The utricle provides critical information about spatiotemporal properties of head movement. It comprises multiple subdivisions whose functional roles are poorly understood. We previously identified four subdivisions in turtle utricle, based on hair bundle structure and mechanics, otoconial membrane structure and hair bundle coupling, and immunoreactivity to calcium-binding proteins. Here we ask whether these macular subdivisions are innervated by distinctive populations of afferents to help us understand the role each subdivision plays in signaling head movements. We quantified the morphology of 173 afferents and identified six afferent classes, which differ in structure and macular locus. Calyceal and dimorphic afferents innervate one striolar band. Bouton afferents innervate a second striolar band; they have elongated terminals and the thickest processes and axons of all bouton units. Bouton afferents in lateral (LES) and medial (MES) extrastriolae have small-diameter axons but differ in collecting area, bouton number, and hair cell contacts (LES ϾϾ MES). A fourth, distinctive population of bouton afferents supplies the juxtastriola. These results, combined with our earlier findings on utricular hair cells and the otoconial membrane, suggest the hypotheses that MES and calyceal afferents encode head movement direction with high spatial resolution and that MES afferents are well suited to signal three-dimensional head orientation and striolar afferents to signal head movement onset. hair cells; turtle; utricle; vestibular afferents THE VESTIBULAR LABYRINTH is poorly understood compared, for example, to the auditory periphery and the retina. This is especially true of the major otoconial organs, the utricle and saccule, which encode linear head accelerations, including head position in gravity space, and transmit this information to the CNS. One important question is how the sensory surface of otoconial organs is organized to transduce the full range of signals generated by head movement. There are two broad possibilities: 1) all regions are equipotential in their ability to encode temporal properties of head movement (e.g., different frequencies, accelerations) or 2) different regions of the macula play distinctive functional roles.Considerable evidence supports the second of these alternatives. The best-known example of regional specializations in otoconial maculae is the ubiquitous division between striola and extrastriola: the striolar region of vertebrate maculae differs from the extrastriola in the structure and molecular composition of the otoconial membrane (OM) (e.g., Goodyear et al. 1994;Lim 1979;Lindeman 1969;Werner 1933; reviewed in Fermin et al. 1998;Lewis et al. 1985), which transmits the mechanical stimuli arising from head movements to receptors (hair cells), and in the structure and physiology of its hair cells (e.g

1. We visualized the central axons of 32 vestibular afferents from the posterior canal by extracellular application of horseradish peroxidase, reconstructed them in three dimensions, and quantified their morphology. Here we compare the descending limbs of central axons that differ in parent axon diameter. 2. The brain stem distribution of descending limb terminals (collaterals and associated varicosities) varies systematically with parent axon diameter. Large-diameter afferents concentrate their terminals in rostral regions of the medial/descending nuclei. As axon diameter decreases, there is a significant shift of terminal concentration toward the caudal vestibular complex and adjacent brain stem. 3. Rostral and caudal regions of the medial/descending nuclei have different labyrinthine, cerebellar, intrinsic, commissural, and spinal connections; they are believed to play different roles in head movement control. Our data help clarify the functions of large- and small-diameter afferents by showing that they contribute differentially to rostral and caudal vestibular complex.

Vestibular bouton afferent terminals in turtle utricle can be categorized into four types depending on their location and terminal arbor structure: lateral extrastriolar (LES), striolar, juxtastriolar, and medial extrastriolar (MES). The terminal arbors of these afferents differ in surface area, total length, collecting area, number of boutons, number of bouton contacts per hair cell, and axon diameter (Huwe JA, Logan CJ, Williams B, Rowe MH, Peterson EH. 113: 2420-2433, 2015). To understand how differences in terminal morphology and the resulting hair cell inputs might affect afferent response properties, we modeled representative afferents from each region, using reconstructed bouton afferents. Collecting area and hair cell density were used to estimate hair cell-to-afferent convergence. Nonmorphological features were held constant to isolate effects of afferent structure and connectivity. The models suggest that all four bouton afferent types are electrotonically compact and that excitatory postsynaptic potentials are two to four times larger in MES afferents than in other afferents, making MES afferents more responsive to low input levels. The models also predict that MES and LES terminal structures permit higher spontaneous firing rates than those in striola and juxtastriola. We found that differences in spike train regularity are not a consequence of differences in peripheral terminal structure, per se, but that a higher proportion of multiple contacts between afferents and individual hair cells increases afferent firing irregularity. The prediction that afferents having primarily one bouton contact per hair cell will fire more regularly than afferents making multiple bouton contacts per hair cell has implications for spike train regularity in dimorphic and calyx afferents. Bouton afferents in different regions of turtle utricle have very different morphologies and afferent-hair cell connectivities. Highly detailed computational modeling provides insights into how morphology impacts excitability and also reveals a new explanation for spike train irregularity based on relative numbers of multiple bouton contacts per hair cell. This mechanism is independent of other proposed mechanisms for spike train irregularity based on ionic conductances and can explain irregularity in dimorphic units and calyx endings.

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