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Introduction: The aim of this study was to determine the effect of reduced availability of dihydrotestosterone (DHT) on the expression of estrogen receptors alpha and beta (ERalpha and ERbeta) in the epididymis in vivo and in vitro. Expression of estrogen receptors (ERs) is interesting because of the fact that the male reproductive system is controlled not only by androgens but also, in a far-reaching and complex manner, by estrogens. Control by estrogens is exercised through activation of ERs widely distributed in the epididymal epithelium. Epididymal epithelial cells contain a 5alpha-reductase (5alpha-red) which catalyzes the irreversible conversion of testosterone (T) into the most potent and chief androgen of the epididymis, dihydrotestosterone, known to maintain and regulate the structure and functions of the epididymis. Two isoforms of the 5alpha-red were identified: type 1 (5alpha-redl) and type 2 (5alpha-red2). 5alpha-reductase type 2 is more widely expressed in the epididymis than 5alpha-redl. DHT deficit was produced by inhibition of 5alpha-red2 using finasteride (Proscar, MSD Sweden), a steroid inhibitor of this enzyme., Material and Methods: The study was performed in the adult, male Wistar rats randomly divided into control (K) and study (Fin56) groups (5 animals in each). Animals in the study group received 5mg finasteride/kg b.w., orally during 56 days (duration of one spermatogenesis). Immunoexpression of ERs was also studied in epididymal epithelial cells cultured with or without finasteride., Results: It was shown that DHT deficiency, both in vivo and in vitro condition, modulated ERs expression in comparison to the epididymis from control rats and to epididymal cells cultured without finasteride. Distribution of ERalpha and ERbeta in epididymal cells changed (from nucleus to cytoplasm) and the level of ERs expression was markedly decreased., Conclusion: The present findings show that the DHT deficiency caused by finasteride altered the expression of ERalpha and ERbeta in the epididymis and possibly may have destabilized the functioning of this organ.

The epididymal epithelial cells of rats in vitro display features of steroidogenic cells. The cells produce and release androgens into the culture medium that are converted to 17beta-estradiol (E2) due to the activity ofcytochrome P450 aromatase. It is probable that steroidogenesis in epididymal epithelial cells of rat is regulated by luteinizing hormone (LH). The aim of the study was to estimate the influence of hCG on the morphology of epididymal epithelial cells and synthesis of E2 by the cells. In conclusion, the hCG stimulation results in changes of the cell morphology, similar to those observed in Leydig cells and increase synthesis of the 17beta-estradiol due to the presence of LH/hCG receptors.

Objective: Microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection (MESA-ICSI) is a viable alternative to the treatment of couple sterility caused by obstructive azoospermia. Because there are structural differences between epididymal and ejaculated spermatozoa and it is not univocally stated, if this fact influences positively or negatively the results of ICSI, it was decided to compare the efficacies of ICSI using both the kinds of spermatozoa at the same setting., Design: Retrospective comparative analysis., Materials and Methods: The MESA group consisted of 27 couples and the control ICSI group of 154 couples--both matched in age, duration of sterility, health status, stimulation protocols and luteal supplementation. The trial was performed at the same period in one center, keeping the same laboratory and personal conditions in both the groups., Results: In the MESA group the fertilization rate was 53.9%, the cleavage rate 87.6% and pregnancy rates 37.0% per microinjection and 43.5% per embryotransfer. In the control group the corresponding values were 50.1%, 90.0%, 28.0% and 31.7%. The differences between the groups were insignificant., Conclusions: It may be concluded, that the structural differences between epididymal and ejaculated spermatozoa do not significantly influence the efficacy of ICSI, however further investigations are still needed.

Studies were performed on cultured epithelial cells of the caput and cauda of epididymis stemming from male rats of inbred Wistar strain. The cultures were conducted on a full medium enriched with 5% fetal calf serum in the presence or without exogenic androgens-T and DHT. The cells were identified by means of immunohistochemical reactions with the use of monoclonal antibodies against cytokeratin and desmin (Fig. 1, 2). All cells in the culture showed positive reaction to cytokeratin. At the same time there was a lack of desmin-positive cells. Through secreting the proteins, glycoproteins, glycolipids, phospholipids and number of other substances the epithelial cells of epididymis create an environment for maturating and storing of spermatozoa in lumen of the duct. Synthesis of these substances is possible thanks to the expression of genes defined for a given zone of epididymis, the expression being mainly regulated by androgen, although a share of estrogens is also evidenced in this process. The cytoplasm of epithelial cells of epididymis fails to reveal the presence of secretory granules, while the mechanism of releasing the secretion still continues to be controversial. There are also some and unverified suggestions about the capability of these cells to synthesize androgens. In connection with what was mentioned above, the objective of the work was to establish the mode of releasing the secretions by cultured epithelial cells of epididymis as well as to determine whether these cells synthesize androgens and if they may be the source of estrogens. Electron-microscopic observations disclosed a rich content of rough endoplasmic reticulum and structures similar to secretory units produced from concentrically arranged membranes encircling cytoplasm fragments in their interior (Fig. 10B, 14, 15A). There were protrusions of cytoplasm on the surface of cells. Released secretion was present between the cells. The apocrine way of releasing was confirmed also by scanning electron microscope. Numerous granular protrusions were released into the intercellular space (Fig. 19). The process of synthesis and release of secretion was androgen-dependent. Cells cultured without addition of exogenic androgens were characterized by disorganization of organelles and reduction of their number, particularly rough endoplasmic reticulum. The surface of cells was prevalently smooth, deprived from protrusions (Fig. 21). Very close neighbourhood, and sometimes a direct contact of lipid droplets and mitochondria with lamellar cristae as well as the presence of smooth endoplasmic reticulum observed in cytoplasm of cultured epithelial cells of epididymis, suggest their similarity to steroidogenic cells (Fig. 11A, 12). This is also indicated by the finding that these cells reveal the presence of active enzymes of the steroidogenesis pathway, 3 beta-HSD and 17 beta-HSD exhibited in histochemical reactions (Fig. 8, 9). RIA determination of hormones in the medium, wherein the epithelial cells had been cultured showed that the said cells synthesized and released DHEA, A and T, but in low and sometimes trace concentrations (Tab. 1-3). Lack of progesterone in medium of the cells on the 3rd and 5th days of culture indicates that the synthesis of testosterone and earlier forms of androgens proceeds using delta 5 metabolites, as it takes place in human testis. The cells' medium on the 3rd and 5th days of culture was found to disclose high concentration of 17 beta-estradiol (E2) (Tab. 4). E2 concentrations were always higher when the cells were grown without the addition of exogenic androgens. In this cases the cytoplasm of the cells displayed depolymerization of microtubules, which enhances the approximation to each other of structures participating in steroidogenesis and translocation of substrates and products of the consecutive stages of steroidogenesis. (ABSTRACT TRUNCATED)