Establishment of the pharmacological sciences to elucidate the signal transduction system.

本研究项目的目的之一是阐明和控制细胞内第二信使系统转导来自细胞外刺激的信号的机制，使用特定的分子探针。另一个是创造新的功能靶向探针，这对研究细胞内机制是有用的。这种药理学方法无疑是强大的，因为各种生物现象是由多个细胞内信使系统组成的网络控制的。通过这项资助下的三年工作，越来越多的特定分子探针已经可用，这使我们能够了解细胞内信使系统。一个很好的例子是KN-62和93，它们是Ca/钙调素依赖性蛋白激酶II的特异性抑制剂，已被广泛用于研究该激酶的功能。我们还成功地纯化了新型的钙/钙调素依赖性激酶。v .我们利用药物获得了神经元中速激肽、GABA和血清素信号转导机制的证据。一氧化氮被证明参与了脑血管扩张的神经控制。从分子生物学的角度，克隆了两种从细胞内储存位点释放Ca^<2+>的Ca通道。所有这些发现表明，迫切需要新的特异性探针来基本了解细胞内信使系统。此外，这些探针可能为创造临床使用的新化合物提供线索。

One of the aims of this research project was to elucidate and control the mechanisms by which the intracellular second messenger systems transduce signals derived from extracellular stimuli, using specific molecular probes. The other was to create novel function-targeted probes which are useful to investigate the intracellular mechanisms. Such a pharmacological approach is undoubtedly powerful since various biological phenomena are controlled by a network of multiple intracellular messenger systems. Through three-year work under this grant, increasing number of specific molecular probes have became available, which enabled us to understand intracellular messenger systems. A good example is KN-62 and 93, specific inhibitors of Ca/calmodulin-dependent protein kinase II,which have been widely used to investigate the functions of the kinase. We also succeeded to purify the novel type of Ca/calmodulin-dependentkinase. V.We obtained evidence concerning signal transduction mechanisms by tachykinin, GABA and serotonin in neurons using pharmacological agents. Nitric oxide was demonstrated to be involved in the neural control of the brain vessel dilation.From an molecular biological approach, two types of Ca channels releasing Ca^<2+>from the intracellular store sites have been cloned. All these findings indicate that novel specific probes are desperately required for fundamental understanding of the intracellular messenger systems. Furthermore, these probes may provide a clue to create new compounds for a clinical use.