MOLECULAR BIOLOGY OF VISION

这项拨款建议进行的研究，目的是深入了解 视觉系统中用于信号转导的机制使用 分子、遗传和生理相结合的方法。这是意料之中的 这些研究的结果将有助于我们理解 生物学中的感觉接收和信息处理基础 系统。 在过去的两年半里，我们集中了我们的研究努力 编码参与光信号转导的蛋白质的基因特征 卡斯卡德。我们分离了NINA基因(亲环素同源)，a 光感受器细胞特异性蛋白激酶C，果蝇的同源物 脊椎动物arrestin，以及光感受器细胞特异性脱氢酶。我们 也继续我们对果蝇视蛋白的分析和遗传 在R1-R6中表达不同视蛋白的转基因果蝇系 感光细胞。这些实验使我们能够检查 不同视蛋白对血管紧张素转运蛋白功能特性的贡献 不同的感光细胞类型。这项提案现在将我们的 对其中两个基因家族的研究工作：opsins和ineaA。 我们将：(1，2)确定光谱特性的分子基础 视蛋白分子。(3)分离编码视蛋白的基因(S)。 果蝇复眼的中央R8感光细胞类，以及 (4)产生缺乏R7和R8视觉色素的突变体，以及 从分子和生理上描述它们的特征。我们还将：(5) 鉴定NINA基因的表达部位，并产生抗体 来对抗它的基因产物。(6)构建高表达转基因株系 NINA，并在细菌(T7)中产生并鉴定了NINA蛋白 表达系统)和果蝇培养细胞(施耐德细胞)。(7) 详细描述了NINA的视觉生理和生物化学 果蝇和野蝇心理表型的敏锐测定 并长期使用CsA治疗。(8)构造和表征热能- 电击诱导的NINA基因--分离温度敏感型的尝试 NINA等位基因。(9)分离NINA基因家族的其他成员，以及 寻找可能的R7细胞特异性同源基因。最后，我们将在中国学习 详细描述那些基于它们的表达模式和 大多数遗传和生化标准都需要进一步研究。

The aim of the research proposed in this grant is to gain insight into the mechanisms used for signal transduction in the visual system using a combined molecular, genetic and physiological approach. It is expected that the results obtained from these studies will help our understanding of the basis of sensory reception and information processing in biological systems. Over the past 2-1/2 years we have concentrated our research efforts characterizing genes encoding proteins involved in the phototransduction cascade. We isolated the ninA gene (cyclophilin-homologue), a photoreceptor-cell specific protein kinase C, a Drosophila homologue of vertebrate arrestin, and a photoreceptor-cell specific dehydrogenase. We have also continued our analysis of Drosophila opsins and genetically engineered Drosophila lines that express different opsins in the R1-R6 photoreceptor cells. These experiments allowed us to examine the contribution of the various opsins to the specific functional properties of the different photoreceptor cell types. This proposal now focus our research efforts on two of these gene families: opsins and ninaA. We will: (1,2) Determine the molecular basis of spectral specificities in the opsin molecule. (3) Isolate the gene(s) encoding the opsin expressed in the central R8 photoreceptor cell class of the Drosophila compound eye, and (4) generate mutants lacking the R7 and R8 visual pigments, and characterize them molecularly and physiologically. We will also: (5) Identify the sites of expression of the ninaA gene, and generate antibodies against its gene product. (6) Generate transgenic lines overexpressing ninaA, and produce and characterize the ninaA protein in bacteria (T7 expression system) and Drosophila culture cells (Schneider cells). (7) Characterize in detail the visual physiology and biochemistry of ninaA flies and determine the psychological phenotype of wild-type flies acutely and chronically treated with CsA. (8) Construct and characterize a heat- shock inducible ninaA gene an attempt to isolate temperature sensitive alleles of ninaA. (9) Isolate other members of the ninaA gene family, and search for a putative R7 cell-specific homolog. Finally, we will study in detail those sequences which based on their patterns of expression and genetic and biochemical criteria most warrant further investigation.