Neuromuscular Transmission in Arteries and Veins

动脉和静脉中的神经肌肉传递

• 批准号: 6728114
• 负责人: Violeta N. Mutafova-Yambolieva
• 金额: $ 27.88万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-10 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6728114
• 关键词: CD38 molecule; adenosine triphosphate; artery; calcium channel; chloride channels; dogs; neuromuscular transmission; neurotransmitter receptor; neurotransmitter transport; norepinephrine; phosphatidylinositol 3 kinase; protein isoforms; protein kinase A; protein kinase C; purinergic receptor; ribose phosphate; vascular smooth muscle nervous control; veins; voltage /patch clamp; western blottings

DESCRIPTION (provided by applicant): The long-term goal of this laboratory is to forward our understanding of vascular neuroeffector mechanisms specifically related to plurichemical neurotransmission. The evidence is substantial that the resistive and capacitative vessels in the splanchnic circulation differ in their mechanical, electrical, and neurochemical responses to sympathetic nerve stimulation. However, the mechanisms by which neuroeffector processes differ in the arteries and veins remain unknown. In Aim 1 we propose to examine the hypothesis that the amount and/or ratio of neurally released norepinephrine (NE) and adenosine 5'-triphosphate (ATP) differ in artery and vein due to different expression levels or functional activity of neuronal Ca2+ channel proteins. Aim 2 is based on our recent observation that activation of postganglionic nerve terminals in mesenteric arteries and veins evokes release of adenosine 5'-diphosphoribose (ADPR), along with ATP, ADP, AMP, adenosine and NE. This represents a novel finding that may lead to identifying new mechanisms of neurovascular control. Arteries and veins show quantitative and qualitative differences in ADPR release. With regard to this, we propose to examine the hypothesis that the formation (primarily CD38-mediated) and release of ADPR or its precursors, NAD+ and cyclic ADPR, are differentially modulated by the activation of protein kinase A (PKA)-dependent pathways in artery and vein. In Aim 3 we hypothesize that electrical responses to nerve stimulation and NE are mediated by adrenoceptor/PI3-kinase/PKC regulation of Ca2+-activated CI- channels, which may have differential prevalence in artery and vein. These studies will involve experiments in canine mesenteric artery and vein to determine the nerve-evoked overflow of nucleotides, nucleosides and NE (HPLC-FLD and ECD techniques); neuronal Ca2+ channel expression and function (Western blotting, neurotransmitter release); PKA/CD38 interactions (immunoblotting, coprecipitation, protein phosphorylation, RT-PCR), as well as mechanisms of NE activation of ICaCI (patch-clamp techniques). This unprecedented combination of approaches should provide important insight into the complex mechanisms governing neural control in arteries and veins. The results may suggest strategies for interrupting or preventing pathological conditions associated with sympathetic hyperactivity and vascular dysfunctions such as hypertension, cardiogenic shock, congestive heart failure, hemorrhage.

描述（由申请人提供）：该实验室的长期目标是转发我们对与多元化学神经传递特别相关的血管神经作用机制的理解。证据证明，斑点循环中的电容性和电容性血管在其机械，电气和神经化学对交感神经刺激的反应上有所不同。但是，动脉和静脉的神经反源过程的机制仍然未知。在AIM 1中，我们建议检验以下假说：神经释放的去甲肾上腺素（NE）和5'-三磷酸腺苷（ATP）的数量和/或比率在动脉和静脉中由于不同的表达水平或神经元Ca2+通道蛋白的功能活性而在动脉和静脉中有所不同。 AIM 2是基于我们最近的观察结果，即肠系膜动脉和静脉中的神经后神经末端的激活唤起了腺苷5'-二磷咖啡（ADPR）的释放，以及ATP，ADP，ADP，AMP，AMP，腺苷，腺苷和Ne。这代表了一个新颖的发现，可能导致识别神经血管控制的新机制。动脉和静脉在ADPR释放中显示出定量和定性差异。关于这一点，我们建议研究以下假设：ADPR或其前体的释放（主要是CD38介导的）或其前体NAD+和环状ADPR是通过蛋白激酶A（PKA）依赖性途径在动脉和静脉中的激活而差异的。在AIM 3中，我们假设对神经刺激和NE的电响应是由Ca2+激活的CI-通道的肾上腺受体/PI3-激酶/PKC调节介导的，这可能在动脉和静脉中具有差异的发生率。这些研究将涉及犬肠系膜动脉和静脉的实验，以确定核苷酸，核苷和NE的神经诱发的溢出（HPLC-FLD和ECD技术）；神经元Ca2+通道表达和功能（蛋白质印迹，神经递质释放）； PKA/CD38相互作用（免疫印迹，共沉淀，蛋白质磷酸化，RT-PCR）以及NE激活的机制（Patch-Clamp Techniques）。这种空前的方法组合应提供对控制动脉和静脉神经控制的复杂机制的重要见解。结果可能表明中断或预防与交感神经多动和血管功能障碍相关的病理状况（例如高血压，心脏病性休克，充血性心力衰竭，出血）。