Open Access Subscription Access
Open Access Subscription Access
The Effect of Pulsatile Hormonal Stimulation on Gene Expression in Cultured Rat Sertoli Cells
FSH and Testosterone (T) are the major endocrine regulators of spermatogenesis. Both these hormones act via testicular Sertoli cells (Sc) to regulate the division and differentiation of male germ cells (Gc). A number of FSH- and/or T-responsive genes, expressed specifically by Sc having critical role in regulating sperm production, have been identified. We have previously reported the efficacy of a novel in vitro administration of pulsatile hormonal treatment protocol (exposure of FSH and T in combination for 30 minutes/3 hr upto 24 hr) on gene expression by cultured rat Sc as compared to that with the conventional constant (for 24 hr) hormonal exposure. In the present study, we further demonstrate that such pulsatile stimulation of hormones to cultured rat Sc show more prominent impact in terms of augmenting the expressions of Sc-specific genes critical for spermatogenesis, like Inhibin-β-B, Androgen binding protein (Abp) and Stem cell factor (Scf) at the 11th hr (hr) of total treatment duration i.e., 24 hr. This report thereby pinpoints the specific treatment regime for evaluating the maximal hormone-induced gene expression in cultured Sc systems. This new protocol of hormonal stimulation (for 11 hr only instead of 24 hr) will significantly improve our current practice of using primary Sc culture systems for detecting hormonal responsiveness or signaling with respect to gene expression in vitro.
FSH, Gene Expression, Sertoli Cells, Spermatogenesis, Testosterone.
- Griswold MD. The central role of Sertoli cells in spermatogenesis. Semin Cell Dev Biol. 1998; 9(4): 411–6.
- Walker WH, Cheng J. FSH and testosterone signaling in Sertoli cells. Reproduction, 2005; 130(1):15–28.
- Kumar TR. Mouse models for gonadotropins: A 15-year saga. Mol Cell Endocrinol. 2007; 260-262:249–54.
- Jonas KC, Oduwole OO, Peltoketo H, Rulli SB, Huhtaniemi I. Mouse models of altered gonadotrophin action: insights into male reproductive disorders. Reproduction, 2014;148(4): R63–70.
- De Gendt K, Verhoeven G, Amieux PS, Wilkinson MF. Research resource: Genome-wide identification of AR-regulated genes translated in Sertoli cells in vivo using the RiboTag approach. Mol Endocrinol. 2014; 28(4):575–91.
- Musnier A, León K, Morales J, Reiter E, Boulo T, Costache V, et al. mRNA-selective translation induced by FSH in primary Sertoli cells. Mol Endocrinol. 2012; 26(4):669–80.
- Majumdar SS, Bhattacharya I. Genomic and post-genomic leads toward regulation of spermatogenesis. Prog Biophys Mol Biol. 2013; 113(3):409–22.
- Bhattacharya I, Gautam M, Sarkar H, Shukla M, Majumdar SS. Advantages of pulsatile hormone treatment for assessing hormone-induced gene expression by cultured rat Sertoli cells. Cell Tissue Res. 2017; 368(2):389–96.
- Welsh MJ, Wiebe JP. Rat sertoli cells: A rapid method for obtaining viable cells. Endocrinology, 1975; 96(3):618–24.
- Majumdar SS, Tsuruta J, Griswold MD, Bartke A. Isolation and culture of Sertoli cells from the testes of adult Siberian hamsters: analysis of proteins synthesized and secreted by Sertoli cells cultured from hamsters raised in a long or a short photoperiod. Biol Reprod. 52(3):658–66.
- Devi YS, Sarda K, Stephen B, Nagarajan P, Majumdar SS. Follicle-stimulating hormone-independent functions of primate Sertoli cells: potential implications in the diagnosis and management of male infertility. J Clin Endocrinol Metab. 2006; 91(3):1062–8.
- Bhattacharya I, Pradhan BS, Sarda K, Gautam M, Basu S, Majumdar SS. A switch in Sertoli cell responsiveness to FSH may be responsible for robust onset of germ cell differentiation during prepubartal testicular maturation in rats. Am J Physiol Endocrinol Metab. 2012; 303(7):E886–98.
- Galdieri M, Zani BM, Monaco L, Ziparo E, Stefanini M. Changes of Sertoli cell glycoproteins induced by removal of the associated germ cells. Exp Cell Res. 1983; 145(1):191–8.
- Fix C, Jordan CL, Cano P, Walker WH. Testosterone activates mitogen-activated protein kinase and the cAMP response element binding protein transcription factor in Sertoli cells. Proc Natl Acad Sci U S A. 2004; 101(30):10919–24.
- Cheng J, Watkins SC, Walker WH. Testosterone activates mitogen-activated protein kinase via Src kinase and the epidermal growth factor receptor in Sertoli cells. Endocrinology. 2007; 148(5):2066–74.
- Viswanathan P, Wood MA, Walker WH. Follicle-stimulating hormone (FSH) transiently blocks FSH receptor transcription by increasing inhibitor of deoxyribonucleic acid binding/differentiation-2 and decreasing upstream stimulatory factor expression in rat Sertoli cells. Endocrinology. 2009; 150(8):3783–91.
- Shupe J, Cheng J, Puri P, Kostereva N, Walker WH. Regulation of Sertoli-germ cell adhesion and sperm release by FSH and nonclassical testosterone signaling. Mol Endocrinol. 2011; 25(2):238–52.
- Mishra J, Gautam M, Dadhich R, Kowtharapu BS, Majumdar SS. Peritubular cells may modulate Leydig cellmediated testosterone production through a nonclassic pathway. Fertil Steril. 2012; 98(5):1308–17.e1.
- Sharpe R. Regulation of spermatogenesis. In: Knobil E, Neill J, editors. The Physiology of Reproduction. 2nd ed. Raven Press, New York; 1994. p. 1363–434.
- Plant TM. 60 YEARS OF NEUROENDOCRINOLOGY: The hypothalamo-pituitary-gonadal axis. J Endocrinol. 2015; 226(2):T41–54.
- Mehats C, Andersen CB, Filopanti M, Jin SLC, Conti M. Cyclic nucleotide phosphodiesterases and their role in endocrine cell signaling. Trends Endocrinol Metab. 2002; 13(1):29–35.
- Usachev Y, Verkhratsky A. IBMX induces calcium release from intracellular stores in rat sensory neurones. Cell Calcium. 1995; 17(3):197–206.
- Bhattacharya I, Gautam M, Majumdar SS. The effect of IBMX and hormones on gene expression by rat Sertoli cells. J Reprod Heal Med. 2015; 1(1):29–40.
Abstract Views: 8
PDF Views: 0