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
  • Animals
  • CRISPR-Cas Systems
  • Cilia/metabolism
  • Cone-Rod Dystrophies/genetics*
  • Disease Models, Animal
  • Eye Proteins/metabolism
  • Gene Knockout Techniques
  • Genetic Predisposition to Disease/genetics*
  • Humans
  • Leber Congenital Amaurosis/genetics*
  • Leber Congenital Amaurosis/metabolism*
  • Leber Congenital Amaurosis/pathology
  • Microtubule-Associated Proteins
  • Phenotype
  • Protein Transport/physiology*
  • Retina/metabolism
  • Retinal Cone Photoreceptor Cells/pathology
  • Retinal Degeneration/genetics
  • Retinal Degeneration/pathology
  • Tumor Suppressor Proteins/genetics
  • Tumor Suppressor Proteins/metabolism
  • Zebrafish/genetics*
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
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping