Birth and Death of Choriocapillaris.

脉络膜毛细血管的出生和死亡。

• 批准号: 7144518
• 负责人: Gerard Anthony Lutty
• 金额: $ 36.47万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7144518
• 关键词: angiopoietins; apoptosis; birth; cell death; cell type; clinical research; complement; cytokine; death; embryo /fetus; eye; family; growth factor; immunocytochemistry; leukocytes; receptor

DESCRIPTION (provided by applicant): The retinal pigment epithelial cells (RPE) and the choriocapillaris (CC) have an intimate relationship in the eye. As nearest neighbors, separated only by Bruch's membrane, they rely on each other and assist each other in accomplishing their critical role of sustaining healthy photoreceptor cells. This proposal explores the relationship between these two cell types from the development or birth of choriocapillaris (5-23 weeks gestation) to the dysfunction or death of either cell type in two different forms of age-related macular degeneration (AMD), geographic atrophy (GA) and exudative AMD. The metabolic status and viability of CC will be evaluated using enzyme and immunohistochemistry (IH) and TUNEL labeling. RPE cell viability and function will be investigated with IH and TUNEL labeling. Growth factors (FGF's, VEGF, PIGF, angiopoietins, erythropoietin, stem cell factor, stromal derived factor-1) that the two cells types produce and use for themselves (autocrine) or their neighbor (paracrine), and their cognate receptors will be investigated by IH and bleaching of the tissue. Tube formation in vitro by fetal human endothelial cells in the presence or absence of fetal RPE will be assessed for paracrine and autocrine effects of growth factors using neutralizing antibodies and siRNA. The contribution of leukocytes and complement to CC death and dysfunction and to CNV in AMD will be investigated by IH to identify cell types, leukocyte numbers (nonspecific esterase, CD45, CD68), leukocyte adhesion molecules (ICAM-1, VCAM-1, P-selectin), and pro-inflammatory (TNFa, IL6, IL8, MCP-1) and angiogenic (FGF's, VEGF, PIGF, angiopoietins) cytokines. The spatial relationship between complement factor H, attack complex, leukocytes and CC viability will also be investigated. Using our alkaline phosphatase flat-embedding technique in which CC vascular density and percent of Bruch's membrane covered with RPE can be quantified, we have established and will continue to investigate the mutualistic symbiotic relationship between CC and RPE. This analysis will permit us to establish how this relationship is disrupted in GA and exudative AMD. The goal of this proposal is to better understand the relationship between these two neighboring cell types and elucidate important factors that could be targeted to maintain or improve this healthy relationship.

描述（由申请人提供）：视网膜色素上皮细胞（RPE）和脉络膜毛细血管（CC）在眼中具有亲密关系。作为仅被Bruch膜隔开的最近的邻居，他们相互依靠，并相互协助完成维持健康的感光细胞的关键作用。该建议探讨了这两种细胞类型之间的关系，从脉络膜毛细血管的发展或出生（妊娠5-23周）到两种不同形式的年龄相关黄斑变性（AMD），地理萎缩（GA）和透明度AMD的两种不同形式的细胞类型的功能障碍或死亡。 CC的代谢状态和生存能力将使用酶和免疫组织化学（IH）和TUNEL标签评估。 RPE细胞的活力和功能将使用IH和TUNEL标记进行研究。生长因子（FGF，VEGF，PIGF，血管生成素，促红细胞生成素，干细胞因子，基质衍生因子1），两种细胞类型为其自身产生和使用（自分泌）或其邻居（旁分泌），以及其同源受体将通过IH和IH进行研究。在存在或不存在胎儿RPE的情况下，将在体外由胎儿人内皮细胞在体外形成，将评估使用中和抗体和siRNA生长因子的旁分泌和自分泌作用。 IH将研究白细胞和补充对CC死亡和功能障碍以及对AMD中CNV的贡献，以识别细胞类型，白细胞数（非特异性酯酶，CD45，CD68），白细胞粘附分子（ICAM-1，iCAM-1，VCAM-1，VCAM-1，P-SELECTIL），和PRAMSIL，PRAMSIN，PRAMENFAMSIN，PRAMSIN，PRAMSIN和PRAMSIN，PRAMSIN，PRAMSIN（PREAM）（PREAM）（PREAM）（PROAM）（PROAM）（proams）rammin和proamm。 MCP-1）和血管生成（FGF，VEGF，PIGF，Angiopoietins）细胞因子。还将研究补体因子H，攻击复合物，白细胞和CC生存能力之间的空间关系。使用我们的碱性磷酸酶扁平化合物技术，其中可以量化CC血管密度和BRUCH膜覆盖的膜百分比，我们已经建立并将继续研究CC和RPE之间的相互共生关系。这种分析将使我们能够确定在GA和渗出性AMD中如何破坏这种关系。该提案的目的是更好地了解这两种相邻细胞类型之间的关系，并阐明可以针对维持或改善这种健康关系的重要因素。