title: Maturation of NaV and KV Channel Topographies in the Auditory Nerve Spike Initiator before and after Developmental Onset of Hearing Function.
abstract: Auditory nerve excitation and thus hearing depend on spike-generating ion channels and their placement along the axons of auditory nerve fibers (ANFs). The developmental expression patterns and native axonal locations of voltage-gated ion channels in ANFs are unknown. Therefore, we examined the development of heminodes and nodes of Ranvier in the peripheral axons of type I ANFs in the rat cochlea with immunohistochemistry and confocal microscopy. Nodal structures presumably supporting presensory spiking formed between postnatal days 5 (P5) and P7, including Ankyrin-G, NaV1.6, and Caspr. These immature nodal structures lacked low-voltage-activated KV1.1 which was not enriched at juxtaparanodes until approximately P13, concurrent with the developmental onset of acoustic hearing function. Anatomical alignment of ANF spike-initiating heminodes relative to excitatory input from inner hair cell (IHC) ribbon synapses continued until approximately P30. High-voltage-activated KV3.1b and KV2.2 were expressed in mutually exclusive domains: KV3.1b was strictly localized to nodes and heminodes, whereas KV2.2 expression began at the juxtaparanodes and continued centrally along the first internode. At spike-initiating heminodes in the distal osseous spiral lamina, NaV1.1 partly overlapped NaV1.6 and ankyrin-G. ANFs displayed KV7.2 and KV7.3 at heminodes, nodes, internodes, and the unmyelinated synaptic terminal segments beneath IHCs in the organ of Corti. In response to sound, spikes are initiated at the heminode, which is tightly coupled to the IHC ribbon synapse ∼20-40 μm away. These results show that maturation of nodal alignment and ion channel content may underlie postnatal improvements of ANF excitability and discharge synchrony.
  
  
  CONCLUSIONS
  Acoustic and electrical hearing depends on rapid, reliable, and precise spike generation in auditory nerve fibers. A limitation of current models and therapies is a lack of information on the identities and topographies of underlying ion channels. We report the developmental profile of the auditory nerve spike generator with a focus on NaV1.1, NaV1.6, KV1.1, KV2.2, KV3.1b, KV7.2, and KV7.3 in relation to the scaffold ankyrin-G. Molecular anatomy of the spike generator matures in the weeks after developmental onset of hearing function. Subcellular positioning of voltage-gated ion channels will enable multicompartmental modeling of auditory nerve responses elicited by afferent chemical neurotransmission from hair cells and modulated by efferent neurotransmitters or evoked by extracellular field stimulation from a cochlear implant.
authors.affiliation: Department of Otolaryngology, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Kyunghee X
authors.initials: KX
authors.lastname: Kim
authors.affiliation: Department of Otolaryngology, Washington University School of Medicine, St. Louis, Missouri 63110 RutherfordM@ent.wustl.edu.
authors.forename: Mark A
authors.identifier: http://orcid.org/0000-0002-2627-6254
authors.initials: MA
authors.lastname: Rutherford
mesh_terms: D000818:Animals; D017487:Ankyrins / Q000235:genetics; D001369:Axons / Q000502:physiology; D003051:Cochlea / Q000166:cytology / Q000254:growth & development / Q000502:physiology; D003056:Cochlear Nerve / Q000254:growth & development / Q000502:physiology*; D005260:Female; D006199:Hair Cells, Auditory, Inner / Q000502:physiology; D006309:Hearing / Q000502:physiology*; D008297:Male; D009412:Nerve Fibers / Q000502:physiology; D009925:Organ of Corti / Q000254:growth & development / Q000502:physiology; D015221:Potassium Channels / Q000502:physiology*; D017729:Presynaptic Terminals / Q000502:physiology; D011901:Ranvier's Nodes / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D015222:Sodium Channels / Q000502:physiology*; D013137:Spiral Lamina / Q000254:growth & development / Q000502:physiology; D013569:Synapses / Q000502:physiology
chemical_list: C490614:Ank3 protein, rat; D017487:Ankyrins; D015221:Potassium Channels; D015222:Sodium Channels
keywords: action potential generation; ankyrin-G; axon initial segment; cochlea; heminode; voltage-gated Na+ and K+ channels