An insight into insulin-like factor 3 regulate its receptor RXFP2 in mouse gubernaculum testis cells

Abstract

The etiology of testicular dysgenesis syndrome is multifactorial and involves abnormalities in the anatomical structures and endocrine factors. Several studies have shown that the abnormal development of the gubernaculum may affect testicular descent, and the insulin-like factor 3 (INSL3) appears to play an important role in development of the gubernaculum have been proved. INSL3 binds its specific receptor (Relaxin family peptide 2, RXFP2), which was highly expressed in gubernaculum, to produce a crucial effect in the first transabdominal descent stage, but its mechanism still remain unclear. In this study, in order to explore how does INSL3 regulate its receptor RXFP2, we cultured mouse gubernaculum testis cells in vitro, which was treated by INSL3, and examined the expression of RXFP2 in mouse gubernaculum testis cells. The results displayed that INSL3 changed RXFP2 expression, and we found that low dose INSL3 can increase RXFP2 expression, the mechanism of above-mentioned might be related with the hormesis of INSL3.

Keywords: INSL3; RXFP2; gubernaculums; mouse.

Figure 1

The staining of HE displayed the culture cells were fibroblast-like and highly homogeneous in morphology (scanning for × 100, × 200, × 400 respectively).

Figure 2

Expression of RXFP2 mRNA after treatment with different concentrations of INSL3 (A, B). RC means relative coefficient = samples value/beta actin value. The INSL3 groups were given 20 ng/ml, 2 ng/ml, 0.2 ng/ml, and 0.02 ng/ml respectively (*P < 0.01 vs. normal group).

Figure 3

The red-tinted region and gray region represent the fluorescence intensity of samples and normal control respectively (A). Protein expression of RXFP2 after different concentrations INSL3 treated (B). FI means fluorescence index = fluorescence intensity of samples-fluorescence intensity of blank control. The INSL3 groups were given 20 ng/ml, 2 ng/ml, 0.2 ng/ml, and 0.02 ng/ml respectively (*P < 0.01 vs. normal group).