Nerve growth factor-induced neurite outgrowth of rat pheochromocytoma PC 12 cells: Dependence on extracellular Mg2+ and Ca2+

Dependence of neurite outgrowth on extracellular Mg2+ and Ca2+ was studied in nerve growth factor-responsive pheochromocytoma PC 12 cells under assay conditions in which neurite formation was independent of both RNA synthesis and protein synthesis. NGF-induced neurite formation occured maximally in the presence of extracellular Mg2+ at concentrations greater than 1.0 mM. However, extracellular Ca2+ alone did not stimulate the neurite formation, and inhibited this process at higher concentrations (greater than 10 mM). These data are consistent with the fact that NGF-mediated neurite extension occured in assay medium containing either 1.0 mM EGTA or 0.5 mM LaCl3. Other divalent cations so far tested proved to be negative, suggesting that this phenomenon appears to be specific to Mg2+. Moveover, quantitative analysis revealed that the length and thickness of neurites formed were controlled by the presence of extracellular Ca2+. Thus, neurites formed at lower concentrations of Ca2+ in the presence of 1.0 mM Mg2+ and NGF were found to be thinner and longer than those formed at higher concentrations of Ca2+, suggesting that Ca2+ and Mg2+ have separate regulatory functions in the formation of neurites of PC 12 cells.