Melanocortin 4 receptor constitutive activity inhibits L-type voltage-gated calcium channels in neurons

Abstract

The melanocortin 4 receptor (MC4R) is a G protein-coupled receptor (GPCR) that is expressed in several brain nuclei playing a crucial role in the regulation of energy balance controlling the homeostasis of the organism. It displays both agonist-evoked and constitutive activity, and moreover, it can couple to different G proteins. Most of the research on MC4R has been focused on agonist-induced activity, while the molecular and cellular basis of MC4R constitutive activity remains scarcely studied. We have previously shown that neuronal N-type voltage-gated calcium channels (Ca_V2.2) are inhibited by MC4R agonist-dependent activation, while the Ca_V subtypes that carry L- and P/Q-type current are not. Here, we tested the hypothesis that MC4R constitutive activity can affect Ca_V, with focus on the channel subtypes that can control transcriptional activity coupled to depolarization (L-type, Ca_V1.2/1.3) and neurotransmitter release (N- and P/Q-type, Ca_V2.2 and Ca_V2.1). We found that MC4R constitutive activity inhibits specifically Ca_V1.2/1.3 and Ca_V2.1 subtypes of Ca_V. We also explored the signaling pathways mediating this inhibition, and thus propose that agonist-dependent and basal MC4R activation modes signal differentially through G_s and G_i/o pathways to impact on different Ca_V subtypes. In addition, we found that chronic incubation with MC4R endogenous inverse agonist, agouti and agouti-related peptide (AgRP), occludes Ca_V inhibition in a cell line and in amygdaloid complex cultured neurons as well. Thus, we define new mechanisms of control of the main mediators of depolarization-induced calcium entry into neurons by a GPCR that displays constitutive activity.

Keywords: G protein; amygdala; calcium channels; electrophysiology; melanocortin receptor.