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The Mouse dc-Electroretinogram

小鼠直流视网膜电图

基本信息

批准号：
6954115
负责人：
NEAL S. PEACHEY
金额：
$ 22.95万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-08-01 至 2007-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6954115
关键词：
chloride channels cone cell electrophysiology electroretinography electrostimulus genetically modified animals high performance liquid chromatography immunocytochemistry laboratory mouse neural degeneration retinal pigment epithelium rhodopsin rod cell transducin visual photoreceptor western blottings

项目摘要

DESCRIPTION (provided by applicant): The retinal pigment epithelium (RPE) is critically involved in many functions required for normal retinal function including the flow of nutrients and waste products between the photoreceptors and the choroidal circulation, the visual cycle, and the phagocytosis of shed outer segment disks. In addition, mutations in RPE-specific genes have been implicated in a wide range of hereditary retinal disease, and this information is being used to develop corresponding mouse models. In the present project, the focus is on the electrical responses generated by the RPE in response to activity of the neural retina. While specific aspects of RPE function may be studied using the electroretinogram (ERG) recorded using dc-coupled amplification, this has not been applied to the mouse. Moreover, fundamental questions remain regarding the origin of the different components of the dc-ERG. In Aim 1, we will study normal wild-type mice, to define the stimulus-response characteristics of the mouse dc-ERG, and the time course over which these develop. In Aim 2, we will test the hypothesis that the mouse dc-ERG is initiated primarily by rod photoreceptor activity, using KO lines for transducin and rhodopsin, and mutants in which the cone population is increased (rd7) or decreased (comas transgenic). In Aim 3, we will use Kir4.1 KO mice to test the hypothesis that this channel is involved in c-wave generation. In Aim 4, we will use cftr KO mice and adenoviral delivery of CFTR constructs to test the hypothesis that the fast oscillation is generated by CFTR activity. In Aim 5, we will use dc-ERG recordings to study the relationship between photoreceptor degeneration and changes in RPE function. First, we will examine how A2E accumulation in abcr-mutant mice alters RPE function. Next, we will test the hypothesis that functional RPE abnormalities occur at early stages using three lines of mice with similar rates of photoreceptor degeneration. In two, the primary defect resides in the photoreceptors (rds/+ and Bouse transgenic mice). In vitilligo mice, a similar rate of photoreceptor degeneration is induced by defect in the RPE. At the completion of this project, we expect to have defined an optimal means for recording the mouse dc-ERG in vivo, to have a thorough understanding of the processes that underlie the different components of the dc-ERG, and a more complete understanding of how these components are affected by disorders of the outer retina.

描述（由申请人提供）：视网膜色素上皮（RPE）在正常视网膜功能所需的许多功能中起关键作用，包括光感受器和脉络膜循环之间的营养物质和废物流动、视觉循环和脱落外节盘的吞噬。此外，rpe特异性基因的突变与多种遗传性视网膜疾病有关，这一信息正被用于开发相应的小鼠模型。在目前的项目中，重点是RPE在响应神经视网膜活动时产生的电反应。虽然RPE功能的特定方面可以通过使用直流耦合放大记录的视网膜电图（ERG）来研究，但这尚未应用于小鼠。此外，关于dc-ERG的不同组成部分的起源仍然存在根本问题。在Aim 1中，我们将研究正常野生型小鼠，以确定小鼠dc-ERG的刺激反应特征，以及这些特征发展的时间过程。在目标2中，我们将测试小鼠dc-ERG主要由杆状光感受器活性启动的假设，使用转导蛋白和视紫红质的KO系，以及锥体种群增加（rd7）或减少（coma转基因）的突变体。在目标3中，我们将使用Kir4.1 KO小鼠来测试该通道参与c波产生的假设。在Aim 4中，我们将使用cftr KO小鼠和腺病毒递送cftr构建物来验证cftr活性产生快速振荡的假设。在Aim 5中，我们将使用dc-ERG记录来研究光感受器变性与RPE功能变化之间的关系。首先，我们将研究A2E在abcr突变小鼠体内的积累如何改变RPE功能。接下来，我们将使用三种具有相似光感受器退化率的小鼠来验证功能性RPE异常发生在早期阶段的假设。在两种情况下，主要缺陷存在于光感受器（rds/+和house转基因小鼠）。在白癜风小鼠中，RPE缺陷诱导的光感受器变性率相似。在这个项目完成后，我们期望能够定义一种最佳的方法来记录小鼠体内dc-ERG，彻底了解dc-ERG不同组成部分的过程，并更全面地了解这些组成部分如何受到外视网膜疾病的影响。