- Title
-
Grxcr1 Promotes Hair Bundle Development by Destabilizing the Physical Interaction between Harmonin and Sans Usher Syndrome Proteins
- Authors
- Blanco-Sánchez, B., Clément, A., Fierro, J., Stednitz, S., Phillips, J.B., Wegner, J., Panlilio, J.M., Peirce, J.L., Washbourne, P., Westerfield, M.
- Source
- Full text @ Cell Rep.
PHENOTYPE:
|
ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions. PHENOTYPE:
|
PHENOTYPE:
|
EXPRESSION / LABELING:
|
|
|
grxcr1 is expressed during early development. Related to Figure 1. (A) grxcr1 is maternally deposited and expressed during early stages of development. RT-PCR for grxcr1 and control ef1rt. hpf: hours post-fertilization. (B) grxcr1 genomic organization diagram. grxcr1 gene is composed of 4 exons. The glutaredoxin domain is indicated in orange. (C) Mosaic image of grxcr1 in situ hybridization at 5 dpf. Red arrow points to a neuromast. Red dashed lines indicate where cross sections were done. Sections through the eye (C’), the forebrain (C’’) and the anterior macula of the inner ear (C’’’). |
Mechanosensation is not affected in grxcr1 mutants. Related to Figure 2. (A) Percentage of larvae responding correctly to the startle response test or with a balance defect. The number of larvae analyzed was as follow: 104 grxcr1+/+, 121 grxcr1+/b1259, 43 grxcr1b1259/b1259, 112 grxcr1+/b1260, 70 grxcr1b1260/b1260. (B) Average intensity of FM1-43 signal within the hair bundles of inner ear hair cells in grxcr1+/+ (n=20 larvae) and grxcr1-/- (n=22 larvae). Incorporation of FM1-43 in the hair bundles of the hair cells of the anterior macula at 5 dpf in grxcr1+/+ (C) and grxcr1-/- (D). (E) Average intensity of FM1-43 signal within the hair bundles of neuromast hair cells in grxcr1+/+ (n=20 larvae) and grxcr1-/- (n=22 larvae). Data were normalized to grxcr1+/+. Data are represented as mean ± SEM. |
Grxcr1 antibody is specific, and anti-Ush1ga and commercial anti-Ush1g antibodies recognize Ush1ga but not Ush1gb. Related to Figures 5 and 6. (A) Schematic of Grxcr1 protein showing the epitope (outlined in red) recognized by the anti-Grxcr1 antibody. (B) MDCK cells were transfected with grxcr1-HA and immuno-labelled using anti-Grxcr1 and anti-HA antibodies. Grxcr1 was detected within transfected cells as shown by the co-localization between Grxcr1 (green) and HA (red). Background at the cell membrane was also observed. (C) MDCK cells were transfected with pcDNA3.1 and immuno-labelled using anti-Grxcr1 and anti-HA antibodies. Only background at the cell membrane could be detected. Immuno-labelling using anti-Grxcr1 antibody in grxcr1+/+ (D) and grxcr1-/- mutant larvae (D’) at 5 dpf. (E) Immuno-labeling using KDEL antibody in Tg[myo6b:kdel-crimson]b1319 larvae at 5 dpf. (F) Schematic of Ush1ga protein showing the epitopes recognized by the anti-Ush1ga antibody (orange line), the mouse anti-Ush1g antibody (red line), the goat anti-Ush1g antibody (blue dotted line) and the rabbit anti-Ush1g antibody (purple line). ANK domains are indicated in green and the SAM domain is shown in pink. (G) MDCK cells were transfected with ush1ga-GFP. Ush1ga was detected within transfected cells as shown by the co-localization between Ush1ga (red) and GFP (green) signals. Background was also observed in some untransfected cells. (H) MDCK cells were transfected with ush1gb-HA. Ush1ga (red) could only be detected at background level in transfected cells (green). (I) MDCK cells were transfected with ush1ga-GFP and immuno-labelled with rabbit anti-Ush1g and anti-GFP antibodies. Ush1ga was detected in transfected cells as shown by the co-localization between Ush1g (red) and GFP (green) signals. (J) MDCK cells were transfected with ush1gb-HA and immuno-labelled with rabbit anti-Ush1g and anti-GFP antibodies. Ush1gb could not be detected in transfected cells. (K) MDCK cells were transfected with ush1ga-GFP and immuno-labelled with goat and mouse anti-Ush1g antibodies and anti-GFP antibody. Ush1ga was detected in transfected cells as shown by the co-localization between Ush1g (red-goat and blue-mouse) and GFP (green) signals. (L) MDCK cells were transfected with ush1gb-HA and immuno-labelled with goat and mouse anti-Ush1g antibodies and anti-GFP antibody. Ush1gb could not be detected in transfected cells. |