An Examination of the Effects and Possible Targets of Nitric Oxide on Olfactory Neurons in the Moth, Manduca Sexta

Abstract

The gaseous messenger, nitric oxide (NO), has emerged as a key component of olfactory systems. Localization and imaging studies in the moth, Manduca sexta, suggest that NO may affect the excitability of olfactory neurons by modifying neuronal membrane properties through sGC-dependent mechanisms. This hypothesis was tested using a multidisciplinary approach, including two types of physiological recording techniques and immunocytochemical analysis of sGC antibody expression in the Manduca brain. The excitability of large populations or individual antennal lobe (AL) neurons was monitored with in vivo physiological recordings while various NO pharmacological agents were bath applied to the brain. To examine possible targets of NO, the binding site of sGC was blocked and the results were compared to NO blockade. Finally, sGC immunocytochemistry was used to also determine possible targets of NO.Two NO synthesis inhibitors and a sGC blocker were potent effectors of resting, baseline activity in the Manduca brain. Blocking NO synthesis caused significant decreases in AL neuron conductance. This conductance decrease led to changes in baseline activity, including the appearance of bursts in some neurons, and increased and decreased firing rates in other neurons. Further, the neurons had a decreased responsiveness and excitability to presynaptic input. Blocking the sGC binding site caused similar effects in most neurons, which indicates that NO likely acts through sGC-dependent signaling to exert its effects in at least a subset of neurons. However, some neurons had different responses to NO and sGC blockade, which indicates that NO may act through other signaling mechanisms in some neurons. Further examination using sGC immunocytochemistry revealed that only about 90% of projection neurons (PNs) and 30% of local neurons (LNs) contained sGC immunoreactivity.The results in this dissertation indicate that NO performs a global function in the antennal lobe to maintain the resting membrane conductance of AL neurons. NO likely exerts its effects through both sGC-dependent and sGC-independent mechanisms. Finally, these results have major implications for odor coding in all species, as NO has been found in the olfactory systems of every animal examined thus far.