Cell and Molecular Biology of Marine Elasmobranchs

描述（由申请人提供）：本提案响应了“非哺乳动物模型和相关研究生物材料的开发”计划。具体来说，该计划公告提到“物种，如鱼类......以及用于非哺乳动物的细胞系和细胞培养模型，其中培养系统尚未充分开发但将被广泛使用。“我们建议从海洋板鳃类动物（鲨鱼和溜冰鞋）中开发细胞系，这些动物在生物医学研究中被广泛利用，作为生理学、内分泌学、毒理学和基因组学的模型。目前，还没有同源的体外系统，研究人员使用这些模型可以测试假设或确认预测在分子水平上，新技术的应用是必要的。驱动该项目的前提是，可以通过采取非常规方法来完成特定细胞类型的体外培养，包括去除或减少血清素培养基组分，并替换细胞类型特异性补充剂，如肽生长因子。我们已经使用这种方法从斑马鱼、河豚（河豚）和其他鱼类物种中产生细胞系。这项工作将利用沙漠山岛生物实验室的独特资源，这是一个致力于研究海洋生物以发现对人类健康具有重要意义的生物机制的研究机构，以及使用角鲨（Squalus acanthias）和小鳐（Raja erinacea）作为研究模型的世界中心。我们建议从（1）角鲨的肝、脑、肾和盐（直肠）腺，以及（2）小鳐的肝和胚胎中培养具有分化能力的、持续增殖的细胞系。我们还建议（3）开发在这些细胞系中进行DNA转染和质粒衍生基因表达的系统。成功将为原创和创新实验提供一个平台，加速推动生物医学研究多个领域的概念的发展，这些领域由大量使用这些模型的研究人员来解决。

DESCRIPTION (provided by applicant): This proposal responds to the Program "Development of Non-mammalian Models and Related Biological Materials for Research." Specifically, the program announcement mentions "Species such as fishes...and cell lines and cell culture models for non-mammalian animals where culture systems are not well developed but would be widely used." We propose to develop cell lines from marine elasmobranchs (sharks and skates), animals that are broadly exploited in biomedical research as models in physiology, endocrinology, toxicology and genomics. Currently there are no homologous in vitro systems by which investigators using these models can test hypotheses or confirm predictions at a molecular level, and application of new technologies is needed. The premise driving this project is that in vitro culture of specialized cell types can be accomplished by taking non-conventional approaches, including removal or reduction of serurnas a medium component and replacement with cell type-specific supplements, such as peptide growth factors. We have used this approach to generate cell lines from zebrafish, pufferfish (fugu), and other fish species. This work will take advantage of the unique resources of the Mount Desert Island Biological Laboratory, a research facility dedicated to the study of marine organisms to discover biological mechanisms of significance to human health, and world center for the use of the dogfish shark (Squalus acanthias) and little skate (Raja erinacea) as research models. We propose to develop differentiation-competent, continuously proliferating cell lines from (1) liver, brain, kidney and salt (rectal) gland of the dogfish shark, and (2) liver and embryo of the little skate. We also propose to (3) develop systems for DNA transfection and plasmid-derived gene expression in these cell lines. Success will provide a platform for original and innovative experiments, accelerating the development of concepts driving multiple areas of biomedical research addressed by the large number of investigators using these models.