Effects of relaxin silencing in a co-culture model of Sertoli and germ cells

• Grant number: 13/07456-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2013
• End date: December 31, 2013
• Field of knowledge: Biological Sciences - Pharmacology - Biochemical and Molecular Pharmacology
• Principal Investigator: Maria de Fátima Magalhães Lazari
• Grantee: Raphael Paulo da Silva
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Relaxin is an insulin-related peptide that interacts with the G-protein coupled receptor RXFP1. The male mice with a relaxin knockout is infertile, and previous studies from our laboratory revealed that relaxin and RXFP1 are expressed in germ cells, and relaxin stimulates the proliferation of Sertoli cells through a mechanism that involves activation of MEK/ERK1/2 and PI3K/AKT pathways. The results suggest that relaxin may play an autocrine/paracrine role to regulate spermatogenesis. To evaluate the role of relaxin in early and late stages of spermatogenesis we have been using a co-culture model of Sertoli and germ cells from 7-8 day-old rats. This model has allowed to determine the effect of exogenous relaxin on proliferation and differentiation of germ cells. The aim of this work is to analyze the effect of silencing relaxin expression in the co-culture. The study will involve the following steps: 1) To establish the best conditions to silence relaxin expression, analyzing the gene expression by real time PCR and the protein expression by immunocytochemistry; 2) The effect of relaxin silencing on ploidy and proportion of germ and Sertoli cells, by flow cytometry; 3) The effect of relaxin silencing on the expression of markers of different phases of spermatogenesis, by flow cytometry and real time PCR. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

PIMENTA, M. T.; FRANCISCO, R. A. R.; SILVA, R. P.; PORTO, C. S.; LAZARI, M. F. M.. Relaxin affects cell organization and early and late stages of spermatogenesis in a coculture of rat testicular cells. ANDROLOGY, v. 3, n. 4, p. 772-786, JUL 2015. (10/10341-1, 10/10274-2, 13/07456-0)