymidis. Fluid from the cauda epididymidis did not promote the fertilising capacity of testicular spermatozoa. These results are discussed in the light of the current understanding of epididymal physiology.

Animals↗

Epididymal cribriform hyperplasia. A variant of normal epididymal histology.

We describe a case of orchiectomy for testicular germ cell neoplasm with a peculiar associated cribriform hyperplasia of the epididymis. The process, which we have termed epididymal cribriform hyperplasia (ECH), is tantalizingly akin to patterns of cribriform ductal carcinoma in situ of the female breast and is characterized by complex arcades and cellular bridges spanning dilated epididymal lumina. The cells lining these interconnecting arches have hyperchromatic nuclei, but lack significant atypia or mitotic activity. A limited search of the Vanderbilt University Medical Center (Nashville, Tenn) surgical pathology files produced 30 cases with evaluable epididymis, 15 of which had some degree of ECH. The ECH occurred in a broad age range with a mean of 40 years. No association of ECH with testicular germ cell neoplasia, adenomatous hyperplasia of the rete testis, or parenchymal atrophy could be documented. We conclude that ECH is a seldom recognized variant of normal epididymal histologic appearance that may be present in up to half of epididymides.

Adolescent↗

[Aspermia of epididymal blockage treated with epididymal tubule vasostomy].

To improve curative effect, we performed aspermia of epididymal blockage on 21 patients with epididymal tubule vasostomy using microsurgical technique between 1985 and 1994. In the 21 patients 20 showed sperm, and 18 increased sperm numbers over 4 x 10(7)/ml postoperation. The wives of 12 patients became reproductive. The selection of 3 different anastomotic ways using microsurgical technique could obviously improve curative effect, according to the dilated degree of epididymal tubule. The selection of anastomotic ways and liquid-passed test of deferential dust was also discussed.

Adult↗

The 26 kD protein recognized on rat cauda epididymal sperm by monoclonal antibody 4E9 has internal peptide sequence that is identical to the secreted form of epididymal protein E.

MAb 4E9, raised against a detergent extract of rat cauda epididymal sperm, recognizes a 26 kD glycoprotein that is found on the plasma membrane of the sperm tail in cauda, but not caput, sperm (Moore et al., 1994). It also recognizes an epididymis-secreted protein that has been shown to be protein E (Xu and Hamilton, 1996). It is felt that the secreted protein becomes associated with sperm, but there has been no biochemical evidence of molecular identity between the secreted and membrane proteins. In this report, the membrane form of the antigen has been purified by reverse phase HPLC. Cyanogen bromide cleavage of the purified protein yielded 3 peptides that were purified, also by reverse phase HPLC. One of the peptides yielded an unambiguous sequence of 34 amino acids that is identical to an internal peptide of the protein found in epididymal fluid. This is the first report showing sequence identity between an epididymis-secreted protein and a protein of the sperm plasma membrane.

Amino Acid Sequence↗

Is micturition disorder a pathogenic factor in acute epididymitis? An evaluation of simultaneous bladder pressure and urine flow in men with previous acute epididymitis.

We evaluated 22 men 22 to 70 years old with previous acute epididymitis by pressure-flow study 3 to 12 months after the inflammation had resolved. Nine healthy men 20 to 62 years old were evaluated as controls. The patients had no symptoms from the lower urinary tract except for 2 men with slight prostatism. The maximum intravesical and maximum voiding pressures were elevated significantly in the patients compared to the controls (p less than 0.05). In most patients and in all of the controls the maximum urinary flow rates were within the normal range according to age. Because of the frequency of high voiding pressures in patients with previous acute epididymitis, this condition may be a pathogenic factor by promoting urethrovasal reflux. The high voiding pressures may be transmitted to the proximal urethra or in cases of a narrow and rigid bladder neck they may produce increased turbulence in the urine stream.

Adult↗