MOLECULAR MECHANISMS IN RETINAL DEGENERATIONS

视网膜变性的分子机制

• 批准号: 2654652
• 负责人: DEBORA B FARBER
• 金额: $ 27.12万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2654652
• 关键词: animal genetic material tag; cone cell; disease /disorder model; dogs; fluorescent in situ hybridization; gene expression; genetic disorder; genetic mapping; genetic strain; human tissue; messenger RNA; molecular cloning; molecular pathology; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; retina degeneration; rod cell

DESCRIPTION: The investigator proposes to isolate cone-specific cDNAs as a means to delineate the function of the visual cascade and cell biology of cone cells in addition to providing candidates for study of the cd dog in specific aim II. To accomplish this, the investigator will use the cd dog, the only available animal model of cone degeneration. Two techniques are proposed to identify mRNA species present in normal dogs, but absent from cd/cd dog retinas. The first is differential display of the two mRNA populations. This will be based on the use of four degenerately tailed oligo dT primers differing only in their last base and 20 10-base upstream primers. Subtractive hybridization will also be used to generate a pool of cone-specific cDNAs, the technique the investigator used to isolate the mouse rd gene. Specific Aim II. The investigator proposes to identify and characterize the gene lesion responsible for the cd dog disease. To accomplish this, differential display of retina mRNAs from pre-degenerate cd/cd and age matched normal dogs and possibly subtractive hybridization of predegenerate cd/cd retinas with age-matched cd/+ retinal cDNAwill be used. The investigator will use GDRDA if the first two techniques are unsuccessful.

描述：研究者建议将锥体特异性cDNA隔离为 描述视觉级联和细胞生物学功能的手段 除了提供研究CD狗的候选者外，锥细胞的 在特定的目标II中。为此，调查人员将使用CD 狗，唯一可用的锥变性动物模型。两种技术 建议识别普通犬中存在的mRNA物种，但不存在 来自CD/CD狗视网膜。第一个是两个mRNA的差分显示 人群。这将基于使用四个脱后的使用 Oligo DT引物仅在其最后一个基部和20个10碱基上有所不同 上游底漆。减法杂交也将用​​于 生成锥体特异性cDNA池，这是研究者的技术 用于分离小鼠RD基因。具体目标II。调查员 建议识别和表征负责的基因病变 用于CD狗疾病。为了实现这一目标，差异显示 来自降级前CD/CD的视网膜mRNA和年龄与正常狗相匹配，并且 可能会减去预元素CD/CD视网膜的杂交 使用年龄匹配的CD/+视网膜CDNAWILL。调查人员将使用 GDRDA如果前两种技术不成功。