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(申请人提供)：在生命的每个阶段，脑血管张力和血流(CBF)的调节是至关重要的。许多新生儿，特别是那些早产儿，在调节大脑血液流动方面存在严重问题。这种失调可能会导致脑室和生发基质出血的严重后果，并伴有长期的神经后遗症。目前的研究试图了解成熟发育如何改变胎儿/早产儿和成人脑血管系统的基本信号转导机制。这个项目是广泛的、多学科的，并使用生理学、细胞、生化和分子方法进行垂直整合。基于几十年的研究结果，我们将检验这一总体假设，即成熟发育与大脑动脉(CA)收缩反应的显著变化有关，继而发生变化的是α1-肾上腺素能受体(1-AR)亚型和/或特定蛋白激酶C亚型(PKC)介导的下游钙依赖和非钙依赖的信号转导通路。一个相关的假说是，与成人的“收缩”表型相比，发育显著改变了1-AR亚型和特定的PKC同工酶介导的原癌基因和代表血管平滑肌“合成”和/或“增殖”表型的基因的表达。四个具体目标如下。1)特定的1-AR亚型和下游效应蛋白在信号转导中的作用是什么？2)特定的PKC亚型、细胞外信号调节激酶(ERKs)、Rho A/Rho激酶和相关的激酶在信号转导中的作用是什么？3)特定的1-AR亚型和PKC亚型在血管发育表型的基因调控中起什么作用？4)目前在这些信号转导和基因调控途径中目前描述较少的其他信号转导蛋白的作用是什么？在绵羊胎儿、新生儿和成年CA中，我们将进行激动剂诱导的收缩和细胞内[Ca2+]测量、免疫印迹、RT-PCR、共聚焦显微镜、流式细胞仪、2D-凝胶-质谱仪、双链RNA或吗啡的基因沉默、基因上调、基因微阵列/通路分析和基因/蛋白质发现。从科学上讲，这些研究将促进我们对脑血管随着胎儿、新生儿和成人的发育而发生表型和功能变化的基本机制的理解。在临床上，这些研究涉及了解胎儿和/或早产儿脑血管张力、压力和血流调节的基础，以及导致脑出血和严重神经后遗症的调节失调。