Novel Chamber for Axonal Glutamate Signal Transduction

轴突谷氨酸信号转导的新型室

• 批准号: 6739385
• 负责人: ANNE MARION TAYLOR
• 金额: $ 2.91万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-12 至 2008-04-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6739385
• 关键词: axon; bioengineering /biomedical engineering; biological signal transduction; biomedical equipment development; cAMP response element binding protein; calmodulin dependent protein kinase; cell differentiation; cell proliferation; confocal scanning microscopy; controlled environment chamber; fluid flow; genetically modified animals; glutamate transporter; immunocytochemistry; laboratory rat; mitogen activated protein kinase; phosphorylation; posttranslational modifications; predoctoral investigator; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): As elaborate and highly polarized cells, neurons of the central nervous system frequently extend their axons over extreme distances. Inherent in this phenomenon, neuronal processes encounter multiple varying extracellular microenvironments. beta-amyloid, glutamate, and neurotrophins are all examples of such microenvironmental exposures that may influence the function or even survival of neurons whose processes encounter them. Studies that examine the impact of these microenvironments on cellular processes and signaling cascades have in large part been hindered by the difficulty in replicating in vitro such microenvironmental exposures. Our lab has recently developed novel chambers using microfabrication and soft lithography techniques that can reliably and reproducibly simulate microenviroments encountered by CNS neurons. Further evidence shows that the neurites isolated are exclusively axons after nine days in vitro. This proposal is designed to further develop this novel multicompartment model for the study of microenvironmental influences over central neurons. In addition, the utility of this model will be examined in a study of the signaling cascades that mediate the phosphorylation of cAMP-responsive element binding protein (CREB) in response to axonally-restricted glutamate exposure. This proposal will test the hypothesis that axonal glutamate exposure induces CREB-phosphorylation via specific receptor-mediated initiation of intracellular signal transduction cascades. In particular, we will focus on potential calcium-mediated activation of Ras-ERK and CaM kinase pathways.

描述(申请人提供)：作为精细和高度极化的细胞，中枢神经系统的神经元经常将它们的轴突延伸到极端距离。在这种现象中，神经元突起会遇到多种不同的细胞外微环境。β-淀粉样蛋白、谷氨酸和神经营养因子都是这种微环境暴露的例子，这些微环境暴露可能会影响其加工过程遇到它们的神经元的功能甚至生存。研究这些微环境对细胞过程和信号级联的影响的研究在很大程度上受到了在体外复制此类微环境暴露的困难的阻碍。我们的实验室最近开发了使用微制造和软光刻技术的新型电离室，可以可靠和可重复地模拟中枢神经系统神经元遇到的微环境。进一步的证据表明，在体外培养9天后，分离的轴突完全是轴突。这项建议旨在进一步发展这一新的多室模型，以研究微环境对中枢神经元的影响。此外，这个模型的实用性将在一项关于介导cAMP反应元件结合蛋白(CREB)磷酸化的信号级联反应的研究中得到检验，以响应轴向受限的谷氨酸暴露。这项提议将检验轴突谷氨酸暴露通过特定受体介导的细胞内信号转导级联启动CREB磷酸化的假说。特别是，我们将关注潜在的钙介导的Ras-ERK和CaM激酶通路的激活。