Genomic structure, cochlear expression, and mutation screening of KCNK6, a candidate gene for DFNA4

J Neurosci Res. 2004 Jan 1;75(1):25-31. doi: 10.1002/jnr.10839.

Abstract

KCNK6 encodes a tandem pore domain potassium channel, TWIK-2, that maps to chromosome 19. Both STS and linkage maps established KCNK6 as a positional candidate gene for DFNA4, a form of autosomal dominant nonsyndromic hereditary hearing loss. Identification and characterization of Kcnk6 expression within the mammalian cochlea established the gene as a functional candidate for DFNA4. Identification of Twik-2 expression in the mouse cochlea was initially established via RT-PCR assay of cochlear RNA. Subsequent immunoblot analysis of cochlear homogenate yielded a distinct 35-kDa band corresponding to the calculated molecular weight of the mouse Twik-2. Immunohistochemical studies localized Twik-2 expression in the cochlea predominantly within the stria vascularis. This vascular tissue borders the cochlear duct and is a critical regulator of potassium concentration in the endolymph. Genomic structure of TWIK-2 was subsequently determined and shown to consist of three coding exons with splice acceptor and donor sites in accordance with the consensus GT-AG rule. Two separate DFNA4 families were screened for KCNK6 sequence alterations. No mutations were found, thus excluding TWIK-2 as the DFNA4 candidate disease gene. Nevertheless, expression of Twik-2 within the stria vascularis suggests a potential role for this protein as one of the terminal components of the potassium ion-recycling pathway that contributes toward its reabsorption into the endolymph.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Northern / methods
  • Blotting, Western / methods
  • Brain / metabolism
  • Carrier Proteins / genetics*
  • Chromosome Mapping / methods
  • Cochlea / metabolism*
  • DNA Mutational Analysis / methods
  • Genomics*
  • Hearing Loss / genetics*
  • Humans
  • Immunohistochemistry / methods
  • Kidney / metabolism
  • Mice
  • Molecular Structure
  • Myosin Heavy Chains
  • Myosin Type II
  • Potassium Channels / genetics*
  • Potassium Channels / metabolism
  • Potassium Channels, Tandem Pore Domain
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction / methods

Substances

  • Carrier Proteins
  • KCNK6 protein, human
  • Kcnk6 protein, mouse
  • MYH14 protein, human
  • Potassium Channels
  • Potassium Channels, Tandem Pore Domain
  • RNA, Messenger
  • Myosin Type II
  • Myosin Heavy Chains