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

描述（由申请人提供）：这个项目的首要目标是进一步了解瘦素在人类脂肪代谢中的作用。瘦素是一种由脂肪细胞分泌的i型细胞因子，与中枢神经系统的Jak/Stat受体结合。瘦素的使用通常会导致食欲下降和储存的脂质动员，但瘦素也对生殖、发育、免疫功能和骨密度有影响。人们对瘦素的研究兴趣非常浓厚（自1994年发现以来，每年约有1000篇论文发表），毫无疑问，这是因为它有可能成为一种对抗肥胖流行的药物。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，如果我们能理解瘦素在“低等”脊椎动物中的功能，我们就能更好地理解和预测它在人类中的功能（类似于鱿鱼轴突、鲨鱼直肠腺等的研究方法）。在此，我们拟研究瘦素对模式生物斑马鱼蛋白表达和发育的影响。该提案的具体目的是：1)验证瘦素对早期斑马鱼胚胎正常发育至关重要的假设；2)验证瘦素功能在哺乳动物和成年斑马鱼之间是保守的假设。将采用一系列技术，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morpholino反义寡核苷酸。拟议研究的结果将有助于更好地了解瘦素在人体中的作用，并为人类肥胖提供可能的治疗方法。公共卫生相关性：本提案研究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提高对人类肥胖的理解和治疗。

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.