PUBLICATION
Molecular analysis and characterization of zebrafish keratocan (zKera) gene
- Authors
- Yeh, L.K., Liu, C.Y., Chien, C.L., Converse, R.L., Kao, W.W., Chen, M.S., Hsieh, F.J., and Wang, I.J.
- ID
- ZDB-PUB-071108-7
- Date
- 2008
- Source
- The Journal of biological chemistry 283(1): 506-517 (Journal)
- Registered Authors
- Wang, I-Jong
- Keywords
- none
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Apoptosis/drug effects
- Base Sequence
- Blotting, Northern
- Blotting, Western
- Caspase 3/metabolism
- Cornea/growth & development
- Cornea/metabolism
- Cornea/ultrastructure
- Gene Expression Regulation, Developmental
- Humans
- Immunohistochemistry
- In Situ Hybridization
- In Situ Nick-End Labeling
- Mice
- Mice, Knockout
- Microscopy, Electron, Transmission
- Molecular Sequence Data
- Phylogeny
- Promoter Regions, Genetic/genetics
- Proteoglycans/genetics
- Proteoglycans/metabolism*
- Reverse Transcriptase Polymerase Chain Reaction
- Sequence Alignment
- Zebrafish/genetics*
- Zebrafish/growth & development
- Zebrafish/metabolism*
- Zebrafish Proteins/classification
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- PubMed
- 17965408 Full text @ J. Biol. Chem.
Citation
Yeh, L.K., Liu, C.Y., Chien, C.L., Converse, R.L., Kao, W.W., Chen, M.S., Hsieh, F.J., and Wang, I.J. (2008) Molecular analysis and characterization of zebrafish keratocan (zKera) gene. The Journal of biological chemistry. 283(1):506-517.
Abstract
Corneal small leucine-rich proteoglycans (SLRPs) play a pivotal role in maintaining corneal transparency and function. In this study, we isolated and characterized the zebrafish (Danio rerio) keratocan (zKera) gene. Human keratocan sequence was used to search zebrafish homologues. The zKera full-length genomic DNA and cDNA were generated via PCR of zebrafish genomic DNA and RT-PCR of total zebrafish eye RNA, respectively. The zKera spanning 3.5 kilobase pairs consists of two exons and one intron, and a TATA-less promoter. The zKera encodes 341 amino acid with 59% identity to its human counterpart and 57% to that of mouse keratocan. Like mouse and chick keratocan, zKera mRNA is selectively expressed in the adult cornea, however, during embryonic development, zKera mRNA is expressed in both the brain and the cornea. Interestingly, it is expressed mainly in corneal epithelium but few in the stroma. A pseudogene was proved by introducing a zKera promoter-driven enhanced green fluorescence protein (EGFP) reporter gene into fertilized zebrafish eggs. Using morpholino-antisense against zKera to knock-down zKera resulted in lethal phenotype due to massive caspase-dependent apoptosis, which was noted by a significant increase of active caspase-3 and caspase-8, in the developing forebrain area including eyes. This is different from mouse, for which keratocan deficient mice are viable. Taken together, our data indicate that mammalian keratocan is conserved in zebrafish in terms of gene structure, expression pattern, and promoter function.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping