Calcitonin gene-related peptide: distribution and effects on spontaneous rhythmic activity in embryonic chick spinal cord

Abstract

Immunohistochemical and in vitro electrophysiological techniques were utilized to examine the distribution and possible role of calcitonin gene-related peptide (CGRP) in the spinal cord of the developing chick. CGRP-like immunoreactivity first appeared in the lateral motor column of the lumbosacral spinal cord at embryonic day 6 followed by the emergence of fiber immunoreactivity in the dorsal horn at embryonic day 11. A rostrocaudal survey of the cervical to lumbosacral spinal cord in embryonic day 18 chick demonstrated robust CGRP-like immunoreactivity at all levels in both putative motor neurons and dorsal horn fibers. Additionally, small immunoreactive lamina VII neurons were observed in sections of lumbosacral cord. In the embryonic day 10 (E10) in vitro reduced spinal cord preparation, bath application of the calcitonin gene-related peptide antagonist human α-CGRP fragment 8-37 decreased the frequency and increased the duration of episodes of spontaneously occurring rhythmic activity. Conversely, application of α or β forms of calcitonin gene-related peptide increased the frequency of the rhythmic episodes. The electrophysiological results suggest there is a constitutive release of calcitonin gene-related peptide contributing to the spontaneous rhythmic activity. Immunohistochemical results from E10 animals suggest that CGRP-like immunoreactive putative motoneurons may be the source of the released CGRP.