AbstractInhibitory synaptic connections mediated by γ‐aminobutyric acid (GABA) play important roles in the neural computation of the brain. To obtain a detailed overview of the neural connections mediated by GABA signals, we analyzed the distribution of the cells that produce and receive GABA in the Drosophila adult brain. Relatively small numbers of the cells, which form clusters in several areas of the brain, express the GABA synthesis enzyme Gad1. On the other hand, many cells scattered across the brain express ionotropic GABAA receptor subunits (Lcch3 and Rdl) and metabotropic GABAB receptor subtypes (GABA‐B‐R1, ‐2, and ‐3). To analyze the expression of these genes in distinct identified cell types, we focused on the antennal lobe, where GABAergic neurons play important roles in odor coding. By combining fluorescent in situ hybridization and immunolabeling against GFP expressed with cell‐type‐specific GAL4 driver strains, we quantified the percentage of the cells that produce or receive GABA for each cell type. GABA was synthesized in the middle antennocerebral tract (mACT) projection neurons and two types of local neurons. Among them, mACT neurons had few presynaptic sites in the antennal lobe, making the local neurons essentially the sole provider of GABA signals there. On the other hand, not only these local neurons but also all types of projection neurons expressed both ionotropic and metabotropic GABA receptors. Thus, even though inhibitory signals are released from only a few, specific types of local neurons, the signals are read by most of the neurons in the antennal lobe neural circuitry. J. Comp. Neurol. 514:74–91, 2009. © 2009 Wiley‐Liss, Inc.