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[Scanning electron and light microscopic studies of the surface epithelium of the rete testis and epididymis in the boar. II. Ductus epididymis].

The entire ductus epididymidis is characterized by pseudostratified columnar epithelium which contain principal and basal cells. Apical cells are found in the proximal segments. Long stereocilia projected from the apical surface of the principal cells. Signs of adjacent apocrine secretory activity are shown. The various segments of the ductus epididymidis present differences in the height of the epithelium, the development of the stereocilia, the appearance of granules in the cytoplasm, the luminal sperm concentration and the height of the peritubular layer.

Animals↗

Direct evidence for the elevated synthesis and secretion of procathepsin L in the distal caput epididymis of boar.

The proteins which are secreted from the restricted part of the epididymis are suggested to sustain sperm maturation. In porcine species, as the potential abilities of sperm for movement and fertilization greatly increase in the corpus epididymis, the secretions in both the caput and corpus epididymis seem to be very important for the sperm maturation. In this study, we have directed our attention to the 40 kDa protein which is detected in the fluid of the distal caput epididymis of boar. It was purified from the porcine cauda epididymal fluid and its cDNA was cloned from the cDNA library of the distal caput epididymis. According to the deduced amino acid sequence, the 40 kDa protein has been identified as procathepsin L. Northern blot analysis showed that the procathepsin L mRNA was most abundant in the distal caput epididymis among the tissues as examined. Consistent with the distribution of the procathepsin L mRNA in the epididymis, the activity of procathepsin L was absent in the fluid of the proximal and mid caput epididymis and first appeared in the distal caput epididymal fluid, whose contents gradually decreased with the passage through the epididymis. These results first appeared in the first distal caput epididymis expresses very high levels of procathepsin L and unusually secretes it into the luminal fluid instead of targeting it to lysosomes. It has been also found that the mRNA of PDGF, which is known to enhance cathepsin L expression in the culture cells, is very high in the mid caput epididymis, which just precedes the site of procathepsin L secretion. This result indicates that PDGF directly regulates the locally restricted expression and secretion of procathepsin L in the epididymis, which is one of the possible mechanisms involved in the functional differentiation in the epididymis.

Amino Acid Sequence↗

Neuronal input from the inferior mesenteric ganglion (IMG) affects sperm transport within the rat cauda epididymis.

Sympathetic nerves emanating from the inferior mesenteric ganglion (IMG) innervate the mammalian epididymis and vas deferens. We have reported previously that surgical removal of the rat IMG results in excessive accumulation of spermatozoa within the cauda epididymis. The objective of the present study was to determine whether sperm accumulation following IMG removal was due to a denervation-induced change in the rate of sperm transport within the cauda epididymis. In these studies epididymal sperm numbers were counted and used as a measure of sperm transport within the epididymis. In order to examine sperm transport within the cauda epididymis specifically, efferent duct ligation (EDL) was used to prevent further entry of testicular spermatozoa into the epididymis. Rats were divided into four treatment groups: sham-operated control, EDL alone for 7 days (EDL + 7), EDL alone for 14 days (EDL + 14), or EDL for 7 days at which time the IMG was removed and the rats were maintained for an additional 7 days (EDL + IMG). Epididymides were homogenized and the number of spermatozoa in the caput and cauda epididymis was counted. In EDL + 7 rats, the caput epididymis was devoid of spermatozoa. The rate of transit of spermatozoa through the cauda epididymis of EDL + 7 rats was approximately 3.43 million/day. The total number of cauda epididymal spermatozoa in EDL + 7 rats was reduced by 20% compared to sham-operated control rats. In EDL + 14 rats, spermatozoa transited the cauda epididymis at a rate of approximately 9.57 million/day, and the total number of spermatozoa in the cauda epididymis was reduced by 73% compared to sham-operated controls. If the IMG was removed 7 days following EDL, spermatozoa transited the cauda epididymis at a rate comparable to that observed in EDL + 7 rats (3.39 million/day). Following IMG denervation of EDL + 7 rats, cauda epididymal sperm numbers were maintained at levels observed in the EDL + 7 rats. These data demonstrate that the transport of spermatozoa through the cauda epididymis is influenced significantly by neuronal input from the IMG.

Animals↗

[Analysis of DNA content of spermatogenic cells in the adult human testis and epididymis by flow cytometry].

OBJECTIVES: To detect the changes of DNA ploidy of spermatogenic cells in testis and epididymis. METHODS: Right epididymides and testes from 15 fertile youth donors who died of accident were collected. Samples of spermatogenic cells in different regions of epididymis (caput, corpus and cauda) and tests were collected. DNA of spermatogenic cells were detected by flow cyctometry (FCM). RESULTS: The haploid(1n), diploid(2n) and tetraploid(4n) spermatogenic cells were existed in different regions of epididymis and testis. The 1n cells increased from (24.87 +/- 7.28)% in testis to (96.33 +/- 1.58)% in epididymis cauda, there were significant differences among regions of testis and epididymis caput, corpus(P < 0.01), and the difference among regions of epididymis corpus and epididymis cauda were also significant(P < 0.05). While the percentages of 2n and 4n cells decreased from (63.07 +/- 8.96)% and (9.43 +/- 3.83)% in tesits to (2.47 +/- 0.93)% and (1.17 +/- 0.95)% in epididymis respectively. There was significant difference of 2n cells between testis and epididymis caput, corpus(P < 0.01), and was also remarkable difference between epididymis corpus and cauda (P < 0.05). There was no difference of 4n cells between testis and epididymis caput(P > 0.05). There were significant difference among regions of epididymis caput, corpus and cauda(P < 0.05). CONCLUSIONS: The percentage of immature spermatogenic cells decreased along with passing through the epididymis.

Adult↗

Angiotensin-converting enzyme in the testis and epididymis: differential development and pituitary regulation of isozymes.

Angiotensin-converting enzyme (ACE, EC 3.14.5.1) is found in particulate fractions of the epididymis but not in soluble epididymal fractions or in the testis of 4-week-old rats. [3H]Captopril autoradiography of testis and epididymis from 4-week-old rats confirms the association of ACE with epididymal ducts but not the testis. ACE appears in the testis between 4 and 6 weeks of age. Soluble ACE is not detectable in the epididymis until 6-7 weeks of age. Within the epididymis, regions closest to the testis develop soluble ACE activity about 1 week before those nearest to the vas deferens. Hypophysectomy of 10 week-old-rats depletes greater than 95% of ACE activity from the testis and soluble fractions of the epididymis, with little change in ACE levels from particulate fractions of the epididymis. [3H]Captopril autoradiography after hypophysectomy reveals luminal and epithelial ACE in the epididymis. The presence of particulate ACE in the epididymis under conditions where there is no testicular ACE indicates that the two forms are synthesized separately. However, soluble ACE from the epididymis might be derived from the membrane-associated ACE of the testis. Such a relationship is supported by the lag of 1 week between the development of ACE in the initial segment of the epididymis and the tail of the epididymis, and by the occurrence of soluble epididymis ACE only in those animals with testicular ACE activity.

Animals↗

Sonographic appearance of the epididymis in pediatric testicular torsion.

OBJECTIVE: The purpose of this study was to determine the frequency of an enlarged epididymis in pediatric patients with testicular torsion and to determine whether an altered epididymis might be a helpful ancillary sonographic sign of testicular torsion. MATERIALS AND METHODS: The sonograms of 50 pediatric patients (age range, neonate-17 years) with testicular torsion were retrospectively reviewed for the size, appearance, and blood flow of the epididymis. Medical records were reviewed for surgical and pathologic findings and to determine whether testicular salvage had been possible. RESULTS: The epididymis was enlarged (maximum dimension, 5.5 cm) in 47 of the 50 patients with acute or late phase torsion and after manual or spontaneous detorsion. The average difference in volume between the ipsilateral epididymis and the contralateral epididymis was 30 cm3, highly significant (p < 0.0001). The shape of the epididymis was altered in 92% of the cases (globular, bilobular, or multilobular). Seventy-three percent showed increased echogenicity and 27% appeared isoechoic. Of those with active torsion, 93% of the epididymides were avascular; 2%, hypovascular; and 5%, hypervascular. After detorsion, 100% of the epididymides (10/10) had blood flow. In patients with testicular loss due to infarction, pathology showed engorgement and enlargement of the epididymis with hemorrhagic infarction. In one patient with a hypervascular epididymis, surgery showed inflammation and erythema of the epididymis. Testicular loss occurred in each type of epididymal flow pattern. CONCLUSION: A markedly enlarged, echogenic, and avascular or hypovascular epididymis is an ancillary sonographic sign in pediatric patients with testicular torsion. A hypervascular enlarged epididymis infrequently occurs (5% of cases) and should not be mistaken for epididymitis. In addition, the return of epididymal blood flow is an ancillary sign of successful testicular detorsion.

Adolescent↗

Androgen-dependent accumulation of carnitine by rat epididymis after injection of [3H]butyrobetaine in vivo.

After i.m. injection of [3H]butyrobetaine into rats, the accumulation of carnitine into the epididymis, prostate gland, seminal vesicles, testis and heart was studied. The concentration of radiolabeled carnitine into the cauda epididymis increased linearly with time up to 72 h after the injection of the precursor, while its level in the prostate and seminal vesicles decreased rapidly. Very low levels of carnitine were found in the testis. Castration reduced the carnitine accumulation by cauda epididymis to 6% of the control levels while treatment of castrated animals with testosterone propionate (500 mug/day) partly restored the carnitine uptake. Similar treatment with 17beta-oestradiol valerate or 17alpha-hydroxyprogesterone had no effect. Surprisingly, cyproterone acetate (5 mg/day) also significantly stimulated carnitine accumulation by the epididymis to a level above that of the castrated controls. Simultaneous injection of both cyproterone acetate and testosterone propionate to castrated animals caused an additive effect of these steroids. This indicated that cyproterone acetate in this system is working as a weak androgen. Treatment of rats with 17beta-oestradiol valerate also reduced carnitine accumulation by the cauda epididymis. This is due to suppression of pituiatry gonadotrophin secretion, since concommitant treatment with testosterone propionate (500 mug/day) caused a normalization of the carnitine uptake. Treatment of intact rats with cyproterone acetate significantly reduced the epididymal weight, but not the carnitine accumulation. 17alpha-Hydroxyprogesterone treatment had no effect either on the epididymal weight or the accumulation of the carnitine. Unilateral orchiectomy reduced the carnitine accumulation by the cauda epididymis to about 40% of that occurring in the non-operated control side. This indicates that the luminal contact between the testis and epididymis or the luminal content of the epididymis itself is of importance for the androgen-dependent metabolic process occurring in the cauda epididymis. Castration or hormone treatment did not change the conversion of butyrobetaine to carnitine, or the carnitine uptake by heart. Carnitine uptake by the testis after [3H]butyrobetaine injection was rather low and this would exclude the possibility of synthesis of carnitine in the testis as a source of epididymal carnitine. Carnitine only accumulated in the cauda epididymis in vivo 4 to 96 h after injection of [3H]butyrobetaine. The presence of radioactively labeled butyrobetaine or methylcholine was not detected.

Animals↗

Expression and regulation of lipocalin-type prostaglandin d synthase in rat testis and epididymis.

Lipocalin-type prostaglandin D synthase (L-PGDS), a bifunctional protein, is expressed in the male reproductive organs of many species. However, the expression and regulation of L-PGDS in rat are still uncertain. The present study investigated the regionalization and regulation of L-PGDS expression in rat testis and epididymis by in situ hybridization and immunohistochemistry under the conditions of sexual maturation, castration, and ethylene dimethane sulfonate (EDS) treatments. In sexually mature rats, L-PGDS mRNA was weakly expressed only in the testicular peritubular cells, whereas L-PGDS immunostaining was highly detected in the Leydig cells by Day 70 postpartum. During sexual maturation, L-PGDS mRNA expression was highly detected in the caput, corpus, and cauda of the epididymis 70 days after birth. Compared with normal L-PGDS expression in adult epididymis, both L-PGDS mRNA expression and protein immunostaining were significantly reduced in the caput, corpus, and cauda epididymis after castration. Testosterone propionate treatment induced a significant increase of L-PGDS expression in the epididymis of castrated rats. Compared with adult rat epididymis, L-PGDS mRNA and protein expression was down-regulated after EDS treatment. Testosterone propionate treatment could induce an increase of L-PGDS mRNA and protein expression in the epididymis of EDS-treated rats. In conclusion, both castration and EDS treatments caused a significant decrease of L-PGDS expression in the epididymis, whereas testosterone propionate treatment could induce an increase of L-PGDS expression in the epididymis of both castrated and EDS-treated rats, indicating that L-PGDS expression in the rat epididymis can be up-regulated by testosterone.

Animals↗

Developmental changes in and hormonal regulation of estrogen and androgen receptors present in the rabbit epididymis.

Both androgen and estrogen receptors (AR and ER) are present in the rabbit epididymis. We have used the sucrose gradient method to examine receptor sedimentation properties, receptor concentration, and distribution of receptors among the caput, corpus, and cauda of the epididymis to determine changes that occur in these parameters as the animals age. The 9S form of the ER is present in all three epididymal segments of the immature rabbit, with the highest concentration occurring in the cauda. The 8.2S form of the AR is also present in all three segments of the immature epididymis, with the highest concentration occurring in the caput. Short-term castration (3 days) leads to an increase in the amount of both AR and ER detected. ER are present in all segments of the immature epididymis at higher concentrations than AR. The functional 9S form of the ER disappears as the animals mature, the result of a tissue-specific protease that our laboratory previously has shown proteolyzes ER to a non-DNA-binding 3.8S form. Long-term castration (3 mo) of adult rabbits results in the reappearance of the 9S form of the ER in all segments of the epididymis. The reappearance of the 9S form of the ER is also seen in animals castrated for 1 mo, but not in those castrated for 2 wk. Administration of testosterone once daily for 2 wk to adult animals castrated for 6 wk results in the disappearance of the 9S form of the ER and the reappearance of the 3.8S form, suggesting that the tissue-specific protease is androgen-dependent. In this way, circulating androgens may play a role in regulating the concentration and form of the ER in the rabbit epididymis. There is little change in the concentration or distribution of AR in the epididymis of adult rabbits castrated for 3 mo as compared to those castrated for 3 days. This implies that circulating androgens are not required for maintenance of AR in the epididymis. Our data demonstrate that there are temporal differences in the presence and concentration of ER and AR in the epididymis and suggest that there is a differential, age-dependent regulation of the development and function of the epididymis by androgens and estrogens.

Aging↗

Immunocytochemical localization of 4-ene steroid 5 alpha-reductase type 1 along the rat epididymis during postnatal development.

Dihydrotestosterone (DHT), the active androgen in many tissues, is synthesized from testosterone by the enzyme 4-ene steroid 5 alpha-reductase (5 alpha-reductase; EC 1.3.1.22). In the epididymis, the maturation and storage of spermatozoa are dependent on the presence of 5 alpha-reduced androgens. The regulation of epididymal 5 alpha-reductase is complex. To date, the regulation of this enzyme has been studied extensively at the level of enzymatic activity and more recently at the mRNA level. Regulation at the level of the protein, however, remains poorly understood. We have raised rabbit polyclonal antibodies to a 24-mer synthetic peptide whose sequence was determined from the predicted amino acid sequence for rat 5 alpha-reductase type 1 to immunolocalize the 5 alpha-reductase type 1 protein in the rat epididymis during postnatal development. Western blot analysis revealed a specific immunoreactive band of 26 kilodaltons in male rat liver, epididymis, and prostate; this apparent molecular size is identical to that obtained when the 5 alpha-reductase type 1 cDNA is expressed in mammalian cells. Furthermore, the relative protein levels, liver > epididymis > prostate, were consistent with the mRNA levels for type 1 rat 5 alpha-reductase. Perfusion-fixed paraffin-embedded epididymal tissue sections were used to immunolocalize type 1 5 alpha-reductase. In the adult rat epididymis, the most intense immunoperoxidase reaction was observed in a discrete lobule of the initial segment of the epididymis. A progressive decrease in staining intensity occurred distally along the tissue to the cauda epididymis. The staining reaction was specific to cytoplasmic elements of epithelial principal cells; no reaction was evident over nuclei. However, specifically in the initial segment, very intense staining was seen in the infranuclear region of the principal cells. In the proximal caput epididymidis, the staining was primarily confined to an oval region above the nuclei, whereas in the remaining epididymal regions, weak staining was seen throughout the cytoplasm. Thus, the intracellular localization of the 5 alpha-reductase type 1 protein changed as one moved down the epididymis. Finally, the pattern of immunolocalization of 5 alpha-reductase type 1 protein was different in the epididymis of rats of different postnatal ages. On day 7, no reactivity was noted; by day 28, a weak apical staining of principal cells was seen throughout the epididymis; by day 47, the adult pattern of staining had been established. Our results revealed that the expression and intracellular localization of the 5 alpha-reductase type 1 protein are both age dependent and epididymal segment specific.

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

Lysosomal integral membrane proteins exhibit region and cell type specific distribution in the epididymis of the adult rat.

The epididymis, a post-testicular site required for maturation and storage of spermatozoa, is actively involved in exocytic and endocytic events, two phenomena likely to depend on the integrity of the lysosomal system. To study the lysosomal system of the epididymis, five monoclonal antibodies, previously characterized as recognizing five distinct lysosomal integral membrane proteins (LIMPs 1-5), were used as molecular probes of lysosome distribution in cells lining the epithelium. Immunocytochemical localization of LIMPs, using biotin-streptavidin immunoperoxidase methodology, was performed on frozen sections of adult rat epididymides and in cell cultures prepared from either the caput or cauda epididymis. In frozen sections, a heterogeneous distribution of the different LIMPs along the length of the epididymis was observed. For example, the distribution of LIMP 1 (35-50 K) was detected in all cells of the caput and quite dramatically in clear cells of the distal caput, corpus, and cauda epididymis, but specifically not in the principal cells of the distal caput, corpus, and cauda. In contrast, LIMP 2 (64-71 K) was present in all cells of the epididymis, except clear cells. LIMPs 4 and 5 (93 K and 93 K) were detected in all epididymal cells, including the clear cells. Finally, whereas the regional and cell type distribution of LIMP 3 (74 K) in the epididymis was identical to that of LIMPs 4 and 5, the nature of the vesicles immunostained was distinct. In cultured cells, the general immunostaining patterns observed in vivo were maintained during the duration of the primary cultures for all five LIMPs. Our results begin to address the molecular heterogeneity of the lysosomal system along the length of the epididymis, and may suggest in part a basis for underlying structural and functional characteristics of the epididymis leading to the sequential maturation of sperm.

Animals↗