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PubMed1997

Effects of chlormadinone acetate and ethinylestradiol treatment on epididymal 5 alpha-reductase activities in patients with prostate cancer.

The difference between in vivo and in vitro inhibitory effects on epididymal 5 alpha-reductase was investigated by using epididymides obtained from patients treated with chlormadinone acetate (6-chloro-3,20-dioxo-4,6-pregnadien-17-yl acetate, CMA) or ethinylestradiol (17 alpha-ethynyl-1,3,5(10)-estratriene-3,17 beta-diol, EE2). In the in vitro study CMA exhibited competitive inhibition, whereas EE2 was a noncompetitive inhibitor of human epididymal 5 alpha-reductase. Their in vitro inhibitory effects were weak compared with the effect of finasteride ((-)-N-tert-butyl-3-oxo-4-aza-5 alpha-androst-1-ene-17 beta carboxamide), a steroidal 5 alpha-reductase inhibitor. The Ki values of CMA, EE2, and finasteride were 1.4 x 10(-5) M, 1.5 x 10(-5) M, and 1.3 x 10(-9) M, respectively. Despite their weak in vitro inhibitory potency, CMA and EE2 strongly inhibited testosterone 5 alpha-reductase in vivo. There were regional differences in inhibitions by CMA and EE2 on human epididymal 5 alpha-reductase activity depending on the site of the epididymis; the efferent ductules, the head, the body or the tail. In vivo administration of CMA reduced epididymal 5 alpha-reductase activity by 49.7% to 89.4%. In vivo administration of EE2 reduced epididymal 5 alpha-reductase activity by 82.7% to 96.3%. The apparent Km values for the enzyme in patients treated with CMA or EE2 and untreated patients did not differ significantly. The Vmax values were significantly decreased in treated patients. These findings suggest that the marked in vivo inhibition of 5 alpha-reductase induced by CMA and EE2 was not related to the direct action of these compounds, but resulted from a reduction in the amount of the enzyme.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed1985

Comparison of growth hormone binding and metabolic response in rat adipocytes of epididymal, subcutaneous, and retroperitoneal origin.

We undertook a comparison of human growth hormone (hGH) binding and metabolic responses in rat adipocytes of epididymal, subcutaneous, and retroperitoneal origin to determine whether the site of fat depot biopsy might affect the response to hGH stimulation. The results showed highest specific binding in epididymal (3.6%), followed by subcutaneous (2.3%) and retroperitoneal adipocytes (1.5%); half-maximal binding was achieved at 14-18 ng/ml hGH for the three sites. Scatchard analysis of the binding data from each site was linear; there was no significant difference in binding affinities (2.1 to 3.3 X 10(9), M-1), but the number of binding sites was statistically higher in epididymal (9.8 X 10(3) as compared to subcutaneous (7.5 X 10(3), P less than 0.05) and retroperitoneal cells (3.3 X 10(3), P less than 0.01). Stimulation with 5 to 2500 ng pituitary hGH produced a dose-related increase in glucose incorporation, with the largest increase in epididymal fat cells (31%, P less than 0.05) followed by subcutaneous cells (18%, P less than 0.05); no significant increase was seen with retroperitoneal cells. Biosynthetic hGH produced a similar pattern of glucose incorporation in the three sites. Addition of hGH antibodies blocked the glucose incorporation in epididymal adipocytes using both pituitary-derived and biosynthetic hGH. It seems clear that this insulin-like effect is caused by hGH, not an insulin-like impurity. We conclude that the number of binding sites, perhaps related to adipose cell size, differs in adipose tissue from different locations and this influences the metabolic response to hGH stimulation.

Adipose Tissue↗
PubMed1989

Gossypol-induced modifications in the microenvironment of rat epididymal spermatozoa.

Gossypol acetic acid (20, 25 or 30 mg/kg/day orally for 5 weeks) decreased epididymal weight in adult Sprague-Dawley rats but the epididymal concentrations of proteins, lactate dehydrogenase and acid phosphatase were unchanged. The concentrations of carnitine, inositol and potassium in epididymal fluid were decreased in a dose-related manner. These modifications were not due to disturbances of Leydig and Sertoli cell functions which were normal. We suggest that the reduction in epididymal secretion results from a decrease in the number of spermatozoa rather than from a direct action of gossypol on the epididymal epithelium.

Animals↗
PubMed1993

Lipid composition of hamster epididymal spermatozoa.

The lipid composition of hamster epididymal spermatozoa was examined. Caput epididymal spermatozoa were isolated by Percoll density gradient centrifugation without contamination by other cells and they had a specific gravity of 1.10-1.12 g cm-3. Caput and cauda epididymal spermatozoa showed little difference in the amounts of total fatty acid and total sterol. However, sterol composition changed markedly during the transit of spermatozoa through the epididymis: the amount of cholesterol decreased, while the amount of desmosterol and cholesta-7,24-dien-3 beta-ol increased. No significant change in fatty acid composition was observed during the transit, although there was a tendency for an increase in chain length. Both 22:5 and 22:6 represented high percentages in fatty acids of hamster spermatozoa. Some difference in lipids was detected between the upper fraction (1.04 g cm-3) and the lower fraction (1.10 g cm-3) obtained by the density gradient centrifugation of cauda epididymal spermatozoa. Total fatty acid content of the upper fraction was 1.4-fold higher than that of the lower fraction, and the percentage of 18:0 was lower in the latter fraction with a higher percentage of 18:2. The total sterol:total phospholipid ratio in hamster cauda epididymal spermatozoa was 0.21.

Animals↗
PubMed1994

Uptake and release of free L-carnitine by boar epididymal spermatozoa in vitro and subsequent acetylation rate.

In the male reproductive tract, very high concentrations (mmol l-1) of free L-carnitine and acetyl-L-carnitine are found in the epididymides, seminal plasma and spermatozoa. It has been reported that the uptake of free L-carnitine by spermatozoa might be related to the epididymal maturation of the sperm membrane, since a greater uptake was found by caput than by cauda spermatozoa in vitro. However, the free L-carnitine concentrations estimated inside the gametes were never greater than those of the surrounding medium. In this study, we investigated the mechanism of transport of free L-carnitine and its ester acetyl-L-carnitine, through the plasma membrane of mature and immature epididymal boar spermatozoa. In vitro, we found a passive diffusion of both compounds to the spermatozoa, whatever the maturation stage. The spermatozoa might progress in the epididymal lumen and accumulate high amounts of free L-carnitine. The active uptake of free L-carnitine occurs only across epididymal mucosa. These results are in agreement with those reported on cells of other organs that exchange pharmacological free L-carnitine concentrations (mmol l-1) by a passive mechanism through the plasma membrane. The acetylation of high amounts of free L-carnitine inside the spermatozoa was found only in caudal spermatozoa. This result suggests that oxidative metabolism (producing acetyl CoA) might be more active in mature cells. The acetyl-L-carnitine added to the incubation medium of boar spermatozoa was hydrolysed. Enzymatic activity of the sperm membrane is low and this may partially explain the low concentrations of acetyl-L-carnitine found in the caudal epididymal plasma.

Acetylation↗
PubMed2005

Sonic hedgehog pathway inhibition alters epididymal function as assessed by the development of sperm motility.

The sonic hedgehog (Shh) signaling pathway plays a role in pattern orientation in the developing embryo and has been shown to be required for development of the prostate and external genitalia. Recent evidence has shown that important elements of the Shh pathway are also expressed in the adult mouse epididymis at both the gene and protein levels. The objective of the present investigation was to refine the expression pattern of Shh in the mouse epididymis and to determine if the Shh pathway is important for epididymal function vis-à-vis sperm maturation. The former was achieved by microarray analysis of Shh expression in all segments of the mouse epididymis, and the latter was determined by 14-day administration of cyclopamine, a Shh pathway inhibitor, followed by a microassay for the activation and duration of cauda epididymal sperm motility. Shh pathway inhibition was monitored by semiquantitative reverse transcriptase-polymerase chain reaction for expression of epididymal Gli1 and Gli3. The Gli family of transcription factors is commonly activated and regulated by Shh pathway activation. Cyclopamine treatment reduced Gli1 expression by 61% and initiation of cauda sperm motility by 50%. Gli3 expression was reduced by approximately 50%. Subsequent cluster analysis using the microarray data on epididymal gene expression highlighted several potential target genes for the Shh pathway, the most prominent of which is prostaglandin D2 synthase. These results indicate that an operating Shh pathway is important in the murine epididymis for the development of sperm motility and implies a role for Shh signaling in adult epididymal function.

Animals↗
PubMed2006

Induction of oxidative stress by organic hydroperoxides in testis and epididymal sperm of rats in vivo.

The present study describes the extent and pattern of oxidative stress induction in testis and epididymal sperm of rats following in vivo exposure to repeated sublethal doses of 2 model pro-oxidants, namely, t-butyl hydroperoxide (tbHP) and cumene hydroperoxide (cHP). Single sublethal (1/40, 1/20, and 1/10 LD(50)) doses of hydroperoxides (HP) administered intraperitoneally to male rats (CFT-Wistar strain) failed to induce any significant increase in malondialdehyde or reactive oxygen species (ROS) levels in testis or epididymal sperm. However, repeated doses for 1 or 2 weeks induced a marked dose-related enhancement of lipid peroxidation (LPO) and ROS levels in both testis and epididymal sperm. Further evidence, such as significant perturbations in both enzymic and nonenzymic antioxidants and enhanced levels of protein carbonyls in testis, suggested induction of oxidative stress. In testis, moderate depletion in reduced glutathione levels and marked diminution in ascorbic acid and alpha-tocopherol content were accompanied by increased activities of various antioxidant enzymes, namely glutathione peroxidase, glutathione-S-transferase, and catalase, in both the HP treatments. Furthermore, significant alterations in the specific activities of testicular enzymes such as LDH-X, G-6-PDH, and SDH indicated altered testicular physiology. Both HP at higher doses induced significant DNA damage (determined by fluorimetric analysis of DNA unwinding assay) in testis and epididymal sperm. Increased total iron levels in testis of HP-treated rats are indicative of the possible involvement of iron-mediated free radical reactions in this model. These findings provide an account of early oxidative damage in testis and epididymal sperm following short-term exposure to HP in vivo, and this model is being further exploited for understanding the consequences of chronic oxidative stress-mediated alterations for the physiology of male reproductive system and its implications for fertility.

Animals↗
PubMed2006

Acute effect of vasectomy on the function of the rat epididymal epithelium and vas deferens.

Persistent infertility after apparently successful vasectomy reversal is common. One possible etiology is epididymal epithelial dysfunction resulting in improper sperm maturation after vasectomy reversal. The epididymal epithelium secretes a number of proteins that are thought to be required for the maturation of sperm. Ligation of the vas deferens during vasectomy may affect the synthesis of some of these proteins. In the present study, the function of the epididymal epithelium was assessed at early times after vasectomy (1, 4, and 7 days) by measuring the level of mRNA of 4 secreted proteins: Crisp-1, clusterin, osteopontin, and transferrin. In addition, the site of synthesis of these proteins was determined by immunocytochemistry. The results demonstrated that the expression of Crisp-1 and clusterin, representative epididymal secretory proteins, was largely unaffected by vasectomy. However, osteopontin mRNA increased in the vas deferens in response to vasectomy. Immunocytochemical localization of osteopontin suggested that both infiltrating immune cells and deferential luminal epithelium were responsible for this up-regulation. Transferrin expression was viewed as a marker for immune cells at the site of injury. However, both the caput epididymis and deferential epithelia were found to express transferrin, in addition to immune cells. In conclusion, there appear to be only minor changes in expression of genes encoding epididymal secretory proteins acutely after vasectomy, but, not surprisingly, there was evidence of an inflammatory response after vasectomy.

Animals↗
PubMed1998

Comparison of the effects of vasectomy and experimental cryptorchidism in rats: II. Epididymal histology.

Vasectomy is a male contraception method that its side effects and reversibility rate are still controversial. Effects of vasectomy on epididymal morphology and function, which plays a crucial role in sperm maturation and storage, have not yet fully described. Therefore the present study was planned to find out the effects of vasectomy on epididymal structure, and comparisons of these effects were also made with the effects of a persistent infertility model, experimental cryptorchidism. To do this, 15 male Swiss albino rats were divided into 3 groups. While 5 animals were sham-operated, the rest were undergone either vasectomy (5 rats) or experimental cryptorchidism (5 rats) procedures. Epididymal tissue samples were collected prepared for light microscopy. Tissue sections were stained using appropriate staining techniques and evaluated under the light microscope. While there were distinct alterations in the epididymal morphology following experimental cryptorchidism, vasectomized rat epididymal features seemed similar to those seen in the sham-operated group. In conclusion, it could be suggested that the vasectomy might be a convenient method for male contraception.

Animals↗
PubMedDate not supplied

Epididymal markers in an andrology clinic.

The ability of the markers alpha-glucosidase, carnitine, and glycerylphosphorylcholine to indicate epididymal obstruction was studied. alpha-Glucosidase was found to be the best discriminant, with 93.4% correct classifications. The possible interrelationship between alpha-glucosidase activity, sperm ATP, and epididymal function was then studied. alpha-Glucosidase correlated positively with the percentage of motile sperm (p = 0.0212) and with the percentage of sperm with good forward progression (p = 0.0374), but correlated negatively with the sperm ATP (p < 0.0500). It was concluded that lower ATP and higher alpha-Glucosidase may be markers of efficient epididymal function. The possible epididymal origin of detached ciliary tufts (DCTs) was studied by determining ATP and alpha-glucosidase activity in patients with DCTs. The mean ATP in patients with DCTs was significantly higher (p = 0.009), but the alpha-glucosidase activity significantly lower (p = 0.0412) than in the control group. These results seem to support the epididymal origin of DCTs.

Adenosine Triphosphate↗
PubMedDate not supplied

Effect of acute androgen withdrawal by GnRH antagonist on epididymal sperm motility and morphology in the cynomolgus monkey.

Hormonal male contraception requires an induction phase before azoospermia and contraceptive safety are achieved. The nature of spermatozoa that may be ejaculated during this induction phase was studied in a nonhuman primate. The GnRH antagonist Cetrorelix was administered daily to five cynomolgus monkeys to induce testicular regression, and the vehicle was given to five control animals. Within 16 days, the antagonist reduced androgens by 80% in the serum and by 50% in the epididymis. Sperm were obtained by mincing different epididymal regions and were examined for morphology (subjectively) and motility (objectively) after removal of the organs 16 and 25 days after continuous treatment. Spermatozoa entering the epididymis of monkeys undergoing regression differed from those of vehicle-treated controls in their greater susceptibility to disruption during preparation for morphological staining. The acquisition of motility by sperm in the epididymides attached to regressing testes occurred in the same epididymal region as controls but did not achieve the median velocities attained by sperm in controls during epididymal passage. Values for most sperm motion parameters developed as in the controls, and, during epididymal passage, sperm developed resistance to stresses encountered during preparation for morphological analysis. These observations suggest that spermatozoa ejaculated before spermatogenesis ceases may be potentially fertilizing because epididymal maturation continues in an androgen-deprived organ. From these preclinical studies, it can be concluded that in men, applying hormonal contraception precautions against pregnancy must be recommended before azoospermia is induced, since the epididymis can partially compensate for poor-quality sperm produced by a regressing testis even when levels of circulating androgens and tissue androgens are low.

Androgen Antagonists↗
PubMed1999

[Experimental vasectomy: comparison of epididymal structure with various surgical techniques].

OBJECTIVE: To analyze the effects of different surgical techniques of vasectomy on the epididymal structure. METHODS: Experimental vasectomies were performed in dogs using the conventional and open-ended techniques. The animals were orchidectomized 1 to 12 months after vasectomy and the epididymal structure was analyzed. RESULTS: The epididymal structure of dogs submitted to the conventional technique showed marked ectasia, reinforced basement membrane, loss of stereocilia in the principal cells, and formation of spermatic granulomas in interstitial tissue. At 12 months the principal cells showed signs of degenerative changes. In the different time periods analyzed in the study, no changes were observed in the epididymal structure of dogs submitted to the open-ended technique. CONCLUSIONS: The changes observed in the principal cells are ascribable to the increased intraluminal pressure produced by ligation of both ends of the vas deferens (conventional technique), since no changes were found in the epididymal structure of dogs submitted to the open-ended technique, at least in the different time periods analyzed in the study.

Animals↗
PubMed1998

Sperm maturation in vitro: co-culture of spermatozoa and epididymal epithelium.

Sperm maturation involves an intimate interaction between spermatozoa and the epididymal epithelium. Aspects of this relationship can be examined by co-incubating epididymal spermatozoa with epididymal epithelium in vitro. Plaques of epididymal epithelium from a variety of species (for example rodents, dogs, humans) can be maintained in culture medium supplemented with growth factors and androgens. When co-incubated with these epithelial cultures, immature epididymal spermatozoa undergo maturation changes that lead to the acquisition of progressive motility, zona binding and, in some instances, fertilizing capacity in vitro. The use of such co-culture techniques for the understanding of sperm maturation in vitro and in vivo is reviewed with reference to recent experiments.

Animals↗
PubMed1998

Role of luminal fluid glycosyltransferases and glycosidases in the modification of rat sperm plasma membrane glycoproteins during epididymal maturation.

It is generally accepted that mammalian spermatozoa undergo biochemical and morphological changes during epididymal transit, collectively termed epididymal maturation. Although many details of the biochemical modification are not fully understood, lectin binding studies from several laboratories strongly suggest that glycan moieties of sperm plasma membrane glycoproteins are extensively modified as spermatozoa transit from the proximal to the distal epididymis. In the present article, we summarize our studies with two sets of glycan modifying enzymes, namely glycosyltransferases (synthetic enzymes) and glycosidases (hydrolytic enzymes) in rat spermatozoa collected from different regions of the epididymis, and similar enzyme activities present in the epididymal luminal fluid. Our data show that the activities of these enzyme are high in the epididymal luminal fluid (> 80% of the total enzyme activities was in the plasma). Evidence presented in this report also demonstrates that: (1) at least one sperm surface glycoprotein (apparent molecular mass of 86 kDa) is fucosylated in vitro when caput spermatozoa are incubated with GDP [14C]fucose; and (2) a peanut agglutinin (PNA)-positive glycoprotein of 135-150 kDa present on plasma membrane of sperm from the caput (but not cauda) epididymidis is degalactosylated by digestion with purified luminal fluid beta-D-galactosidase. Taken together, these results strongly suggest a role for glycoprotein modifying enzymes in the modification of sperm surface glycoproteins during epididymal maturation.

Animals↗
PubMedDate not supplied

Epididymal epithelial cells cultured in vitro prolong the motility of bovine sperm.

It is well known that the epididymis is an excellent environment to maintain sperm viability. Therefore, we used different sections of bovine epididymis (caput, corpus, and cauda) to develop epithelial cell culture monolayers to identify factors that will increase sperm survival in the freezing-thawing process. Each epididymal section was dissected and treated with collagenase to obtain epithelial cell clusters. The cells were cultured in RPMI-1640 medium with 10% serum at 38.5 degrees C. A confluent monolayer was obtained after 5-7 days in culture and preliminary characterization using cytokeratin antibody indicated that the cell culture contained 85%-95% of epithelial cells. These cellular cultures were tested for their ability to maintain motility of epididymal and frozen-thawed spermatozoa. Washed spermatozoa were added to obtain a final dilution of 1 x 10(6) spermatozoa/mL. The motility of frozen-thawed spermatozoa was also recorded after incubation in conditioned media. Our results show that cocultures of spermatozoa and epididymal cell monolayers for 24 and 48 hours were beneficial for maintaining epididymal and frozen-thawed sperm motility (36.0% and 20.4%) compared with spermatozoa cultured with fibroblast cells or in the absence of a cell monolayer (0%; P < .01). The conditioned medium provides favorable conditions for sperm motility. Results with conditioned medium on maintenance of frozen-thawed sperm motility suggest that epididymal cells in vitro secrete beneficial factors that prolong the sperm survival.

Animals↗
PubMed2000

Epididymal protein secretion and its androgenic control in wall lizards Hemidactylus flaviviridis (Ruppell).

The total protein level in different segments of epididymis of normal lizard exhibited noticeable increase from early February to late March of a same reproductive phase. Comparison among the protein level of different epididymal segments showed insignificant variation from anterior to posterior part in early February but in late March, the protein level in posterior segment was appreciably higher than in anterior and middle segments. Further, testosterone-induced epididymal protein did not exhibit any significant quantitative variation among different regions. The electrophoretic pattern of luminal fluid from different epididymal regions of normal lizard showed 28 protein bands without any marked regional difference. However, only 16 protein bands could be demonstrated in the epididymal fluid of any region. Unlike molecular size, isoelectric focussing of testosterone induced epididymal proteins revealed that three regions of epididymis differ in their nature of protein. The number of proteins having alkaline pH range in anterior and middle regions were 4 and 3, respectively which increased upto 6 in posterior region.

Androgens↗
PubMed2001

Effect of lindane on antioxidant enzymes in epididymis and epididymal sperm of adult rats.

AIM: To find out the changes induced by lindane on the antioxidant enzymes in epididymis and epididymal sperm of adult rats. METHODS: Adult male rats were orally administered lindane at a dose of 5.0 mg/kg body weight per day for 30 days. At the end of the treatment, the rats were sacrificed. The epididymis was removed and weighed and sperm were collected for sperm count, motility and biochemical studies. A 1% homogenate of epididymis was prepared and used for biochemical estimations. RESULTS: In lindane-treated rats, there were significant reductions in the epididymal weight, epididymal sperm count and motility compared with the controls. Significant decreases in the superoxide dismutase (SOD), catalase, glutathione reductase and glutathione peroxidase activities and significant increases in the H2O2 generation and lipid peroxidation were also observed in the epididymis and epididymal sperm of lindane-treated rats. CONCLUSION: Lindane decreases the levels of antioxidant enzymes in the epididymis and epididymal sperm of adult rats thereby inducing oxidative stress.

Age Factors↗
PubMed2002

Epididymal cribriform hyperplasia with nuclear atypia in p53 homozygous knockout mice on a mixed 129/Sv-FVB/N background.

Epididymal cribriform hyperplasia (ECH) is a variant of normal epididymal histologic features in men, and has also been reported in rats, mice, dogs, cats, and bulls. The epididymal change has been associated with aging, testicular atrophy, cryptorchidism, and germ cell tumors. Epididymal cribriform hyperplasia was observed in p53 homozygous knockout mice on a mixed 129/Sv-FVB/N background, but not in wild-type or heterozygous mice. The aim of the study reported here was to determine the prevalence and characterize the morphologic, immunohistochemical, and ultrastructural features of ECH in these mice. Epididymal cribriform hyperplasia was present in 88% (72/82) of male mice ranging in age from seven to 65 weeks. The lesion was characterized microscopically by epithelial cells with atypical hyperchromatic nuclei, vacuolization, intratubular lumina formation, infrequent apoptosis, and rare mitotic figures. In contrast to germ cells, the cells of ECH did not express alpha-fetoprotein, carcinoembryonic antigen, or S-100. Ultrastructurally, the cells were pleomorphic with stereocilia at their apical borders and within intratubular lumina, and were supported by a basement membrane. Although 14% (10/72) of mice had concomitant testicular neoplasia, ECH did not appear to be a preneoplastic change. Investigators using these mice for modeling human disease should be aware of the background prevalence of this lesion.

Age Factors↗