New tools for studying osteoarthritis genetics in zebrafish

被引:64
作者
Mitchell, R. E. [1 ]
Huitema, L. F. A. [3 ]
Skinner, R. E. H. [1 ]
Brunt, L. H. [1 ]
Severn, C. [1 ]
Schulte-Merker, S. [3 ,4 ]
Hammond, C. L. [1 ,2 ]
机构
[1] Univ Bristol, Dept Biochem, Bristol BS8 1TD, Avon, England
[2] Univ Bristol, Dept Physiol & Pharmacol, Bristol BS8 1TD, Avon, England
[3] KNAW & UMC, Hubrecht Inst, NL-3584 CT Utrecht, Netherlands
[4] Wageningen Univ, EZO, NL-6700 AP Wageningen, Netherlands
关键词
Zebrafish; Gene expression; Cartilage; Bone; Model; Osteoarthritis; NUCLEOTIDE EXCHANGE FACTOR; DANIO-RERIO; OSTEOGENESIS-IMPERFECTA; DEVELOPING SKELETON; SIGNALING PATHWAYS; CELL-MIGRATION; BONE-FORMATION; RETINOIC ACID; EXPRESSION; GDF5;
D O I
10.1016/j.joca.2012.11.004
中图分类号
R826.8 [整形外科学]; R782.2 [口腔颌面部整形外科学]; R726.2 [小儿整形外科学]; R62 [整形外科学(修复外科学)];
学科分类号
摘要
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, Co19a2, 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 col10al 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. (C) 2012 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:269 / 278
页数:10
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