Localization and expression of clarin-1, the Clrn1 gene product, in auditory hair cells and photoreceptors

Marisa L Zallocchi, Daniel T. Meehan, Duane Delimont, Charles Askew, Suneetha Garige, Michael Anne Gratton, Christie A. Rothermund-Franklin, Dominic E Cosgrove

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The Usher syndrome 3A (CLRN1) gene encodes clarin-1, which is a member of the tetraspanin family of transmembrane proteins. Although identified more than 6 years ago, little is known about its localization or function in the eye and ear. We developed a polyclonal antibody that react with all clarin-1 isoforms and used it to characterize protein expression in cochlea and retina. In the cochlea, we observe clarin-1expression in the stereocilia of P0 mice, and in synaptic terminals present at the base of the auditory hair cells from E18 to P6. In the retina, clarin-1 localizes to the connecting cilia, inner segment of photoreceptors and to the ribbon synapses. RT-PCR from P0 cochlea and P28 retina show mRNAs encoding only isoforms 2 and 3. Western blots show that only isoform 2 is present in protein extracts from these same tissues. We examined clarin-1 expression in the immortomouse-derived hair cell line UB/OC-1. Only isoform 2 is expressed in UB/OC-1 at both mRNA and protein levels, suggesting this isoform is biologically relevant to hair cell function. The protein co-localizes with microtubules and post-transgolgi vesicles. The subcellular localization of clarin-1 in hair cells and photoreceptors suggests it functions at both the basal and apical poles of neurosensoriepithelia.

Original languageEnglish (US)
Pages (from-to)109-120
Number of pages12
JournalHearing Research
Volume255
Issue number1-2
DOIs
StatePublished - Sep 1 2009

Fingerprint

Auditory Hair Cells
Protein Isoforms
Cochlea
Retina
Genes
Proteins
Usher Syndromes
Stereocilia
Photoreceptor Cells
Messenger RNA
Tissue Extracts
Cilia
Presynaptic Terminals
Microtubules
Synapses
Ear
Western Blotting
Cell Line
Polymerase Chain Reaction
Antibodies

Keywords

  • Clarin-1
  • Connecting cilia
  • Ribbon synapse
  • Stereocilia
  • Usher syndrome

ASJC Scopus subject areas

  • Sensory Systems

Cite this

Localization and expression of clarin-1, the Clrn1 gene product, in auditory hair cells and photoreceptors. / Zallocchi, Marisa L; Meehan, Daniel T.; Delimont, Duane; Askew, Charles; Garige, Suneetha; Gratton, Michael Anne; Rothermund-Franklin, Christie A.; Cosgrove, Dominic E.

In: Hearing Research, Vol. 255, No. 1-2, 01.09.2009, p. 109-120.

Research output: Contribution to journalArticle

Zallocchi, Marisa L ; Meehan, Daniel T. ; Delimont, Duane ; Askew, Charles ; Garige, Suneetha ; Gratton, Michael Anne ; Rothermund-Franklin, Christie A. ; Cosgrove, Dominic E. / Localization and expression of clarin-1, the Clrn1 gene product, in auditory hair cells and photoreceptors. In: Hearing Research. 2009 ; Vol. 255, No. 1-2. pp. 109-120.
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abstract = "The Usher syndrome 3A (CLRN1) gene encodes clarin-1, which is a member of the tetraspanin family of transmembrane proteins. Although identified more than 6 years ago, little is known about its localization or function in the eye and ear. We developed a polyclonal antibody that react with all clarin-1 isoforms and used it to characterize protein expression in cochlea and retina. In the cochlea, we observe clarin-1expression in the stereocilia of P0 mice, and in synaptic terminals present at the base of the auditory hair cells from E18 to P6. In the retina, clarin-1 localizes to the connecting cilia, inner segment of photoreceptors and to the ribbon synapses. RT-PCR from P0 cochlea and P28 retina show mRNAs encoding only isoforms 2 and 3. Western blots show that only isoform 2 is present in protein extracts from these same tissues. We examined clarin-1 expression in the immortomouse-derived hair cell line UB/OC-1. Only isoform 2 is expressed in UB/OC-1 at both mRNA and protein levels, suggesting this isoform is biologically relevant to hair cell function. The protein co-localizes with microtubules and post-transgolgi vesicles. The subcellular localization of clarin-1 in hair cells and photoreceptors suggests it functions at both the basal and apical poles of neurosensoriepithelia.",
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