RETINAL MICROVASCULAR CELLS, MATRIX, & OCULAR DISORDERS

鉴于众所周知，微血管细胞起着至关重要的作用 在糖尿病视网膜病变及糖尿病视网膜病变中的作用 早产，调控疾病的分子和细胞机制 它的起源仍然不甚明朗。探讨调控这些因素的机制 视网膜血管增殖性疾病、抗体和重组DNA探针 将在模型系统中使用。结果将揭示这一角色 细胞-基质、生长因子-基质和细胞-细胞相互作用 视网膜微血管分化与病理学。视网膜 发育中分离的微血管内皮细胞和周细胞 出生后的视网膜将单独和一起培养。刚性或 可机械变形的衬底，已由 视网膜血管细胞和之前由我们小组显示的 差异调节血管细胞生长和收缩表型， 将会被评估。具体地说，我们将描述分子 调控视网膜周细胞发育和分化的机制 以及视网膜微血管内皮细胞对损伤的反应。 为此，我们将使用下列综合方法：(I)抗体 利用收缩蛋白亚型特异性和生长性进行定位 实验室制备和鉴定的因子特异性抗体，(Ii) 生物标记的定量免疫沉淀和荧光光谱分析 来自发育、分化和损伤的亚细胞组分 细胞群体，(III)Northern印迹和转录径流分析 损伤的内皮细胞，生长受阻和生长受激 从新生儿和成人视网膜中纯化的周细胞和(Iv)PCR，同源性 Lambda Zap II基因文库的探测和抗体筛选 使用旨在揭示遗传元素的探针从视网膜周细胞中分离出 调节视网膜微血管生肌的测定。结果： 这些跨学科的研究将有助于确定增长是否 因子-基质、基质-细胞和/或细胞-细胞相互作用调节 顺式DNA调控元件和反式作用因子的表达 负责控制视网膜微血管的生长、分化 以及与糖尿病相关的难以控制的血管增殖 视网膜病变。

Whereas it is well established that microvascular cells play crucial roles in the etiology of diabetic retinopathy and retinopathy of prematurity, the molecular and cellular mechanisms regulating disease inception remain obscure. To approach the mechanisms regulating these retinal vasoproliferative disorders, antibody and recombinant DNA probes will be used in model systems. Results will reveal the role that cell-matrix, growth factor-matrix and cell-cell interactions play in retinal microvascular differentiation and pathology. Retinal microvascular endothelial cells and pericytes isolated from developing and post-natal retinas will be cultured alone and together. Rigid or mechanically deformable substrates, which have been synthesized by retinal vascular cells and previously shown by our group to differentially regulate vascular cell growth and contractile phenotype, will be assessed. Specifically, we will characterize the molecular mechanisms regulating retinal pericyte development and differentiation as well as the retinal microvascular endothelial cell response to injury. To this end, we will use the combined approaches of: (i) antibody localization using contractile protein isoform-specific and growth factor-specific antibodies prepared and characterized in the lab, (ii) quantitative immunoprecipitation and fluorographic analysis of biolabeled subcellular fractions derived from developing, differentiated and injured cell populations, (iii) Northern blot and transcription run-off analyses of injured endothelial cells, growth-arrested and growth-stimulated pericytes purified from neonatal and adult retinas and (iv) PCR, homology probing and antibody screening of lambda Zap II cDNA libraries derived from retinal pericytes using probes designed to reveal genetic elements regulating retinal microvascular myogenic determination. Results of these interdisciplinary studies will help to determine whether growth factor-matrix, matrix-cell and/or cell-cell interactions modulate the expression of cis DNA regulatory elements and trans-acting factors responsible for controlling retinal microvascular growth, differentiation and the unmanageable vasoproliferation associated with diabetic retinopathy.