Investigating Neural Processing of Cerebrovascular Dynamics via Calcium Imaging of Vascular Cells and Neurons, and by Optogenetic Vascular Pertubation, In Vivo

通过血管细胞和神经元的钙成像以及体内光遗传学血管微管研究脑血管动力学的神经处理

• 批准号: 10458534
• 负责人: Eric M Klein
• 金额: $ 2.37万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10458534
• 关键词: Address; Adenosine; Adenosine A1 Receptor; Affect; Alzheimer' s Disease; Anatomy; Area; Arteries; Astrocytes; Automobile Driving; Basic Science; Blood Vessels; Blood capillaries; Blood flow; Calcium; Caliber; Cations; Cells; Central Nervous System Diseases; Cerebrovascular Circulation; Cerebrovascular system; Characteristics; Chlorides; Clinical Research; Communication; Data; Disease; Endothelium; Environment; Event; Frequencies; Functional Magnetic Resonance Imaging; Glutamates; Halorhodopsins; Homeostasis; Hyperemia; Hypertension; Image; Ischemia; Knowledge; Lead; Location; Maps; Mediating; Medicine; Mentorship; Multiple Sclerosis; Nature; Nerve Block; Neuraxis; Neuroglia; Neurons; Neurosciences; Neurosciences Research; Outcome; Parkinson Disease; Pharmacology; Population; Pump; Reporting; Research; Research Training; Role; Science; Sensory; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Somatosensory Cortex; Source; Stereotyping; System; Tactile; Testing; Training; Universities; Writing; antagonist; arteriole; base; brain endothelial cell; calcium indicator; cerebrovascular; constriction; experimental study; glutamatergic signaling; hemodynamics; in vivo; in vivo two-photon imaging; insight; neural circuit; neuronal cell body; neurovascular; optogenetics; prevent; programs; receptor; relating to nervous system; response; tactile stimulation; therapy development; two-photon

PROJECT SUMMARY Understanding neural-vascular communication is vital to clinical and basic research. Perivascular neuron (PVN) activity can drive cerebral blood vessel dynamics. However, the impact of vascular events on neural activity has been only sparsely investigated. Our lab has found that a population of PVNs in primary somatosensory cortex (SI) encode cerebrovascular activity in vivo. However, the nature of this encoding, and its anatomical organization, is untested. Vessel-to-PVN signaling may support vascular homeostasis and rich communication across systems. These signals are relevant for research using blood flow to map neural activity (e.g., fMRI). Investigating perturbations of this signaling may elucidate mechanisms of cerebrovascular disfunction (e.g., as in ischemia, Parkinson’s Disease, and M.S.). To analyze PVN encoding of vascular activity, I will use in vivo two-photon imaging of neural and vascular cells, and optogenetics to perturb vessels and analyze the PVN response. In Aim I, I will test the hypothesis that vascular-encoding PVNs occur commonly in SI, and their activity is organized by cortical layer and vascular compartment, by expressing calcium indicators (jRGECO1a) in neurons and (GCaMP6f) in vascular endothelia to image their activity simultaneously. My preliminary data identified spatially distinct calcium events in the vascular signal that predict subsequent PVN activity. In this paradigm, the frequency of vessel responsive PVNs will be categorized by their stereotyped activity and anatomical location. Preliminary data in our lab has also shown that selective optogenetic vascular drive can modulate PVN activity. In Aim II, I will test the hypothesis that PVNs driven by optogenetically evoked vascular diameter changes will also be organized anatomically by their activity, that and their response to endogenous vascular events will parallel their response to optogenetic vascular drive. I will optogenetically constrict SI blood vessels by driving endothelial channelrhodopsin, dilate them with smooth muscle halorhodopsin, and evoke natural tactile driven functional hyperemia, to analyze the responses of PVNs expressing GCaMP6s. In Aim III, I will test the hypothesis that PVN responses to optogenetically driven vascular activity can be pharmacologically perturbed by TRPV4 and adenosine A1 receptor antagonists, but that they are likely unaffected by blocking glutamatergic signaling. I will test this prediction by evoking PVN responses to optogenetic vascular activity as in Aim II, and by exposing SI cortex to receptor antagonists. Training Environment: This project will take place over three years in the Brown University Neuroscience Graduate Program under the mentorship of Dr. Christopher Moore. The Research Training Plan includes didactic professional, technical, and science writing training, as well as hands-on technical seminars.

项目摘要 了解神经血管沟通对于临床和基础研究至关重要。血管周神经元 （PVN）活性可以驱动大脑血管动力学。但是，血管事件对中性的影响 活动仅经过稀疏研究。我们的实验室发现，主要的PVN人数 体感皮质（SI）在体内编码脑血管活性。但是，这种编码的本质及其 解剖组织未经测试。血管到PVN信号传导可能支持血管稳态和富含的血管稳态 跨系统的通信。这些信号与使用血流来绘制中性活性的研究有关 （例如fMRI）。调查这种信号的扰动可能阐明脑血管的机制 函数（例如，缺血，帕金森氏病和M.S.）。 为了分析血管活性编码的PVN，我将使用神经和血管的体内两光子成像 细胞和光遗传学以扰动血管并分析PVN反应。在目标I中，我将检验假设 该血管编码PVN通常发生在SI中，其活性由皮层和血管组织 隔室，通过表达神经元中的钙指标（JRGECO1A）和（GCAMP6F）在血管内皮中（GCAMP6F） 简单地对其活动进行图像。我的初步数据确定了在空间上不同的钙事件 预测随后的PVN活性的血管信号。在此范式中，血管响应PVN的频率 将根据其陈规定型活动和解剖位置进行分类。我们实验室中的初步数据也有 表明选择性光遗传性血管驱动可以调节PVN活性。在AIM II中，我将检验假设 由光遗传学诱发的血管直径变化驱动的PVN也将通过解剖学组织 他们的活动，以及对内源血管事件的反应将平行它们对光学遗传的反应 血管驱动。我将通过驾驶内皮通道旋转蛋白来束缚Si血管，扩张 它们具有平滑肌甲状腺素，并引起自然触觉驱动的功能充血，以分析 表达GCAMP6的PVN的响应。在AIM III中，我将检验PVN对 trpv4和腺苷A1接收器可以在光源性驱动的血管活性上扰动 对抗者，但可能不会因阻断谷氨酸能信号传导而受到影响。我将通过 像AIM II一样，唤起对光遗传学血管活性的PVN反应，并通过将Si Cortex暴露于受体中 对手。 培训环境：该项目将在布朗大学进行三年 克里斯托弗·摩尔博士的心态下的神经科学研究生课程。研究培训计划 包括教学专业，技术和科学写作培训以及动手技术半手。