CALCIUM-DEPENDENT REGULATION OF THE ENKEPHALIN PHENOTYPE BY NEURONAL-ACTIVITY DURING EARLY ONTOGENY

Genetic components of the neuronal phenotype are regulated by epigenetic factors-trophic molecules and neuronal activity-during neurodifferentiation. Developing neurons in dissociated cultures of embryonic mouse spinal cord show spontaneous electrical activity for two days with tetrodotoxin (TTX) causes virtually complete down-regulation of preproenkephalin A gene transcripts in embryonic spinal cord cultures. This TTX-induced down-regulation is fully reversed upon reinitiation of neuronal activity (removal of TTX from cultures). This reversible, tetrodotoxin-induced down-regulation of enkephalin mRNA is confined to a restricted period of early neurodevelopment (days 7 to 21 in culture). Since depolarization triggers calcium entry through voltage-activated calcium channels, we have investigated the involvement of calcium in the mechanism of this activity- and age-dependent regulation of preproenkephalin A expression. The selective activation of the L-type of voltage-sensitive calcium channels by a dihydropyridine derivative [(+) 202-791] prevented this TTX-induced down-regulation without reducing methionine enkephalin secretion. This effect was observed only when the drug was applied to electrically active cultures, prior to the addition of TTX. Simultaneous application of (+) 202-791 and TTX, or pretreatment with TTX, failed to prevent TTX-induced down-regulation. Thus, activity-dependent phenotypic plasticity of met-enkephalinergic neurons in spinal cord is: 1) maximum at an early age of neuronal development (< 10 days in culture) and becomes less apparent in old cultures (> 30 days); 2) reversible throughout; and 3) mediated by calcium entry through L-type channels.