Rapid eye movement sleep induction by vasoactive intestinal peptide infused into the oral pontine tegmentum of the rat may involve muscarinic receptors.

In rats, rapid eye movement sleep can be induced by microinjection of either the cholinergic agonist carbachol or the neuropeptide vasoactive intestinal peptide into the oral pontine reticular nucleus. Possible involvement of cholinergic mechanisms in the effect of vasoactive intestinal peptide was investigated using muscarinic receptor ligands. Sleep-waking cycles were analysed after infusion into the oral pontine reticular nucleus of vasoactive intestinal peptide (10 ng in 0.1 microl), carbachol (20 ng), atropine (200 ng) and pirenzepine (50, 100 ng), performed separately or in combination at 15-min intervals. The increase in rapid eye movement sleep due to the combined infusion of vasoactive intestinal peptide and carbachol (+58.7+/-4.6% for 8 h, P<0.05) was not significantly different from that induced by each compound separately. The enhancement of rapid eye movement sleep by vasoactive intestinal peptide was totally prevented by infusion of atropine, but not pirenzepine, a relatively selective M1 antagonist. On their own, none of the latter two compounds affected the sleep-waking cycle. Quantitative autoradiographic studies using [3H]quinuclidinyl benzylate (1 nM) and pirenzepine (0.5 microM) indicated that muscarinic receptors correspond to pirenzepine-insensitive binding sites in the oral pontine reticular nucleus. In vitro, vasoactive intestinal peptide (1-100 nM) significantly increased (+30-40%) the specific binding of [3H]quinuclidinyl benzylate to the oral pontine reticular nucleus in rat brain sections. This effect appeared to be due to an increased density, with no change in affinity, of pirenzepine-insensitive binding sites in this area. These data suggest that pirenzepine-insensitive muscarinic binding sites are involved in the induction of rapid eye movement sleep by vasoactive intestinal peptide at the pontine level in the rat.