ZFIN ID: ZDB-FIG-130815-33
Nishiwaki et al., 2013 - The BH3-Only SNARE BNip1 Mediates Photoreceptor Apoptosis in Response to Vesicular Fusion Defects. Developmental Cell   25(4):374-387 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Protruding-mouth

Fig. 4 The Syntaxin-18 cis-SNARE Complex Is Required for BNip1-Dependent Apoptosis(A) Molecular mechanism underlying vesicular fusion to the ER. Interactions among syntaxin-18 (green), BNip1 (blue), Use1 (yellow), and Sec22b (red) initiate the fusion of transport vesicles to the ER membrane. Three accessory proteins, Zw10, NAG, and RINT1, play a role in the tethering of transport vesicles. After membrane fusion occurs, β-SNAP binds to the cis-SNARE complex and recruits NSF. NSF promotes disassembly of the cis-SNARE complex. In the absence of β-SNAP or NSF activity, the syntaxin-18 cis-SNARE complex accumulates. MOs against syntaxin-18, Use1, Sec22b, and Sly1 inhibit the formation of the syntaxin-18 cis-SNARE complex, even in the absence of β-SNAP or NSF activity. (B) TUNEL staining (magenta) of 72 hpf WT, nsf-a mutant, and nsf-a mutant retinas injected with misMO-BNip1a/b, and a mixture of MO-BNip1a and MO-BNip1b. Nuclei were counterstained with SYTOX Green (green). Arrowheads indicate apoptosis in the ONL. (C) Zpr1 labeling (green) of 84 hpf WT, nsf-a mutant, nsf-a mutant retinas injected with misMO-BNip1a/b, and a mixture of MO-BNip1a and MO-BNip1b. Arrowheads indicate rescued photoreceptors. (D and E) Percentage of TUNEL-positive (D) and zpr1-positive areas (E) relative to the total retinal area in the experiments shown in (B) and (C), respectively. Green and black bars indicate the means and SDs, respectively. **p < 0.01. (F) Zpr1 labeling (green) of coa mutant and coa mutants injected with MO-syntaxin-18, MO-Use1, MO-Sec22ba, MO-Sly1, their 5misMOs, and MO-syntaxin 5al. Arrowheads indicate rescued photoreceptors. (G) Percentage of zpr1-positive area relative to the total retinal area in the experiments shown in (F). Green and black bars indicate the means and SDs, respectively (**p < 0.01). The numbers of retinal sections used in the experiments shown in (D), (E), and (G) and p values for the t test are shown in Table S2. Scale bars, 50 μm. See also Figure S5.

Gene Expression Details No data available
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
zpr-1 IHC WT standard conditions Protruding-mouth retinal cone cell
IHC napbbrw76b/rw76b standard conditions Protruding-mouth retinal cone cell
IHC napbbrw76b/rw76b + MO1-scfd1 standard conditions Protruding-mouth retinal cone cell
IHC napbbrw76b/rw76b + MO1-sec22ba standard conditions Protruding-mouth retinal cone cell
IHC napbbrw76b/rw76b + MO1-stx18 standard conditions Protruding-mouth retinal cone cell
IHC napbbrw76b/rw76b + MO1-stx5al standard conditions Protruding-mouth retinal cone cell
IHC napbbrw76b/rw76b + MO1-use1 standard conditions Protruding-mouth retinal cone cell
IHC nsfast53/st53 standard conditions Protruding-mouth retinal cone cell
IHC nsfast53/st53 + MO1-bnip1a + MO1-bnip1b standard conditions Protruding-mouth retinal cone cell
Phenotype Details
Fish Conditions Stage Phenotype
napbbrw76b/rw76b standard conditions Protruding-mouth retinal cone cell decreased amount, abnormal
napbbrw76b/rw76b + MO1-scfd1 standard conditions Protruding-mouth retinal cone cell decreased amount, abnormal
napbbrw76b/rw76b + MO1-sec22ba standard conditions Protruding-mouth retinal cone cell decreased amount, abnormal
napbbrw76b/rw76b + MO1-stx18 standard conditions Protruding-mouth retinal cone cell decreased amount, abnormal
napbbrw76b/rw76b + MO1-stx5al standard conditions Protruding-mouth retinal cone cell decreased amount, abnormal
napbbrw76b/rw76b + MO1-use1 standard conditions Protruding-mouth retinal cone cell decreased amount, abnormal
nsfast53/st53 standard conditions Protruding-mouth photoreceptor cell apoptotic, abnormal
Protruding-mouth retina apoptotic process increased occurrence, abnormal
Protruding-mouth retinal cone cell decreased amount, abnormal
nsfast53/st53 + MO1-bnip1a + MO1-bnip1b standard conditions Protruding-mouth photoreceptor cell apoptotic, abnormal
Protruding-mouth retina apoptotic process increased occurrence, abnormal
Protruding-mouth retinal cone cell decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 25(4), Nishiwaki, Y., Yoshizawa, A., Kojima, Y., Oguri, E., Nakamura, S., Suzuki, S., Yuasa-Kawada, J., Kinoshita-Kawada, M., Mochizuki, T., and Masai, I., The BH3-Only SNARE BNip1 Mediates Photoreceptor Apoptosis in Response to Vesicular Fusion Defects, 374-387, Copyright (2013) with permission from Elsevier. Full text @ Dev. Cell