MOLECULAR CHARACTERIZATION OF THE RHODOPSIN GENE

视紫红质基因的结构特征与其相关性的研究 光信号转导调控其表达的分子机制 而对光感受器细胞的存活是重要的神经生物学 人们对这些问题知之甚少。总体目标 这五年的研究计划是表征分子结构 以及小鼠视紫红质基因的功能。这包括对 基因和环境对其表达的调节(光与 因子及其控制机制的研究进展 正常小鼠和遗传性痴呆小鼠发育过程中的表达 视觉缺陷。视蛋白和48K(S抗原， Arrestin)以非协调方式由光和/或 动物所处的明/暗循环。此外， 若干光感受器特异蛋白在视杆中的定位 内段和外段是瞬变的，并且依赖于光。这个 MRNAs浓度变化与细胞周期变化的关系 光感受器特异蛋白与发育，基因， 而光线将由西方和北方的分析和 免疫细胞化学。小鼠视蛋白基因已被克隆并 测序，并证明了推测的调控序列。顺应时代 参与基因转录的核苷酸序列和反式作用因子 光介导的反应将使用DNA延迟分析来鉴定 和转基因研究。这些基因的确切染色体位置 小鼠视紫红质和48K将通过Southern分析确定 中国仓鼠-小鼠体细胞基因组DNA的限制性内切酶分析 细胞杂交和来自重组近交系的小鼠。该结构 这些基因的表达将在发生突变的小鼠身上进行检测 它们映射在它们的轨迹上或附近。该病毒的核苷酸序列 将确定视紫红质基因的等位基因形式并 视紫红质基因的定性和定量活性 仅为这些基因确定，并与突变(RD)相结合 和rds)，导致遗传性视网膜变性和转录 以及转录后对视蛋白基因表达的影响， 在感光细胞死亡之前。分子结构 而小鼠视紫质基因的五个转录本的功能将 在正常小鼠和突变小鼠身上检测到。由此生成的数据 项目可能会增加我们对正常流程的理解 人类的光转导和/或遗传性的分子病理学 视网膜变性。

The relationship of the structural features of the rhodopsin gene and the molecular mechanisms regulating its expression, to phototransduction and to the survival of photoreceptor cells are important neurobiological problems about which relatively little is known. The overall objective of this 5 year research plan is to characterize the molecular structure and function of the mouse rhodopsin gene. This includes studies on the regulation of its expression by genetic and environmental (light vs. dark) factors and the investigation of the mechanisms controlling its expression during development of normal mice and of mice with inherited visual defects. The amounts of mRNA for opsin and 48k (S-antigen, Arrestin) are regulated in a non-coordinate manner by light and/or the light/dark cycle to which the animal is exposed. In addition, the localization of a number of photoreceptor specific proteins in the rod inner and outer segments is transient and dependent on light. The relationship between the changes in concentration of mRNAs and photoreceptor specific proteins with respect to development, genotype, and light will be determined by Western and Northern analysis and by immunocytochemistry. The mouse opsin gene has been cloned and sequenced, and putative regulatory sequence demonstrated. Cis-acting nucleotide sequences and trans-acting factors which participate in the light mediated responses will be identified using DNA retardation assays and transfection studies. The exact chromosomal loci for the genes for mouse rhodopsin and 48k will be determined using Southern analysis of restriction digests of genomic DNA from Chinese hamster-mouse somatic cell hybrids and from recombinant inbred lines of mice. The structure and expression of these genes will be examined in mice with mutations which map at or near to their loci. The nucleotide sequences of identified allelic forms of the rhodopsin gene will be determined and the qualitative and quantitative activity of the rhodopsin gene determined for these genes alone, and in combination with mutations (rd and rds) which cause inherited retinal degeneration and transcriptional and post transcriptional effects on the expression of the opsin gene, prior to the death of the photoreceptor cells. The molecular structure and function of the five transcripts from the mouse rhodopsin gene will be determined in normal and mutant mice. The data generated by this project may increase our understanding of the normal process of phototransduction in man and/or the molecular pathology of inherited retinal degeneration.