PUBLICATION

Knocking out lca5 in zebrafish causes cone-rod dystrophy due to impaired outer segment protein trafficking

Authors
Qu, Z., Yimer, T.A., Xie, S., Wong, F., Yu, S., Liu, X., Han, S., Ma, J., Lu, Z., Hu, X., Qin, Y., Huang, Y., Lv, Y., Li, J., Tang, Z., Liu, F., Liu, M.
ID
ZDB-PUB-190729-7
Date
2019
Source
Biochimica et biophysica acta. Molecular basis of disease   1865(10): 2694-2705 (Journal)
Registered Authors
Han, Shanshan, Huang, Yuwen, Hu, Xuebin, Li, Jingzhen, Liu, Fei, Liu, Mugen, Liu, Xiliang, Lu, Zhaojing, Qin, Yayun, Qu, Zhen, Yu, Shanshan
Keywords
CRISPR/Cas9, Ift88, LCA5, Photoreceptor degeneration, Zebrafish
MeSH Terms
  • Cilia/metabolism
  • Gene Knockout Techniques
  • Retinal Degeneration/genetics
  • Retinal Degeneration/pathology
  • Phenotype
  • CRISPR-Cas Systems
  • Eye Proteins/metabolism
  • Zebrafish/genetics*
  • Protein Transport/physiology*
  • Retina/metabolism
  • Animals
  • Retinal Cone Photoreceptor Cells/pathology
  • Disease Models, Animal
  • Genetic Predisposition to Disease/genetics*
  • Humans
  • Cone-Rod Dystrophies/genetics*
  • Leber Congenital Amaurosis/genetics*
  • Leber Congenital Amaurosis/metabolism*
  • Leber Congenital Amaurosis/pathology
  • Tumor Suppressor Proteins/genetics
  • Tumor Suppressor Proteins/metabolism
  • Microtubule-Associated Proteins
(all 22)
PubMed
31348989 Full text @ BBA Molecular Basis of Disease
Abstract
Leber congenital amaurosis (LCA) is the most serious form of inherited retinal dystrophy that leads to blindness or severe visual impairment within a few months after birth. Approximately 1-2% of the reported cases are caused by mutations in the LCA5 gene. This gene encodes a ciliary protein called LCA5 that is localized to the connecting cilium of photoreceptors. The retinal phenotypes caused by LCA5 mutations and the underlying pathological mechanisms are still not well understood. In this study, we knocked out the lca5 gene in zebrafish using CRISPR/Cas9 technology. An early onset visual defect is detected by the ERG in 7 dpf lca5-/- zebrafish. Histological analysis by HE staining and immunofluorescence reveal progressive degeneration of rod and cone photoreceptors, with a pattern that cones are more severely affected than rods. In addition, ultrastructural analysis by transmission electron microscopy shows disordered and broken membrane discs in rods' and cones' outer segments, respectively. In our lca5-/- zebrafish, the red-cone opsin and cone α-transducin are selectively mislocalized to the inner segment and synaptic terminal. Moreover, we found that Ift88, a key component of the intraflagellar transport complex, is retained in the outer segments. These data suggest that the intraflagellar transport complex-mediated outer segment protein trafficking might be impaired due to lca5 deletion, which finally leads to a type of retinal degeneration mimicking the phenotype of cone-rod dystrophy in human. Our work provides a novel animal model to study the physiological function of LCA5 and develop potential treatments of LCA.
Genes / Markers
Figures
No images available
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Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
zf3156
    Small Deletion
    1 - 1 of 1
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    Human Disease / Model
    Human Disease Fish Conditions Evidence
    Leber congenital amaurosislca5zf3156/zf3156standard conditionsTAS
    1 - 1 of 1
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    Sequence Targeting Reagents
    Target Reagent Reagent Type
    lca5CRISPR2-lca5CRISPR
    1 - 1 of 1
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    Fish
    1 - 2 of 2
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    Antibodies
    Orthology
    Engineered Foreign Genes
    No data available
    Mapping
    No data available