Effects of Follicle-Stimulating Hormone on Intermediate Filaments and Cell Division of Sertoli Cells of Fetal Rat Testis in Culture.

Abstract

ABSTRACT. The present study was aimed to examine the effects of follicle-stimulating hormone (FSH) on cell division of Sertoli cells from rat fetal testes and on the kinetics of 2 kinds of intermediate filaments, cytokeratin and vimentin, which comprise the cytoskeleton of Sertoli cells. Testes from rat fetuses of different ages (from day 15 to day 17 of gestation ) were cultured for 48 hr, with or without added FSH. for the presence of sperm in the vaginal smear. The day on which sperm was detected was counted as day 0 of gestation. Fetuses of 15-17 days of age were used for culture of their testes. These fetuses were removed from maternal uteri under aseptic conditions and transferred to sterile petri dishes containing Hanks' balanced salt solution (HBSS) with penicillin (100 units/ml) and streptomycin (100 µg/ml). Their testes were removed and rinsed with HBSS. Each testis was placed on the surface of a Millipore filter in the medium (Medium RPMI 1640; Gibco-BRL, U.S.A.) containing 0.2% sodium bicarbonate. Three series of culture were prepared. One was added with no FSH as control (group C). The second was added with 0.5 µg/ml FSH (Sigma, 1 mg equivalent to 1 unit shown by Steelman and Pohley [18]) as group FSH 0.5. The third was added with 5 µg/ml FSH as group FSH 5. The Millipore filter rested on a stainless-steel grid in a culture dish (Nunc, Roskilde, Denmark). All the cultures were incubated at 37˚C in a humidified atmosphere of 5% CO 2 and 95% O 2 for 48 hr. All of the above procedures were performed under sterile conditions.After incubation, the cultures were immediately frozen in liquid nitrogen. Frozen sections (8 µm) were air-dried for a few min, fixed in acetone at 20˚C for 10 min and allowed to be air-dried. The indirect immunofluorescence with amplification by the biotin-streptavidin system was used. Sections were incubated with monoclonal antibody to cytokeratin (clone Lu5, Boeringer Mannheim GmbH, Germany) or vimentin (clone V 9, Biodesign International, U.S.A.) and visualized with rhodamine X isothiocyanate (XRITC) or fluorescein isothiocyanate (FITC), respectively.To measure the proliferative activity of Sertoli cells in cultured testes, in another series of experiments, 10 µM 5-bromo-2'-deoxyuridine (BrdU) was added to the culture medium after 24 hr-incubation and the incubation was continued for 2 hr. Then, each explant was transferred Many studies have been made on differentiation and development of rat testes during the fetal period. In the formation of seminiferous tubule in the rat [20] and mouse [3], several mesenchyme-derived cells initially surround primordial germ cells to create a tube-like shape in the gonadal primordium. These mesenchyme-derived cells are considered to differentiate into Sertoli cells thereafter [7,8].It has been reported that intermediate filaments, cytokeratin and vimentin, are simultaneously found in Sertoli cells during the fetal period [15]. Cytokeratin disappears on 14 days after birth. Rat Sertoli cells show active cell division and proliferation during t...

Bovine Sertoli Cells Colonize and Form Tubules in Murine Hosts Following Transplantation and Grafting Procedures

“…Vimentin is an intermediate filament protein in Sertoli cells, and its synthesis is regulated by follicle‐stimulating hormone (FSH; DePhilip and Kierszenbaum, 1982; Sasaki et al, 1998) and androgen (Wang et al, 2006). The Non‐POU‐domain‐containing, octamer binding protein (NONO) forms a complex with the androgen receptor and functions as a coactivator for the receptor (Kuwahara et al, 2006).…”

Involvement of calcium-dependent mechanisms in T3-induced phosphorylation of vimentin of immature rat testis

“…However, nongenomic actions of thyroid hormones are now widely described and characterized by rapid responses, since they occur through plasma membrane transport systems (Incerpi et al, 1999b;Davis et al, 2000Davis et al, , 2002Silva et al, 2002;Volpato et al, 2004). Although short-term effects of FSH and steroid hormones on vimentin have been described in Sertoli cells (Spruill et al, 1983;Sasaki et al, 1998;Show et al, 2003), little is known about short-term effects of thyroid hormones on cytoskeletal proteins in testicular cells. In the present study we have shown short-term effects of the thyroid hormone T 3 on the in vitro phosphorylation of cytoskeletal-associated vimentin in immature rat testis.…”

Effects of secretome on cisplatin-induced testicular dysfunction in rats

“…Vimentin and cytokeratin, which are intermediate filaments, were observed in this present study. In normal conditions, during the developmental stage, vimentin was detected in the Sertoli cells of both fetuses and adults [ 23 - 26 ]. The maturation of Sertoli cells was supported by the presence of vimentin in the cells.…”

“…However, an extremely strong vimentin immunoreactivity was observed in the spermatozoa, Sertoli cells, and Leydig cells 1 week after the second secretome injection at high dose. Vimentin was observed in the spermatogenic, Sertoli, and Leydig cells because vimentin supported and controlled the cell structure and function of fetuses or adults [ 26 ]. Very strong vimentin immunoreactivity indicated a regenerative process.…”