Rescue of Chga knockout phenotype by CHGA-BAC transgene

通过 CHGA-BAC 转基因拯救 Chga 敲除表型

• 批准号: 7191652
• 负责人: SUCHETA M VAINGANKAR
• 金额: $ 10.67万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-15 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7191652
• 关键词: Anabolism; Bacterial Artificial Chromosomes; Biogenesis; Blood Pressure; CHGA gene; Catecholamines; Cause of Death; Cessation of life; Characteristics; Cholinergic Agents; Chromaffin granule; Chromogranin A; Coronary heart disease; Developed Countries; Developing Countries; Diabetes Mellitus; Drosophila pros protein; Employee Strikes; Endopeptidases; Genes; Genetic Polymorphism; Heart Diseases; Human; Hypertension; In Vitro; Knock-out; Knockout Mice; Laboratories; Mus; Nicotine; Nicotinic Agonists; Patients; Peptide Hydrolases; Peptides; Phenotype; Process; Regulation; Risk Factors; Single Nucleotide Polymorphism; Smoke; Testing; Tobacco; Tobacco use; Transgenes; Transgenic Organisms; Variant; Vesicle; cholinergic; chromogranin A (344-364); desensitization; in vivo; mouse model; mutant; prenatal; response

DESCRIPTION (provided by applicant): Chromogranin A (CGA) is co-stored with catecholamines in chromaffin granules and co-released in response to sympathoadrenal stimulation. From CGA pro-protein several biologically active peptides including catecholamine release-inhibitory peptide catestatin are derived. Nicotine, the primary component of tobacco, stimulates secretion of catecholamines, augments biosynthesis of CGA, and, as a nicotinic cholinergic agonist, promotes desensitization of catecholamine secretion. In developed countries, tobacco use is the largest single cause of premature death and heart disease the most common cause of death. Studies have demonstrated that smoking, diabetes and hypertension are major risk factors for coronary heart disease. Catestatin level is diminished in hypertensive patients. Chga knockout (Ko) mice have been generated in this laboratory. Their striking phenotypic characteristics include elevated systolic blood pressure (SBP), decreased catecholamines levels, loss of chromaffin granules and partial prenatal lethality. The proposed study specifically aims to: 1. Rescue the Chga-Ko phenotype by introducing the human CHGA gene using bacterial artificial chromosome (BAC) transgenic approach. 2. Sequencing of the human CHGA gene, revealed three non-synonymous SNPs (single nucleotide polymorphism) in the catestatin sequence. The proposed study will analyze the differences in regulation of SBP, catecholamine levels and nicotine cholinergic responses between the wild type and mutant variants of CGA. Variant catestatin peptides will be synthesized and administered into Chga -/- mice. The 3 mutant CHGA-BAC mouse models (Gly364Ser, Pro370Leu, and Arg374GIn) will also be generated to explore the significance of such polymorphisms. 3. Finally, the CGA variants will be assessed for processing by proteases in vitro and in vivo.

描述（由申请人提供）：嗜铬颗粒蛋白A （CGA）与儿茶酚胺共同储存在嗜铬颗粒中，并在交感肾上腺刺激下共同释放。从CGA前蛋白衍生出多种生物活性肽，包括儿茶酚胺释放抑制肽catestatin。尼古丁是烟草的主要成分，刺激儿茶酚胺的分泌，增加CGA的生物合成，并且作为一种烟碱能激动剂，促进儿茶酚胺分泌的脱敏。在发达国家，烟草使用是过早死亡的最大单一原因，心脏病是最常见的死亡原因。研究表明，吸烟、糖尿病和高血压是冠心病的主要危险因素。高血压患者睾酮水平降低。Chga基因敲除（Ko）小鼠已在本实验室产生。其显著的表型特征包括收缩压升高，儿茶酚胺水平降低，染色质颗粒丢失和部分产前死亡率。建议研究的具体目的是：