Intrasomatic Injection of Radioactive Precursors for Studying Transmitter Synthesis in Identified Neurons of Aplysia californica

We introduced radioactive precursors directly into identified neurons of Aplysia californica. (3)H-Choline and L-(3)Htryptophan were injected with pressure into nerve cell bodies to study synthesis of acetylcholine and serotonin. We confirmed the cholinergic nature of R2, L10, and L11, identified neurons of the abdominal ganglion. Cells in the LD cluster (which contains motor neurons to the heart and gill) also converted most of the injected choline into acetylcholine. Neurons in the RB cluster (which contains an excitatory motor neuron to the heart) and the two metacerebral cells of the cerebral ganglion converted injected tryptophan to serotonin. No cell studied could convert both choline to acetylcholine and tryptophan to serotonin. Pressure permits rapid injection of precursors, from small amounts to amounts large enough to saturate intracellular synthetic pathways. In contrast to the results with injection, we found far less synthesis of acetylcholine and serotonin in identified nerve cell bodies when ganglia were incubated in the presence of the radioactive precursors.