Pharmacological analysis of tonic activity in motoneurons during stick insect walking.

Stick insect middle leg (mesothoracic) motoneurons receive tonic excitatory input during front leg stepping on a treadmill. We studied the pharmacology of this excitatory input to the motoneurons during single-legged treadmill walking (in situ). During bath application of drugs restricted to the mesothoracic ganglion, activity in motoneurons contralateral to the stepping front leg was recorded from neuropilar processes. Application of the cholinergic antagonist atropine reduced the tonic depolarization amplitude. These results were compared with findings in acutely dissociated motoneuron cell bodies (in vitro) under whole cell voltage-clamp conditions. The presence of an acetylcholine-induced current in situ was supported by the finding of an acetylcholine evoked biphasic inward current with a sustained component that could be blocked by atropine. In situ the tonic depolarization was generally increased by application of the neuro-modulator octopamine and decreased by its antagonist mianserin. In vitro, however, octopamine reduced the inward current evoked by acetylcholine application to motoneurons. Intracellular application of bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) into motoneurons in situ revealed a dependence of the tonic depolarization on Ca(2+) and application of the membrane-permeable cAMP analogue 8-bromo-cAMP increased the tonic depolarization. In contrast, 8-bromo-cAMP reduced the inward current evoked by acetylcholine application to motoneurons in vitro. We conclude that during walking, acetylcholine contributes to mediating the tonic depolarization possibly by acting on atropine-sensitive receptors on motoneurons. Octopamine that is released during walking increases the tonic depolarization. This increase, however, is not based on modulation of cholinergic action on motoneurons but rather on effects on premotor neurons. Both, Ca(2+) and cAMP are likely second messengers involved in mediating the tonic depolarization, but whereas Ca(2+) acts in motoneurons, cAMP does not appear to mediate a cholinergic depolarization in motoneurons.