bioimagesUnknownOluwatobi OgunThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201404016JCB

Shown is an immunohistochemistry analysis conducted at 3 days post-fertilization of neuromasts from whole-mount zebrafish embryos of clarin-1 morphants co-injected with a truncated clarin-1 cRNA. Hair cells were immunostained for pcdh15a (red) and counter-stained for F-actin (green). The truncated version of clarin-1 transcript does not rescue the morphant phenotype.

Shown is an immunohistochemistry analysis conducted 4 days post-fertilization of the anterior crista from whole-mount zebrafish larvae. Hair cells were immunostained for clarin-1 (red) and counter-stained for F-actin (green). Clarin-1 localizes below the cuticular plate and at the fonticulus.

Shown is an immunohistochemistry analysis conducted at 2 days post-fertilization of anterior macula from whole-mount transgenic (bnr3c:mGFP) zebrafish, clarin-1 morphant embryos co-injected with clarin-1 cRNA. Hair cells were immunostained for the pre-synaptic marker ribeye b (red) and the post-synapitc marker MAGUK (green). Transgenic brn3c fish express a membrane-targeted GFP (white) in ear and neuromast hair cells and show normal distribution of ribeye b puncta when co-injected with clarin-1 cRNA.

Shown is an immunohistochemistry analysis conducted at 3 days post-fertilization of neuromasts from whole-mount control-injected transgenic (bnr3c:mGFP) zebrafish embryos. Hair cells were immunostained for the pre-synaptic marker ribeye b (red) and the post-synaptic marker MAGUK (green). Bnr3c transgenic fish express a membrane-targeted GFP (white) in ear and neuromast hair cells and show basal distribution of ribeye b puncta.

Shown is an immunohistochemistry analysis conducted 4 days post-fertilization of neuromasts from whole-mount zebrafish larvae. Hair cells were immunostained for clarin-1 (red) and the transition zone marker cc2d2a (green). Clarin-1 localizes at the most basal region of this sub-ciliary compartment.

Shown is an immunohistochemistry analysis conducted at 3 days post-fertilization of neuromasts from whole-mount transgenic (bnr3c:mGFP) zebrafish, clarin-1 morphant embryos. Brn3c transgenic fish were microinjected with the clarin-1 morpholino mixture and immunostained for the pre-synaptic marker ribeye b (red) and the post-synaptic marker MAGUK (green). Bnr3c transgenic fish express a membrane-targeted GFP (white) in ear and neuromast hair cells. Clarin-1 morphants show an increase in the number of ribeye b puncta.

Shown is an immunohistochemistry analysis conducted at 2 days post-fertilization of anterior macula from whole-mount control-injected transgenic (bnr3c:mGFP) zebrafish embryos. Hair cells were immunostained for the pre-synaptic marker ribeye b (red) and the post-synaptic marker MAGUK (green). Bnr3c transgenic fish express a membrane-targeted GFP (white) in ear and neuromast hair cells and show basal distribution of ribeye b puncta.

Shown is an immunohistochemistry analysis conducted 4 days post-fertilization of neuromasts from whole-mount zebrafish larvae. Hair cells were immunostained for clarin-1 (red) and counter-stained with phalloidin for F-actin (green). Clarin-1 localizes below the cuticular plate and at the point of insertion of the kinocilium.

Shown is an immunohistochemistry analysis conducted at 2 days post-fertilization of anterior macula from whole-mount transgenic (bnr3c:mGFP) zebrafish, clarin-1 morphant embryos. Inner ear hair cells were immunostained for the pre-synaptic marker ribeye b (red) and the post-synaptic marker MAGUK (green). This transgenic fish line expresses membrane-targeted GFP (white) in ear and neuromast hair cells. Clarin-1 morphants show accumulation of ribeye b puncta at the apical aspect of the hair cells.

Shown is an immunohistochemistry analysis conducted at 3 days post-fertilization of neuromasts from whole-mount transgenic (bnr3c:mGFP) zebrafish, clarin-1 morphant embryos co-injected with a truncated clarin-1 cRNA. Hair cells were immunostained for the pre-synaptic marker ribeye b (red) and the post-synaptic marker MAGUK (green). Transgenic brn3c fish express a membrane-targeted GFP (white) in ear and neuromast hair cells. The truncated version of the clarin-1 transcript does not rescue the morphant phenotype in the transgenic bnr3c zebrafish line.

Shown is an immunohistochemistry analysis conducted 3 days post-fertilization of hair cells from whole-mount zebrafish clarin-1 morphant embryos. Hair cells were immunostained for pcdh15a (red) and counter-stained for F-actin (green). Clarin-1 morphants lack Pcdh15a expression at the apical aspect of neuromast hair cells.

Shown is an immunohistochemistry analysis conducted 3 days post-fertilization of hair cells from whole-mount control-injected zebrafish embryos. Hair cells were immunostained for pcdh15a (red) and counter-stained for F-actin (green).

Shown is an immunohistochemistry analysis conducted 1 day post-fertilization of whole-mount zebrafish embryos. Hair cells were immunostained for clarin-1 (red) and counter-stained for F-actin with phalloidin (green). Clarin-1 is present in both anterior and posterior macular hair cells.

Shown is an immunohistochemistry analysis conducted at 3 days post-fertilization of hair cells from whole-mount embryos of clarin-1 morphants co-injected with clarin-1 cRNA. Hair cells were immunostained for pcdh15a (red) and counter-stained for F-actin (green). Pcdh15a apical expression is restored in the cRNA injected morphants.

Shown is an immunohistochemistry analysis conducted at 2 days post-fertilization of anterior macula from whole-mount transgenic (bnr3c:mGFP) zebrafish, clarin-1 morphant embryos co-injected with a truncated clarin-1 cRNA. Hair cells were immunostained for the pre-synaptic marker ribeye b (red) and the post-synaptic marker MAGUK (green). Bnr3c transgenic fish express a membrane-targeted GFP (white) in ear and neuromast hair cells. The truncated version of the clarin-1 transcript does not rescue the morphant phenotype in the transgenic bnr3c zebrafish line.

Shown is an immunohistochemistry analysis conducted at 3 days post-fertilization of neuromasts from whole-mount transgenic (bnr3c:mGFP) zebrafish, clarin-1 morphant embryos co-injected with clarin-1 cRNA. Hair cells were immunostained for the pre-synaptic marker ribeye b (red) and the post-synaptic marker MAGUK (green). Transgenic brn3c fish express a membrane-targeted GFP (white) in ear and neuromast hair cells and show normal distribution of ribeye b puncta when co-injected with clarin-1 cRNA.

bioimagesFigure 8 - FFigure 8 - CImage 627190 (Figure 6 - K)Image 627193 (Figure 8 - I)Image 627191 (Figure 8 - F)Image 627157 (Figure 1 - I-B)Figure 9 - LImage 627160 (Figure 2 - H)Figure 9 - FImage 627192 (Figure 8 - C)Image 627159 (Figure 1 - I-F)Figure 9 - IImage 627158 (Figure 1 - II-D)Figure 9 - CImage 627196 (Figure 9 - C)Figure 1 - II-DImage 627188 (Figure 6 - G)Figure 8 - LFigure 8 - IImage 627198 (Figure 9 - L)Image 627189 (Figure 6 - I)Figure 6 - KFigure 6 - IImage 627162 (Figure 6 - C)Image 627195 (Figure 9 - F)Image 627197 (Figure 9 - I)Figure 1 - I-BFigure 2 - HFigure 6Figure 9Figure 1 - I-FFigure 8Figure 6 - CImage 627194 (Figure 8 - L)Figure 1Figure 6 - GFigure 2Oluwatobi OgunMarisa ZallocchifalseClarin-1 acts as a modulator of mechanotransduction activity and presynaptic ribbon assemblyClarin-1 is a four-transmembrane protein expressed by hair cells and photoreceptors. Mutations in its corresponding gene are associated with Usher syndrome type 3, characterized by late-onset and progressive hearing and vision loss in humans. Mice carrying mutations in the clarin-1 gene have hair bundle dysmorphology and a delay in synapse maturation. In this paper, we examined the expression and function of clarin-1 in zebrafish hair cells. We observed protein expression as early as 1 d postfertilization. Knockdown of clarin-1 resulted in inhibition of FM1-43 incorporation, shortening of the kinocilia, and mislocalization of ribeye b clusters. These phenotypes were fully prevented by co-injection with clarin-1 transcript, requiring its C-terminal tail. We also observed an in vivo interact2014-11-03T11:26:22Z2018-11-29T11:26:22Z2018-11-29T11:26:22ZS-JCBD-20140401610.1083/jcb.201404016