Subproject 3 – Early effects of estrogen on smooth muscle cell behavior: the role of GPR30 membrane receptor

Participants: Sérgio Luis Felisbino (Unesp-Botucatu) and Hernandes F Carvalho

Estrogen has been shown to exert strong effects on male reproduction. Estrogen action is exerted through its receptor (ERalpha and ERbeta). However, more recently, early effects of estrogen on mammalian cells were observed an estrogen receptor was found at the cell membrane. The former orphan receptor GPR30 was identified as the functional transmembrane receptor for estrogen. The purpose of this work is to characterize the early effects of estrogen on prostatic smooth muscle cells, characterize the presence and variations in GPR30 at the mRNA and protein levels, inhibit GPR30 expression by siRNA and monitor cytoskeletal modifications in real time by using smooth muscle cells isolated from the transgenic actin-GFP mouse. Physiological parameters of the cells will be monitored by the use of CARS and FLIM in parallel with migratory behavior of smooth muscle cells in culture.