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New tools for studying osteoarthritis genetics in zebrafish

机译：研究斑马鱼骨关节炎遗传学的新工具

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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. ? 2012 Osteoarthritis Research Society International.
机译：目的：越来越多的证据表明，骨关节炎（OA）具有很强的遗传成分，而骨关节炎的某些变化概括了软骨内骨化的发展过程。由于斑马鱼是遗传研究和发育生物学的公认模型，因此我们的目标是建立幼虫斑马鱼中许多OA易感基因的时空表达模式，从而为功能研究这些基因在OA中的作用提供平台。设计：我们从Ensembl基因组浏览器中鉴定了Mcf2l，Gdf5，PthrP / Pthlh，Col9a2和Col10a1的斑马鱼同源物。产生了针对这些基因的标记探针，并在野生型斑马鱼幼虫上进行了原位杂交。此外，我们通过修饰含有col2a1和col10a1全长启动子的细菌人工染色体（BAC）生成了转基因报告基因系。结果：首次显示了Mcf2l的时空表达模式。此外，我们显示了所有六个推定的OA基因在斑马鱼幼虫发育过程中动态表达，并且都在发育中的骨骼系统中表达。此外，我们证明我们为col2a1和col10a1生成的转基因报告基因可用于可视化体内软骨细胞肥大。结论：在这项研究中，我们描述了六个OA易感基因在斑马鱼幼虫中的表达模式，以及在成熟的不同阶段标记软骨细胞的两个新转基因品系的产生。此外，所使用的工具证明了斑马鱼模型对确定为在OA中起作用的基因的功能研究的实用性。 ？ 2012年国际骨关节炎研究学会。

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