Regulation of Calcitonin Gene-Related Peptide Expression in Vitro

Abstract

Calcitonin-gene related peptide is among a group of peptides whose expressions are down-regulated following peripheral nerve damage. It is known that this is probably due to deprivation of some target-derived neurotrophic factors, mainly of nerve growth factor though positive effect of other factors, for example that of leukemia inhibitory factor on galanin has also been demonstrated. In this study, the effects of leukemia inhibitory factor and nerve growth factor on calcitonin gene related peptide expression in cultured dorsal root ganglion explants and in their outgrowing axons were examined. Lumbar dorsal root ganglia with short pieces of peripheral nerves were removed from adult mice and explanted into collagen gels. They were covered with RPMI 1640 culture medium and left in an incubator for 2 days after which they were fixed. These whole mount preparations with outgrowing axons were stained with an antibody against calcitonin gene related peptide. Following microscopic examination and imaging, sections were cut from the cultured ganglia as well as from some freshly taken normal ones and they were also stained to determine calcitonin gene related peptide immunoreactivity in the primary sensory neurons. The results demonstrated that besides the positive effect of nerve growth factor on the expression of this peptide in outgrowing axons, leukemia inhibitory factor also supported the expression of calcitonin gene related peptide in the primary sensory neurons of adult mouse lumbar dorsal root ganglia and in their outgrowing axons in vitro. When the time course of changes in calcitonin gene related peptide expression in dorsal root ganglia and the up-regulation of leukemia inhibitory factor at the site of a peripheral nerve injury in vivo are considered together, this novel finding may lead to new explanations for the changes in neuropeptide expression following axotomy. (C) 2002 Elsevier Science B.V. All rights reserved.