CONTROL OF CELL DIVISION IN THE OCULAR LENS

晶状体细胞分裂的控制

• 批准号: 2444243
• 负责人: John R. Reddan
• 金额: $ 18.32万
• 依托单位: OAKLAND UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-09-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2444243
• 关键词: DNA replication; SDS polyacrylamide gel electrophoresis; atomic absorption spectrometry; biomarker; cataract; cell cycle; cell growth regulation; cell line; epithelium; growth factor; growth media; human tissue; immunofluorescence technique; lens; lens proteins; organ culture; transmission electron microscopy; western blottings

Cataract is the world's leading cause of blindness, the most frequently performed surgery on the elderly in the U.S., and is the largest item in Medicare expenses. Our long-term goal is to identify changes in the human lens epithelium that precede cataract formation and to identify therapeutic agents that prevent cataract formation. The epithelium is responsible for the formation of the lens, has a major transport function, provides cells that produce lens fibers and is the primary locus of enzymes and other compounds that protect the lens from oxidative insult. The major factor limiting experimentation on human lens epithelial cells (LECs) is that there are insufficient numbers of cells to initiate detailed biochemical and physiological experiments. Well characterized human LEC lines would be particularly useful for studying the control of crystallin gene expression. We will determine the conditions required to generate cell lines from normal and cataractous human lenses and will obtain detailed baseline information on growth and biochemical properties of human LECs. We will compare the biochemical and metabolic properties of lines established from normal and cataractous lenses with these parameters in immortalized human LECs transfected with AD12-SV40. Metabolic and transport properties in cultured human lECs will be compared with those in intact human lenses. We will determine the specific growth factor(s) that trigger mitosis in the epithelium of human lenses in organ culture thus permitting comparison with tissue culture studies. Aims are to 1) Determine the culture conditions required for the long-term growth of human LECs from normal lenses and frog tags obtained from human cataractous lenses. 2) Characterize the differentiated state of primary and long-term cultures of human LECs using biochemical markers (such as alphaA-crystallin, alphaB-crystallin, beta crystallin and gamma-crystallin) and ultrastructural markers (such as nuclear dissolution, loss of cytoplasmic organelles, and formation of gap junctions). 3) Determine the levels of selected biochemical parameters in newly established lens epithelial liens and in transfected human LECs. 4) Determine the conditions required to initiate DNA synthesis and mitosis in human lenses in organ culture.

白内障是世界上致盲的主要原因， 在美国为老年人做手术，是世界上 医疗费用。 我们的长期目标是确定人类 在白内障形成之前的透镜上皮细胞，并鉴定治疗性 预防白内障形成的药物。 上皮细胞负责 透镜的形成具有主要的运输功能，为细胞提供 产生透镜纤维，并且是酶和其他酶的主要场所。 保护透镜免受氧化损伤的化合物。 的主要因素 对人透镜上皮细胞（LEC）的限制性实验是 细胞数量不足以启动详细的生化反应 和生理学实验。 良好表征的人类LEC系将是 特别适用于研究晶体蛋白基因表达的调控。 我们将确定产生细胞系所需的条件， 正常和白内障的人类晶状体并将获得详细的基线 关于人LEC的生长和生化特性的信息。 我们将比较品系的生化和代谢特性 根据正常和白内障晶状体建立， 用AD 12-SV 40转染的永生化人LEC。 代谢和 将培养的人lEC中的转运性质与 完整的人类晶状体。 我们将确定特定的生长因子， 在器官培养中触发人晶状体上皮细胞的有丝分裂， 允许与组织培养研究进行比较。 目的是1）确定长期培养所需的培养条件 来自正常晶状体的人LEC的生长和来自人晶状体的青蛙标签 白内障晶状体 2)描述初级的分化状态 以及使用生物化学标记物（例如， α A-晶状体蛋白、α B-晶状体蛋白、β-晶状体蛋白和γ-晶状体蛋白） 和超微结构标志物（如核溶解、细胞核丢失、细胞核内的蛋白质丢失）。 细胞质细胞器和间隙连接的形成）。 3)确定 新建立的透镜中选定生化参数的水平 上皮细胞和转染的人LEC中。 4)确定 启动人类晶状体中DNA合成和有丝分裂所需的条件 器官培养。