Figure 4.
Young male rats express AR45 protein in membrane lipid rafts. Young rats (3 mo old) were either gonadectomized (A) or gonadally intact (B). Micropunches of substantia nigra tissue were collected, and the membrane was isolated and then separated into nine fractions using a sucrose gradient to examine lipid rafts. Primary antibodies targeting AR45 (AR-C19 antibody), G proteins Gαq and Gαo, and lipid raft markers (caveolin-1 and flotillin) were used. AR45 expression was observed only in lipid raft fractions, as evidenced by caveolin-1 and flotillin expression. Hormone status did not alter AR45 expression in lipid rafts. Gαq and Gαo was observed throughout the membrane, regardless of hormone status. No protein expression of Gαi1-3 and Gαs G proteins or full-length (100-kDa) AR was observed in the substantia nigral membranes of young male rats (data not shown). GDX, gonadectomy; intact, gonadally intact. n = 3 per treatment group.