A short splice form of Xin-actin binding repeat containing 2 (XIRP2) lacking the xin repeats is required for maintenance of stereocilia morphology and hearing function

Shimon P. Francis, Jocelyn F. Krey, Evan S. Krystofiak, Runjia Cui, Sonali Nanda, Wenhao Xu, Bechara Kachar, Peter Barr-Gillespie, Jung Bum Shin

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Approximately one-third of known deafness genes encode proteins located in the hair bundle, the sensory hair cell’s mechanoreceptive organelle. In previous studies, we used mass spectrometry to characterize the hair bundle’s proteome, resulting in the discovery of novel bundle proteins. One such protein is Xin-actin binding repeat containing 2 (XIRP2), an actin-cross-linking protein previously reported to be specifically expressed in striated muscle. Because mutations in other actin-cross-linkers result in hearing loss, we investigated the role of XIRP2 in hearing function. In the inner ear, XIRP2 is specifically expressed in hair cells, colocalizing with actin-rich structures in bundles, the underlying cuticular plate, and the circumferential actin belt. Analysis using peptide mass spectrometry revealed that the bundle harbors a previously uncharacterized XIRP2 splice variant, suggesting XIRP2’s role in the hair cell differs significantly from that reported in myocytes. To determine the role of XIRP2 in hearing, we applied clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-mediated genome-editing technology to induce targeted mutations into the mouse Xirp2 gene, resulting in the elimination of XIRP2 protein expression in the inner ear. Functional analysis of hearing in the resulting Xirp2-null mice revealed high-frequency hearing loss, and ultrastructural scanning electron microscopy analyses of hair cells demonstrated stereocilia degeneration in these mice. We thus conclude that XIRP2 is required for long-term maintenance of hair cell stereocilia, and that its dysfunction causes hearing loss in the mouse.

Original languageEnglish (US)
Pages (from-to)1999-2014
Number of pages16
JournalJournal of Neuroscience
Volume35
Issue number5
DOIs
StatePublished - 2015

Fingerprint

Stereocilia
Hearing
Actins
Maintenance
Inner Ear
Proteins
Hearing Loss
Mass Spectrometry
Clustered Regularly Interspaced Short Palindromic Repeats
High-Frequency Hearing Loss
Mutation
Striated Muscle
Deafness
Proteome
Organelles
Electron Scanning Microscopy
Muscle Cells

Keywords

  • CRISPR
  • Hair bundle
  • Hair cell
  • Hearing
  • Stereocilia
  • XIRP2

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

A short splice form of Xin-actin binding repeat containing 2 (XIRP2) lacking the xin repeats is required for maintenance of stereocilia morphology and hearing function. / Francis, Shimon P.; Krey, Jocelyn F.; Krystofiak, Evan S.; Cui, Runjia; Nanda, Sonali; Xu, Wenhao; Kachar, Bechara; Barr-Gillespie, Peter; Shin, Jung Bum.

In: Journal of Neuroscience, Vol. 35, No. 5, 2015, p. 1999-2014.

Research output: Contribution to journalArticle

Francis, Shimon P. ; Krey, Jocelyn F. ; Krystofiak, Evan S. ; Cui, Runjia ; Nanda, Sonali ; Xu, Wenhao ; Kachar, Bechara ; Barr-Gillespie, Peter ; Shin, Jung Bum. / A short splice form of Xin-actin binding repeat containing 2 (XIRP2) lacking the xin repeats is required for maintenance of stereocilia morphology and hearing function. In: Journal of Neuroscience. 2015 ; Vol. 35, No. 5. pp. 1999-2014.
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abstract = "Approximately one-third of known deafness genes encode proteins located in the hair bundle, the sensory hair cell’s mechanoreceptive organelle. In previous studies, we used mass spectrometry to characterize the hair bundle’s proteome, resulting in the discovery of novel bundle proteins. One such protein is Xin-actin binding repeat containing 2 (XIRP2), an actin-cross-linking protein previously reported to be specifically expressed in striated muscle. Because mutations in other actin-cross-linkers result in hearing loss, we investigated the role of XIRP2 in hearing function. In the inner ear, XIRP2 is specifically expressed in hair cells, colocalizing with actin-rich structures in bundles, the underlying cuticular plate, and the circumferential actin belt. Analysis using peptide mass spectrometry revealed that the bundle harbors a previously uncharacterized XIRP2 splice variant, suggesting XIRP2’s role in the hair cell differs significantly from that reported in myocytes. To determine the role of XIRP2 in hearing, we applied clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-mediated genome-editing technology to induce targeted mutations into the mouse Xirp2 gene, resulting in the elimination of XIRP2 protein expression in the inner ear. Functional analysis of hearing in the resulting Xirp2-null mice revealed high-frequency hearing loss, and ultrastructural scanning electron microscopy analyses of hair cells demonstrated stereocilia degeneration in these mice. We thus conclude that XIRP2 is required for long-term maintenance of hair cell stereocilia, and that its dysfunction causes hearing loss in the mouse.",
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AU - Krey, Jocelyn F.

AU - Krystofiak, Evan S.

AU - Cui, Runjia

AU - Nanda, Sonali

AU - Xu, Wenhao

AU - Kachar, Bechara

AU - Barr-Gillespie, Peter

AU - Shin, Jung Bum

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N2 - Approximately one-third of known deafness genes encode proteins located in the hair bundle, the sensory hair cell’s mechanoreceptive organelle. In previous studies, we used mass spectrometry to characterize the hair bundle’s proteome, resulting in the discovery of novel bundle proteins. One such protein is Xin-actin binding repeat containing 2 (XIRP2), an actin-cross-linking protein previously reported to be specifically expressed in striated muscle. Because mutations in other actin-cross-linkers result in hearing loss, we investigated the role of XIRP2 in hearing function. In the inner ear, XIRP2 is specifically expressed in hair cells, colocalizing with actin-rich structures in bundles, the underlying cuticular plate, and the circumferential actin belt. Analysis using peptide mass spectrometry revealed that the bundle harbors a previously uncharacterized XIRP2 splice variant, suggesting XIRP2’s role in the hair cell differs significantly from that reported in myocytes. To determine the role of XIRP2 in hearing, we applied clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-mediated genome-editing technology to induce targeted mutations into the mouse Xirp2 gene, resulting in the elimination of XIRP2 protein expression in the inner ear. Functional analysis of hearing in the resulting Xirp2-null mice revealed high-frequency hearing loss, and ultrastructural scanning electron microscopy analyses of hair cells demonstrated stereocilia degeneration in these mice. We thus conclude that XIRP2 is required for long-term maintenance of hair cell stereocilia, and that its dysfunction causes hearing loss in the mouse.

AB - Approximately one-third of known deafness genes encode proteins located in the hair bundle, the sensory hair cell’s mechanoreceptive organelle. In previous studies, we used mass spectrometry to characterize the hair bundle’s proteome, resulting in the discovery of novel bundle proteins. One such protein is Xin-actin binding repeat containing 2 (XIRP2), an actin-cross-linking protein previously reported to be specifically expressed in striated muscle. Because mutations in other actin-cross-linkers result in hearing loss, we investigated the role of XIRP2 in hearing function. In the inner ear, XIRP2 is specifically expressed in hair cells, colocalizing with actin-rich structures in bundles, the underlying cuticular plate, and the circumferential actin belt. Analysis using peptide mass spectrometry revealed that the bundle harbors a previously uncharacterized XIRP2 splice variant, suggesting XIRP2’s role in the hair cell differs significantly from that reported in myocytes. To determine the role of XIRP2 in hearing, we applied clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-mediated genome-editing technology to induce targeted mutations into the mouse Xirp2 gene, resulting in the elimination of XIRP2 protein expression in the inner ear. Functional analysis of hearing in the resulting Xirp2-null mice revealed high-frequency hearing loss, and ultrastructural scanning electron microscopy analyses of hair cells demonstrated stereocilia degeneration in these mice. We thus conclude that XIRP2 is required for long-term maintenance of hair cell stereocilia, and that its dysfunction causes hearing loss in the mouse.

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