Alteration of association of agonist-activated renal D1A dopamine receptors with G proteins in proximal tubules of the spontaneously hypertensive rat

BACKGROUND: Defective D1A dopamine receptor-G protein coupling has been identified in renal proximal tubules of the spontaneously hypertensive rat (SHR). OBJECTIVE: To determine whether association of D1A dopamine receptors with the alpha subunits of G proteins in kidney of SHR is normal. METHODS: We analyzed the association of agonist-activated [1251]-labeled D1A dopamine receptors in kidneys of SHR and the normotensive Wistar-Kyoto (WKY) rat through immunoprecipitation, using highly specific antipeptide antibodies directed against alpha subunits of G proteins. RESULTS: We have shown for the first time that the D1A receptors of renal proximal tubules are associated with the adenylyl cyclase inhibitory G proteins G(i)alpha. The association of WKY rat proximal tubule D1A receptors with Gi1alpha and Gi2alpha in the presence of agonist is significantly (P<0.01) greater (2.4-fold and 3.1-fold greater, respectively) than it is without agonist D1A receptors of WKY rat also exhibit (twofold greater) association with G(s)alpha, consistently with the ability of these receptors to mediate stimulation of adenylyl cyclase. The WKY rat D1A receptors do not associate either with G(o)alpha or with G(q)alpha. The D1A receptors of SHR proximal tubule membranes appear to be resistant to activation by agonist and do not associate with G(s)alpha, G(o)alpha and any of the subunits of G(i)alpha. However, the SHR D1A sites exhibit a modestly (1.7-fold) greater association with G(q)alpha, which was not statistically significant. The differences among associations of the D1A receptors of WKY rat and SHR with these Galpha proteins may be important in understanding renal dopaminergic functions in normal and pathophysiologic states.