Nitric oxide modulates a swimmeret beating rhythm in the crayfish.

Abstract

The modulatory effects of nitric oxide (NO) and cAMP on the rhythmic beating activity of the swimmeret motor neurones in the crayfish were examined. Swimmerets are paired appendages located on the ventral side of each abdominal segment that show rhythmic beating activity during forward swimming, postural righting behaviour and egg ventilation in gravid females. In isolated abdominal nerve cord preparations, swimmeret motor neurones are usually silent or show a continuous low-frequency spiking activity. Application of carbachol, a cholinergic agonist, elicited rhythmic bursts of motor neurone spikes. The co-application of L-arginine, the substrate for NO synthesis with carbachol increased the burst frequency of the motor neurones. The co-application of the NO donor SNAP with carbachol also increased the burst frequency of the motor neurones. By contrast, co-application of a NOS inhibitor, L-NAME, with carbachol decreased beating frequency of the motor neurones. These results indicate that NO may act as a neuromodulator to facilitate swimmeret beating activity. The facilitatory effect of L-arginine was cancelled by co-application of the soluble guanylate cyclase (sGC) inhibitor ODQ suggesting that NO acts by activating sGC to promote the production of cGMP. Application of L-arginine alone or membrane-permeable cGMP analogue 8-Br-cGMP alone did not elicit rhythmic activity of motor neurones, but co-application of 8-Br-cGMP with carbachol increased bursting frequency of the motor neurones. Furthermore, application of the membrane-permeable cAMP analogue CPT-cAMP alone produced rhythmic bursting of swimmeret motor neurones, and the bursting frequency elicited by CPT-cAMP was increased by co-application with L-arginine. Co-application of the adenylate cyclase inhibitor SQ22536 ceased rhythmic bursts of motor neurone spikes elicited by carbachol. These results suggest that a cAMP system enables the rhythmic bursts of motor neurone spikes and that a NO-cGMP signaling pathway increases cAMP activity to facilitate swimmeret beating.