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快速改编斑马鱼和青鳉的分子资源以建立河口/海洋模型。

Rapid adaptation of molecular resources from zebrafish and medaka to develop an estuarine/marine model.

作者信息

Chen Xueping, Li Li, Wong Chris Kong Chu, Cheng Shuk Han

机构信息

Centre for Marine Environmental Research and Innovative Technology (MERIT), Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong SAR, China.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2009 May;149(4):647-55. doi: 10.1016/j.cbpc.2009.01.009. Epub 2009 Jan 23.

Abstract

Many estuary and coastal waters are highly threatened by heavy anthropogenic pollutants. Oryzias melastigma, also called O. dancena, a marine medaka that showed sensitive response to hypoxia and estrogenic endocrine disruptors in previous studies, is becoming a sentinel species for marine ecotoxicology studies. However, the lack of strong molecular foundation and knowledge of early developmental stages hampers its practical applications. Combining our research strength on zebrafish embryos, this study revealed both morphological and molecular (at mRNA and protein levels) development of embryos of this emergent model. Whole mount immunostaining technique specific for O. melastigma was successfully developed based on zebrafish standard protocols. We demonstrated that 17 out of 61 primary antibodies, which were previously tested in zebrafish, showed specific immunoreactivity with O. melastigma. These antibodies clearly illustrated the embryonic development of target tissues (principally neurons) in this medaka. Additionally, partial cDNA fragments of 11 organ-specific marker genes were isolated according to genomic resources of zebrafish, Japanese medaka and other fishes. Of the 11 genes, 8 are widely used as organ markers and their expression patterns were remarkably similar to their homologues in zebrafish and Japanese medaka. The expression profiles of the remaining 3 genes in fish are reported for the first time. These molecular markers (17 antibodies and 11 mRNA probes) can be used as responsive indicators in environmental toxicity evaluation. Moreover, this study brought forward and demonstrated the advantage of transferring techniques and resources from one model to another to hasten the research of interest.

摘要

许多河口和沿海水域受到严重人为污染物的高度威胁。黑背青鳉,也被称为丹氏青鳉,一种在先前研究中对缺氧和雌激素内分泌干扰物表现出敏感反应的海洋青鳉,正成为海洋生态毒理学研究的指示物种。然而,缺乏坚实的分子基础以及对早期发育阶段的了解阻碍了其实际应用。结合我们在斑马鱼胚胎方面的研究优势,本研究揭示了这种新兴模型胚胎的形态和分子(mRNA和蛋白质水平)发育情况。基于斑马鱼标准方案成功开发了针对黑背青鳉的全胚胎免疫染色技术。我们证明,先前在斑马鱼中测试的61种一抗中有17种与黑背青鳉表现出特异性免疫反应。这些抗体清晰地展示了这种青鳉中靶组织(主要是神经元)的胚胎发育。此外,根据斑马鱼、日本青鳉和其他鱼类的基因组资源分离出了11个器官特异性标记基因的部分cDNA片段。在这11个基因中,有8个被广泛用作器官标记,它们的表达模式与斑马鱼和日本青鳉中的同源物非常相似。其余3个基因在鱼类中的表达谱首次被报道。这些分子标记(17种抗体和11种mRNA探针)可作为环境毒性评估中的反应指标。此外,本研究提出并证明了将技术和资源从一种模型转移到另一种模型以加速相关研究的优势。

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