Cell and Molecular Biology of Marine Elasmobranchs

描述(申请人提供)：本提案响应项目“非哺乳类动物模型及相关生物研究材料的开发”。具体而言，节目公告提到了“鱼类等物种……以及非哺乳类动物的细胞系和细胞培养模式，这些地方的培养系统并不发达，但会得到广泛应用。”我们建议从海洋弹簧鱼(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，使用这些模型的研究人员还没有相应的体外系统来检验假设或在分子水平上确认预测，需要应用新的技术。该项目的前提是，特殊细胞类型的体外培养可以通过采取非常规方法来完成，包括去除或减少血清作为培养基组分，并替换为特定细胞类型的补充剂，如多肽生长因子。我们已经用这种方法从斑马鱼、河豚(河豚)和其他鱼类中产生了细胞系。这项工作将利用沙漠山生物实验室的独特资源，该实验室是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)和小溜冰(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)小冰鲨的肝脏和胚胎中培养出分化能力强、持续增殖的细胞系。我们还建议(3)在这些细胞系中建立DNA转染和表达质粒源基因的系统。Success将为原创和创新的实验提供一个平台，加速概念的发展，推动大量使用这些模型的研究人员解决生物医学研究的多个领域。

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.