CAMP-DEPENDENT PROTEIN KINASES IN NEURONAL MODULATION

神经元的特性可以被调节性递质改变， 这一过程有助于神经回路的可塑性。我们的龙德 M 目的是描述细胞内信号转导途径如何 利用cAMP参与神经元调节。获取的信息 将特别有助于我们对学习和记忆的理解， 心脏肌肉的活动，生物节律的起源， e 肽类激素分泌机制。我们的发现将有助于设计 药物干预这些过程。 具体地说，分子生物学技术将被用于 e cAMP依赖性蛋白激酶（cAPKs）在调节 神经递质改变了机械感觉神经元的特性， 简单的反射行为软体动物Aaplasia californica将被用于 这些研究，因为它将有可能将我们的研究结果与正在进行的 行为和电生理研究，这些研究已经很好地 先进首先，将阐明Aaplasia cAPK的多样性程度， D 通过检测编码调节和调节基因的cDNA和基因组DNA片段， 酶的催化亚基。接下来，介绍 参与感觉细胞调节的酶，特别是促进 感觉运动突触，将通过显微注射每种多肽 （通过表达获得）进入感觉神经元，并监测 这些细胞和相关运动神经元的电生理特性 细胞和亚细胞位置和浓度的检查 相应的mRNA和多肽的研究也将进行， 然后进行实验，以确定是否形式，水平或位置， 在突触调制过程中被改变，如果是这样，是通过什么机制改变的？ 最后，生理上重要的形式的分子特性， 将检查失活菌cAPK，包括鉴定底物， D 磷酸化对它们的影响。 利用这些信息，现有的短期和长期调制模型 将进行严格评估，如有必要，将设计新的模型 和测试。

The properties of neurons can be altered by modulatory transmitters, a process which contributes to the plasticity of neural circuits. Our long-te m goal is to delineate how the intracellular signal transduction pathway utilizing cAMP is involved in neuronal modulation. The information acquired will contribute especially to our understanding of learning and memory, and also to the action of heart muscle, the origin of biological rhythms, and t e mechanism of peptide hormone secretion. Our findings will aid in the design of pharmacological agents to intervene in these processes. Specifically, the techniques of molecular biology will be used to investiga e the roles played by cAMP-dependent protein kinases (cAPKs) when modulatory neurotransmitters alter the properties of mechanosensory neurons involved i simple reflex behaviors. The mollusk Aplysia californica will be used in these studies because it will be possible to relate our findings to ongoing behavioral and electrophysiological investigations, which are already well advanced. First, the extent of diversity of Aplysia cAPK will be investigat d by examining cDNAs and genomic DNA fragments encoding the regulatory and catalytic subunits of the enzyme. Next, the forms and properties of the enzyme involved in sensory cell modulation, in particular facilitation of sensorimotor synapses, will be determined by microinjecting each polypeptid (obtained by expression) into sensory neurons and monitoring the electrophysiological properties of these cells and associated motor neurons An examination of the cellular and subcellular locations and concentrations of the corresponding mRNAs and polypeptides will also be carried out, followed by experiments to determine whether the forms, levels or locations are altered during synaptic modulation, and if so by what mechanisms? Finally, the molecular properties of the physiologically important forms of Aplysia cAPK will be examined, including the identification of substrates a d the effects phosphorylation has on them. Using this information, existing models for short- and long-term modulation will be critically evaluated and, if necessary, new models will be devised and tested.