Neural activity-dependent modulation of cortical microvascular restoration

皮质微血管修复的神经活动依赖性调节

• 批准号: 10370395
• 负责人: ANDRE OBENAUS
• 金额: $ 55.46万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-15 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10370395
• 关键词: Action Potentials; Acute; Animals; Arteries; Astrocytes; Blood Vessels; Blood capillaries; Blood flow; Brain; Brain Injuries; Cell physiology; Cells; Cerebrovascular Circulation; Cerebrum; Coupled; Data; Dextrans; Elements; Endothelial Cells; Endothelium; Fluorescent Dyes; Generations; Genetic; Growth; Head; Histologic; Image; Imaging technology; Injury; Label; Longitudinal Studies; Magnetic Resonance Imaging; Measurement; Metabolism; Methods; Microscope; Modeling; Molecular; Monitor; Motor Cortex; Movement; Mus; Neuraxis; Neurological outcome; Neurons; Outcome; Pattern; Perfusion; Pericytes; Phase; Physiological Processes; Plasma; Play; Process; Recovery; Reporter; Research; Resolution; Role; Rotarod Performance Test; Signal Transduction; Site; Speed; Stains; Synapses; Testing; Therapeutic Intervention; Time; Tissue Preservation; Tissues; Vascular Endothelial Growth Factors; Vascular remodeling; Vascularization; Veins; behavior test; beta catenin; cell type; cerebral microvasculature; clinically translatable; endothelial stem cell; excitatory neuron; foot; improved; in vivo; in vivo imaging; inhibitory neuron; injury and repair; innovation; light weight; migration; miniaturize; motor function recovery; mouse model; multidisciplinary; neovascularization; nervous system development; neural circuit; neural stimulation; novel; optogenetics; quantitative imaging; recruit; relating to nervous system; repaired; response; restoration; serial imaging; temporal measurement; therapeutically effective; two-photon; vasculogenesis; venule

Project Summary / Abstract Blood vessels, from arteries to capillaries to venules and then to veins, contribute to fundamental physiological processes. However, the vascular responses for repair and restoration of microvascular networks after cortical brain injury are poorly understood, including how neuronal activity influences these processes. A major barrier to research is the poor accessibility of micro-vessels in the brain and associated technical difficulties. To overcome this barrier, we propose to use innovative imaging technologies that we have co-developed to investigate micro-vessel formation and re-growth in response to focal cortical injury. We have used light-weight head-mounted, miniaturized microscopes (“miniscopes”) to dynamically image the vasculature and associated cells with high spatial and temporal resolution. We will use cortical injury models by applying a controlled moderate impact to the mouse motor cortex. Combining in vivo longitudinal miniscope and 2-photon imaging, histological “vessel painting” and perfusion-weighted magnetic resonance imaging (PWI MRI), we aim to achieve a deeper understanding of microvascular restoration following cortical injury. We will apply targeted optogenetic stimulation of excitatory neurons and specific inhibitory neurons to modulate microvascular repair in early and late phases of vessel re-growth. Our guiding hypothesis is that microvascular restoration and remodeling after cortical injury are regulated by vascularization sequences and cellular processes that are similarly observed in normal vasculogenesis during central nervous system development. In Aim 1, we will identify the time course and spatial pattern of vascular regrowth, and blood flow dynamics after focal cortical injury. Vascular networks and blood flow are visualized with fluorescent-labeled dextrans for in vivo imaging for quantitative measurements. In Aim 2, we will determine the role of endothelial cells in new blood vessel sprouting and the establishment of functional microvascular by imaging Tie2-Cre reporter mice during the first two weeks post- injury. We will also examine the influence of astrocytes and pericytes in vascular re-growth. In Aim 3, we will test the hypothesis that optogenetic stimulation of specific neuron types in a temporally controlled manner facilitates and enhances microvasculature restoration for post-injury repair. We will also examine if and how targeted modulation of neural activities modulate Wnt/ß-catenin and VEGF signaling mechanisms that are critical for micro-vessel re-growth. Behavioral testing will assess the outcomes of the optogenetic treatment. We have strong preliminary data that supports the premise for the proposed research for all aims. The proposed research will advance our understanding of the cellular and molecular mechanisms underlying cortical microvascular restoration and how neural stimulation enhances vascular network formation.

项目概要/摘要 血管，从动脉到毛细血管，再到小静脉，然后到静脉，有助于基础生理学 然而，皮质后微血管网络修复和恢复的血管反应。 人们对脑损伤知之甚少，包括神经活动如何影响这些过程。 研究的一个难题是大脑中微血管的可及性较差以及相关的技术困难。 为了克服这一障碍，我们建议使用我们共同开发的创新成像技术 研究针对局灶性皮质损伤的微血管形成和再生长我们使用了轻型。 头戴式微型显微镜（“微型显微镜”）对脉管系统和相关的动态成像 我们将通过应用受控的方法来使用具有高空间和时间分辨率的细胞。 结合体内纵向显微镜和 2 光子成像，对小鼠运动皮层产生中等影响。 组织学“血管绘画”和灌注加权磁共振成像（PWI MRI），我们的目标是实现 我们将应用靶向光遗传学来更深入地了解皮质损伤后的微血管修复。 刺激兴奋性神经元和特异性抑制性神经元以调节早期和晚期的微血管修复 我们的指导假设是微血管修复和重塑。 皮质损伤受到血管化序列和细胞过程的调节，这在 在目标 1 中，我们将确定中枢神经系统发育过程中的正常血管发生。 局灶性皮质损伤后血管再生的空间模式和血流动力学。 使用荧光标记的葡聚糖可视化血流，进行体内成像以进行定量 在目标 2 中，我们将确定内皮细胞在新血管萌芽中的作用以及 在治疗后的前两周内通过对 Tie2-Cre 报告小鼠进行成像来建立功能性微血管 我们还将研究星形胶质细胞和周细胞对血管再生的影响。 检验特定神经元类型的光遗传学刺激的假设 促进和增强损伤后修复的微血管恢复我们还将检查是否以及如何进行。 神经活动的靶向调节调节至关重要的 Wnt/ß-连环蛋白和 VEGF 信号传导机制 行为测试将评估光遗传学治疗的结果。 强有力的初步数据支持所有目标的拟议研究的前提。 将增进我们对皮质微血管潜在细胞和分子机制的理解 恢复以及神经刺激如何增强血管网络的形成。