A CRYSTALLIN MUTATION WITH ABNORMAL ASTROCYTES AND RETINAL VESSELS

晶状体蛋白突变导致星形胶质细胞和视网膜血管异常

• 批准号: 7350844
• 负责人: DEBASISH SINHA
• 金额: $ 41万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7350844
• 关键词: Address; Adult; Affect; Antibodies; Apoptosis; Astrocytes; Axon; Axonal Transport; Base Pairing; Binding; Biochemistry; Biological Assay; Biology; Blood Vessels; Blood flow; Bromodeoxyuridine; Cataract; Cell physiology; Cells; Chromosomes, Human, Pair 10; Coculture Techniques; Colchicine; Communication; Complement; Crystallins; Deposition; Development; Dyes; Electrical Resistance; Endothelial Cells; Endothelium; Evans blue stain; Exons; Eye; Genes; Genetic; Glial Cell Proliferation; Glial Fibrillary Acidic Protein; Goals; Growth; Heterozygote; Homozygote; Hour; Human; In Situ Nick-End Labeling; Injection of therapeutic agent; Intermediate Filaments; Label; Measures; Microaneurysm; Molecular; Mutation; Names; Neurons; Optic Nerve; Phenotype; Photography; Play; Process; Proteins; Rattus; Retina; Retinal; Retinal Ganglion Cells; Role; Signal Pathway; Sprague-Dawley Rats; Staining method; Stains; Structure; Surface; System; Techniques; Testing; Thick; Tight Junctions; Vascular Permeabilities; Vimentin; Yeasts; anterior chamber; base; cell motility; fetal; insight; migration; mutant; nestin protein; novel; research study; retina blood vessel structure; time use; tool; yeast two hybrid system

DESCRIPTION (provided by applicant): We have discovered a spontaneous mutation in the Sprague Dawley rat with an unusual eye phenotype that we have named Nuc1. The mutation behaves as a single semi-dominant locus with a viable homozygote and an intermediate phenotype in the heterozygotes. The mutation causing Nuc1 is a 27 base pair insertion in exon 6 of the ¿A3/A1 crystallin gene on chromosome 10. In addition to Nuc1 several human mutations in ¿A3/A1 crystallin are known, all of which cause dominant cataract. In homozygous Nuc1 rats the fetal intraocular vessels persist even after development of the retinal vessels. During early post-natal development these rats also have a much thicker retina than normal with an excess number of vessels. As the Nuc1 homozygote rat matures, we find evidence of microaneurysm formation, intravascular deposits and blockage of blood flow inside some vessels. We have found that in the retina, ¿A3/A1 is expressed only in astrocytes and that in the Nuc1 homozygotes the astrocytes are morphologically abnormal. The purpose of this study is to address the possibility that the normal functioning of the retinal astrocytes is compromised as a consequence of the Nuc1 mutation. It is now accepted that astrocytes play a major role in the establishment of a functional retinal vasculature, however, the cellular and molecular mechanisms involved in this process remain elusive. Establishing Nuc1 as a genetic tool will provide a unique system in which to study the biology of astrocytes, in particular their interactions with retinal ganglion and endothelial cells. Our goal for the proposed studies is to test our hypothesis that expression of mutant ¿A3/A1 crystallin affects astrocyte function, leading to improper organization and function of the retinal vasculature in the Nuc1 rat. To test this hypothesis, the following specific aims are proposed: SPECIFIC AIM 1: To characterize and compare the structure, sub-cellular localization and protein interactions of ¿A3/A1 crystallin protein in retinal astrocytes from Nuc1 and wild type rats. SPECIFIC AIM 2: To investigate if the proliferation or migration of astrocytes is disrupted by expression of mutant ¿A3/A1 crystallin SPECIFIC AIM 3: To demonstrate the effect of astrocytes expressing mutant ¿A3/A1 on retinal vasculature. Despite rapid progress made in understanding the development of the retinal vasculature, many questions remain to be answered about the mechanisms and signaling pathways regulating vascular development. We believe that studies on the Nuc1 rat will provide new insights into the cellular and molecular mechanisms that regulate vascular development including the molecular interactions among neurons, astrocytes and endothelial cells.

描述（通过应用证明）：我们在Sprague大鼠中发现了一个异常的眼睛表型，我们具有D nuc1 nuc1是27个碱基对插入的外显子6中的插入A3/A1晶体基因在10号染色体上。除了NUC1外，还具有几种人类突变。 A3/A1晶体都是已知的，所有这些都会引起纯合核1大鼠的主体成熟，我们发现某些血管内的微型神经瘤形成的证据。 A3/A1仅在星形胶质细胞中表达，并且在星形胶质细胞中是形态学。脉络膜，涉及该过程的细胞和分子SM，尤其是与拟议的研究相互作用的是测试我们的假设，即A3/A1结晶蛋白星形胶质细胞功能，导致NUC1大鼠视网膜Vascularture的组织和功能不正确，以下特定目的是支撑的：特定的目标1：以表征和比较结构，亚细胞定位和蛋白质相互作用»来自NUC1和野生型大鼠的视网膜星形胶质细胞中的A3/A1晶体蛋白是对突变体的讨论。 A3/A1晶体特异性目标3：证明表达突变体的有效量A3/A1在视网膜脉络膜上进行了介绍的机理和信号的血管发育。