Signaling pathway downstream of GABA(A) receptor in the growth cone

The growth cone is responsible for axonal elongation and pathfinding by responding to various modulators for neurite growth, including neurotransmitters, although the sensor mechanisms are not fully understood. Among neurotransmitters, GABA is most likely to demonstrate activity in vivo because GABA and the GABA(A) receptor appear even in early stages of CNS development. We investigated the GABA(A) receptor-mediated signaling pathway in the growth cone using isolated growth cones (IGCs). Both the GABA(A) binding site and the benzodiazepine modulatory site were enriched in the growth cone membrane. In the intact IGC, GABA induced picrotoxin-sensitive Cl- flux (not influx but efflux) and increased the intracellular Ca2+ concentration in a picrotoxin- and verapamil-sensitive manner. Protein kinase C (PKC)-dependent phosphorylation of two proteins identified as GAP-43 and MARCKS protein was enhanced in the intact IGC stimulated by GABA, resulting in the release of MARCKS protein and GAP-43 from the membrane. Collectively, our results suggest the following scheme: activation of the functional GABAA receptor localized in the growth cone membrane --> Cl- efflux induction through the GABA(A)-associated Cl- channel --> Ca2+ influx through an L-type voltage-sensitive Ca2+ channel --> Ca2+-dependent phosphorylation of GAP-43 and MARCKS protein by PKC.