MICROARRAY ANALYSIS OF RETINAL DEGENERATION IN MICE

描述(由申请人提供)： 神经退行性疾病的特征是有丝分裂后神经元不可阻挡的变性，并且在很大程度上难以治疗。这些疾病的一个重要子集，视网膜色素变性(RP)，影响终末分化的神经元群体，杆状光感受器，并导致失明。与大多数其他神经退行性疾病不同，RP是由特征明确的单基因缺陷引起的，并影响神经系统的解剖可及部分。这一建议的中心假设是，尽管有不同的遗传病因，但神经元退化的机制集中在共同的关键途径上。识别这些通路将促进对神经元内稳态的了解，也是开发有效治疗方法的先决条件。这项拟议的研究将结合两种策略，在多种不同基因的视网膜变性小鼠模型中识别促进光感受器死亡的基因。首席调查员(PI)已经在RD-1小鼠身上进行了初步的cDNA阵列实验，并建立了早期光感受器死亡过程中分子事件的工作模型。使用RD-1作为基准模型，PI将：1)评估在另外三个不同的遗传模式的RP(rds小鼠、tulpl小鼠和视紫红质突变小鼠)中是否出现类似的表达模式，以及2)在RD-1小鼠中消除在RD-1小鼠中过度表达的选定基因，并将这些影响对RD-1视网膜表型和基因表达模式的影响联系起来。PI已经确定了三个基因[caspase-3、肿瘤坏死因子受体1(TNFR1)和早期生长反应-1(EGR-1)]作为这一方法的候选基因，并对caspase-3进行了初步实验。通过整合这两种策略的数据，PI将能够在一个或多个RP模型中建立促进光感受器凋亡的单个基因之间的因果关系。所进行的培训和研究将扩展和扩大候选人之前在兽医学、解剖病理学和光感受器障碍的分子遗传学方面的研究生培训，导致真正的科学独立。培训将在耶鲁大学医学院进行，指导人员是神经生物学和眼科教授科林·巴恩斯特布尔博士，他是一位国际公认的分子生物学家，使用小鼠的视网膜发育和退化模型。

DESCRIPTION (provided by applicant): Neurodegenerative diseases are characterized by inexorable degeneration of post-mitotic neurons and are largely refractory to therapy. One important subset of these diseases, retinitis pigmentosa (RP), affects a terminally differentiated neuronal population, the rod photoreceptor, and leads to blindness. In contrast to the majority of other neurodegenerative diseases, RP is caused by well-characterized single gene defects, and affects an anatomically accessible portion of the nervous system. The central hypothesis in this proposal is that despite differing genetic etiologies, mechanisms of neuronal degeneration converge upon common critical pathways. Identification of such pathways would advance understanding of neuronal homeostasis and is a prerequisite for developing effective therapies. The proposed research will combine two strategies to identify genes promoting photoreceptor death in multiple genetically distinct mouse models of retinal degeneration. The Principal Investigator (PI) has performed preliminary cDNA array experiments in the rd-1 mouse and has generated a working model of molecular events during early photoreceptor death. Using rd-1 as a benchmark model, the PI will: 1) assess whether similar expression patterns occur in three additional, genetically distinct models of RP (the rds mouse, the tulpl mouse and the rhodopsin mutant mouse) and, 2) eliminate, in the rd-1 mouse, selected genes which are overexpressed in rd-1, and correlate the effects upon the rd-1 retinal phenotype and gene expression pattern. The PI has identified three genes [caspase-3, tumor necrosis factor receptor 1 (TNFR1), and early growth response-1 (EGR-1)], as candidates for this approach, and has performed preliminary experiments with caspase-3. By integrating data from both strategies, the PI will be able to establish cause and effect relationships between individual genes that promote photoreceptor apoptosis in one or more models of RP. The training and research undertaken will extend and broaden the candidate's previous training in veterinary medicine, anatomic pathology, and graduate training in molecular genetics of photoreceptor disorders, leading to true scientific independence. Training will be undertaken at the Yale University School of Medicine under the mentorship of Dr. Colin Barnstable, Professor of Neurobiology and Ophthalmology, who is an internationally recognized molecular biologist using mouse models of retinal development and degeneration.