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PubMed1996

Cases of human infertility are associated with the absence of P34H an epididymal sperm antigen.

The interaction of spermatozoa with the zona pellucida is a critical step of fertilization. Specific sperm surface proteins involved in this process can be added or modified during epididymal transit. We have previously described a 34-kDa human epididymal sperm protein (P34H) that we proposed to be involved in sperm-zona pellucida interaction. In this study, Western blot analysis were performed to determine the level of P34H protein present on the spermatozoa of 16 men with idiopathic infertility. These levels were compared with the amount of P34H protein found in men of proven fertility. In addition, a sperm-zona pellucida binding assay was performed with spermatozoa from fertile and infertile men. Spermatozoa obtained from different semen samples from a given individual had similar P34H levels. However, the amount of P34H varied from one man to another. Nine of 16 infertile men had a P34H level that was less than 30% of the normal value based on a population of fertile men, while the remaining 7 males were in the normal range. Sperm from infertile subjects with a normal P43H determination bound to zonae pellucidae as efficiently as those from controls. However, spermatozoa from subjects with a low amount of P34H exhibited a dramatic diminution in their ability to interact with zonae pellucidae. Our results show that the quantity of the epididymal protein P34H varied from one male to another and that low levels of this epididymal sperm protein are associated with certain cases of idiopathic infertility. Results are discussed with regard to the function of human epididymis in sperm maturation.

Adult↗
PubMed1998

Shedding of a rat epididymal sperm protein associated with infertility induced by ornidazole and alpha-chlorohydrin.

The protein composition of epididymal fluid and sperm extracts of rats treated with the nitroimidazole compound ornidazole was investigated by two-dimensional gel electrophoresis. Epididymal luminal fluid from the corpus and cauda regions of male animals rendered infertile by ornidazole treatment contained a prominent protein (contraception-associated protein 1, CAP1) with a molecular mass of approximately 25 kDa and an isoelectric point (pI) of 5.8; it was not found in fluids, but was present in sperm, from fertile vehicle-fed rats. Infrared matrix-assisted laser desorption/ionization mass spectrometry indicated that the molecular mass of CAP1 was 20420+/-120 daltons. Analysis of 17 amino acids demonstrated 49% homology to a diuretic hormone from an insect (Acheta domesticus). Densitometric quantitation of CAP1 on silver-stained gels indicated its presence in greater amounts in cauda than in corpus fluid from treated animals, whereas fluid from the rete testis lacked CAP1. In vitro incubations of tissue from the caput, corpus, and cauda epididymidal regions with [35S]methionine gave no hint that CAP1 was a secretion product of the epididymal epithelium. The absence of CAP1 from luminal fluid obtained from the sperm-depleted corpus epididymidis of efferent duct-ligated ornidazole-fed rats suggested a spermatozoal origin. CAP1 was present in spermatozoa from the caput epididymidis but not from the rete testis in control animals. Less CAP1 was present in detergent extracts of cauda sperm from ornidazole-treated rats than in sperm from control animals, suggesting a contraceptive-related displacement of protein from sperm to fluid. The association of ornidazole- and alpha-chlorohydrin-induced infertility with the presence of CAP1 in epididymal fluid, probably originating from spermatozoa, suggests a critical role for this protein in fertilization.

Animals↗
PubMed1999

Immunolocalization of CRES (Cystatin-related epididymal spermatogenic) protein in the acrosomes of mouse spermatozoa.

The CRES (cystatin-related epididymal spermatogenic) protein is a member of the cystatin superfamily of cysteine protease inhibitors and exhibits highly restricted expression in the reproductive tract. We have previously shown that CRES protein is present in elongating spermatids in the testis and is synthesized and secreted by the proximal caput epididymal epithelium. The presence of CRES protein in developing germ cells and in the luminal fluid surrounding maturing spermatozoa prompted us to examine whether CRES protein is associated with spermatozoa. In the studies presented, indirect immunofluorescence, immunogold electron microscopy, and Western blot analysis demonstrated that CRES protein is localized in sperm acrosomes and is released during the acrosome reaction. Interestingly, while the 19- and 14-kDa CRES proteins were present in testicular and proximal caput epididymal spermatozoa, the 14-kDa CRES protein was the predominant form present in mid-caput to cauda epididymal spermatozoa. Furthermore, following the ionophore-induced acrosome reaction, CRES protein localization was similar to that of proacrosin/acrosin in that it was detected in the soluble fraction as well as associated with the acrosome-reacted spermatozoa. The presence of CRES protein in the sperm acrosome, a site of high hydrolytic and proteolytic activity, suggests that CRES may play a role in the regulation of intraacrosomal protein processing or may be involved in fertilization.

Acrosin↗
PubMed2000

Identification of a hamster epididymal region-specific secretory glycoprotein that binds nonviable spermatozoa.

Even though the epididymis produces an environment promoting sperm maturation and viability, some sperm do not survive transit through the epididymal tubule. Mechanisms that segregate the epididymal epithelium and/or the viable sperm population from degenerating spermatozoa are poorly understood. We report here the identification and characterization of HEP64, a 64-kDa glycoprotein secreted by principal cells of the corpus and proximal cauda epididymidis of the hamster that specifically binds to and coats dead/dying spermatozoa. The HEP64 monomer contains approximately 12 kDa carbohydrate and, following chemical deglycosylation, migrates as a approximately 52-kDa polypeptide. Both soluble (luminal fluid) and sperm-associated HEP64 are assembled into disulfide-linked high molecular weight oligomers that migrate as a doublet band of 260/280 kDa by nonreducing SDS-PAGE. In the epididymal lumen, HEP64 is concentrated into focal accumulations containing aggregates of structurally abnormal or degenerating spermatozoa, and examination of sperm suspensions reveals that HEP64 forms a shroudlike coating surrounding abnormal spermatozoa. The HEP64 glycoprotein firmly binds degenerating spermatozoa and is not released by either nonionic detergent or high salt extraction. Electron microscopic immunocytochemistry demonstrates that HEP64 localized to an amorphous coating surrounding the abnormal spermatozoa. The potential mechanisms by which this epididymal secretory protein binds dead spermatozoa as well as its possible functions in the sperm storage function of the cauda epididymidis are discussed.

Acrosome↗
PubMed2002

Rabbit epididymal secretory proteins. I. Characterization and hormonal regulation.

Analyses of samples of luminal fluid from the rete testis, distal efferent ducts, and epididymal regions 2-5 and 8 revealed that 91% of the fluid leaving the testis is reabsorbed by the efferent ducts, 79% of the remainder is reabsorbed proximal to epididymal regions 4 and 5, and there is a net secretion of fluid into the duct caudally. There is a net reabsorption by the efferent ducts of 73% of the protein leaving the testis and then a net secretion along the epididymis. SDS-PAGE of the luminal fluids indicated that four new protein bands that were not present in blood appeared in the efferent ducts, 5 in epididymal regions 1-5, 6 in regions 6 and 7, and one in region 8. Two bands in samples from the efferent ducts were absent caudally, and one band present in region 7 was absent in region 8. The rates of incorporation of (35)S-methionine into minced duct in vitro varied among regions when expressed per milligram of wet weight of tissue (region 2-5 > region 7 > region 6 > region 1 > region 8 > ductuli efferentes), and orchidectomy had little effect on the rates. Incorporation into four proteins that were secreted in vitro (M(r) 38 000, 20 000, 15 000, and 13 000) was reduced or abolished by orchidectomy and restored by testosterone therapy. The secretion of three proteins (M(r) 52 000, 23 000, and 22 000) was reduced or abolished by orchidectomy and not restored by testosterone therapy. SDS-PAGE of detergent extracts of sperm indicated that five proteins were lost and nine were gained during epididymal transit. Seven of the proteins gained were about the same molecular weight as proteins secreted by the epididymis (M(r) 94 000, 52 000, 38 000, 36 000, 22 000, 20 000, and 13 000) and were analyzed using N-terminal amino acid microsequencing.

Amino Acids↗
PubMed2003

The efficacy of repeat percutaneous epididymal sperm aspiration procedures.

PURPOSE: The development of intracytoplasmic sperm injection spawned new methods of sperm retrieval for men with obstructive azoospermia who did not want to undergo reconstruction of the seminal tract. There is a wide array of different procedures that may be performed in these cases, for instance percutaneous epididymal sperm aspiration (PESA) and microepididymal sperm aspiration. However, concerns regarding the presence of sperm in a second PESA attempt due to possible fibrosis have been suggested by many authors. We evaluate if it is worthwhile to repeat percutaneous epididymal sperm aspiration. MATERIALS AND METHODS: The records of 20 patients (23 attempts) who underwent repeat PESA from January 1996 to September 2000 for assisted reproductive technique purposes were reviewed. In all patients the repeat procedure was performed on the same side as the previous PESA. Data were collected on patient age, presence of motile sperm during PESA, epididymal side, pregnancies and abortion rates. RESULTS: Mean patient age +/- SD was 32.4 +/- 5.6 years. One patient was excluded from our analysis due to lack of information on the chart regarding the side of the procedure. Repeat PESA was performed in the right epididymis in 12 attempts and in the left in 10. Of the remaining 19 patients 14 (73.68%) did not and 5 (26.3%) have sperm in the epididymal fluid. In these 5 patients 8 repeat PESA procedures were performed (3 procedures in 1 and motile sperm was always found (8 of 22 attempts, 36.4%). Three patients achieved pregnancy with the motile sperm retrieved from the repeat PESA (3 of 8 repeat attempts, 37.5%). No abortions were detected. CONCLUSIONS: More than a third of repeat PESA attempts resulted in the presence of motile sperm. Before performing testicular sperm aspiration or extraction in patients who have undergone previous PESA without achieving pregnancy, repeat PESA may be done. Further attempts should be added in the future to confirm these results.

Adult↗
PubMed2004

The epididymal slide technique: a tension-free anastomosis in a vasoepididymostomy.

PURPOSE: Vasoepididymostomy is a technically demanding procedure that requires excellent positioning of the abdominal vas deferens at the epididymal tubule as well as a tension-free anastomosis. We describe a novel technique of securing the vas deferens to the testicle that allows manipulation of the epididymis and ensures a tension-free anastomosis. MATERIALS AND METHODS: After the vas deferens and epididymis are isolated in the usual fashion the vas is brought to the lateral portion of the testicle. A stay stitch is placed on the abdominal vas. This stay stitch is then placed through the tunica albuginea immediately posterior to the corpus or cauda epididymis. This allows the epididymis to slide in cephalad and caudad fashion prior to vasoepididymostomy and allows a tension-free anastomosis. RESULTS: We have used this technique routinely for vasoepididymostomy without any associated complications and have found it to be technically feasible in our last 50 cases. CONCLUSIONS: Although it is simple, the epididymal slide technique allows minimal to no distortion of the epididymal tubule, secure and safe fixation of the vas deferens, flexibility in choosing an appropriate epididymal tubule and tailoring of tension at the conclusion of the procedure. This technique is now used routinely at our center to allow a tension-free anastomosis during vasoepididymostomy and it has been technically feasible in all of our last 50 cases.

Anastomosis, Surgical↗
PubMed2004

Epididymal metastasis from carcinoma of the prostate.

Epididymal Metastasis from a primary carcinoma of the prostate gland is a rare but recognised phenomena. We describe a case of such metastasis which, unlike previous reports, presents as a painful epididymal mass. Therefore it is important for urologists to consider epididymal metastasis as part of the differential diagnosis in a patient with known carcinoma of the prostate and a tender epididymal mass.

Aged↗
PubMed1980

Effects of an intra-epididymal and intra-scrotal copper device on rat spermatozoa.

The effects of implantation of an intra-epididymal (IECD) and intra-scrotal (ISCD) copper device on rat testicular and epididymal spermatozoa and the copper contents in its various reproductive tissues were studied and compared with another group of animals bearing the copper device and fed supplemental ascorbic acid. The motility of cauda epididymal spermatozoa in vitro in the presence of different concentrations of copper ions was also investigated. The results revealed that the in vitro decrease in motility was more rapid with increasing concentrations of copper sulphate solutions and the addition of ascorbic acid inhibited the motility of the spermatozoa much more rapidly than the copper sulphate solution alone, probably due to the strong acidic properties of ascorbic acid which caused a rapid fall in pH. In the copper device bearing rats, the count, motility, fertilizing capacity, and metabolism of epididymal spermatozoa were reduced which has been correlated with alterations in their morphology. An accumulation of copper occurred in several reproductive tissues, but recovery to almost normal levels was obtained by feeding ascorbic acid to copper implanted rats. The data supports the view that ascorbic acid had a beneficial effect in copper toxicity. Although the intraepididymal copper implantation was found to be a more effective male contraceptive device than the intra-scrotal, further work is necessary in terms of reversibility of the effects and easy removal of the device.

Animals↗
PubMed1994

Gene expression in the dog epididymis: a model for human epididymal function.

The physiology of the epididymis is an integral part of the maturation process by which human spermatozoa acquire the ability to reach and fertilize an oocyte. Because of the high degree of species specificity exhibited by the epididymal proteins involved in sperm maturation, we have assessed tissue from several alternative species for their suitability as a model for human epididymal physiology. Of these, the dog appears to offer an appropriate system. Northern hybridization using cDNA probes specific for human epididymal genes established that, irrespective of dog breed, the canine equivalents of the epididymis-specific HE1, HE4 and HE5 mRNAs were expressed highly in the canine epididymis. cDNA cloning and sequencing confirmed that the canine gene products, CE1, CE4 and CE5 were indeed true structural homologues of their human counterparts. Finally, tissue culture conditions were established wherein all three specific canine genes remained up-regulated after 5 days of culture. Thus, the prerequisite criteria for the development of a system which models human epididymal physiology are to a large degree fulfilled by this canine culture system.

Amino Acid Sequence↗
PubMed1995

Functional and structural characteristics of human epididymal spermatozoa retrieved by transcutaneous aspiration.

The present paper describes the development of a simple, repeatable technique for transcutaneous aspiration of epididymal spermatozoa in patients with inoperable excretory azoospermia. Some functional and structural parameters of the retrieved spermatozoa were also evaluated. Epididymal spermatozoa from patients and ejaculated spermatozoa from fertile donors showed similar percentages of acrosome-reacted cells after incubation for 6 or 15 h in capacitating medium. Penetration of zona-free hamster oocytes, expressed as the percentage of penetrated ova, was significantly lower (p < 0.01) for spermatozoa from patients than for ejaculated spermatozoa from fertile donors. The percentage of epididymal spermatozoa with stable nuclei in sodium dodecyl sulphate was lower (p < 0.001) than for ejaculated spermatozoa from fertile donors. A positive linear correlation (r = 0.74, p < 0.005) was evident between the oocyte penetration rate and the percentage of spermatozoa with stable nuclei. In conclusion the percutaneous retrieval of epididymal spermatozoa in excretory azoospermia allows collection of motile spermatozoa, which can be evaluated for their structural and functional characteristics before repetition of the technique for retrieval of spermatozoa for assisted fertilization techniques.

Adult↗
PubMedDate not supplied

Effect of adrenalectomy and corticosterone replacement on epididymal carbohydrate metabolism--studies on mature male rats.

The effect of adrenalectomy and corticosterone replacement on epididymal enzymes involved in obligatory steps of glycolysis and pentose phosphate pathway were studied along with serum hormonal profiles. Adrenalectomy was found to elevate serum prolactin while the gonadotropins and testosterone were unaltered. In caput epididymal tissue enzymes of the pentose phosphate pathway. Glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities were increased after adrenalectomy. However, in corpus epididymal tissue the key enzymes viz. hexokinase, 6-phosphofructokinase and pyruvatekinase of the glycolytic pathway were elevated leaving the pentose phosphate pathway unaffected. Adrenalectomy was also found to favour glycolysis of the epididymal spermatozoa. The possible direct effect of prolactin is discussed to explain the enzymatic changes in epididymis. Corticosterone replacement was found to maintain the enzyme activities along with serum prolactin and corticosterone at control levels. In conclusion, it is suggested that the adrenalectomy induced changes in enzyme activities could be due to the direct effect of prolactin.

Adrenalectomy↗
PubMedDate not supplied

Vesicles in rat epididymal fluid. Existence of two populations differing in ultrastructure and enzymatic composition.

Glycosidase activity is very high in rat epididymal fluid as a consequence of the secretory capacity of the epithelium. The mechanism of this secretion is, so far, unknown. Membrane-bound vesicles with activity of beta-galactosidase and N-acetyl-beta-D-glucosaminidase were previously isolated by us from rat epididymal fluid. We report here the existence of two populations of epididymal vesicles separated by centrifugation in a sucrose gradient. They were found to differ in isopicnic equilibrium, size, ultrastructure, and enzymatic activity. Seven days after castration the protein content and specific activities of both enzymes were found decreased in the fractions containing the vesicles. A role in enzyme secretion by the epididymal epithelium is suggested for each vesicle population.

Acetylglucosaminidase↗
PubMedDate not supplied

Chromatin condensation in cat spermatozoa during epididymal transit as studied by aniline blue and acridine orange staining.

Chromatin stability and DNA-resistance to acidic denaturation was evaluated by acidic aniline blue and acridine orange staining of cat sperm from different regions of the epididymis. The results were related to conventional sperm parameters. The percentage of aniline blue-stained spermatozoa (persisting histones) decreased significantly from the caput to the cauda region (31.8% and 7.8%, respectively; P<0.0001). The percentage of stained heads of cauda sperm was much lower in populations of morphologically normal forms than in those with abnormal forms (4.1% and 13.8%, respectively, P<0.0001). Among spermatozoa with abnormalities, the percentage of stained heads was significantly higher in cells with head abnormalities than in sperm with only tail abnormalities (87.1% and 10.3%, respectively; P>0.0001). With acridine orange fluorescence staining, 86.5% of cauda epididymal region (51.1%) to the cauda epididymal region (86.5%). The percentage of cauda epididymal sperm with normal condensed chromatin was neither linked to testicular sperm count, motility nor to age of the cats. The parameter of chromatin condensation and stability can be a valuable index of sperm quality, reflecting the possible disorders of spermatogenesis and epididymal sperm maturation, frequently observed in feline species.

Acridine Orange↗
PubMed1999

Regulation of anion secretion by cyclo-oxygenase and prostanoids in cultured epididymal epithelia from the rat.

1. The role of cyclo-oxygenase (COX) in the regulation of anion secretion (measured as short- circuit current, Isc) in cultured epididymal epithelia from immature rats was investigated. 2. COX inhibitors attenuated the increase of anion secretion caused by bradykinin (LBK) but had no effect on that caused by PGE2, suggesting that prostaglandin synthesis mediates the secretory response of the tissues to LBK. 3. The apparent IC50 values for indomethacin, piroxicam and L-745,337 in inhibiting the LBK-induced Isc were 0.14, 1.34 and 15.7 microM, respectively. This order of potency: indomethacin > piroxicam > L-745,337 >> DFU suggests the involvement of the COX-1 isozyme in the mediation of the secretory response to LBK. 4. Among the COX products (prostaglandins, thromboxane and prostacyclins) tested, only PGE2 and, to a much lesser extent, PGF2alpha stimulated anion secretion by cultured rat epididymal epithelia. 5. The effect of PGE2 was mimicked by 11-deoxyl PGE1, a specific prostaglandin E (EP)2/4 receptor agonist, but not by sulprostone, a specific EP1/3 receptor agonist, indicating that cyclic AMP-coupled EP2/4 receptors are involved in the LBK-stimulated anion secretion. 6. A reverse transcriptase-polymerase chain reaction study detected the expression of COX-1 and COX-2 mRNA in intact rat epididymis and in cultured epididymal epithelia. The expression of COX-1 mRNA was reduced by LBK by 44 %. 7. Immunohistochemical studies demonstrated the presence of COX-1 immunoreactivity in the basal cells of the intact rat epididymis. By comparison, COX-2 immunoreactivity was detected in the apical pole of the principal cells. 8. The role of COX in the formation of the epididymal microenvironment and the implication of long term administration of non-steroidal anti-inflammatory drugs (NSAIDs) on male fertility are discussed.

Animals↗
PubMed2006

Gene deletion reveals roles for annexin A1 in the regulation of lipolysis and IL-6 release in epididymal adipose tissue.

In this study, epididymal adipose tissue from male annexin 1 (ANXA1)-null and wild-type control mice were used to explore the potential role of ANXA1 in adipocyte biology. ANXA1 was detected by Western blot analysis in wild-type tissue and localized predominantly to the stromal-vascular compartment. Epididymal fat pad mass was reduced by ANXA1 gene deletion, but adipocyte size was unchanged, suggesting that ANXA1 is required for the maintenance of adipocyte and/or preadipocyte cell number. Epididymal tissue from wild-type mice responded in vitro to noradrenaline and isoprenaline with increased glycerol release, reduced IL-6 release, and increased cAMP accumulation. Qualitatively similar but significantly attenuated responses to the catecholamines were observed in tissue from ANXA1-null mice, an effect that was not associated with changes in beta-adrenoceptor mRNA expression. Lipopolysaccharide (LPS) also stimulated lipolysis in vitro, but its effects were muted by ANXA1 gene deletion. By contrast, LPS failed to influence IL-6 release from wild-type tissue but stimulated the release of the cytokine from tissue from ANXA1-null mice. ANXA1 gene deletion did not affect glucocorticoid receptor expression or the ability of dexamethasone to suppress catecholamine-induced lipolysis. It did, however, augment IL-6 expression and modify the inhibitory effects of glucocorticoids on IL-6 release. Collectively, these studies suggest that ANXA1 supports aspects of adipose tissue mass and alters the sensitivity of epididymal adipose tissue to catecholamines, glucocorticoids, and LPS, thereby modulating lipolysis and IL-6 release.

Adipocytes↗
PubMed1988

Dietary influence on the insulin function in the epididymal fat cell of the Wistar rat. I. Effect of type of fat.

For at least 3-4 weeks, young male Wistar rats were fed semisynthetic diets containing sunflowerseed oil, palm oil, olive oil, linseed oil, cocoa butter and coconut oil as dietary fat. The type of dietary fat had little effect on body weight, epididymal fat pad weight and on the diameter of fat cells isolated from the epididymal fat pads, but the rats fed the linseed oil-containing diet had a lower epididymal fat pad weight. The fatty acid composition of the triacylglycerol fractions of the fat pads correlated well with those of the dietary fats. The correlations with the fatty acid composition of the phospholipids of the fat pads were less pronounced. High responses to insulin in the epididymal fat cells were obtained with sunflowerseed oil, linseed oil and olive oil, whereas low responses were found for cocoa butter, palm oil or coconut oil. Rather than the amount of polyunsaturated fatty acids or the ratio of polyunsaturated to saturated fatty acids, the amount of saturated fatty acids with 12, 14 or 16 carbon atoms appeared to be the most important parameter in determining the maximal insulin response. A negative correlation was found between the amount of saturated fatty acids in the diet and the extent of insulin response. The modulating effects of the dietary type of fat on the response to insulin cannot be fully explained by changes in the number of insulin receptors on the fat cell surface as determined by insulin binding but must, at least partially, be ascribed to postreceptor effects.

Adipose Tissue↗
PubMed1976

Exogenous hypercortisolism and epididymal fat cell count in young rats.

The influence of two different grades of exogenous hypercortisolism on body weight, epididymal fat pad weight and the total number of fat cells in epididymal fat pads was investigated in young, growing rats. Cortisol, 4-5 mg/kg/day orally from the 7th to the 9th week, reduced body weight gain, whereas epididymal fat pad weight and fat cell content did not differ from those of the control rats. Cortisone acetate, 2.5 mg per 100 g, given subcutaneously for 2 weeks to rats 4-11 week of age caused in the young rat (4-6 weeks) a partial inhibition of the normal increase in body weight, whereas in the young-adult rat (6 weeks and older) an actual decrease of body weight was seen. At both dose levels and - with respect to the higher dose level- in all age groups studied, the weight and fat cell content of the epididymal fat pad were not changed by the cortisone (cortisol) treatment.

Adipose Tissue↗