Drosophila model to clarify the pathological significance of OPA1 in autosomal dominant optic atrophy
- Brain Research Institute, Niigata University, Japan
- School of Life Science and Technology, Tokyo Institute of Technology, Japan
- Research Initiatives and Promotion Organization, Yokohama National University, Japan
- Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University, Japan
- Department of Gene Function and Phenomics, National Institute of Genetics, Japan
- Division of Ophthalmology and Visual Science, Graduate School of Medical and Dental Sciences, Niigata University, Japan
Figures
Figure 1 with 1 supplement
Effect of dOPA1 knockdown on mitochondrial density and size in retinal axons.
(A) Schematic illustration of the Drosophila visual system from a dorsal view, highlighting the arrangement of photoreceptor neurons. The rectangular area indicated in (B) represents a single R7/8 …
Figure 1—figure supplement 1
Analysis of trafficking defects in axons of dOPA1 knockdown flies.
Schematic illustration of the Drosophila visual system around 24 hr after puparium formation. The cell bodies of photoreceptors (magenta) are found in the retina; type 1–6 photoreceptors project …
Figure 2
Elevated reactive oxygen species (ROS) and mitophagy activity levels in the dOPA1 knockdown.
(A, B) Mitochondrial ROS levels in a set of the R7/8 retinal axon visualized using Mito-GFP (green) driven by GMR-Gal4 and MitoSOX (magenta) to specifically detect superoxide, an indicator of …
Figure 3 with 2 supplements
Effect of dOPA1 knockdown on photoreceptor neurodegeneration in Drosophila.
Schematic of the visual system in Drosophila. A set of R7/8 photoreceptors project their axons to the second optic ganglion medulla. The rectangular prism indicated in B–E represents the area of the …
Figure 3—figure supplement 1
The impact on the retinal axons of another RNAi line in Drosophila OPA1.
(A) The images show the representations of retinal axons labeled with an anti-Chaoptin antibody (gray). Control and dOPA1 RNAi (VDRC 106290) from VDRC 2 weeks after eclosion. Scale bar: 50 µm. (B) …
Figure 3—figure supplement 2
Evaluation of the impact on Drosophila compound eyes due to knockdown of dOPA1.
(A) Bright-field microscopic images of representative fly eyes expressing 40D-UAS (control), RNAi #1 (BDSC 32358), and RNAi #2 (VDRC 106290) using the GMR-Gal4 driver. (B) Graph representing the P …
Figure 4
Role of dOPA1 in the retinal axon according to mutant analysis.
(A–C) Representations of retinal axons labeled with an anti-Chaoptin antibody (gray), 1 day after eclosion. The images show Control (A), dOPA1s3475 somatic mosaic flies (B), and dOPA1s3475 somatic …
Figure 5 with 1 supplement
Verification of the pathological significance of disease mutations in hOPA1.
(A) Schematic illustration of the hOPA1 gene construct with HA and myc tags in the UAS-based vector. hOPA1 includes a mitochondrial import sequence (MIS) cleaved by mitochondrial processing …
Figure 5—figure supplement 1
Effects of hOPA1 variants on COXII expression and autophagy in the context of dOPA1 knockdown.
(A) Western blot analysis to show the expression of COXII upon expression of hOPA1 variants in the context of dOPA1 knockdown. (B) Quantification of COXII protein levels. Control (n=3), luciferase …
Figure 6 with 3 supplements
Loss-of-function (LOF) or dominant-negative (DN) effects of disease mutations in hOPA1.
(A) Impact of each human OPA1 variant on the axon number in the optic nerve of Drosophila as quantified using MeDUsA (method for the quantification of degeneration using fly axons). (B) Expression …
Figure 6—figure supplement 1
Pairwise alignment of GTPase, Middle, and GED domains of hOPA1 and dOPA1.
(A–C) The amino acid sequences of hOPA1 ([NM_015560.3]) and dOPA1 ([NM_166040.2]) were aligned using EMBOSS Needle. GTPase domain (A), Middle domain (B), and GED domain (C). Each percentage value …
Figure 6—figure supplement 2
Comparative analysis of mitochondrial reactive oxygen species (ROS) levels, COXII expression, and autophagy between hOPA1 wild-type and the variants expression under dOPA1 knockdown conditions.
(A) Impact on mitochondrial ROS levels upon expression of wild-type hOPA1 and its variants under conditions of dOPA1 knockdown. Control (n=16 optic lobes), hOPA1_WT (n=20 optic lobes), hOPA1_2708-271…
Figure 6—figure supplement 3
The effect of a dominant-negative mutation in dOPA1.
(A) The images show the representations of retinal axons labeled with an anti-Chaoptin antibody (gray). Control and the expression of a presumably GTPase-negative form of dOPA1K273A 1 week after …
Additional files
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Supplementary file 1
The genotypes of the Drosophila used in the experiments.
Detailed information about the specific genotypes of Drosophila used in each experiment described in the manuscript, including the relevant genetic constructs, mutations, and background strains.
- https://cdn.elifesciences.org/articles/87880/elife-87880-supp1-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/87880/elife-87880-mdarchecklist1-v1.pdf
