Generating a sensitive and reliable reporter system for studying INSL3-RXFP2 interactions

摘要

Insulin-like peptide 3 (INSL3), a peptide hormone, is primarily expressed in the testis and ovaries. It regulates the inguinal canal translocation of fetal gonad during development through activation of its receptor, RXFP2. Agonists and antagonists of RXFP2 are potential novel medications for contraceptive approaches or infertility treatments. However identification of effective RXFP2 agonists and antagonists was impeded due to the lack of a reliable system to study INSL3-RXFP2 interactions. Here we established such a reporter system named HEK293T-RXFP2 by stable transfection of HEK293T cells with a controllably expressing RXFP2. Compared to transiently transfected cells which were commonly used by previous studies, we found that our HEK293T-RXFP2 cells provided a high sensitive and reliable system to examine the response of RXFP2 to its ligands, as demonstrated by their greater signaling differences of the pEC50 and Emax values when stimulated by various ligands. Our HEK293T-RXFP2 cells can be used not only to study the functional response of RXFP2, but also to functionally screen the modulators of RXFP2 signaling.