Fiber Cell Junctions in Normal and Cataractous Lenses

描述（由申请人提供）：人类核性年龄相关性白内障是 在北美最常见的需要矫正手术的白内障类型， 仍然是全世界失明的主要原因。在这场竞争中， 继续申请的主要目标是确定和表征 在人体细胞核内产生散射中心的精确细胞变化。 近年来，在确定 纤维细胞的改变，可能会产生增加的光散射内 人类透镜核，以及对衰老本质的理解， 正常人晶状体中的致密化和细胞间通讯。的 拟议的研究将建立在以前的成功， 方法，并将采用各种现代电子显微镜技术 包括扫描和透射电子显微镜、低温显微镜、3D 断层扫描、傅立叶分析、荧光和明场光学显微镜， 激光扫描和双光子共聚焦显微镜。我们建议将 细胞质的分子组织和特化的 正常晶状体中的接合处。重点将放在包含 MEP/水通道蛋白0，其被假设为提供水通道 运输并作为粘附连接。分离自不同细胞的膜 将检查人类晶状体的区域以确定 组织使用荧光和金颗粒标记和表征 粘合剂接合处。在连接和细胞质内的变化， 核性白内障形成过程中的纤维细胞将被记录，特别是 我们最近描述的多层体是潜在的散射中心。 我们期望这项工作将有助于更好地了解 目前白内障形成的假设和策略，可以减缓或 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.