In vitro studies of axonally-regulated Schwann cell genes during Wallerian degeneration.

Wallerian degeneration in vivo is associated with marked downregulation of myelin protein genes such as P(o) and upregulation of other genes such as nerve growth factor receptor (NGF-R), glial fibrillary acidic protein (GFAP) and neural cell adhesion molecule (N-CAM). This study examines the expression of these genes during Wallerian degeneration in vitro and how manipulating Ca2+ affects this response. Small explants of sciatic nerve from normal young adult rats cultured for five days show similar reversal of the myelinating phenotype as found in vivo. If Ca++ is removed from the culture medium through the addition of EGTA, expression of the nerve growth factor receptor and glial fibrillary acidic protein genes is inhibited but downregulation of the P(o) gene still occurs. Explants cultured in medium containing EGTA are still capable of expressing nerve growth factor receptor if the medium is replaced by one containing Ca2+. Supplementation of normal medium with drugs modulating Ca2+, such as Bepridil which blocks the Na+Ca2+ exchanger or compound 48/80 which inhibits calmodulin, also prevent the expression of the nerve growth factor receptor gene during Wallerian degeneration in vitro. Treatment of the cervical sympathetic trunk with Bepridil leads to loss of the nerve growth factor receptor immunoreactivity which is normally present. The results indicate that Ca2+ may play a role in the expression of the nerve growth factor receptor gene during Wallerian degeneration and provide some indication that this effect may be directly on the Schwann cell rather than operating indirectly via the axon.