Fiber Cell Junctions in Normal and Cataractous Lenses

描述(申请人提供)：人类核老年性白内障是 北美最常见的需要矫正手术的白内障类型和 仍然是全世界导致失明的主要原因。在这场竞争中 继续应用程序的主要目标是识别和表征 在人类细胞核内产生散射中心的精确细胞变化。 近些年来，在表征 纤维细胞的变化，可能导致内部光散射增加 人类晶状体的细胞核，以及对衰老本质的理解， 正常人晶状体的致密化和细胞间通讯。这个 拟议的研究将建立在使用形态学的先前成功的基础上 采用并将采用各种现代电子显微镜技术 包括扫描和透射式电子显微镜、低温显微镜、3D 断层扫描、傅里叶分析、荧光和明场光学显微镜以及 激光扫描和双光子共聚焦显微镜。我们建议以文件形式记录 细胞质的分子组织和特异体的分布 正常晶状体中的接合处。重点将放在包含以下内容的交叉点 MEP/Aquaporin0被认为是提供水的途径 运输和作为粘连的连接点。从不同来源分离的膜 人类晶状体的区域将被检查以确定分子 使用荧光和金粒子标记的组织和表征 粘性接合处。结合部和细胞质内的变化 核性白内障形成过程中的纤维细胞将被记录下来，特别是 我们最近描述的多层天体是潜在的散射中心。 我们期待这项工作将有助于更好地了解 目前关于白内障形成的假设以及可能减缓或 预防老年人白内障的进展。

DESCRIPTION (provided by applicant): Human nuclear age-related cataract is the most common type of cataract requiring corrective surgery in North America and remains a leading cause of blindness throughout the world. In this competing continuation application the main goal is to identify and characterize the precise cellular changes that produce scattering centers within human nuclei. Significant progress has been made in recent years in characterizing the alterations to fiber cells that may produce increased light scattering within human lens nuclei, as well as to an understanding the nature of aging, compaction and intercellular communication in the normal human lenses. The proposed research will build on the previous successes using morphological approaches and will employ a variety of modern electron microscopy techniques including scanning and transmission electron microscopy, cryo microscopy, 3D tomography, Fourier analysis, fluorescent and brightfield light microscopy and laser scanning and two-photon confocal microscopy. We propose to document the molecular organization of the cytoplasm and the distribution of specialized junctions in normal lenses. Emphasis will be given to junctions containing MEP/Aquaporin0 which have been hypothesized to provide a pathway for water transport and serve as adhering junctions. Membranes isolated from different regions of human lenses will be examined to determine the molecular organization using fluorescent and gold particle labeling and to characterize the adhesive junctions. Changes in the junctions and within the cytoplasm of fiber cells during nuclear cataract formation will be documented, especially the multilamellar bodies we described recently as potential scattering centers. We expect that this work will contribute to a better understanding of the current hypotheses of cataract formation and to strategies that could slow or prevent the progression of cataracts in the elderly.