PUBLICATION
The role of cldnh during the early retinal development in zebrafish
- Authors
- Lu, J., Liu, R., Miao, A., Chen, X., Xiao, W., Wang, Y., Cao, D., Pan, J., Li, L., Luo, Y.
- ID
- ZDB-PUB-200901-9
- Date
- 2020
- Source
- Experimental Eye Research 200: 108207 (Journal)
- Registered Authors
- Keywords
- Claudin-3, Claudin-h, Hyaloid vessels, Neural retina, Retinal vessels, Zebrafish
- MeSH Terms
-
- Animals
- Blood-Retinal Barrier/physiology*
- Blotting, Western
- Claudins/biosynthesis
- Claudins/genetics*
- Gene Expression Regulation, Developmental*
- Models, Animal
- RNA/genetics*
- Retina/growth & development
- Retina/metabolism*
- Zebrafish
- Zebrafish Proteins/biosynthesis
- Zebrafish Proteins/genetics*
- PubMed
- 32866532 Full text @ Exp. Eye. Res.
Citation
Lu, J., Liu, R., Miao, A., Chen, X., Xiao, W., Wang, Y., Cao, D., Pan, J., Li, L., Luo, Y. (2020) The role of cldnh during the early retinal development in zebrafish. Experimental Eye Research. 200:108207.
Abstract
Claudin-3, an integral component of tight junction, has recently been shown to be expressed in retinal ganglion cells, retinal pigment cells, and retinal vascular endothelial cells. However, the role of claudin-3 in the development of the neural retina and its vessels remains undefined. This study aimed to investigate the role of zebrafish claudin-h (cldnh), the closest ortholog of mouse and human claudin-3, in the development of the neural retina and its vessels. Cldnh levels in green fluorescent protein transgenic zebrafish were genetically manipulated by cldnh morpholino oligonucleotide (MO) and cldnh mRNA to investigate gene function. The expression of cldnh was analyzed using polymerase chain reaction and immunofluorescence staining. The altered morphological, cellular and molecular events in the cldnh MO-morphant eyes were detected using hematoxylin-eosin staining, fluorescent dye injection, confocal in vivo imaging, BrdU labeling, TUNEL assay, RNA sequencing, and Western blot. We demonstrated that the cldnh protein was expressed in the neural retina and the hyaloid vessel which is the predecessor of the retinal vessel in zebrafish. Cldnh knockdown delayed lamination of the neural retina and reduced its thickness, which might be associated with the downregulation of the retinal development-related genes of atoh7, pcdh17, crx, neurod1, insm1a, sox9b and cdh11, and the upregulation of the cell cycle and apoptosis-associated genes of tp53, cdkn1a and casp8. Cldnh knockdown also reduced the density and interrupted the lumenization of the hyaloid vessels, which might be owing to the downregulation of the vessel formation-related genes of hlx1 and myl7. In conclusion, cldnh was required for the normal development of the neural retina and its vessels in zebrafish, providing a basis for elucidating its role in the pathogenesis of retinal vascular or inflammatory diseases.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping