Mechanotransduction and hyperpolarization-activated currents contribute to spontaneous activity in mouse vestibular ganglion neurons

The hyperpolarization-activated, cyclic nucleotide–sensitive current, Ih, is present in vestibular hair cells and vestibular ganglion neurons, and is required for normal balance function. We sought to identify the molecular correlates and functional relevance of Ih in vestibular ganglion neurons. Ih is carried by channels consisting of homo- or heteromeric assemblies of four protein subunits from the Hcn gene family. The relative expression of Hcn1–4 mRNA was examined using a quantitative reverse transcription PCR (RT-PCR) screen. Hcn2 was the most highly expressed subunit in vestibular neuron cell bodies. Immunolocalization of HCN2 revealed robust expression in cell bodies of all vestibular ganglion neurons. To characterize Ih in vestibular neuron cell bodies and at hair cell–afferent synapses, we developed an intact, ex vivo preparation. We found robust physiological expression of Ih in 89% of cell bodies and 100% of calyx terminals. Ih was significantly larger in calyx terminals than in cell bodies; however, other biophysical characteristics were similar. Ih was absent in calyces lacking Hcn1 and Hcn2, but small Ih was still present in cell bodies, which suggests expression of an additional subunit, perhaps Hcn4. To determine the contributions of hair cell mechanotransduction and Ih to the firing patterns of calyx terminals, we recorded action potentials in current-clamp mode. Mechanotransduction currents were modulated by hair bundle defection and application of calcium chelators to disrupt tip links. Ih activity was modulated using ZD7288 and cAMP. We found that both hair cell transduction and Ih contribute to the rate and regularity of spontaneous action potentials in the vestibular afferent neurons. We propose that modulation of Ih in vestibular ganglion neurons may provide a mechanism for modulation of spontaneous activity in the vestibular periphery.

Survival of the Vestibular Nerve after Labyrinthectomy in the Cat

Temporal bone studies in cat, monkey, and man demonstrate that the cell bodies of the primary vestibular neurons located in Scarpa's ganglion persist after labyrlnthectomy. However, it is not known whether the centrally directed axon process of deafferented vestibular neurons survive or degenerate after labyrinthectomy. If the central axon were to persist, then the primary vestibular neuron could influence vestibular compensation or produce symptoms of vestibular dysfunction. In the present study the temporal bones and brain stem of four cats were prepared for light microscopic examination with hematoxylin-eosin, silver, and trichrome connective tissue stains. Cell counts within Scarpa's ganglion were performed. After labyrinthectomy, many intact axons were demonstrated in the brain stem, a finding that correlated with survival of neurons in Scarpa's ganglion. This study provides anatomic evidence that primary vestibular neurons that survive labyrinthectomy may retain their central axon processes. The persistence of this neural pathway and data from behavioral studies in the cat suggest that vestibular neurons may affect vestibular compensation after labyrinthectomy. Deafferented vestibular neurons may play a role in human vestibular compensation and dysfunction.