Light-induced effects of a fluorescent voltage-sensitive dye on neuronal activity in the crab stomatogastric ganglion

Optical imaging being one of the cutting-edge methods for the investigation of neural activity, it is very important to understand the mechanisms of how dye molecules work and the range of side effects that they may induce. In particular, it is very important to reveal potential toxic effects and effects impairing the functioning of the investigated neural system. Here, we investigate the effects of illumination in the presence of the commonly used di-4-ANEPPS voltage-sensitive dye on the rhythmic motor pattern generated by the pyloric central pattern generator in the crab stomatogastric nervous system, a model system for motor pattern generation. We report that the dye allows long recording sessions with little bleaching and no obvious damage to the pyloric rhythm. Yet, exciting illumination induced a temporary and reversible change in the phase relationship of the pyloric motor neurons and a concomitant speed-up of the rhythm. The effect was specific to the excitation wavelength of di-4-ANEPPS and only obtained when the neuropile and cell bodies were illuminated. Thus, di-4-ANEPPS acts as a photo-switch that causes a quick and reversible change in the phase relationship of the motor neurons, but no permanent impairment of neuronal function. It may thus also be used as a means to study the maintenance of phase relationships in rhythmic motor patterns