Extracellular ATP neurotransmission and nicotine sex-specifically modulate habenular neuronal activity in adolescence.

The recent increase in the use of nicotine products by teenagers has revealed an urgent need to better understand the impact of nicotine on the adolescent brain. Here, we sought to examine the actions of extracellular adenosine triphosphate (ATP) as a neurotransmitter and to investigate whether ATP and nicotinic signaling interact during adolescence. With the GRAB ATP sensor, we first demonstrated that nicotine induces extracellular ATP release in the medial habenula (MHb), a brain region involved in nicotine aversion and withdrawal. Using patch clamp electrophysiology, we then demonstrated that activation of the ATP receptors, P2X or P2Y 1 , increases the neuronal firing of cholinergic neurons. Surprisingly, contrasting interactive effects were observed with nicotine exposure. For the P2X receptor, activation had no observable effect on acute nicotine-mediated activity, but during abstinence after ten days of nicotine exposure, co-exposure to nicotine and the P2X agonist potentiated neuronal activity in female, but not male, neurons. For P2Y 1 signaling, a potentiated effect of the agonist and nicotine was observed with acute exposure, but not following extended nicotine exposure. These data reveal a complex interactive effect between nicotinic and ATP signaling in the adolescent brain and provide mechanistic insights into extracellular ATP signaling with sex-specific alterations of neuronal responses based on prior drug exposure. Significance Statement In these studies, it was discovered that nicotine induces extracellular ATP release in the medial habenula and subsequent activation of the ATP purinergic receptors increases habenular cholinergic neuronal firing in the adolescent brain. Interestingly, following extended nicotine exposure, nicotine was found to alter the interplay between purinergic and nicotinic signaling in a sex-specific manner. Taken together, these studies provide a novel understanding for the role of extracellular ATP in mediating habenular activity and reveal how nicotine exposure during adolescence alters these signaling mechanisms, which has important implications given the high incidence of e-cigarette/vape use by youth.