Localization, Interactions, And Functions of Synaptotagmins in the Pituitary

突触结合蛋白在垂体中的定位、相互作用和功能

• 批准号: 8185499
• 负责人: Edwin R Chapman
• 金额: $ 39.07万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8185499
• 关键词: Ablation; Anterior Pituitary Gland; Binding; Biological; Biological Process; Blood Circulation; Cell membrane; Cells; Co-Immunoprecipitations; Corticotropin; Coupling; D Cells; Data; Defect; Dense Core Vesicle; Dependence; Electric Capacitance; Electron Microscopy; Endocrine; Endocytosis; Exocytosis; Fluid Balance; Follicle Stimulating Hormone; Frequencies; Gene Targeting; Genes; Genetic; Gold; Growth; Homologous Gene; Hormones; Immunoassay; Immunofluorescence Immunologic; In Vitro; Knock-out; Knockout Mice; Light; Liposomes; Luteinizing Hormone; Measurement; Measures; Mediating; Mental disorders; Metabolic Diseases; Metabolism; Mus; Nerve; Neurologic; Neurons; Organelles; Oxytocin; PC12 Cells; Peptides; Phosphatidylserines; Physiological; Physiology; Pituitary Gland; Pituitary Hormones; Population; Posterior Pituitary Gland; Preparation; Prolactin; Property; Protein Family; Protein Isoforms; Proteins; Psyche structure; Reproduction; Reproductive Health; Role; Running; S-nitro-N-acetylpenicillamine; SNAP receptor; Somatotropin; Stimulus; Stress; Structure; Synaptic Vesicles; System; Testing; Thyrotropin; Tracer; Transgenic Mice; Vasopressins; Vesicle; Work; biological adaptation to stress; improved; nervous system disorder; neurotransmitter release; novel; offspring; patch clamp; peptide hormone; programs; research study; response; sensor; synaptotagmin; synaptotagmin I; synaptotagmin IV; synaptotagmin VII; syntaxin; target SNARE proteins; voltage

DESCRIPTION (provided by applicant): Synaptotagmin 1, a protein found on synaptic vesicles, serves as a Ca2+ sensor in neurotransmitter release. This protein has 16 homologues, and while these other proteins are widely thought to serve related functions essentially nothing is known about where they are or what they do. Preliminary data presented in this application shows that several synaptotagmin isoforms are abundant in the posterior and/or anterior pituitary, glands that release peptide hormones that control essentially all vital aspects of mammalian physiology. Prior to these new preliminary results, the pituitary had been established as a preparation amenable to quantitative biophysical study of how peptide-containing vesicles fuse with the plasma membrane to release their content into the circulation. Thus, the finding of novel synaptotagmin isoforms in the pituitary offers a unique opportunity to test their function in detail and elucidate their precise roles in excitation-secretion coupling. This project will study the pituitary synaptotagmin isoforms with complementary approaches, investigating their binding properties in vitro, and their localization and physiological function in the pituitary. The functional studies employ a strategy of using transgenic mice for gene ablation of specific synaptotagmin isoforms, and gene targeting to identify specific cell populations that express those isoforms for electrical recording. By performing capacitance recording in identified cells and nerve terminals we will determine how syt isoforms regulate exocytosis in terms of Ca2+ sensitivity and fusion pores. The experiments proposed here will test the functions of synaptotagmins 4, 7, 9, 10, 11, and 12 in the release of hormones that control growth, metabolism, stress, fluid balance, and reproduction. PUBLIC HEALTH RELEVANCE: These experiments will shed light on the function of a class of proteins with broad roles in neurological, mental, and endocrine function and thus contribute to improving treatment of neurological disorders and mental illness. By illuminating the mechanisms of control of the pituitary hormones oxytocin, vasopressin, growth hormone, adrenocorticotropin, thyroid stimulating hormone, follicle stimulating hormone, luteinizing hormone, and prolactin, these studies will improve our understanding of growth defects, abnormal stress responses, metabolic disorders, and reproductive health.

描述（由申请人提供）：突触结合蛋白1是一种在突触囊泡上发现的蛋白质，在神经递质释放中充当Ca 2+传感器。这种蛋白质有16个同源物，虽然这些其他蛋白质被广泛认为具有相关功能，但基本上对它们的位置或功能一无所知。本申请中提供的初步数据显示，几种突触结合蛋白同种型在垂体后叶和/或垂体前叶中丰富，垂体后叶和/或垂体前叶是释放肽激素的腺体，肽激素基本上控制哺乳动物生理学的所有重要方面。在这些新的初步结果之前，垂体已被确定为一种适合定量生物物理研究的制剂，研究含肽囊泡如何与质膜融合以将其内容物释放到循环中。因此，在垂体中发现新的突触结合蛋白亚型提供了一个独特的机会，以测试其功能的细节，并阐明其在兴奋分泌耦合的确切作用。本计画将以互补的方法研究垂体突触结合蛋白的异构体，探讨其在体外的结合特性，以及其在垂体中的定位与生理功能。功能研究采用了一种策略，使用转基因小鼠的基因消融特定的synaptotagmin亚型，基因靶向，以确定特定的细胞群，表达这些亚型的电记录。通过在确定的细胞和神经末梢进行电容记录，我们将确定syt亚型如何调节胞吐作用的Ca 2+敏感性和融合孔。本文提出的实验将测试突触结合蛋白4、7、9、10、11和12在控制生长、代谢、应激、体液平衡和生殖的激素释放中的功能。 公共卫生关系：这些实验将揭示一类蛋白质的功能，这些蛋白质在神经，精神和内分泌功能中具有广泛的作用，从而有助于改善神经系统疾病和精神疾病的治疗。通过阐明垂体激素催产素、加压素、生长激素、促肾上腺皮质激素、促甲状腺激素、促卵泡激素、促黄体激素和催乳素的控制机制，这些研究将提高我们对生长缺陷、异常应激反应、代谢紊乱和生殖健康的理解。