Optical methods can be utilized to map the location and activity of putative motor neurons and interneurons during rhythmic patterns of activity in the buccal ganglion of Aplysia

We sought to develop a map of the locations of neurons that are active during patterned activity in the buccal ganglion of Aplysia using optical techniques. Staining ganglia with a voltage-sensitive absorbance dye (JPW 1124) did not prevent them from generating patterned activity similar to that observed before staining, in response to shock of the esophageal nerve. Absorbance changes were monitored with a 124-element photodiode array, while extracellular electrodes monitored activity of the 6 buccal nerves. Optical and extracellular spikes were grouped with the aid of a template matching program; a total of 120 distinct units were detected in one 15 s recording. Optical signals (83 units) were found in the region of the ganglion containing mainly large neurons. Of these, 13 were detected on both optical and extracellular electrode recordings, suggesting that they might be motor neurons, while 25 of the optically detected neurons appeared not to be correlated with extracellular activity, suggesting that they might be interneurons. It was not possible to determine whether the remaining 45 optically identified units did or did not have correlated nerve activity. The ganglionic locations of putative motor neurons corresponded to the locations of large neurons identified by backfilling nerves of other buccal ganglia, and were consistent with the locations of putative motor neurons found in two other ganglia studied using optical methods. Thus, optical methods have generated a map of the locations and activity patterns of putative motor neurons and interneurons in the buccal ganglion that may be involved in the generation of rhythmic patterns.