Intracellular analysis of the innervation of a crayfish sensory interneuron by regenerating afferents.

Abstract

Mechanoreceptors of the crayfish tail fan have peripheral somata and send their axons to the last (sixth) abdominal ganglion via five bilateral pairs of nerve roots (R1-R5). Comparisons were made between normal crayfish and regenerate preparations in which R4 had been cut and directed back to an extensively denervated sixth abdominal ganglion; 8 to 15 weeks postoperatively, an identified target interneuron (A) in this ganglion was impaled, and its response to water currents, electrical excitation of R4, and stimulation of individual sensory hairs supplying axons to R4 was studied along with several other properties of the pre- and post-synaptic neurons. Normal levels of excitability in A to R4 stimulation were achieved within six weeks as judged by extracellular criteria. Subsequent intracellular analysis revealed that few differences exist between regenerated and normal inputs: probability of (re-) connection, unitary EPSP amplitude and time course distributions, resting membrane potentials, and critical firing levels were comparable in the two groups; input impedance, however, may have been lower in regenerates. Compound electrically elicited EPSPs were similar in amplitude, rise time, and half amplitude width, but differed slightly in latency to onset (regenerates greater than normals). This was accounted for by differences in conduction time to the ganglion in regenerates, and central delay estimates suggest that connections in both groups are monosynaptic. The body root (R1) providing input to A that remained intact in the regenerate preparations increased in efficacy; over 12 postoperative weeks the response of A to R1 activation by water drops steadily increased and at 12 weeks unitary increments to ascending electrical stimulation of R1 were significantly larger than in normals. The response to giant interneuron activation demonstrated that recurrent inhibitory inputs were normal in regenerates. In addition, synaptic depression, normally responsible for behavioral habituation in this system, was comparable across groups. Further, protection from habituation was observed in both normals and regenerates if R4 stimulation was preceded by giant interneuron activation, thus indicating that normal presynaptic inhibitory inputs to the regenerated afferent terminals have also successfully regenerated.(ABSTRACT TRUNCATED AT 400 WORDS)