GABAergic Neurons as Putative Neurochemical Substrate Mediating Aversive Effects of Nicotine

Nicotine, the main addictive component of tobacco smoke, has both rewarding and aversive properties. Recent studies have suggested that GABAergic neurons, one of the main neurochemical components of the reward-addiction circuitry, may also play a role in the aversive responses to nicotine. In the present study of transgenic mice expressing Green Fluorescent Protein (GFP) in Glutamate Decarboxylase 67 (GAD67) neurons, we hypothesized that a subpopulation of GABAergic neurons in the Ventral Tegmental Area (VTA) are the targets of aversive doses of nicotine in the CNS. We tested this hypothesis using c-Fos immunohistochemical techniques to identify GAD67-GFP positive cells within the VTA, that are activated by a single intraperitoneal (i.p.) injection of a low (40 ug/kg) or a high (2 mg/kg) dose of nicotine. We also assessed the anatomical location of GAD67-GFP positive cells with respect to tyrosine hydroxylase (TH) Immunoreactive (IR) dopaminergic cells in VTA. Consistent with our previous studies low- and high-dose nicotine both induced c-Fos activation of various intensities at multiple sites in VTA. Double labeling of c-Fos activated cells with GAD67-GFP positive cells identified a subpopulation of GABAergic neurons in Substantia Nigra Compact part Medial tier (SNCM) that were activated by high- but not by low-dose nicotine. Of 217 GABAergic cells counted at this site, 48.9% exhibited nicotine induced c-fos immunoreactivity. GAD67-GFP positive cells in other regions of VTA were not activated by the nicotine doses tested. Double labeling of GAD67-GFP positive cells with TH IR cells showed that the GABAergic neurons that were activated by high-dose nicotine were located in close proximity to the dopaminergic neurons of substantia nigra compact part and VTA. Dose-dependent activation of GAD67-GFP positive neurons in SNCM, by a nicotine dose known to produce aversive responses, implies that GABAergic neurons at these sites may be an important component of the nicotine aversive circuitry.