Cerebral Artery Alpha1 Adrenergic and PKC Regulatory Mechanisms

脑动脉 Alpha1 肾上腺素能和 PKC 调节机制

• 批准号: 8811457
• 负责人: Ravi Goyal
• 金额: $ 30.77万
• 依托单位: LOMA LINDA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8811457
• 关键词: Adrenergic Agents; Adrenergic Receptor; Adult; Agonist; Americas; Biochemical; Birth Rate; Blood; Blood Vessels; Blood flow; Brain; Brain Injuries; Calcium; Caring; Cerebral hemisphere hemorrhage; Cerebrovascular Circulation; Cerebrum; Child; Clinical; Confocal Microscopy; Core-Binding Factor; Country; Development; Disabled Persons; Disease; Double-Stranded RNA; Expenditure; Extracellular Signal Regulated Kinases; Face; Family; Fetus; Flow Cytometry; Gel; Gene Expression; Gene Expression Regulation; Gene Proteins; Gene Silencing; Genes; Health; Healthcare; Heart Diseases; Hypoxia; Impairment; Infant; Intraventricular; Isoenzymes; L-Type Calcium Channels; Life; Live Birth; Lung diseases; MAPK3 gene; Mass Spectrum Analysis; Measurement; Mediating; Membrane; Messenger RNA; Metabolism; Molecular; Neurologic; Newborn Infant; Pathway Analysis; Pathway interactions; Perinatal subependymal hemorrhage; Phenotype; Phosphotransferases; Physiological; Population; Potassium; Pregnancy; Premature Birth; Prematurity of fetus; Prevalence; Protein Isoforms; Protein Kinase C; Proteins; Proto-Oncogenes; Regulation; Research; Resources; Reverse Transcriptase Polymerase Chain Reaction; Rho-associated kinase; Role; Science; Secondary to; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Smoke; Staging; Stream; Testing; Transduction Gene; United States; Up-Regulation; Vascular Smooth Muscle; Western Blotting; Woman; Work; adrenergic; base; cerebral artery; cerebrovascular; clinically relevant; density; disability; extracellular; fetal; handicapping condition; large-conductance calcium-activated potassium channels; maternal stress; multidisciplinary; premature; prenatal; pressure; programs; protein activation; receptor; response; rho; therapeutic target

DESCRIPTION (provided by applicant): At every stage of life the regulation of cerebral vascular tone and blood flow (CBF) is of vital importance. Many newborn infants, particularly those that are premature, have serious problems in the regulation of blood flow to their brains. This dysregulation may have serious consequences with intraventricular and germinal matrix hemorrhage with long-term neurological sequelae. The present studies seek to understand whereby maturational development alters fundamental signal transduction mechanisms in the cerebrovasculature of the fetus/premature newborn and the adult. This project is broadly based, multidisciplinary, and vertically integrated using physiologic, cellular, biochemical, and molecular approaches. Based on several decades of research findings, we shall test the overall hypothesis that maturational development is associated with significant changes in cerebral artery (CA) contractile responses secondary to altered alpha1-adrenergic-receptor (1-AR) subtype and/or specific protein kinase C isoform (PKC)-mediated downstream Ca2+-dependent and Ca2+-independent signal transduction pathways. An associated hypothesis is that development significantly alters 1-AR-subtype- and specific PKC isozyme-mediated expression of proto-oncogenes and genes representing vascular smooth muscle "synthetic" and/or "proliferative" phenotypes, as compared to adult "contractile" phenotype. Four Specific Aims are as follows. 1) What is the role of specific 1-AR subtypes and downstream effector proteins in signal transduction? 2) What is the role of specific PKC isoforms, extracellular signal regulated kinases (ERKs), Rho A/Rho kinases, and related kinases in signal transduction? 3) What is the role of specific 1-AR subtypes and PKC isoforms in gene regulation of developing vascular phenotypes? 4) What is the role of other signal transduction proteins presently poorly described in these signal transduction and gene regulation pathways? In ovine fetal, newborn, and adult CA, we will perform agonist-induced contractility and intracellular [Ca2+] measurements, Western immunoblots, RT- PCR, confocal microscopy, flow cytometry, 2D-gel-mass spectroscopy, gene silencing by double stranded RNA or morpholinos, gene upregulation, gene microarray/pathway analysis, and gene/protein discovery. Scientifically, the studies will advance our understanding of basic mechanisms whereby cerebral vessels change phenotypically and functionally with development from fetus, to newborn, to adult. Clinically, the studies relate to understanding the basis of the regulation of cerebral vascular tone, pressure, and blood flow in the fetus and/or premature newborn infant, and its dysregulation that results in intracerebral hemorrhage and serious neurologic sequelae.

描述（由申请人提供）：在生命的每个阶段，脑血管张力和血流（CBF）的调节至关重要。许多新生婴儿，尤其是那些早产的婴儿，在调节流向其大脑的血液中有严重的问题。这种失调可能会在脑室室内和生发基质出血带有长期神经系统后遗症。本研究旨在了解成熟发展会改变胎儿/早产新生儿和成人脑脑腔中的基本信号转导机制。该项目是基于生理，细胞，生化和分子方法的广泛基于多学科和垂直整合的。根据几十年的研究结果，我们应检验总体假设，即成熟发展与脑动脉（CA）收缩反应的显着变化有关，其继发于改变α1-肾上腺素能受体受体（1-AR）亚型和/或特定的蛋白质激酶CINase Cisoform（PKC）同工型（PKC）介导的下流式CA2+-CA2+依赖性CA2+依赖性和CA2+依赖性和Ca2+ipperdeddected+ipperdeddected+ipperded+ipperdedected+依赖性和Ca2+依赖性。一个相关的假设是，与成人“收缩”表型相比，发展的开发显着改变了1- ar-囊型和特定的PKC同工酶介导的原始基因和代表血管平滑肌“合成”和/或“增殖”表型的基因，与成人“收缩”表型相比。四个具体目标如下。 1）特定1-AR亚型和下游效应蛋白在信号转导中的作用是什么？ 2）特定PKC同工型，细胞外信号调节激酶（ERK），RHO A/RHO激酶和相关激酶在信号转导中的作用是什么？ 3）特异性1-AR亚型和PKC同工型在发育中的血管表型调节中的作用是什么？ 4）目前在这些信号转导和基因调节途径中描述的其他信号转导蛋白的作用是什么？ In ovine fetal, newborn, and adult CA, we will perform agonist-induced contractility and intracellular [Ca2+] measurements, Western immunoblots, RT- PCR, confocal microscopy, flow cytometry, 2D-gel-mass spectroscopy, gene silencing by double stranded RNA or morpholinos, gene upregulation, gene microarray/pathway analysis, and gene/protein发现。从科学上讲，这些研究将提高我们对基本机制的理解，从而随着胎儿的发育，到新生儿再到成人的发育，在表型和功能上改变了脑血管。从临床上讲，研究与了解胎儿和/或早产新生婴儿的脑血管张力，压力和血流的基础有关，以及导致脑内出血和严重神经系统后遗症的失调。