GABA transporters in Drosophila melanogaster: molecular cloning, behavior, and physiology.

Abstract

Molecular cloning of GABA transporter-homologous cDNAs from a Drosophila melanogaster head-specific library was accomplished using a conserved oligomer from a highly conserved domain within the mammalian GABA transporters. Partial DNA sequencing of these cDNAs demonstrated homology with the mammalian transporters, indicating these are ancient, evolutionarily conserved molecules. Although the Drosophila cDNAs had distinct restriction enzyme patterns, they recognized the same locus in Drosophila genomic DNA, suggesting that the multiple isoforms might arise via alternative splicing. Antibodies specific for the mammalian GABA transporters GAT 1, GAT 2 and GAT 3 recognized non-overlapping and developmentally distinct patterns of expression in Drosophila neuronal tissues. Treatment of larval instars with nipecotic acid, a generalized GABA reuptake inhibitor, revealed specific, dose-dependent alterations in behavior consistent with the presence of multiple transporter molecules with differing affinities for this drug. Synaptic current recordings revealed that nipecotic acid treated larvae have an increase in latency jitter of evoked quantal release, resulting in a broader average excitatory junctional current which was manifested in a broader EJP. These results imply that alterations in the development of the CNS occur if GABAergic neurotransmission is potentiated during development. The data suggest that, as in mammals, there are multiple GABA transporters in Drosophila whose expression is differentially regulated.