Anti-clarin-1 AAV-delivered ribozyme induced apoptosis in the mouse cochlea

Aarnisalo, Antti A.; Pietola, Laura; Joensuu, Jaana; Isosomppi, Juha; Aarnisalo, Piia; Dinculescu, Astra; Lewin, Alfred S.; Flannery, John G.; Hauswirth, William W.; Sankila, Eeva‐Marja; Jero, Jussi

doi:10.1016/j.heares.2007.03.004

Cited by 13 publications

(6 citation statements)

References 30 publications

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“…Exposure of the cranial base and middle ear bulla was performed through a ventral paramedian incision as described previously [6]. Lentivirus vectors were introduced surgically into the inner ear of the mice by injecting the vector directly through the round window membrane (RWM) [7]. Pure lentivirus vector (1 ml) was injected into the inner ear with a glass capillary (inner diameter 0.5 mm, SM100F-15, Harvard apparatus, tip diameter 5 mm) with the help of a micromanipulator (Narishige, Japan) and a preparation microscope (Olympus SZX12).…”

Section: Animal Surgery and Transduction Of Mice With Lentivirus Vectorsmentioning

confidence: 99%

HOX-GFP and WOX-GFP lentivirus vectors for inner ear gene transfer

Pietola

Aarnisalo

Joensuu

et al. 2008

Acta Oto-Laryngologica

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HOX-GFP and WOX-GFP expression was restricted to the lining cells of the scala tympani and scala vestibuli. No GFP expression was seen in the organ of Corti or the spiral ganglion. Aminoglycoside treatment had no effect on the expression of these vectors. The distant spread of lentivirus vectors was minimal; only the liver of one animal showed some GFP expression. Inflammatory reaction caused by these vectors was mild. Few inflammatory cells were found in the perilymphatic space of the cochlea and in the vestibular organ.

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Section: Animal Surgery and Transduction Of Mice With Lentivirus Vectorsmentioning

confidence: 99%

HOX-GFP and WOX-GFP lentivirus vectors for inner ear gene transfer

Pietola

Aarnisalo

Joensuu

et al. 2008

Acta Oto-Laryngologica

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“…The Clarin (CLRN) proteins belong to a superfamily of small integral proteins with four alpha‐helical transmembrane domains, which also includes Tetraspanins, Connexins, Claudins, Occludins and calcium channel gamma subunit‐like proteins (Adato et al , ; Aarnisalo et al , ). In humans, the CLRN family comprises three proteins encoded by the paralogous genes CLRN1 , CLRN2 and CLRN3 , which contain no known functional domains apart from their four transmembrane domains and a C‐terminal class‐II PDZ‐binding motif (PBM type II) (Fig A).…”

Section: Introductionmentioning

confidence: 99%

Clarin‐2 is essential for hearing by maintaining stereocilia integrity and function

et al. 2019

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Hearing relies on mechanically gated ion channels present in the actin‐rich stereocilia bundles at the apical surface of cochlear hair cells. Our knowledge of the mechanisms underlying the formation and maintenance of the sound‐receptive structure is limited. Utilizing a large‐scale forward genetic screen in mice, genome mapping and gene complementation tests, we identified Clrn2 as a new deafness gene. The Clrn2clarinet/clarinet mice (p.Trp4* mutation) exhibit a progressive, early‐onset hearing loss, with no overt retinal deficits. Utilizing data from the UK Biobank study, we could show that CLRN2 is involved in human non‐syndromic progressive hearing loss. Our in‐depth morphological, molecular and functional investigations establish that while it is not required for initial formation of cochlear sensory hair cell stereocilia bundles, clarin‐2 is critical for maintaining normal bundle integrity and functioning. In the differentiating hair bundles, lack of clarin‐2 leads to loss of mechano‐electrical transduction, followed by selective progressive loss of the transducing stereocilia. Together, our findings demonstrate a key role for clarin‐2 in mammalian hearing, providing insights into the interplay between mechano‐electrical transduction and stereocilia maintenance.

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“…Alternative splicing of the mouse Clrn1 gene can result in the potential expression of three distintc protein isoforms: Isoforms 1 (27.9 kDa) and 2 (25.8 kDa), both contain four-transmembrane domains, and isoform 3 (19.2 kDa) that contains only two (Adato et al, 2002). Based on sequence homology, clarin-1 belongs to a large hyperfamily of small integral proteins with four transmembrane domains (TM4SF) that includes two more clarins, tetraspanins, connexins, claudins and calcium channel gamma subunit-like proteins (CACNGS) (Adato et al, 2002, Aarnisalo et al, 2007). Members of this hyperfamily are involved in a variety of processes from cell sorting and signaling to cell-cell interaction and scaffolding functions (Hemler, 2001, Berditchevski, 2001, Adato et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2009

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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 immortomousederived 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 sub-cellular 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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Anti-clarin-1 AAV-delivered ribozyme induced apoptosis in the mouse cochlea

Cited by 13 publications

References 30 publications

HOX-GFP and WOX-GFP lentivirus vectors for inner ear gene transfer

HOX-GFP and WOX-GFP lentivirus vectors for inner ear gene transfer

Clarin‐2 is essential for hearing by maintaining stereocilia integrity and function

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

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