CAMP-DEPENDENT PROTEIN KINASES IN NEURONAL MODULATION

神经元的特性可以通过调制传递器改变，一种 有助于神经回路可塑性的过程。我们的长特 M 目标是描绘细胞内信号转导通路是如何 CAMP的利用参与了神经元的调制。所获得的信息 将特别有助于我们对学习和记忆的理解 也与生物节律的起源--心肌的活动有关，以及 E 多肽类激素的分泌机制。我们的发现将有助于设计 药理药剂干预这些过程。 具体地说，将使用分子生物学技术来研究 E CAMP依赖的蛋白激酶(CAPK)在调控中的作用 神经递质改变机械感觉神经元的特性 简单的反射行为。加州海兔软体动物将用于 这些研究是因为可以将我们的发现与正在进行的 行为和电生理检查，已经很好了 高级。首先，将对海兔的多样性程度进行调查 D 通过检测编码调控基因和基因片段的cDNA和基因组DNA片段 酶的催化亚基。下一步， 参与感觉细胞调节的酶，特别是促进 感觉运动性突触，将通过微量注射每个多肽来确定 (通过表达获得)进入感觉神经元并监测 这些细胞及其相关运动神经元的电生理特性 细胞和亚细胞的位置和浓度的检查 也将进行相应的RNA和多肽的研究， 然后进行实验，以确定形状、级别或位置 在突触调节过程中被改变，如果是这样的话，是通过什么机制改变的？ 最后，生理上重要的形式的分子性质 将检查Aplysia CAPK，包括底物的鉴定 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 将进行严格的评估，如有必要，将设计新的模型 并进行了测试。

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.