PUBLICATION

New tools for studying osteoarthritis genetics in zebrafish

Authors
Mitchell, R.E., Huitema, L.F., Skinner, R.H., Brunt, L.H., Severn, C., Schulte-Merker, S., and Hammond, C.L.
ID
ZDB-PUB-121205-18
Date
2013
Source
Osteoarthritis and Cartilage   21(2): 269-278 (Journal)
Registered Authors
Brunt, Lucy, Hammond, Chrissy, Huitema, Leonie, Schulte-Merker, Stefan
Keywords
zebrafish, gene expression, cartilage, bone, model, osteoarthritis
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Chondrocytes/pathology
  • Chromosomes, Artificial, Bacterial/genetics
  • Collagen Type II/genetics
  • Collagen Type II/metabolism
  • Collagen Type II/physiology
  • Collagen Type IX/genetics
  • Collagen Type IX/metabolism
  • Collagen Type IX/physiology
  • Collagen Type X/genetics
  • Collagen Type X/metabolism
  • Collagen Type X/physiology
  • Disease Models, Animal*
  • Gene Expression Regulation, Developmental/physiology*
  • Genetic Predisposition to Disease/genetics*
  • Growth Differentiation Factor 5/genetics
  • Growth Differentiation Factor 5/physiology
  • Guanine Nucleotide Exchange Factors/genetics
  • Guanine Nucleotide Exchange Factors/physiology
  • Hypertrophy/genetics
  • Osteoarthritis/genetics*
  • Osteoarthritis/physiopathology*
  • Parathyroid Hormone-Related Protein/genetics
  • Parathyroid Hormone-Related Protein/physiology
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/physiology*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
  • Zebrafish Proteins/physiology
PubMed
23159952 Full text @ Osteoarthritis Cartilage
Abstract

Objective

Increasing evidence points to a strong genetic component to osteoarthritis (OA) and that certain changes that occur in osteoarthritic cartilage recapitulate the developmental process of endochondral ossification. As zebrafish are a well validated model for genetic studies and developmental biology, our objective was to establish the spatiotemporal expression pattern of a number of OA susceptibility genes in the larval zebrafish providing a platform for functional studies into the role of these genes in OA.

Design

We identified the zebrafish homologues for Mcf2l, Gdf5, PthrP/Pthlh, Col9a2, and Col10a1 from the Ensembl genome browser. Labelled probes were generated for these genes and in situ hybridisations were performed on wild type zebrafish larvae. In addition, we generated transgenic reporter lines by modification of bacterial artificial chromosomes (BACs) containing full length promoters for col2a1 and col10a1.

Results

For the first time, we show the spatiotemporal expression pattern of Mcf2l. Furthermore, we show that all six putative OA genes are dynamically expressed during zebrafish larval development, and that all are expressed in the developing skeletal system. Furthermore, we demonstrate that the transgenic reporters we have generated for col2a1 and col10a1 can be used to visualise chondrocyte hypertrophy in vivo.

Conclusion

In this study we describe the expression pattern of six OA susceptibility genes in zebrafish larvae and the generation of two new transgenic lines marking chondrocytes at different stages of maturation. Moreover, the tools used demonstrate the utility of the zebrafish model for functional studies on genes identified as playing a role in OA.

Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes