Effect of Ethanol on DARPP‐32 Phosphorylation in Transgenic Mice That Express Human Type VII Adenylyl Cyclase in Brain

Abstract

Dopamine and cyclic adenosine monophosphate-regulated phosphoprotein of molecular weight 32 kDa (DARPP-32) is a bidirectional signaling protein found in dopaminergically innervated brain areas. The characteristics and direction of DARPP-32 effects are regulated by phosphorylation of this protein. Phosphorylation of DARPP-32 on threonine-34 (T34) is regulated through the activation of dopamine (D1) receptors and stimulation of adenylyl cyclase (AC) and protein kinase A activity and by calcineurin. Phosphorylation of DARPP-32 on threonine-75 (T75) is regulated by cyclin-dependent kinase 5 and protein phosphatase 2A. DARPP-32 has been implicated in the motivational effects of ethanol. The authors characterized transgenic mice that overexpress an ethanol-sensitive isoform of AC (AC7) in brain by measuring basal and ethanol-modulated DARPP-32 phosphorylation. Phosphorylated and total DARPP-32 were measured by immunoblotting in brain areas associated with the motivational and anxiolytic effects of ethanol (nucleus accumbens, striatum, and amygdala). AC7 transgenic mice had higher basal levels of T34 DARPP-32 than wild-type mice in striatum and amygdala, whereas basal levels of T75 DARPP-32 did not differ between wild-type and transgenic mice. Ethanol administration increased T34 DARPP-32 in nucleus accumbens and amygdala (but not in the striatum) of wild-type and transgenic mice (with a greater effect in amygdala of transgenic mice than wild-type mice). Ethanol administration increased T75 DARPP-32 in amygdala of only the wild-type mice and in nucleus accumbens and striatum of both the transgenic and wild-type mice. The effect of ethanol on the balance of DARPP-32 phosphorylation, especially in amygdala of wild-type versus transgenic mice, may contribute to differential motivational effects of ethanol in these animals.