Nobue Kanematsu1, Ikuyo Nakajima2, Kiyonori Haga3, Madoka Suto1. 1. Department of Animal Physiology and Nutrition. 2. Department of Animal Products Research, and. 3. Department of Research Planning and Coordination, National Institute of Livestock and Grassland Science, Tsukuba, Japan.
Abstract
Aims: There is increasing concern about the effect of environmental toxins and endocrine disrupters on human spermatogenesis but ethical considerations prohibit direct research. Therefore, we developed a method of isolating mature pig Sertoli cells and studying the levels of mRNA of transferrin and inhibin, markers of Sertoli cell function, using real-time polymerase chain reaction analysis. Methods: Sertoli cells from mature Meishan boars were isolated and cultured. The mRNA was isolated from the cells after 3 days of treatment with follicle-stimulating hormone (FSH), testosterone and β-estradiol. The amounts of transferrin and inhibin mRNA were analyzed by real-time polymerase chain reaction and the relative level of mRNA was calculated. Results: FSH tended to increase the levels of transferrin and inhibin mRNA, as did the combinations of FSH and the steroids. The effect of testosterone and β-estradiol alone on the level of mRNA was less than that of FSH. Beta-estradiol treatment resulted in a dose-dependent decrease in mRNA. Conclusion: The Sertoli cell culture used in the present study appears to have a normal secretory function because the mRNA levels of both markers increased after FSH treatment. The suppressive effect of β-estradiol on inhibin mRNA level suggests that β-estradiol has an effect on the function of Sertoli cells. (Reprod Med Biol 2005; 4: 259 -264).
Aims: There is increasing concern about the effect of environmental toxins and endocrine disrupters on human spermatogenesis but ethical considerations prohibit direct research. Therefore, we developed a method of isolating mature pig Sertoli cells and studying the levels of mRNA of transferrin and inhibin, markers of Sertoli cell function, using real-time polymerase chain reaction analysis. Methods: Sertoli cells from mature Meishan boars were isolated and cultured. The mRNA was isolated from the cells after 3 days of treatment with follicle-stimulating hormone (FSH), testosterone and β-estradiol. The amounts of transferrin and inhibin mRNA were analyzed by real-time polymerase chain reaction and the relative level of mRNA was calculated. Results:FSH tended to increase the levels of transferrin and inhibin mRNA, as did the combinations of FSH and the steroids. The effect of testosterone and β-estradiol alone on the level of mRNA was less than that of FSH. Beta-estradiol treatment resulted in a dose-dependent decrease in mRNA. Conclusion: The Sertoli cell culture used in the present study appears to have a normal secretory function because the mRNA levels of both markers increased after FSH treatment. The suppressive effect of β-estradiol on inhibin mRNA level suggests that β-estradiol has an effect on the function of Sertoli cells. (Reprod Med Biol 2005; 4: 259 -264).
Authors: K Miyamoto; Y Hasegawa; M Fukuda; M Nomura; M Igarashi; K Kangawa; H Matsuo Journal: Biochem Biophys Res Commun Date: 1985-06-14 Impact factor: 3.575
Authors: D M de Kretser; J J Buzzard; Y Okuma; A E O'Connor; T Hayashi; Shyr-Yeu Lin; J R Morrison; K L Loveland; M P Hedger Journal: Mol Cell Endocrinol Date: 2004-10-15 Impact factor: 4.102