ASSESSMENT OF RETINAL FUNCTION IN HEALTH AND DISEASE FROM MOUSE TO HUMAN

从小鼠到人类的健康和疾病中的视网膜功能评估

基本信息

批准号：
9088931
负责人：
Frans Vinberg
金额：
$ 9.48万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9088931
关键词：
Affect Age related macular degeneration Animal Model Animal Testing Area Argon Blindness Burn injury Cell physiology Cells Cessation of life Clinical Trials Collaborations Cone Data Developed Countries Devices Diabetes Mellitus Diabetic Retinopathy Disease Disease model Dissection Electrophysiology (science)Electroretinography Environment Euthanasia Eye Eye diseases Family suidae Functional disorder Genetic Glaucoma Health Human Individual Lasers Lateral Light Link Macular degeneration Measures Mediating Methodology Methods Molecular Muller&apos s cell Mus Mutation Neuroglia Neurons Noise Opsin Peripheral Pharmaceutical Preparations Pharmacology Photoreceptors Positioning Attribute Primates Protocols documentation Retina Retinal Retinal Diseases Retinitis Pigmentosa Rhodopsin Rodent Safety Shapes Signal Transduction Streptozocin Techniques Time Vertebrate Photoreceptors Vision Wild Type Mouse base cell type design fovea centralis ganglion cell in vivo injured macula mouse model multi-electrode arrays novel patch clamp public health relevance research study response scale up screening

项目摘要

DESCRIPTION (provided by applicant): Leading causes of blindness in developed countries include macular degeneration, glaucoma and diabetic retinopathy which are all manifested as dysfunction and degeneration of specific cell types in the retina. The molecular mechanisms and pathophysiology of these diseases are not well understood. Widely used techniques to study the pathophysiology of retinal diseases include In Vivo electroretinogram (ERG) and single cell electrophysiology. In Vivo ERG can assess the overall health state of the retina but is limited in providing quantitative information about the cellular and molecular origin of the functional deficits in a diseased retina. Single cell recordings, on the other hand, provide quantitative data from individual cells but are challenging, offer only limited recording time and are not easily scaled up to assess the function and signaling across the whole retina. The objective of this proposal is to advance techniques and methodology to dissect the function of photoreceptor, bipolar and Müller-glial cells in the intact mouse, primate, pig and ultimately human retinas. I wll first develop methodology to quantitatively assess the intrinsic functional "state" of these cells y using Ex Vivo ERG from isolated wild-type mouse retinas (K99). Ex Vivo ERG provides information about the average functional state of all cells across the lateral axis of the retina. However, some of the retinal diseases affect retina only locally (e.g. DR) or target primarily ganglion cells (e.g. glaucoma) that will not necessarily be observable in the Ex Vivo ERG signal. One objective of this proposal is to develop a novel device combining Ex Vivo ERG and multi-electrode array (MEA) methods. This device will be used to assess the local function of photoreceptors, bipolar, Müller-glial and ganglion cells across the whole wild-type mouse retinas (K99). Ex Vivo ERG (K99) will be applied to determine how the function of rod and cone bipolar, Müller-glial and ganglion cells are affected in the mouse models of retinitis pigmentosa (RP, P23H rhodopsin mutation) and diabetic retinopathy (DR, Streptozotocin-induced diabetes) and MEA-ERG device will be used to assess the local function of the retina that has been focally injured by laser (K99 and R00). These experiments will advance the understanding of pathophysiology of these diseases known to affect primarily outer and inner retina, respectively. The methodology developed for mouse retinas will be used to establish protocols to dissect the function of photoreceptors, bipolar and Müller-glial cells in primate, pig, and ultimately human donor retinas. I will first develop the recording protocols to obtain viable responses from primate retinas dissected from eye balls enucleated immediately following the euthanasia (K99). Then, in collaboration with Dr. Hanneken, we will determine the acceptable time frame between death and enucleation by using pig eyes (R00). Finally, based on the primate and pig experiments we will design and conduct recordings to assess function of photoreceptor, bipolar and Müller glia cells in a macula and peripheral regions of the human retina (R00).

描述（通过应用证明）：在发达国家，青光眼和糖尿病的糖尿病症状是疾病中特定细胞类型的功能障碍和变性。限制提供了量化的信息性信息和分子Igin，另一方面，单细胞记录中的功能缺陷。来自单个细胞，但充满挑战，只有有限的时间，并且不被释放以评估整个视网膜的函数。，最终使用分离的野生型小鼠视网膜的人类视网膜y的功能“状态”（K99）仅在局部（例如DR）或目标的一级神经节细胞（例如青光眼）中，不一定在事实ERG信号中观察到。 - 用于评估光感受器，双极，müller-glial和神经节细胞的局部功能，整个野生型小鼠视网膜（K99），müller-glial and Ganglion细胞都在视网膜炎的小鼠模型中p23h的视紫红素突变）和糖尿病性视网膜病（DR，链霉菌素诱导的糖尿病）和MEA-RG设备将降临，以评估已被激光局部受伤的视网膜的局部离子（K99和R00）。对于现有的视网膜和小鼠的Interner，将使用甲基甲基甲化物来剖析光感受器的功能，并最终将人类供体视网膜。在安乐死之后，视网膜从in核中解剖（K99）。人类视网膜中的细胞（R00）。