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Evidence of protein secretion by cultured pachytene spermatocytes.
No full textPachytene spermatocytes isolated from immature rat testis were cultured for 6 h in the presence of [35S]methionine and the macromolecules present in the culture medium were analyzed by one-dimensional and two-dimensional gel electrophoresis. The electrophoretic profiles obtained showed a limited number of polypeptides, some of them consisting of families of spots with the same molecular weight but a different isoelectric point. The reproducibility of the results and the unaltered metabolic activity of the cells during culture time, indicate that the macromolecules present in the medium do not represent degradative products of the cells. Part of the detected macromolecules are considered secreted proteins since the addition of monensin to the cells induces their disappearance from the culture medium
Retinoid modulation of plasminogen activator production in rat sertoli cells
No full textTissue type (t) and urokinase type (u) plasminogen activators (PAs) have been shown to be secreted by Sertoli cells in the seminiferous tubules in a cyclic fashion and to be dependent upon FSH stimulation or upon the presence of adjacent spermatogenic cells. In the present study we have analyzed the production of PAs by retinoid-treated rat Sertoli cells. In addition, because retinoids modulate the response of Sertoli cells to FSH either potentiating or antagonizing its action, we have investigated a possible modulation of FSH-stimulated PA production. Under basal conditions, Sertoli cells, isolated from prepubertal rats, secrete predominantly uPA. A significant dose-dependent inhibition of uPA activity was observed after treatment with retinol, while no significant effect was detected upon tPA secretion. When Sertoli cells were cultured in the presence of 0.25 mu M retinol, a significant inhibition of uPA activity was evident after 16 h of treatment and reached approximately 80% after 48 h of treatment. The analysis of the mRNA levels revealed that retinol induces an inhibition of the steady-state levels of uPA mRNA without affecting those of tPA. Moreover, retinol affected uPA mRNA levels by increasing mRNA turnover. The effect of retinoids on Sertoli cells isolated from older animals was less evident, possibly due to the reduced production of uPA with the increase of age of the donor animals. Our results on the effect of retinoids upon Sertoli cell uPA production reinforce the importance of retinoids in the control of postnatal testis development
Retinol increases synthesis and secretion of Sertoli cell mannose-containing glycoproteins.
No full textSertoli cell isolated from prepuberal rats were cultured in a chemically defined medium in the presence or in the absence or retinol. In the two experimental conditions the incorporation of 3H-mannose into cellular glycoproteins was measured. The results obtained demonstrated a 3 fold increase of mannose incorporation into cellular glycoproteins after retinol addition without any qualitative variation of their electrophoretic profile. Sertoli cell secretory activity increased of approximately 50% after retinol administration; the electrophoretic analysis showed that two of the secretory products were selectively enhanced by the vitamin addition
Secretion of androgen binding protein by Sertoli cells is influenced by contact with germ cells.
No full textSertoli cells were cultured alone or with germ cells to evaluate the effect of the association with germ cells on the secretory activity of Sertoli cells. Secretion of androgen-binding protein, which is specifically secreted by Sertoli cells, was measured under several experimental conditions. The following experimental models were utilized: 1) cultures of explants of seminiferous epithelium from prepubertal animals in which germ cells adherent to Sertoli cells are present (Sertoli cell enriched cultures); 2) monolayers formed only by Sertoli cells, obtained by removing germ cells from Sertoli cell enriched cultures, and 3) cocultures of Sertoli cell only cultures and germ cell populations at defined stages of differentiation. The results obtained indicated that FSH-induced ABP secretion was greatly reduced in Sertoli cell only cultures as compared to enriched Sertoli cell cultures, and that this difference was stable throughout the first eight days of culture. In addition, cocultures of Sertoli cell only cultures with germ cells induced an increase of ABP when cocultured germ cells were at differentiation stages, such as pachytene spermatocytes, which are able to recognize and firmly adhere to the Sertoli cell monolayers. Cocultures with round spermatids, which do not adhere to Sertoli cells, did not increase the amount of FSH-induced ABP production. The addition of nongerminal cells such as lymphocytes and fibroblasts were also not effective in stimulating ABP secretion. Surface interaction between Sertoli cells and cocultured germ cells seemed to be necessary for this FSH-induced ABP production.(ABSTRACT TRUNCATED AT 250 WORDS
Cellular Retinoic-acid-binding-protein and Retinol-binding-protein Messenger-rna Expression In the Cells of the Rat Seminiferous Tubules and Their Regulation By Retinoids
No full textThe levels of the mRNA corresponding to the intracellular binding proteins for retinoic acid and retinol (CRABP1 and CRBP1, respectively) were studied in primary cultures of somatic and germ cells of the rat seminiferous tubules. We show that the CRABP1 mRNA is expressed in Sertoli and germ cells and a single molecular species of mRNA is detected. CRBP1 mRNA is detected in Sertoli and peritubular cells. The regulation of the expression of both genes by retinoids was studied in Sertoli cells. CRABP1 mRNA levels are not affected by either retinoic acid or retinol, whereas both compounds positively regulate CRBP1 mRNA synthesis in a dose-dependent manner. A fivefold increase in CRBP1 mRNA levels was observed 32 - 48 h after addition of either agent. These results demonstrate that in Sertoli cells the expression of CRABP1 is not affected by retinoids, similar to the situation observed in vivo and in other in-vitro cultures. CRBP1-gene expression is, instead, induced and the variations in CRBP1-mRNA levels may regulate the intracellular concentrations of retinoids, as a response to changes in the vitamin-A nutritional status
Morphological and biochemical effects of lonidamine on cultured Sertoli cells.
No full textLonidamine, an antispermatogenic compound derivative of indazole-3-carboxylic acid, administered to rat Sertoli cells induced morphological changes of the cells which rapidly shrinked forming cytoplasmic projections. Some biochemical parameters of Sertoli cells measured after drug administration were their aromatizing ability, protein synthesis, and ribonucleic acid synthesis. The amount of estradiol produced by treated cells was significantly lower than the amount produced by control cells, whereas protein and ribonucleic acid synthesis were not affected by the drug. Our results demonstrate the change of a specific metabolic parameter of Sertoli cells induced by this antispermatogenic compound
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