Abstract

Calcium-binding proteins are essential for buffering intracellular calcium concentrations, which are critical for regulating cellular processes involved in neuronal computations. One such calcium-binding protein, calretinin, is present in many neurons of the central nervous system as well as those which innervate cranial sensory organs, although often with differential distributions in adjacent cellular elements. Here, we determined the presence and distribution of calretinin-immunoreactivity in the peripheral vestibular and auditory system of ranid frogs. Calretinin-immunoreactivity was observed in ganglion cells innervating the basilar and amphibian papilla, and in a subpopulation of ganglion cells innervating the saccular epithelium. In contrast, none of the ganglion cells innervating the lagena, the utricle, or the three semicircular canals were calretinin-immunopositive, suggesting that this calcium-binding protein is a marker for auditory but not vestibular afferent fibers in the frog. The absence of calretinin in vestibular ganglion cells corresponds with the lack of type I hair cells in anamniote vertebrates, many of which in amniotes are contacted by the neurites of large, calyx-forming calretinin-immunopositive ganglion cells. In the sensory epithelia of all endorgans, the majority of hair cells were strongly calretinin-immunopositive. Weakly calretinin-immunopositive hair cells were distributed in the intermediate region of the semicircular canal cristae, the central part of the saccular macula, the utricular, and lagenar striola and the medial part of the amphibian papilla. The differential presence of calretinin in the frog vestibular and auditory sensory periphery might reflect a biochemical feature related to firing patterns and frequency bandwidths of self-motion versus acoustic stimulus encoding, respectively.

Highlights

  • Calcium-binding proteins regulate cellular calcium homeostasis and control neuronal processes, such as excitability thresholds, response dynamics, synaptic transmission mechanisms, and indirectly influence neuronal susceptibility to impairments (Baimbridge et al, 1992; Fairless et al, 2019; Schwaller, 2002)

  • In contrast to the selective labeling of afferent fibers, the majority of hair cells in all endorgans were strongly CaR-immunopositive, with variations related to epithelial location and cell morphology

  • The selective CaR-immunopositivity of all papilla and a subpopulation of saccular ganglion cells suggests that CaR is a marker for non-vestibular afferent fibers, which transmit sound information (Lewis et al, 1982a,b)

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Summary

INTRODUCTION

Calcium-binding proteins regulate cellular calcium homeostasis and control neuronal processes, such as excitability thresholds, response dynamics, synaptic transmission mechanisms, and indirectly influence neuronal susceptibility to impairments (Baimbridge et al, 1992; Fairless et al, 2019; Schwaller, 2002). Vestibular afferents of, e.g., ranid frogs (Honrubia et al, 1981, 1989; Baird and Lewis, 1986; Baird and Schuff, 1994; Reichenberger and Dieringer, 1994; Straka and Dieringer, 2004) form a broad spectrum of fibers with correlations between resting discharge regularity, response dynamics, fiber diameter, and regional innervation of the sensory epithelium similar to the morpho-physiology of vestibular afferents in amniote vertebrates (Goldberg, 2000) This structural and functional similarity prompted us to study the distribution of CaRimmunoreactivity of inner ear neuronal elements in frogs with the hypothesis that CaR has a cell type-specific differential expression pattern

MATERIALS AND METHODS
Immunocytochemical Procedures
RESULTS
DISCUSSION
ETHICS STATEMENT
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