Growth properties of larval and adult locust neurons in primary cell culture.

Abstract

We developed a cell culture system for thoracic neurons of fifth instar or adult locusts (Locusta migratoria) in order to obtain maximum visualization of cellular morphology and direct access to the neurons for electrophysiological analysis. The dissociated neurons regenerated new neurites in a serum-free defined culture medium, in which they remained viable for up to 3 weeks. Viability of the cells was confirmed by intracellular recordings demonstrating active membrane properties and action potentials. While the morphology of the cultured neurons is distinct from their in vivo counterparts, they retained some cellular surface properties and markers related to transmitter metabolism. Two factors influencing cellular morphology in vitro were identified in Locusta: 1) the presence of a primary neurite stump, and 2) membrane contacts between cells. Dissociated neurons of the locust species Schistocerca gregaria grown in a hemolymph-enriched medium showed a marked reduction in branching patterns and a tenfold increase in neurite length compared to neurons growing in a medium without hemolymph. This culture system could prove useful for identifying the action of hemolymph-derived growth factors.