Structure and Function of the Cyclic Nucleotide Phosphodiesterases

戴维斯博士建议分离cDNA和基因组克隆 包含几种已知老鼠的结构信息 磷酸二酯酶（PDE）。 两个这样的基因组克隆已经 利用克隆的果蝇基因， 杂交探针 一旦知道了老鼠的基因序列， 将选择特定区域用于原位探针， mRNA在大鼠脑中的定位，并用于表征 处理过的信息的形式。 高滴度抗血清将 同时制备大鼠免疫细胞化学 个脑袋 最后，将检测小鼠/仓鼠杂交细胞系 用探针来确定 同源小鼠PDE基因。 自从萨瑟兰发现环核苷酸以来， 在调节细胞生长中起着重要作用。 几乎所有生物体的新陈代谢。 结果导致 调节产生（环化酶）和降解的酶 （偏微分方程）这些化合物一直是一个很大的主题， 重要性 大量直接和间接的证据， 指出了不同偏微分方程在各种 哺乳动物大脑的功能（例如，视觉和 内存）。 然而，到目前为止，关于这方面的信息还很少。 PDE基因的数量，关于它们的调节，以及关于 它们的产物在大脑中的定位和最终命运。 拟议的工作将提供所需的工具， 的该项目海外

Dr. Davis proposes to isolate cDNA and genomic clones containing structural information for the several known rat phosphodiesterases (PDEs). Two such genomic clones have already been identified using the cloned drosophila gene as a hybridization probe. Once the rat sequences are known, isozyme specific regions will be selected for probes for in situ localization of mRNAs in the rat brain, and for characterization of processed forms of the messages. High titer antisera will also be prepared and then used immunocytochemistry of the rat brain. Finally, mouse/hamster hybrid cell lines will be tested with the probes to establish the chromosomal locations of the cognate mouse PDE genes. Since their discovery by Sutherland, cyclic nucleotides have been shown to play a central role in the regulation of cell metabolism in virtually all organisms. As a result, the regulation of the enzymes that produce (cyclases) and degrade (PDEs) these compounds has continued to be a subject of great importance. Considerable evidence, both direct and indirect, points to important roles for the different PDEs in various functions of the mammalian brain (for example, vision and memory). As yet, however, there is little information about the number of PDE genes, about their regulation, and about the localization and eventual fate of their products in the brain. The proposed work will provide the tools needed to provide this information.