Relaxin affects cell organization and early and late stages of spermatogenesis in a coculture of rat testicular cells

Author(s)

Pimenta, M. T. ^[1] ; Francisco, R. A. R. ^[1] ; Silva, R. P. ^[1] ; Porto, C. S. ^[1] ; Lazari, M. F. M. ^[1]

Affiliation

^[1] Univ Fed Sao Paulo, Sect Expt Endocrinol, Dept Pharmacol, Escola Paulista Med, BR-04044020 Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: ANDROLOGY; v. 3, n. 4, p. 772-786, JUL 2015.
• Web of Science Citations: 3

Abstract

Relaxin and its receptor RXFP1 are co-expressed in Sertoli cells, and relaxin can stimulate proliferation of Sertoli cells. In this study, we investigated a role of relaxin in spermatogenesis, using a short-term culture of testicular cells of the rat that allowed differentiation of spermatogonia to spermatids. Sertoli, germ, and peritubular myoid cells were the predominant cell types in the culture. Sertoli and germ cells expressed RXFP1. Cultures were incubated without (control) or with 0.5% fetal bovine serum (FBS) or 100ng/mL H2 relaxin (RLN) for 2days. Cell organization, number, and differentiation were analyzed after 2 (D2), 5 (D5) or 8 (D8) days of culturing. Although the proportion of germ cells decayed from D2 to D5, the relative contribution of HC, 1C, 2C, and 4C germ cell populations remained constant in the control group during the whole culture. RLN did not affect the proportion of germ cell populations compared with control, but increased gene and/or protein expression of the undifferentiated and differentiated spermatogonia markers PLZF and c-KIT, and of the post-meiotic marker Odf2 in D5. RLN favored organization of cells in tubule-like structures, the arrangement of myoid cells around the tubules, arrangement of c-KIT-positive spermatogonia at the basal region of the tubules, and expression of the cell junction protein -catenin close to the plasma membrane region. Knockdown of relaxin with small interfering RNA (siRNA) reduced expression of -catenin at the cell junctions, and shifted its expression to the nucleus. We propose that relaxin may affect spermatogenesis by modulating spermatogonial self renewal and favoring cell contact. (AU)

• FAPESP's process: 10/10341-1 - Characterization of the relaxin role on proliferation and differentiation of germ cells in different stages of development
• Grantee: Maristela Taliari Pimenta
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 10/10274-2 - Analysis of the role of relaxin in early and late stages of spermatogenesis, using co-cultures of Sertoli and germ cells
• Grantee: Maria de Fátima Magalhães Lazari
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/07456-0 - Effects of relaxin silencing in a co-culture model of Sertoli and germ cells
• Grantee: Raphael Paulo da Silva
• Support Opportunities: Scholarships in Brazil - Scientific Initiation