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）关键参与正常视网膜功能所需的许多功能，包括光感受器和脉络膜循环之间的营养物和废物的流动、视觉循环以及脱落的吞噬作用外段磁盘。此外，RPE特异性基因的突变与多种遗传性视网膜疾病有关，这些信息正被用于开发相应的小鼠模型。在本项目中，重点是 RPE 响应神经视网膜活动而产生的电反应。虽然可以使用直流耦合放大记录的视网膜电图 (ERG) 研究 RPE 功能的特定方面，但这尚未应用于小鼠。此外，关于 dc-ERG 不同成分的起源仍然存在基本问题。在目标 1 中，我们将研究正常野生型小鼠，以确定小鼠 dc-ERG 的刺激响应特征，以及这些特征发展的时间过程。在目标 2 中，我们将使用转导蛋白和视紫红质的 KO 系以及视锥细胞群增加 (rd7) 或减少（comas 转基因）的突变体来测试小鼠 dc-ERG 主要由视杆光感受器活性启动的假设。在目标 3 中，我们将使用 Kir4.1 KO 小鼠来测试该通道参与 c 波生成的假设。在目标 4 中，我们将使用 cftr KO 小鼠和腺病毒递送 CFTR 构建体来测试快速振荡是由 CFTR 活性产生的假设。在目标 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 abcr 突变小鼠中 A2E 的积累如何改变 RPE 功能。接下来，我们将使用光感受器变性率相似的三系小鼠来测试功能性 RPE 异常在早期发生的假设。其中两种，主要缺陷在于光感受器（rds/+ 和 Bouse 转基因小鼠）。在白癜风小鼠中，RPE 缺陷会导致类似的光感受器变性。在该项目完成时，我们期望定义一种记录小鼠体内 dc-ERG 的最佳方法，以全面了解 dc-ERG 不同组成部分的过程，并更全面地了解这些成分如何受到外视网膜疾病的影响。