CERKL gene knockout disturbs photoreceptor outer segment phagocytosis and causes rod-cone dystrophy in zebrafish

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E-pub ahead of print

DOI

  • Shanshan Yu
  • Chang Li
  • Lincoln Biswas
  • Xuebin Hu
  • Fei Liu
  • James Reilly
  • Xiliang Liu
  • Ying Liu
  • Yuwen Huang
  • Zhaojing Lu
  • Shanshan Han
  • Lei Wang
  • Jing Yu Liu
  • Tao Jiang
  • Xinhua Shu
  • Fulton Wong
  • Zhaohui Tang
  • Mugen Liu

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Original languageEnglish
JournalHuman Molecular Genetics
Early online date7 Apr 2017
DOIs
StateE-pub ahead of print - 7 Apr 2017

Abstract

In humans, CERKL mutations cause widespread retinal degeneration: early dysfunction and loss of rod and cone photoreceptors in the outer retina and, progressively, death of cells in the inner retina. Despite intensive efforts, the function of CERKL remains obscure and studies in animal models have failed to clarify the disease mechanism of CERKL mutations. To address this gap in knowledge, we have generated a stable CERKL knockout zebrafish model by TALEN technology and a 7bp deletion in CERKL cDNA that caused the premature termination of CERKL. These CERKL-/- animals showed progressive degeneration of photoreceptor outer segments (OSs) and increased apoptosis of retinal cells, including those in the outer and inner retinal layers. Additionally, we confirmed by immunofluorescence and western-blot that rod degeneration in CERKL-/- zebrafish occurred earlier and was more significant than that in cone cells. Accumulations of shed OSs in the interphotoreceptor matrix were observed by transmission election microscopy (TEM). This suggested that CERKL may regulate the phagocytosis of OSs by the retinal pigment epithelium (RPE). We further found that the phagocytosis-associated protein MERTK was significantly reduced in CERKL-/- zebrafish. Additionally, in ARPE-19 cell lines, knockdown of CERKL also decreased the mRNA and protein level of MERTK, as well as the ox-POS phagocytosis. We conclude that CERKL deficiency in zebrafish may cause rod-cone dystrophy, but not cone-rod dystrophy, while interfering with the phagocytosis function of RPE associated with down-regulation of the expression of MERTK.

Keywords

  • genetics, gene knockout, photoreceptor, zebrafish, retinal degeneration