Voltage-activated currents in identified giant interneurons isolated from adult crickets gryllus bimaculatus

The electrophysiological properties of cultured giant interneurons isolated from the terminal ganglion of adult crickets (Gryllus bimaculatus) were investigated using whole-cell patch-clamp techniques. To allow for unequivocal identification of these interneurons in cell culture, a protocol for fast and selective labeling of their cell bodies was established. Prior to cell dissociation, the giant interneurons were backfilled through their axons in situ with a fluorescent dye (dextran tetramethylrhodamine). In primary cell cultures, the cell bodies of giant interneurons were identified among a population of co-cultured neurons by their red fluorescence. Action potentials were recorded from the cell bodies of the cultured interneurons suggesting that several types of voltage-activated ion channels exist in these cells. Using voltage-clamp recording techniques, four voltage-activated currents were isolated and characterized. The giant interneurons express at least two distinct K+ currents: a transient current that is blocked by 4-aminopyridine (4x10(-3 )mol l-1) and a sustained current that is partially blocked by tetraethylammonium (3x10(-2 )mol l-1) and quinidine (2x10(-4 )mol l-1). In addition, a transient Na+ current sensitive to 10(-7 )mol l-1 tetrodotoxin and a Ca2+ current blocked by 5x10(-4 )mol l-1 CdCl2 have been characterized. This study represents the first step in an attempt to analyze the cellular and ionic mechanisms underlying plasticity in the well-characterized and behaviorally important giant interneuron pathway in insects.