LITHIUM REGULATION OF BRAIN PROTEIN KINASE C SUBSTRATES

锂对脑蛋白激酶 C 底物的调节

• 批准号: 3430346
• 负责人: Robert H. Lenox
• 金额: $ 4.22万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-07-01 至 1994-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3430346
• 关键词: biological models; brain cell; cell line; enzyme activity; enzyme substrate; genetic transcription; genetic translation; hippocampus; isozymes; laboratory mouse; lithium; model design /development; molecular cloning; neuropharmacology; phorbols; phosphoproteins; posttranslational modifications; protein kinase C

We have recently reported that chronic lithium profoundly alters the intracellular concentration of the rat. This reduction in a Myristoylated Alanine Rich C Kinase Substrate (MARCKS) protein appears to parallel the time course of action of lithium in the clinical treatment of bipolar disorder. Accumulating evidence from our laboratory and others implicates PKC in the long term action of lithium in the brain as well as the regulation of some of its major phosphoprotein substrates, including MARCKS. In order to move this research to the next phase of examining our hypotheses regarding the mechanism of action of lithium in regulating this PKC substrate, we must initiate the development of a physiologically relevant cell system, amenable to sophisticated experimental approaches and devoid of the limitations of in vivo models. In this Small Grant proposal we will establish a new cell model system using immortalized hippocampal cells in culture, and take advantage of current cellular heterogeneity to initiate a subcloning strategy to isolate and stabilize phenotypic properties as a valuable resource for our future studies. For example, while we have identified MARCKS expression in an early passage of these cells, we anticipate the development of subclones with differential levels of expression of this phosphoprotein. Inasmuch as lithium's action in brain appears to depend upon limited access to inositol, we will develop analogous conditions for lithium exposure in our cell system. We will replicate our in vivo findings in these hippocampal cells and examine transcriptional, translational and posttranslational mechanisms by which lithium alters the regulation and expression of MARCKS in the hippocampus. These studies will provide the necessary foundation to pursue more major funding in the future to examine the possibility that the action of lithium on MARCKS is shared by other known major PKC substrates unique to brain. In addition, we will then be in an excellent position to test our hypothesis that alterations in MARCKS and/or the other phosphoproteins may play a critical role in the action of chronic lithium on the regulation of muscarinic receptors coupled to phospholipase C in key regions of the brain.

我们最近报道称，长期锂盐会深刻改变 大鼠细胞内浓度。 这减少了 肉豆蔻酰化丙氨酸富集 C 激酶底物 (MARCKS) 蛋白出现 平行锂在临床中的作用时间过程 双相情感障碍的治疗。 从我们的实验室积累证据 等人认为 PKC 与锂在大脑中的长期作用有关 以及对其一些主要磷蛋白底物的调节， 包括马克斯。 为了将这项研究推进到下一阶段 检查我们关于锂作用机制的假设 调节这种 PKC 底物，我们必须开始开发 生理相关的细胞系统，适合复杂的 实验方法，不受体内模型的限制。 在这个小额资助提案中，我们将建立一个新的细胞模型系统 使用培养中的永生化海马细胞，并利用 当前的细胞异质性启动亚克隆策略 分离和稳定表型特性作为宝贵的资源 我们未来的学习。 例如，虽然我们已经确定了 MARCKS 在这些细胞的早期传代中表达，我们预计 具有不同表达水平的亚克隆的发育 磷蛋白。 因为锂在大脑中的作用似乎取决于 在肌醇的获取有限的情况下，我们将开发类似的条件 我们的细胞系统中的锂暴露。 我们将在体内复制我们的 在这些海马细胞中发现并检查转录， 锂改变的翻译和翻译后机制 海马MARCKS的调控和表达。 这些 学习将为追求更多专业提供必要的基础 未来的资金，以审查该行动的可能性 MARCKS 上的锂与其他已知的主要 PKC 基材共享，是独一无二的 到大脑。 此外，我们将处于有利的位置来测试 我们的假设是 MARCKS 和/或其他的改变 磷蛋白可能在慢性锂的作用中发挥关键作用 与磷脂酶 C 偶联的毒蕈碱受体的调节 大脑的关键区域。