Characteristics of regenerating horizontal semicircular canal afferent and efferent fibers in the toadfish, Opsanus tau

The horizontal semicircular canal nerve of the toadfish, Opsanus tau, was transected and allowed to regenerate. The time course, morphometrics, and projection patterns of regenerating afferent and efferent vestibular fibers were determined. Nerve transections were performed both pre- and postganglionically, and regeneration was assessed in afferent and efferent fibers by bulk labeling the peripheral axons of the horizontal semicircular canal nerve with biocytin after nerve regrowth. Afferent fibers regrew through the transection site within 14 days and projected to all vestibular nuclei within 3 weeks. Bouton and branch number, axon length, surface area, volume, fiber diameter, and internodal distance were quantified for afferent fibers from eight sites within the vestibular nuclei, and axon number and soma size was quantified for the efferent fibers. Extensive regeneration was seen within 5 weeks of transection in all nuclei, and most morphometric parameters approached or exceeded control levels within 10 weeks. Regeneration appeared to recapitulate morphogenesis with an initial overproduction of boutons and branch points followed by elimination of presumably superfluous structures. Internodal distance remained significantly shorter in regenerating afferent axons than in control fish throughout the 15-week observation period. Efferent fibers also were observed to regenerate. Efferent axon number, diameter, and soma size were indistinguishable from those in controls from 3 weeks posttransection through week 15. Electrophysiological recordings from the horizontal canal nerve during mechanical stimuli of the canal confirmed that the regenerated axons transmitted normal signals. The return of normal equilibrium and behavior coincided with the projection of afferent fibers into the central vestibular nuclei, indicating that functional connections had been reestablished.