Acute and Chronic Afferent Engagement: Sympathetic and End Organ Responses

项目总结/摘要-总体 这是一个新的应用程序项目赠款的NHLBI由三名研究人员位于 宾夕法尼亚州赫尔希市的宾夕法尼亚州立大学医学院。所有调查人员都高度 多产的科学家所有项目和核心领导人都是合作的，所有人都将从中受益匪浅 程序.我们认为该应用程序满足PPG的标准，因为：1）有一个统一的主题，即 运动加压反射（EPR）在外周动脉疾病（PAD）中加重， 有严重的心血管后果。两种肢体缺血模型将被用来探讨这一点 反射：（a）人类受试者的PAD（项目1）;和（B）大鼠的后肢动脉闭塞（项目2和3）; 2） 项目之间存在概念协同作用，将导致心血管领域的重要进展 生理学和病理生理学; 3）该PPG具有广泛定义的目标，可以在5 4）许多提出的想法和概念是以前的直接结果 项目负责人之间的互动。Sinoway博士将担任PPG项目总监（PD）。他 丰富的科学和管理经验，担任NIH资助的临床和 宾夕法尼亚州立大学转化科学研究所，以及2004年至2016年PPG的PD。在项目1中，Sinoway博士 将研究人类受试者运动加压反射增强对心血管的影响 PAD和间歇性跛行他将检查强化反射对周围神经的影响， 和心脏循环。在项目2中，李博士将研究促炎细胞因子在改变 后肢动脉闭塞后血压对后肢收缩的反应。他还将研究如何 促炎细胞因子改变这些动物DRG神经元中的信号转导。在项目3中，博士。 考夫曼将研究背根神经节神经元上的EP 4、ASIC 3和P2 X3受体在加重背根神经节细胞凋亡中的作用。 后肢动脉闭塞啮齿动物后肢收缩的升压反射。两个核心实验室 一个行政（核心A），将监督所有行政职能和一个转染核心 实验室（核心B）将设计有效的siRNA序列，并提供编码这些序列的cDNA克隆 “短发夹”核苷酸序列，进行DRG（L4-L5）的体内和体外转染， 腓肠肌，并采用定量RT-PCR（mRNA水平）验证蛋白质敲低， 蛋白质印迹法（蛋白质定量）。

PROJECT SUMMARY/ABSTRACT – OVERALL This is a new application of a program project grant to the NHLBI by three investigators located within the College of Medicine of the Pennsylvania State University in Hershey, Pennsylvania. All investigators are highly productive scientists. All Project and Core Leaders are collaborative and all will greatly benefit from this program. We believe this application fulfills the criteria for a PPG because: 1) there is a unifying theme namely that the exercise pressor reflex (EPR) is accentuated in peripheral artery disease (PAD) and this accentuation has significant cardiovascular consequences. Two models of limb ischemia will be employed to explore this reflex: (a) PAD in human subjects (Project 1); and (b) hindlimb arterial occlusion in rats (Projects 2 and 3); 2) conceptual synergy exists between the projects that will lead to important advances in cardiovascular physiology and pathophysiology; 3) this PPG has broadly defined goals that can be accomplished within the 5 year funding period; and 4) many of the ideas and concepts proposed are a direct result of previous interactions between the project leaders. Dr. Sinoway will be the PPG Program Director (PD). He has substantial scientific and administrative experience, serving as the PD of the NIH funded Clinical and Translational Science Institute at Penn State, and PD of a PPG from 2004 to 2016. In Project 1, Dr. Sinoway will examine the cardiovascular consequences of an accentuated exercise pressor reflex in human subjects with PAD and intermittent claudication. He will examine the impact the accentuated reflex has on the peripheral and cardiac circulations. In Project 2, Dr. Li will examine the role pro-inflammatory cytokines play in altering the blood pressure response to hindlimb contraction after hindlimb arterial occlusion. He will also examine how pro-inflammatory cytokines alter signal transduction in DRG neurons of these animals. In Project 3, Dr. Kaufman will examine the role EP4, ASIC3 and P2X3 receptors on DRG neurons play in accentuating the pressor reflex to hindlimb contraction in rodents with hindlimb arterial occlusion. Two core laboratories are proposed; an Administrative (Core A) that will oversee all administrative functions and a Transfection Core Laboratory (Core B) that will design effective siRNA sequences and provide cDNA clones that code for these `short hairpin' nucleotide sequences, perform the in vivo and in vitro transfection of DRG (L4-L5) and gastrocnemius muscle, and verify protein knockdown employing quantitative RT-PCR (mRNA levels) and Western blotting (protein quantification).