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Analysis of the role of relaxin in early and late stages of spermatogenesis, using co-cultures of Sertoli and germ cells

Abstract

Relaxin is a member of the family of insulin-related peptides, which interacts with a G-protein coupled receptor, RXFP1. Disruption of relaxin gene in males causes infertility, and studies from our laboratory have shown that relaxin and RXFP1 co-localize in Sertoli cells, RXFP1 is present in germ cells and relaxin induces Sertoli cell proliferation through a mechanism that seems to involve activation of ERK1/2. These data suggest that relaxin may play an autocrine and/or paracrine role to regulate spermatogenesis. The aim of the present project is to evaluate the role of the hormone in early and late stages of spermatogenesis. The studies will be conducted in a co-culture of Sertoli and germ cells from rats in different stages of development (7, 20 and 60 days after birth), and it will be analyzed: a) the role of relaxin on proliferation and diferentiation of gonocytes and spermatogonias, differentiation of round spermatids to spermatozoa, using specific markers of proliferation and diferentiation, by flow cytometry, real time PCR, Western blot and immunofluorescence; b) the effect of silencing relaxin receptors on proliferation and differentiation of germ cells; b) the effect of relaxin on activation of transcription factor CREMt, which is important for the differentiation of round spermatids, by chromatin immunoprecipitation assay; c) the effect of relaxin on Sertoli cell production of c-kit, GDNF and caderins, which are important factors for spermatogenesis, by real time PCR, Western blot and immunofluorescence. (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)
NASCIMENTO, ALINE R.; MACHERONI, CARLA; LUCAS, THAIS F. G.; PORTO, CATARINA S.; LAZARI, MARIA F. M. Crosstalk between FSH and relaxin at the end of the proliferative stage of rat Sertoli cells. Reproduction, v. 152, n. 6, p. 613-628, DEC 2016. Web of Science Citations: 5.
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. Web of Science Citations: 3.

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