Development of Teleost Intermuscular Bones Undergoing Intramembranous Ossification Based on Histological-Transcriptomic-Proteomic Data
Development of Teleost Intermuscular Bones Undergoing Intramembranous Ossification Based on Histological-Transcriptomic-Proteomic Data
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基于组织学-转录组-蛋白质组数据进行膜内骨化的硬骨鱼肌间骨的发育
DOI：
10.3390/ijms20194698
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发表时间：
2019-09
影响因子：
5.6
通讯作者：
Ze-Xia Gao
中科院分区：
文献类型：
--
作者：
Chun-Hong Nie;Shi-Ming Wan;Yu-Long Liu;Han Liu;Wei-Min Wang;Ze-Xia Gao
肌间骨（Intermuscular bones, IBs）存在于低等硬骨鱼类中，其发育的分子机制尚不清楚。本研究通过不同的染色方法和比较蛋白质组学研究了双头巨鲷IB发育的组织学结构和蛋白质组学，包括IB发育的四个关键阶段（s1 - IB尚未在尾部出现；s2 -几个小IB在尾部开始骨化；s3 - IB迅速出现；s4 -所有IB形态成熟）。阿利新蓝和茜素红S染色结果显示，IBs从S2到S4逐渐形成，经历膜内骨化，未经历软骨期。采用等压标签相对和绝对定量（iTRAQ）方法共鉴定了3368个蛋白。功能注释显示，不同阶段差异表达的蛋白涉及钙、MAPK、Wnt、TGF-β和破骨细胞通路，这些通路在骨形成和分化中起关键作用。从S2到S4，与软骨细胞相关的3种蛋白（胶原9α1、stat1、tnc）没有明显变化；然而，与成骨细胞和骨形成和分化相关的蛋白（entpd5、casq1a、pvalb、anxa2a、anxa5）在S1到S2以及S3和S4的表达水平明显较高。这进一步表明IBs的发展并没有经历软骨阶段。TGF-β和Wnt通路的抑制剂在斑马鱼（sp7/eGFP）上进行了测试，结果表明这两种抑制剂都能显著延缓IB的发展。该研究提供了对硬骨鱼IB骨化模式的全面了解，有助于进一步阐明硬骨鱼IB发育的分子机制。
Intermuscular bones (IBs) specially exist in lower teleost fish and the molecular mechanism for its development remains to be clarified. In this study, different staining methods and comparative proteomics were conducted to investigate the histological structure and proteome of IB development in Megalobrama amblycephala, including four key IB developmental stages (S1—IBs have not emerged in the tail part; S2—several small IBs started to ossify in the tail part; S3—IBs appeared rapidly; S4—all the IBs appeared with mature morphology). Alcian blue and alizarin red S stained results indicated that IBs were gradually formed from S2 to S4, undergoing intramembranous ossification without a cartilaginous phase. A total of 3368 proteins were identified by using the isobaric tags for relative and absolute quantitation (iTRAQ) approach. Functional annotation showed that proteins which were differentially expressed among stages were involved in calcium, MAPK, Wnt, TGF-β, and osteoclast pathways which played a critical role in bone formation and differentiation. Three proteins (collagen9α1, stat1, tnc) associated with chondrocytes did not exhibit significant changes through S2 to S4; however, proteins (entpd5, casq1a, pvalb, anxa2a, anxa5) which associated with osteoblasts and bone formation and differentiation showed significantly a higher expression level from S1 to S2, as well as to S3 and S4. These further demonstrated that development of IBs did not go through a cartilaginous phase. The inhibitors of TGF-β and Wnt pathways were tested on zebrafish (sp7/eGFP) and the results indicated that both inhibitors significantly delayed IB development. This study provides a comprehensive understanding of the IB ossification pattern, which will help further elucidate the molecular mechanisms for IB development in teleosts.
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影响因子：
3.3
作者：
Long, Meng;Zhao, Juan;Li, Aihua
通讯作者：
Li, Aihua
DOI：
--
发表时间：
2008
期刊：
Chinese Journal of Zoology
影响因子：
--
作者：
Keming Zhong
通讯作者：
Keming Zhong
影响因子：
3.3
作者：
Sophia Fox AJ;Bedi A;Rodeo SA
通讯作者：
Rodeo SA
影响因子：
2.7
作者：
Kague E;Roy P;Asselin G;Hu G;Simonet J;Stanley A;Albertson C;Fisher S
通讯作者：
Fisher S
影响因子：
2.7
作者：
Florbela A. Vieira;M. Thorne;K. Stueber;M. Darias;Richard Reinhardt;Melody S. Clark;Enric Gisbert
通讯作者：
Florbela A. Vieira;M. Thorne;K. Stueber;M. Darias;Richard Reinhardt;Melody S. Clark;Enric Gisbert