Neuromuscular regeneration by buccal motoneuron B15 after peripheral nerve crush in Aplysia californica.

Abstract

1. We studied regeneration of neuromuscular connections by identified buccal motoneuron B15 after axotomy produced by crushing nerve 4; the intact contralateral nerve 4 served as control. Electrophysiological recordings, intracellular dye injections, and light and electron microscopy were used to characterize the nature and time course of neuromuscular reinnervation as well as the fate of the isolated distal stump of the motor axon. 2. Axonal outgrowth or sprouting in the form of numerous "regenerites" occurred from the proximal stump of the transected B15 axon, and these regenerites projected through the crush site along the length of the nerve to innervate target muscles at the periphery. 3. Reinnervation of one of the target muscles, the accessory radula closer (I5), was first detected 3 wk after nerve crush. Neuromuscular excitatory postsynaptic potentials measured in individual I5 muscle fibers were initially small and approached control amplitudes by 8 wk postlesion. Newly regenerated neuromuscular synapses displayed facilitation and depression to repeated B15 stimulation with properties similar to those of control synapses, even at early times postlesion. 4. Reinnervation of other buccal muscles by B15, such as I4, appeared slightly delayed relative to that observed for I5. No evidence of abnormal or enlarged fields of innervation were observed, and as in control preparations, regenerated neuromuscular connections were strictly limited to muscles ipsilateral to the B15 cell body. 5. Physiological evidence suggested that the distal axon stumps of B15, although isolated from their cell bodies, survive for several weeks after axotomy. In addition, several large axon profiles indicative of motor axons were seen in cross-sections of nerve 4 taken close to the muscle and distal to the crush site, indicating survival of distal axon stumps. 6. When B15 was selectively stimulated, the newly formed regenerites failed to fire the distal axon stump of B15, demonstrating that the regenerites do not reinnervate the distal stump. 7. Degeneration of axons in nerve 4 distal to the crush site was observed in cross-sections of the nerve at 8 wk postlesion; using ultrathin sections we found cellular debris in individual axon profiles as well as large acellular masses within nerve 4, the latter likely representing the concretion of many axons. Additional evidence for such degenerative changes appeared in the form of autofluorescing spherical bodies or "spheroids" both in individual axons and the nerve distal to the crush site.(ABSTRACT TRUNCATED AT 400 WORDS)