Novel Chamber for Axonal Glutamate Signal Transduction

轴突谷氨酸信号转导的新型室

• 批准号: 6897929
• 负责人: ANNE MARION TAYLOR
• 金额: $ 2.66万
• 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 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6897929
• 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（由申请人提供）：作为精致和高度极化的细胞，中枢神经系统的神经元经常将其轴突延伸到极端距离上。在这种现象中，神经元过程遇到了多个不同的细胞外微环境。 β-淀粉样蛋白，谷氨酸和神经营养蛋白都是这种微环境暴露的例子，可能会影响过程遇到的神经元的功能甚至存活。研究这些微环境对细胞过程和信号级联反应的影响的研究很大程度上是由于难以在体外复制这种微环境暴露的困难而阻碍。我们的实验室最近使用微型制动和软光刻技术开发了新的腔室，这些技术可以可靠，可重复地模拟CNS神经元遇到的微观视力。进一步的证据表明，分离的神经突在体外九天后仅是轴突。该提案旨在进一步开发这种新型的多区域模型，用于研究中央神经元的微环境影响。此外，该模型的实用性将在对介导cAMP响应元件结合蛋白（CREB）磷酸化的信号传导级联反应中进行研究，以响应轴突限制的谷氨酸暴露。该提案将检验以下假设：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反应引起CREB磷酸化。特别是，我们将专注于Ras-ERK和CAM激酶途径的潜在钙介导的激活。