Characterization of leptin's effects in early developing and adult zebrafish

项目描述（申请人提供）：本项目的主要目的是进一步了解瘦素在人体脂肪代谢中的作用。瘦素是由脂肪细胞分泌的I型细胞因子，其结合中枢神经系统中的Jak/Stat受体。瘦素给药通常导致食欲下降和储存脂质的动员，然而瘦素也对生殖、发育、免疫功能和骨密度有影响。人们对瘦素的研究兴趣一直很大（自1994年发现以来，每年约有1000篇出版物），这无疑是因为瘦素作为一种对抗肥胖流行病的药物的潜力。由于瘦素的作用是复杂的，该领域的领导者呼吁用进化的方法来理解瘦素的功能。也就是说，如果我们能够理解瘦素在“低级”脊椎动物中的功能，我们就可以更好地理解和预测它在人类中的功能（类似于鱿鱼轴突，鲨鱼直肠腺等的方法）。在这里，我们建议研究瘦素的蛋白质表达和模式生物斑马鱼的发育的影响。 该提案的具体目的是：1）检验瘦素对早期斑马鱼胚胎的正常发育至关重要的假设，2）检验瘦素功能在哺乳动物和成年斑马鱼之间保守的假设。一套技术，包括重组蛋白生产，免疫印迹，免疫细胞化学，原位杂交，高速数字摄像，和吗啉代反义寡核苷酸将被采用。这些研究的结果将使人们更好地理解瘦素在人类中的作用，并为人类肥胖提供可能的治疗方法。公共卫生相关性：这项提案调查瘦素对斑马鱼发育和成年生理的影响。这些实验的结果可能会导致对人类肥胖的理解和治疗的改善。

DESCRIPTION (provided by applicant): The over-arching objective of this project is to further our understanding of leptin's role in fat metabolism of humans. Leptin is a type-I cytokine secreted by adipocytes that binds to Jak/Stat receptors in the central nervous system. Leptin administration typically leads to reduction in appetite and mobilization of stored lipid, however leptin also has effects on reproduction, development, immune function, and bone density. Intense research interest has been focused on leptin (~1000 publications/year since its discovery in 1994), undoubtedly because of its potential as a drug to combat the obesity epidemic. Because leptin's actions are complex, leaders in the field have called for an evolutionary approach to understanding leptin's function. That is, if we can understand leptin's function in 'lower' vertebrates, we can better understand and predict its function in humans (similar to the approach of squid axon, shark rectal gland, etc.). Here we propose to study leptin's effects on protein expression and development in the model organism zebrafish. The specific aims of the proposal are: 1) to test the hypothesis that leptin is essential for normal development in early zebrafish embryos and 2) to test the hypothesis that leptin function is conserved between mammals and adult zebrafish. A suite of techniques, including recombinant protein production, immunoblotting, immunocytochemistry, in situ hybridization, high-speed digital videography, and morpholino antisense oligonucleotides will be employed. Results from the proposed studies should lead to greater understanding of leptin's role in humans and to possible therapies for human obesity. PUBLIC HEALTH RELEVANCE: This proposal investigates leptin's effects on zebrafish development and adult physiology. Results from these experiments may lead to improved understanding of and therapy for human obesity.