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.

项目总结 了解神经-血管沟通对于临床和基础研究是至关重要的。血管周围神经元 下丘脑室旁核(PVN)活动可以驱动脑血管动力学。然而，血管事件对神经的影响 对这一活动的调查很少。我们的实验室发现，在初选中有一群PVN 躯体感觉皮层(SI)在体内编码脑血管活动。但是，这种编码的性质及其 解剖组织，是未经检验的。血管到PVN信号转导可能支持血管内稳态和RICH 跨系统的通信。这些信号与使用血流映射神经活动的研究相关 (例如，功能磁共振成像)。研究这一信号的扰动可能有助于阐明脑血管疾病的机制。 功能障碍(例如，脑缺血、帕金森氏病和多发性硬化症)。 为了分析PVN编码的血管活动，我将使用体内神经和血管的双光子成像 细胞，以及扰乱血管和分析PVN反应的光遗传学。在《目标1》中，我将检验这一假设 血管编码PVN在SI中普遍存在，其活动由皮质层和血管组成 通过在神经元上表达钙指示剂(JRGECO1a)和在血管内皮细胞表达钙指示剂(GCaMP6f) 以同时显示他们的活动。我的初步数据确定了在空间上不同的钙事件 预测后续PVN活动的血管信号。在这个范例中，血管反应性PVN的频率 将根据他们刻板的活动和解剖位置进行分类。我们实验室的初步数据也 表明选择性光发生血管驱动可以调节PVN的活性。在《目标2》中，我将检验这一假设 由光基因诱发的血管直径变化驱动的室旁核在解剖学上也将由 他们的活动，以及他们对内源性血管事件的反应将与他们对光遗传的反应类似 血管驱动。我将通过驱动内皮通道视紫红质来光遗传收缩SI血管，扩张 将其与平滑肌卤视紫红质，并诱发自然触觉驱动的功能性充血，以分析 表达GCaMP6s的PVN的免疫应答在Aim III中，我将测试PVN对 TRPV4和腺苷A1受体可从药理上干扰光遗传驱动的血管活动 拮抗剂，但它们可能不受阻断谷氨酸能信号的影响。我将通过以下方式验证这一预测 像在AIM II中一样，通过将SI皮层暴露于受体来激发PVN对光遗传血管活动的反应 对抗者。 培训环境：该项目将在布朗大学进行，为期三年 克里斯托弗·摩尔博士指导下的神经科学研究生项目。研究培训计划 包括教授专业、技术和科学写作的培训，以及实际操作的技术研讨会。