Does Flotillin play a role in lipid raft organization of the GnRH receptor and its ability to transduce an intracellular signal?
Phillips, Adam Paul, author
Mrdutt, Mary Megan, author
Clay, Colin McKeown, author
Gonadotropin-Releasing Hormone (GnRH) and its subsequent signaling through the GnRH Receptor (GnRH-R) is critical for gonadal development and control of reproduction function. The GnRH-R is a member of the G-protein coupled receptor (GPCR) superfamily and is localized to specialized low-density areas on the cell membrane termed lipid rafts. These raft domains are implicated in GPCR coupled signaling by spatially organizing receptors and their associated signaling proteins to specific domains in the plasma membranes of cells. These raft domains appear to play an important role in the organization of GnRH-R and the signaling of GnRH to MAPkinase. Flotillin-1 is a protein thought to be intricately involved in the organization of rafts and the trafficking of proteins to raft domains. To examine the potential role of flotillin in GnRH signaling, gonadotrope derived αT3-1 cells were transfected with a specific siRNA for Flotillin-1 with the long-term goal of assessing the impact of Flotillin-1 deficiency on GnRHR trafficking to lipid rafts and signaling to intracellular targets including extracellular signal regulated kinase (ERK). Objectives: to use si-GLO Red to determine the transfection efficiency of Flotillin-1 siRNA; to use siRNA technology to knockdown Flotillin-1 expression; to determine the effect of Flotillin-1 knockdown on GnRH-R signaling.