MOLECULAR MECHANISMS IN RETINAL DEGENERATIONS

描述：研究者建议分离视锥细胞特异性cDNA， 一种描述视觉级联和细胞生物学功能的方法 除了为CD犬的研究提供候选者之外， 具体目标二。为了实现这一点，研究者将使用cd 狗，唯一可用的视锥细胞变性的动物模型。两种技术 提出了识别mRNA的物种存在于正常狗，但缺乏 来自CD/CD狗视网膜。第一种是两种mRNA的差异显示 人口。这将是基于使用四个退化尾 oligo dT引物仅在其最后一个碱基和20个10-碱基上不同 上游引物。消减杂交也将用于 产生一个视锥细胞特异性cDNA库，这项技术 用于分离小鼠RD基因。具体目标二。研究者 提出了识别和表征基因损伤负责 关于CD狗的疾病为了实现这一点， 视网膜mRNAs来自前退化cd/cd和年龄匹配的正常狗， 前变性CD/CD视网膜与 将使用年龄匹配的CD/+视网膜cDNA。研究者将使用 如果前两种技术不成功，则使用GDRDA。

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.