ROLE OF Nna1 IN RETINAL AND CEREBELLAR NEURODEGENERATION

Nna1 在视网膜和小脑神经变性中的作用

基本信息

批准号：
7486853
负责人：
ALBERT R LA SPADA
金额：
$ 25.38万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-09-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7486853
关键词：
Age-Months Alleles Amino Acids Antibodies Apoptosis Autophagocytosis Bacterial Artificial Chromosomes Biochemical Biological Assay Brain Stem Candidate Disease Gene Carboxypeptidase Cell Fractionation Cells Cessation of life Characteristics Chromosomes, Human, Pair 13 Cleaved cell Defect Degenerative Disorder Development Disease Drosophila genus Electron Microscopy Follow-Up Studies Future Genes Goals Homologous Gene Homozygote Human Immunohistochemistry Libraries Life Male Sterility Maps Measurable Mind Molecular Moths Mouse Protein Mus Mutation Nature Necrosis Nerve Degeneration Neurologic Neurologic Mutants Mice Neurons Pathway interactions Pattern Peptide Hydrolases Phenotype Photoreceptors Population Process Proteins Purkinje Cells Recombinants Research Personnel Resources Retina Retinal Retinal Diseases Retinal Photoreceptors Role Sequence Analysis Southern Blotting System Testing Thalamic structure Time Tissues Transgenic Organisms Western Blotting Yeasts Zinc base cell type day in vivo insight loss of function neuron loss olfactory bulb programs transgene expression yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Despite significant advances in our understanding of degenerative diseases of the brainstem and retina, the pathways that underlie most of these disease processes remain undefined. Furthermore, the basis for cell-type specific degeneration and neuron cell death in many important neurological and retinal diseases is unknown. With these issues in mind, we hypothesized that delineation of the molecular basis of the neurological mouse mutant, Purkinje cell degeneration (pcd), would yield insights into how and why certain types of neurons degenerate. pcd is an autosomal recessive mutation that produces a non-developmental degeneration of cerebellar Purkinje cells and retinal photoreceptors. To determine the basis of the degeneration in pcd, we mapped the pcd gene defect to a 1 cM region on chromosome 13 and then identified mutations in the Nna1 gene in two alleles of pcd. Nna1 is a predicted 1218 amino acid protein that has a zinc carboxypeptidase domain with highly conserved orthologues in human (NNA1) and Drosophila (NnaD). Additional homologues of Nna1 exist in mouse, human, moth, and worms, all of which retain the carboxypeptidase domain. Using the results and resources that we have generated, we now wish to advance our understanding of how loss of Nna1 function leads to cell-type specific neurodegeneration in pcd, and thereby determine if Nna1's pathway of action has implications for other diseases. To accomplish these goals, we will begin by confirming that Nna1 is the causal gene by attempting transgenic rescue of the entire pcd phenotype and of one selected neuronal population, namely Purkinje cells. We will seek the mutations responsible for remaining pcd alleles. We will study the expression characteristics of the Nna1 gene product, and will define the nature of neuronal death in pcd mice. We will evaluate Nna1 for carboxypeptidase activity and will determine if retention of carboxy-peptidase activity is required for successful transgenic rescue. To identify Nna1's pathway of action, we will use lines of Drosophila that carry NnaD loss-of-function alleles to perform modifier screens to identify NnaD interacting genes. We will characterize Nna1 interacting proteins detected in a yeast two-hybrid screen, and will compare the results of these two screens to guide future studies.

描述（由申请人提供）：尽管我们对脑干和视网膜退行性疾病的理解取得了重大进展，但大多数这些疾病过程背后的途径仍然不明确。此外，许多重要的神经系统和视网膜疾病中细胞类型特异性变性和神经元细胞死亡的基础尚不清楚。考虑到这些问题，我们假设，对神经小鼠突变体浦肯野细胞变性（pcd）的分子基础的描述，将有助于深入了解某些类型的神经元如何以及为何退化。 pcd 是一种常染色体隐性突变，会导致小脑浦肯野细胞和视网膜感光细胞发生非发育性变性。为了确定 PCD 变性的基础，我们将 PCD 基因缺陷定位到 13 号染色体上的 1 cM 区域，然后鉴定了 PCD 两个等位基因中 Nna1 基因的突变。 Nna1 是一种预测的 1218 个氨基酸蛋白，具有锌羧肽酶结构域，在人类 (NNA1) 和果蝇 (NnaD) 中具有高度保守的直系同源物。 Nna1 的其他同源物存在于小鼠、人类、飞蛾和蠕虫中，所有这些都保留了羧肽酶结构域。利用我们产生的结果和资源，我们现在希望加深对 Nna1 功能丧失如何导致 PCD 中细胞类型特异性神经变性的理解，从而确定 Nna1 的作用途径是否对其他疾病有影响。为了实现这些目标，我们将首先通过尝试对整个 PCD 表型和一个选定的神经元群体（即浦肯野细胞）进行转基因拯救来确认 Nna1 是致病基因。我们将寻找导致剩余 PCD 等位基因的突变。我们将研究 Nna1 基因产物的表达特征，并定义 PCD 小鼠神经元死亡的性质。我们将评估 Nna1 的羧肽酶活性，并将确定成功的转基因拯救是否需要保留羧肽酶活性。为了确定 Nna1 的作用途径，我们将使用携带 NnaD 功能丧失等位基因的果蝇品系进行修饰筛选，以识别 NnaD 相互作用基因。我们将表征在酵母双杂交筛选中检测到的 Nna1 相互作用蛋白，并将比较这两个筛选的结果以指导未来的研究。