The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic system leads to the rewarding properties of alcohol. The mesolimbic DA system, which plays an important role in regulating reward and addiction, consists of DA neurons in the midbrain ventral tegmental area (VTA) that innervate the nucleus accumbens (NAc). It is believed that VTA DA neurons are inhibited by local gamma-aminobutyric acid (GABA) interneurons that express connexin-36 (Cx36) gap junctions (GJs). We have previously demonstrated that blocking Cx36 GJs suppresses electrical coupling between VTA GABA neurons and reduces ethanol intoxication and consumption suggesting that electrical coupling between mature VTA GABA neurons underlies the rewarding properties of ethanol. The aim of this study was to further investigate the role of VTA GABA neurons expressing Cx36 GJs in regulating DA neuron activity and release and mediating ethanol effects on VTA GABA neurons. To this end, we customized a Designer Receptor Exclusively Activated by Designer Drugs (DREADDs) viral vector to target VTA GABA neurons expressing Cx36 GJs in order to chemogenetically modulate their activity. In order to more conclusively demonstrate the role of this sub population of VTA GABA neurons in regulating DA neural activity and release we used electrophysiology to characterize the electrical changes that occur in VTA DA and GABA neurons when Cx36-expressing VTA GABA cells were selectively activated. In addition, we evaluated the effects of activation of VTA GABA neurons on brain stimulation reward and alcohol consumption in ethanol naive and dependent mice. Results indicate that there are two populations of GABA neurons in the VTA, one that is GAD65+/Cx36+ and one that is GAD67+/Cx36-. Activation of Cx36+ VTA GABA neurons by clozapine-n-oxide (CNO) in mice injected with Gq DREADD activated VTA DA neurons and subsequent DA release in the NAc, suggesting that Cx36-containing GABA neurons are inhibiting non-Cx36 GABA neurons to disinhibit DA neurons. In hM3Dq animals, CNO administration provided a rewarding stimulus in the conditioned pace preference paradigm, and reduced consumption in the drink-in-the-dark ethanol consumption paradigm in dependent and naïve mice. A better understanding of the circuitry of the mesolimbic DA system is key to understanding the mechanisms that lead to addiction and may ultimately lead to improved therapies for substance abuse.

The prevailing view is that enhancement of dopamine (DA) transmission in the mesolimbic system leads to the rewarding properties of alcohol. The mesolimbic DA system, which plays an important role in regulating reward and addiction, consists of DA neurons in the midbrain ventral tegmental area (VTA) that innervate the nucleus accumbens (NAc). It is believed that VTA DA neurons are inhibited by local gamma-aminobutyric acid (GABA) interneurons that express connexin-36 (Cx36) gap junctions (GJs). We have previously demonstrated that blocking Cx36 GJs suppresses electrical coupling between VTA GABA neurons and reduces ethanol intoxication and consumption suggesting that electrical coupling between mature VTA GABA neurons underlies the rewarding properties of ethanol. The aim of this study was to further investigate the role of VTA GABA neurons expressing Cx36 GJs in regulating DA neuron activity and release and mediating ethanol effects on VTA GABA neurons. To this end, we customized a Designer Receptor Exclusively Activated by Designer Drugs (DREADDs) viral vector to target VTA GABA neurons expressing Cx36 GJs in order to chemogenetically modulate their activity. In order to more conclusively demonstrate the role of this sub population of VTA GABA neurons in regulating DA neural activity and release we used electrophysiology to characterize the electrical changes that occur in VTA DA and GABA neurons when Cx36-expressing VTA GABA cells were selectively activated. In addition, we evaluated the effects of activation of VTA GABA neurons on brain stimulation reward and alcohol consumption in ethanol naive and dependent mice. Results indicate that there are two populations of GABA neurons in the VTA, one that is GAD65+/Cx36+ and one that is GAD67+/Cx36-. Activation of Cx36+ VTA GABA neurons by clozapine-n-oxide (CNO) in mice injected with Gq DREADD activated VTA DA neurons and subsequent DA release in the NAc, suggesting that Cx36-containing GABA neurons are inhibiting non-Cx36 GABA neurons to disinhibit DA neurons. In hM3Dq animals, CNO administration provided a rewarding stimulus in the conditioned pace preference paradigm, and reduced consumption in the drink-in-the-dark ethanol consumption paradigm in dependent and naïve mice. A better understanding of the circuitry of the mesolimbic DA system is key to understanding the mechanisms that lead to addiction and may ultimately lead to improved therapies for substance abuse.