Release of monoamines and nitric oxide is involved in the modulation of hyperpolarization-activated inward current during acute thalamic hypoxia

Using slices of the dorsal lateral geniculate nucleus, it has been shown that, in the presence of excitatory and inhibitory amino acid antagonists, brief periods of hypoxia (3-4 min of 95% N-2/5% CO2) induce in thalamocortical neurons an increase in instantaneous input conductance (GN) accompanied by an inward shift in baseline holding current (I-BH) These effects have been suggested to be mediated, at least in part, by a positive shift in the voltage-dependence of the hyperpolarization-activated, mixed Na+/K+ current (I-h) and a change in its activation kinetics which transforms it into an almost instantaneously activated current. In this study, using the whole-cell patch-clamp technique, the contribution of an increased Ca2+-dependent transmitter release to the hypoxic response of thalamocortical neurons was further investigated using (i) blockers of calcineurin, a Ca2+/calmodulin-activated phosphatase that selectively regulates Ca2+-dependent release, and (ii) antagonists of neurotransmitters that are known to modulate I-h Thalamocortical neurons (n = 23) recorded with electrodes filled with calcineurin autoinhibitory fragment (30-250 mu M), a membrane impermeable blocker of calcinuerin, showed no difference either in resting, or in the hypoxia-induced changes in, G(N), I-BH and I-h, when compared to thalamocortical cells patched with electrodes that did not contain calcineurin autoinhibitory fragment. In contrast, in 18 of these neurons recorded with calcineurin autoinhibitory fragment-filled electrodes, bath application either of cyclosporin-A (20 mu M) or tacrolimus (50-100 mu M), two membrane permeable blockers of calcineurin, abolished the effects of hypoxia on G(N), I-BH, and I-h Separate application of noradrenaline, serotonin, histamine and nitric oxide antagonists produced only a small depression of the hypoxic response, while concomitant bath application of these antagonists decreased the hypoxia-induced changes in G(N) and I-BH by 55 and 42%, respectively (n = 12). Concomitant bath application of 8-bromo-adenosine-3'5'-cyclicmonophosphate and 8-bromo-guanosine-3'5'-cyclicmonophosphate (both 1 mM), which are known to mediate the action of these transmitters on I-h, increased G(N) (40%), decreased I-h time-constant of activation (30%) and significantly occluded (50%) the hypoxia-induced effect on G(N) and I-BH Thalamocortical neurons (n = 6) patched with electrodes filled with 8-bromo-adenosine-3'5'-cyclicmonophosphate and 8-bromo-guanosine-3 '5' -cyclicmonophosphate (both 1 mM) showed a larger G(N) than the one recorded with the standard internal solution, and a significant depression of the hypoxia-induced changes in G(N) and I-BH These results indicate that during acute thalamic hypoxia an increased release of noradrenaline, serotonin, histamine and nitric oxide is responsible for transforming I-h into an instantaneously activating current via cyclic AMP- and cyclic GMP-mediated mechanisms. (C) 2000 IBRO. Published by Elsevier Science Ltd.