Transferrin and inhibin mRNA in mature pig Sertoli cells

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)

Development and Validation of Endogenous Reference Genes for Expression Profiling of Medaka (Oryzias latipes) Exposed to Endocrine Disrupting Chemicals by Quantitative Real-Time RT-PCR

“…RPL-7 is an integral part of the large subunit of ribosome (von Mikecz et al, 1999). It has been found constantly expressed in some mammalian tissues and cells at stages of development, and has been used to normalize mammalian gene expression in many studies (Huot et al, 2001;Kanematsu et al, 2005;Muroya et al, 2005;Sun et al, 2004). We cloned the complete mRNA sequence of the RPL-7 from medaka (Fig.…”

“…However, it has been reported that these genes were unstably expressed by EDCs exposure (Farnell and Ing, 2003;Hsu and Frankel, 1987;Kamata et al, 2005;Larkin et al, 2003). Ribosomal protein L7 (RPL-7) is a housekeeping gene that has been recently used to normalize gene expression in mammals (Huot et al, 2001;Muroya et al, 2005;Sun et al, 2004), and has been adopted as an internal control in studying mature pig sertoli cells and mouse uterus treated with EDCs (Das et al, 1998;Kanematsu et al, 2005). However, RPL-7 has never been used as endogenous reference in lower vertebrate, such as in fish which are widely used in EDCs study.…”