Detection of Nitric Oxide Release from Single Neurons in the Pond Snail, Lymnaea stagnalis

Abstract

Multiple film-coated nitric oxide sensors have been fabricated using Nafion and electropolymerized polyeugenol or o-phenylenediamine on 30-microm carbon fiber disk electrodes. This is a rare study that utilizes disk electrodes rather than the widely used protruding tip microelectrodes in order to measure from a biological environment. These electrodes have been used to evaluate the differences in nitric oxide release between two different identified neurons in the pond snail, Lymnaea stagnalis. These results show the first direct measurements of nitric oxide release from individual neurons. The electrodes are very sensitive to nitric oxide with a detection limit of 2.8 nM and a sensitivity of 9.46 nA microM-1. The sensor was very selective against a variety of neurochemical interferences such as ascorbic acid, uric acid, and catecholamines and secondary oxidation products such as nitrite. Nitric oxide release was measured from the cell bodies of two neurons, the cerebral giant cell (CGC) and the B2 buccal motor neuron, in the intact but isolated CNS. A high-Ca2+/high-K+ stimulus was capable of evoking reproducible release. For a given stimulus, the B2 neuron released more nitric oxide than the CGC neuron; however, both cells were equally suppressed by the NOS inhibitor l-NAME.