The role of carboxypeptidase A6 in development

羧肽酶 A6 在发育中的作用

• 批准号: 7931898
• 负责人: Peter Jonathan Lyons
• 金额: $ 5.38万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7931898
• 关键词: Adult; Affect; Affinity Chromatography; Amino Acids; Axon; Biological Models; Brain; C-terminal; Carboxypeptidase; Cell Line; Cells; Confocal Microscopy; Cranial Nerves; DNA; Defect; Development; Diagnosis; Down-Regulation; Embryo; Enzyme Inhibition; Enzymes; Extracellular Matrix; Extracellular Matrix Proteins; Eye; Fellowship; Future; Genes; Goals; Immigration; In Situ Hybridization; Kallmann Syndrome; Kinetics; Lateral; Lead; Life; Light; Location; Mammalian Cell; Mass Spectrum Analysis; Mediating; Mesoderm; Mesoderm Cell; Methods; Microinjections; Microscopic; Microscopy; Modeling; Molecular; Mus; Muscle; Mutation; Nature; Nervous system structure; Neurons; Patients; Pattern; Peptide Hydrolases; Peptides; Physiological; Plasmids; Population; Post-Translational Protein Processing; Post-Translational Regulation; Process; Proliferating; Property; Proteins; Proteolysis; Publishing; Recipe; Reporter; Research; Role; Schizophrenia; Site; Specificity; Staging; Substrate Specificity; Syndrome; System; Systems Analysis; Techniques; Testing; Tissues; Tracer; Transgenic Organisms; Type 1 Duane Retraction Syndrome; Up-Regulation; Zebrafish; abducens nerve; axon guidance; axonal guidance; base; cell motility; hindbrain; in vivo; inhibitor/antagonist; knock-down; migration; nerve supply; nervous system development; nervous system disorder; neuronal guidance; olfactory bulb; overexpression; precursor cell; promoter; protein distribution; research study; skill acquisition; skills; subventricular zone; success; tool; zebrafish development

DESCRIPTION (provided by applicant): During development and throughout adult life certain populations of neurons are proliferating, migrating to their sites of action, and extending axons towards appropriate targets. These processes are regulated by many post-translational modifications, including proteolysis. Defects in these processes have been found to result in neurological disorders such as schizophrenia and Kallmann syndrome. A protease, carboxypeptidase A6 (CPA6), involved in cleavage of C-terminal hydrophobic amino acids, has been implicated in the mechanisms of cranial nerve development and axon guidance towards its target muscle. Specifically, a patient with a mutation disrupting CPA6 has been diagnosed with Duane syndrome, a defect in the innervation of the lateral rectus eye muscle by the abducens nerve. CPA6 is expressed in the lateral rectus eye muscle during development as well as in the adult olfactory bulb. This restricted expression in the adult olfactory bulb further implicates CPA6 in neuronal migration, as the olfactory bulb is one of the few locations of adult neuronal migration. The goal of this project is to investigate the role of CPA6 in regulating neuronal migration and axon guidance in the developing brain. First, the enzymatic properties of purified CPA6 will be assessed and substrates of CPA6 will be identified through established peptidomics techniques. The distribution of CPA6 will be characterized in detail throughout the development of the zebrafish using in situ hybridization, while the distribution of CPA6 in the mouse will be pursued by a collaborator. Finally, the genetically and microscopically tractable zebrafish system will be employed to assess the in vivo role of CPA6. Specific inhibitors as well as morpholino-mediated knockdown and plasmid- based overexpression of CPA6, followed by microscopic and physiological analysis, will be used to assess the role of CPA6 in neuronal migration and axonal guidance in vivo. This research will increase our understanding of post-translational regulation of neuronal guidance mechanisms during development. It will also shed light on the molecular defects leading to Duane syndrome, a syndrome affecting as much as 0.1% of the population.

描述(由申请人提供)：在发育期间和整个成年生活中，某些神经元群体正在增殖，迁移到它们的作用部位，并向适当的靶点延伸轴突。这些过程受许多翻译后修饰的调控，包括蛋白质分解。这些过程中的缺陷被发现会导致神经疾病，如精神分裂症和卡尔曼综合征。C端疏水氨基酸的裂解过程中涉及到一种名为羧肽酶A6(CPA6)的蛋白酶，它参与了脑神经发育和轴突导向靶肌肉的机制。具体地说，一名携带CPA6基因突变的患者已被诊断为杜安综合征，这是一种外展神经支配眼外直肌的缺陷。CPA6在发育过程中的眼外肌和成年嗅球中表达。这种在成年嗅球中的限制性表达进一步表明CPA6参与了神经元的迁移，因为嗅球是少数几个成年神经元迁移的位置之一。该项目的目标是研究CPA6在调节发育中的大脑中神经元迁移和轴突引导方面的作用。首先，将评估纯化的CPA6的酶性质，并通过建立的肽组学技术鉴定CPA6的底物。CPA6的分布将通过原位杂交在斑马鱼的整个发育过程中得到详细描述，而CPA6在小鼠体内的分布将由合作者进行研究。最后，将利用遗传和微观上易驯化的斑马鱼系统来评估CPA6在体内的作用。CPA6的特异性抑制剂以及吗啡介导的敲除和基于质粒的过度表达，以及随后的显微镜和生理分析，将被用来评估CPA6在体内神经元迁移和轴突引导中的作用。这项研究将增加我们对发育过程中神经元引导机制的翻译后调节的理解。它还将揭示导致杜安综合征的分子缺陷，杜安综合征影响多达0.1%的人口。