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Volume 17, Number 1, Issue of January 1, 1997 pp. 363-371
Copyright ©1997 Society for Neuroscience

Electrophysiological and Immunocytochemical Evidence for a cGMP-Mediated Inhibition of Subfornical Organ Neurons by Nitric Oxide

Received Aug. 30, 1996; revised Oct. 11, 1996; accepted Oct. 21, 1996.

Matthias Rauch¹, Herbert A. Schmid¹, Jan deVente², and Eckhart Simon¹

¹ Max-Planck-Institut für Physiologische und Klinische Forschung, W. G. Kerckhoff-Institut, 61231 Bad Nauheim, Germany, and ² University of Limburg, Maastricht, The Netherlands

The activation of neurons in the subfornical organ (SFO) by angiotensin II (AngII) is well established and is widely regarded as the basis for the AngII-induced increase in water intake. Application of the nitric oxide (NO) donor sodium nitroprusside (SNP) led to an inhibition of the spontaneous electrical activity in 96% of the neurons sensitive for SNP (n = 50). In addition, the firing rate in 60% of the neurons inhibited by SNP decreased in response to superfusion with the natural substrate of the NO synthase (NOS) L-arginine whereas 70% increased their frequency after application of the NOS blocker N^G-monomethyl-L-arginine (L-NMMA; n = 10). The inhibitory effect of SNP could be mimicked by application of membrane-permeable 8-Br-cGMP. The presence of nNOS, the neuronal isoform of NOS, was demonstrated immunocytochemically and using the NADPH-diaphorase technique on SFO slices. Using a highly selective antibody against cGMP in formaldehyde-fixed tissue, the NO donors SNP, 3-morpholinosydnonimine (SIN-1), and S-nitroso-N-acetyl-DL-penicillamine (SNAP) caused a strong increase in cGMP formation when applied under the same conditions as used for the electrophysiological recordings. These electrophysiological results suggest an important role for NO in SFO-mediated responses and offer a plausible explanation for the in vivo-observed opposite effects of AngII and NO on water intake.

Key words: sodium nitroprusside; nitric oxide; thirst; drinking; neuron; osmoregulation; rat; electrophysiology; subfornical organ; angiotensin II

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