Calcitonin gene-related peptide regulates gene transcription in primary afferent neurons

Although primary afferent neurons express receptors for calcitonin gene-related peptide (CGRP), understanding of the cellular effects of these receptors is limited. We determined that CGRP receptors regulate gene transcription in primary afferent neurons through a cyclic AMP (cAMP)-dependent pathway. CGRP increased cAMP in neonatal dorsal root ganglion (DRG) neurons in a concentration-dependent manner that was blocked by the receptor antagonist CGRP_8-37. The response to CGRP also occurred in adult DRG cells. In contrast, CGRP did not alter the concentration of free intracellular calcium in neonatal or adult DRG neurons. Immunohistochemical data showed that one downstream effect of the cAMP signaling pathway was phosphorylation of cAMP response element binding (CREB) protein, suggesting that CGRP regulates gene expression. This interpretation was supported by evidence that CGRP increased CRE-dependent gene transcription in neurons transiently transfected with a CRE-luciferase DNA reporter construct. The effect of CGRP on gene transcription was inhibited by H89, myristoylated-protein kinase A inhibitor _14-22-amide and U0126, indicating that protein kinase A and mitogen-activated protein kinase/extracellular receptor kinase kinase are enzymes that mediate effects of CGRP on gene transcription. Therefore, CGRP receptors may regulate expression of proteins by primary afferent neurons during development and in response to tissue-damaging stimuli.