Structural and Functional imaging with Multiphoton Microscopy in Alzheimer's Mice

使用多光子显微镜对阿尔茨海默病小鼠进行结构和功能成像

基本信息

批准号：
7332658
负责人：
Kishore V Kuchibhotla
金额：
$ 3万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7332658
关键词：
Adult Affect Alzheimer&apos s Disease Amyloid beta-Protein Animal Model Animals Area Axon Biological Neural Networks Bolus Infusion Brain Calcium Cells Chronic Cultured Cells Cyclophosphamide/Fluorouracil/Prednisone Dendrites Deposition Development Disease Disease Marker Disruption Dyes Environment Exhibits Fluorescence Resonance Energy Transfer Fluorescent Dyes Fluorescent Probes Functional Imaging Functional disorder Gene Transfer Gene Transfer Techniques Genetic Goals Homeostasis Image Incidence Individual Induced Mutation Injection of therapeutic agent Lead Learning Life Location Mediating Memory Microscopy Modality Modeling Molecular Monitor Mus Mutation Neurites Neurobiology Neurons Neurophysiology - biologic function Neurosciences Pathogenesis Pharmacology Population Preparation Principal Investigator Process Property Proteins Publishing Recovery Reporter Resolution Role Scientist Senile Plaques Staining method Stains Structure System Techniques Testing Therapeutic Time Transgenic Mice Transgenic Organisms Vertebral column Virus Diseases base in vivo insight mouse model neuronal cell body novel presenilin programs relating to nervous system research study response small molecule submicron

项目摘要

DESCRIPTION (provided by applicant): In recent years, multiphoton microscopy has been used to gain a better understanding of the pathophysiology of Alzheimer's Disease (AD) in intact live mouse brains. We will use transgenic mouse models that develop senile plaques, a dominant marker of the disease, to investigate the functional and structural consequences of these plaques on neurons in vivo. Neuronal function can be investigated with multiphoton microscopy by monitoring the concentration of calcium within a given cell. Intracellular calcium increases exponentially during neuronal activation and can be detected using calcium-sensitive fluorescent probes. These probes increase their brightness, shift their excitation/emission spectra or engage in Fluorescence Resonance Energy Transfer (FRET) to denote a change in calcium concentration. The ability to observe in real-time the functional effects of senile plaques on neuronal activity will provide a novel marker for testing therapeutics with unprecedented spatial and temporal resolution. To achieve this goal we will develop a gene transfer technique that will allow us to introduce a FRET-based, calcium-sensitive genetic construct directly into the adult mouse brain. The protein encoded by this construct robustly fills soma and neuritic processes. This will permit us to investigate homeostatic alterations in calcium concentration caused by plaque deposition. Dynamic calcium transients can also be monitored using this probe, providing the ability to investigate neuronal activation with spine-level resolution in vivo. We also aim to adapt newly published techniques in bulk loading of functional small-molecule dyes for use in adult, transgenic mice. With a large ensemble of neurons stained with such calcium-sensitive indicators, we will determine the functional consequences of plaques on neighboring cells with single-cell resolution. By combining new imaging modalities, disease neurobiology, and systems-level neuroscience, we hope to provide important and unique insight into the pathogenesis of Alzheimer's Disease. SUMMARY: To date, scientists do not understand how networks of individual cells in a living brain malfunction as Alzheimer's Disease progresses. We will use animal models to determine how neural networks are affected and whether specific therapies can lead to recovery of normal function.

描述（由申请人提供）：近年来，多光子显微镜已被用于更好地了解完整活体小鼠大脑中阿尔茨海默病（AD）的病理生理学。我们将使用产生老年斑（该疾病的主要标志物）的转基因小鼠模型来研究这些斑块对体内神经元的功能和结构影响。可以通过多光子显微镜监测给定细胞内的钙浓度来研究神经元功能。细胞内钙在神经元激活过程中呈指数增加，可以使用钙敏感荧光探针进行检测。这些探针会增加亮度、改变激发/发射光谱或参与荧光共振能量转移 (FRET) 以表示钙浓度的变化。实时观察老年斑对神经元活动的功能影响的能力将为以前所未有的空间和时间分辨率测试治疗提供新的标记。为了实现这一目标，我们将开发一种基因转移技术，使我们能够将基于 FRET 的钙敏感基因构建体直接引入成年小鼠大脑中。该构建体编码的蛋白质有力地填充了体细胞和神经突。这将使我们能够研究斑块沉积引起的钙浓度稳态变化。还可以使用该探针监测动态钙瞬变，从而能够在体内以脊柱水平分辨率研究神经元激活。我们还旨在采用新发表的技术来批量加载功能性小分子染料，以用于成年转基因小鼠。通过用这种钙敏感指示剂染色的大量神经元，我们将以单细胞分辨率确定斑块对邻近细胞的功能影响。通过结合新的成像方式、疾病神经生物学和系统级神经科学，我们希望为阿尔茨海默病的发病机制提供重要而独特的见解。摘要：迄今为止，科学家们还不了解随着阿尔茨海默病的进展，活体大脑中的单个细胞网络如何发生故障。我们将使用动物模型来确定神经网络如何受到影响以及特定疗法是否可以恢复正常功能。