PUBLICATION

Rapamycin relieves the cataract caused by ablation of Gja8b through stimulating autophagy in zebrafish

Authors
Ping, X., Liang, J., Shi, K., Bao, J., Wu, J., Yu, X., Tang, X., Zou, J., Shentu, X.
ID
ZDB-PUB-210123-4
Date
2021
Source
Autophagy   17(11): 3323-3337 (Journal)
Registered Authors
Zou, Jian
Keywords
3-MA, GJA8, Gja8b, autophagy, cataract, lens, organelle degradation, rapamycin
MeSH Terms
  • Lens, Crystalline/abnormalities
  • Lens, Crystalline/growth & development
  • Lens, Crystalline/physiology
  • Animals
  • Zebrafish Proteins/antagonists & inhibitors*
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/physiology
  • Cataract/genetics*
  • Cataract/pathology
  • Cataract/physiopathology
  • Sirolimus/pharmacology*
  • Adenine/analogs & derivatives
  • Adenine/pharmacology
  • Mutation
  • Autophagy/drug effects*
  • Autophagy/genetics*
  • Autophagy/physiology
  • Animals, Genetically Modified
  • Zebrafish/genetics*
  • Zebrafish/physiology*
  • Connexins/antagonists & inhibitors*
  • Connexins/genetics*
  • Connexins/physiology
(all 23)
PubMed
33472493 Full text @ Autophagy
Abstract
Macroautophagy/autophagy is known to be important for intracellular quality control in the lens. GJA8 is a major gap junction protein in vertebrate lenses. Mutations in GJA8 cause cataracts in humans. The well-known cataractogenesis mechanism is that mutated GJA8 leads to abnormal assembly of gap junctions, resulting in defects in intercellular communication among lens cells. In this study, we observed that ablation of Gja8b (a homolog of mammalian GJA8) in zebrafish led to severe defects in organelle degradation, an important cause of cataractogenesis in developing lens. The role of autophagy in organelle degradation in lens remains disputable. Intriguingly, we also observed that ablation of Gja8b induced deficient autophagy in the lens. More importantly, in vivo treatment of zebrafish with rapamycin, an autophagy activator that inhibits MAPK/JNK and MTORC1 signaling, stimulated autophagy in the lens and relieved the defects in organelle degradation, resulting in the mitigation of cataracts in gja8b mutant zebrafish. Conversely, inhibition of autophagy by treatment with the chemical reagent 3-MA blocked these recovery effects, suggesting the important roles of autophagy in organelle degradation in the lens in gja8b mutant zebrafish. Further studies in HLE cells revealed that GJA8 interacted with ATG proteins. Overexpression of GJA8 stimulated autophagy in HLE cells. These data suggest an unrecognized cataractogenesis mechanism caused by ablation of Gja8b and a potential treatment for cataracts by stimulating autophagy in the lens. Abbreviations: 3-MA: 3-methyladenine; ATG: autophagy related; AV: autophagic vacuoles; Dpf: days post fertilization; GJA1: gap junction protein alpha 1; GJA3: gap junction protein alpha 3; GJA8: gap junction protein alpha 8; Hpf: hours post fertilization; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; PtdIns3K: class III phosphatidylinositol 3-kinase; WT: wild type.
Genes / Markers
Figures
Figure Gallery (8 images)
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Expression
No data available
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
zko933a
    Insertion
    1 - 1 of 1
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    Human Disease / Model
    Human Disease Fish Conditions Evidence
    cataractTAS
    1 - 1 of 1
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    Sequence Targeting Reagents
    Target Reagent Reagent Type
    gja8bCRISPR1-gja8bCRISPR
    1 - 1 of 1
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    Fish
    1 - 2 of 2
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    Antibodies
    Name Type Antigen Genes Isotypes Host Organism
    Ab1-mippolyclonal
      Rabbit
      Ab1-prkczpolyclonalIgGRabbit
      Ab1-tjp1monoclonalIgG1Mouse
      Ab4-map1lc3bpolyclonal
        Rabbit
        Ab14-hspa5monoclonal
          IgG2aMouse
          zl-1monoclonal
            IgG1Mouse
            1 - 6 of 6
            Show
            Orthology
            No data available
            Engineered Foreign Genes
            No data available
            Mapping
            No data available