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Evidence That the Swim Afferent Neurons of Tritonia diomedea Are Glutamatergic

Department of Cell Biology and Anatomy, The Chicago Medical School, Rosalind Franklin University of Medicine and Science, 3333 Green Bay Road, North Chicago, Illinois 60064

^* To whom correspondence should be addressed. E-mail: william.frost{at}rosalindfranklin.edu

The escape swim response of the marine mollusc Tritonia diomedea is a well-established model system for studies of the neural basis of behavior. Although the swim neural network is reasonably well understood, little is known about the transmitters used by its constituent neurons. In the present study, we provide immunocytochemical and electrophysiological evidence that the S-cells, the afferent neurons that detect aversive skin stimuli and in turn trigger Tritonia's escape swim response, use glutamate as their transmitter. First, immunolabeling revealed that S-cell somata contain elevated levels of glutamate compared to most other neurons in the Tritonia brain, consistent with findings from glutamatergic neurons in many species. Second, pressure-applied puffs of glutamate produced the same excitatory response in the target neurons of the S-cells as the naturally released S-cell transmitter itself. Third, the glutamate receptor antagonist CNQX completely blocked S-cell synaptic connections. These findings support glutamate as a transmitter used by the S-cells, and will facilitate studies using this model system to explore a variety of issues related to the neural basis of behavior.

Abbreviations: CNQX, 6-cyano-7-nitroquinoxaline-2,3-dione • DMSO, dimethyl sulfoxide • PBS, phosphate buffered saline • EPSP, excitatory postsynaptic potential • NMDA, N-methyl-D-aspartate • Pl 5–8, pleural cells 5–8