GENETICS AND BIOCHEMISTRY OF THE LENS IN CELL CULTURE

细胞培养中晶状体的遗传学和生物化学

• 批准号: 3261515
• 负责人: ROBERT L CHURCH
• 金额: $ 13.37万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-01-01 至 1988-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3261515
• 关键词: basement membrane; cataract; chromatography; chromosomes; cytogenetics; electrofocusing; electron microscopy; enzyme linked immunosorbent assay; epithelium; flow cytometry; gel electrophoresis; genetic mapping; genetic regulation; histochemistry /cytochemistry; human tissue; hybrid cells; hybridomas; immunochemistry; isozymes; lens proteins; monoclonal antibody; tissue /cell culture

For proper functioning of the lens, elasticity and clarity are of paramount importance. These characteristics are highly dependent on the integrity of the lens capsule basement membrane and the lens fiber cell crystallins and cell membranes. The system for their synthesis and maintenance is known to be delicate, however, and can often be disrupted in many diseases, generally leading to cataracts. We intend to continue our studies of the normal lens capsule basement membrane and the crystallins produced by the lens epithelium and fiber cells, and investigate how these proteins are affected by cataracts. We will establish cell lines from human normal and cataractous lens epithelium, as well as with appropriate mouse models in monolayer tissue culture. This will provide us with large quantities of biosynthetic products for direct biochemical and immunological study without the need for harsh extraction procedures. By use of somatic cell hybridization, we will investigate the genetic control of basement membrane and crystallin biosynthesis, and carry out gene mapping of the chromosomes responsible for the proteins and their processing enzymes. Furthermore, we will produce hybrids between normal x diseased, and diseased x diseased lens cells to investigate the mechanisms and controls that function in the biosynthesis of lens proteins. These data will help define the true nature of the molecules of the lens and further help to establish a biochemical basis for symptoms seen in the disease states. These studies will provide information about the biochemical and genetic regulation of lens proteins, specifically basement membrane and crystallins. The control of normal basement membrane and crystallin synthesis by cells is an important aspect of normal growth and development, and studies involved with understanding how such controls work, as well as the chromosomal localizations of the various lens proteins, will aid in our understanding of errors that occur in various lens proteins, will aid in our understanding of errors that occur in various hereditary and acquired disorders of the lens, such as cataract.

为了使透镜正常工作，弹性和清晰度是至关重要的 重要性 这些特征高度依赖于 透镜囊基膜和透镜纤维细胞晶体蛋白， 细胞膜 已知用于它们的合成和维持的系统 然而，它是脆弱的，在许多疾病中经常会被破坏， 通常导致白内障。 我们打算继续研究 正常透镜囊基底膜和晶状体囊膜产生的晶体蛋白 透镜上皮细胞和纤维细胞，并研究这些蛋白质是如何 受白内障的影响。 我们将从正常人和 白内障透镜上皮，以及适当的小鼠模型， 单层组织培养 这将为我们提供大量的 用于直接生物化学和免疫学研究的生物合成产品 而不需要苛刻的提取程序。 利用体细胞 杂交，我们将调查遗传控制的基底膜 和晶状体蛋白的生物合成，并进行染色体的基因定位 负责蛋白质和它们的加工酶。 而且我们 会产生正常x患病和患病x患病的杂交种 透镜细胞研究机制和控制功能， 透镜蛋白的生物合成。 这些数据将有助于确定 并进一步帮助建立一个生物化学 在疾病状态下观察到的症状的基础。 这些研究将提供有关生物化学和遗传学的信息。 调节透镜蛋白，特别是基底膜， 晶体蛋白。 正常基底膜和晶体蛋白的调控 细胞合成是正常生长和发育的一个重要方面， 以及了解这种控制如何工作的研究，以及 各种透镜蛋白的染色体定位，将有助于我们的研究。 对各种透镜蛋白中发生的错误的理解，将有助于 我们对各种遗传性和后天性错误的理解 透镜疾病，如白内障。