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

Yu, S., Li, C., Biswas, L., Hu, X., Liu, F., Reilly, J., Liu, X., Liu, Y., Huang, Y., Lu, Z., Han, S., Wang, L., Liu, J.Y., Jiang, T., Shu, X., Wong, F., Tang, Z., Liu, M.
Human molecular genetics   26(12): 2335-2345 (Journal)
Registered Authors
Han, Shanshan, Huang, Yuwen, Hu, Xuebin, Li, Chang, Liu, Fei, Liu, Mugen, Liu, Xiliang, Liu, Ying, Lu, Zhaojing, Yu, Shanshan
MeSH Terms
  • Animals
  • Cell Line
  • Down-Regulation
  • Gene Knockout Techniques/methods
  • Humans
  • Mutation
  • Phagocytosis/genetics
  • Phosphotransferases (Alcohol Group Acceptor)/genetics*
  • Phosphotransferases (Alcohol Group Acceptor)/metabolism*
  • Photoreceptor Cells
  • RNA, Messenger/metabolism
  • Receptor Protein-Tyrosine Kinases/metabolism
  • Retina/metabolism
  • Retinal Cone Photoreceptor Cells/metabolism
  • Retinal Degeneration/genetics
  • Retinal Pigment Epithelium/metabolism
  • Retinitis Pigmentosa/metabolism
  • Zebrafish/genetics
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism*
28398482 Full text @ Hum. Mol. Genet.
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.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes