Adenosine Postsynaptically Modulates Supraoptic Neuronal Excitability

Department of Cell Biology and Neuroscience, University of California, Riverside, California Submitted 9 December 2003; accepted in final form 6 September 2004 Effects of adenosine on the excitability of supraoptic nucleus neurons were investigated in whole cell patch-clamp experiments conducted in horizontal slices of rat hypothalamus. Adenosine (10–100 µM) inhibited all neurons tested by reducing or abolishing spontaneous or evoked discharge. Large hyperpolarizations were seen, averaging -6.08 ± 0.83 mV below resting membrane potential, and action potential durations were significantly reduced by 134 ± 41 µs in the presence of 100 µM adenosine. The A 1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 1 µM) blocked these effects, whereas the A 1 agonists N 6 -cyclopentyladenosine (CPA) and N 6 -cyclohexyladenosine (CHA) mimicked the actions of adenosine. A 2 receptor contributions to excitability were assessed by application of an A 2 agonist, carboxamidoadenosine (CPCA). This resulted in membrane depolarizations (3.56 ± 0.65 mV) and maintenance of firing. The presence of endogenous adenosine in the slice was revealed by both the application of the adenosine uptake inhibitor dilazep (1–100 µM), which resulted in a strong inhibition of firing activity, and the application of DPCPX, which induced firing in cells silenced by negative current injection. We tested for postsynaptic actions of adenosine by blocking G protein activation via GDP- -S infusion into recorded neurons. Under these conditions, the adenosinergic inhibition of firing and reduction of spike duration were blocked, suggesting the effects were mediated by postsynaptic adenosine receptors. That the effects on excitability could be due to direct activation of adenosine A 1 receptors on supraoptic neurons was further explored immunocytochemically via the co-labeling of magnocellular neurons with polyclonal antibodies raised against the A 1 receptors. It is concluded that adenosine, acting at postsynaptic A 1 receptors, exhibits a powerful inhibitory influence on supraoptic magnocellular activity and is an important endogenous regulator of magnocellular neuroendocrine function. Address reprint requests and other correspondence to: T. A. Ponzio (E-mail: todd.ponzio{at}email.ucr.edu )