Endothelial-neuronal interactions during development

描述(由申请人提供)： 我们的假设是，神经营养受体之间存在复杂的、动态的相互作用 和“血管”受体信号在神经元和中枢神经内皮细胞在出生后 发育，在慢性亚致死性缺氧时受到干扰。在此授权期内， 使用各种方法，我们建议：1.表征 神经营养(NT)受体(BDNF受体)和血管内皮生长因子和血管生成素受体(VEGFR1& 2，Tie2)在体内和体外常氧和低氧条件下对中枢神经系统(CNS)神经元和内皮的影响 低氧状态。2.描述相互作用和涉及的信号通路 NT(TrkB&p75NGF)和VEGF(VEGFR1&R2)受体在神经元中的相互作用 在常氧和低氧条件下体外培养。具体来说，我们将确定：VEGFR的影响 NT受体的信号转导及大脑神经元的信号转导；NT的作用 信号对脑神经元中血管内皮生长因子受体表达和信号的影响 神经降压素和血管内皮生长因子(以及血管内皮生长因子和神经降压素)受体在神经元上的顺序结合 增殖、存活和分化。3.描述相互作用和信号传递 NT(TrkB&p75NGF)和VEGF(VEGFR1&R2)受体参与的通路 常氧和低氧条件下体外培养脑内皮细胞的结合。 具体地说，我们将确定VEGFR信号对NT受体(TrkB& P75NGF及其配体在脑微血管内皮细胞中的表达 NT信号(通过BDNF)对脑内血管内皮生长因子受体表达和信号的影响 微血管内皮细胞；以及序贯NT和血管内皮生长因子(和血管内皮生长因子)的作用 NT)受体参与内皮细胞的增殖、存活和分化。4. 神经元与脑微血管内皮细胞共培养中NT、VE-GF受体和配体的表达模式及相互作用 常氧和低氧条件下培养的星形胶质细胞、神经元、RBE4细胞和星形胶质细胞 条件。5.调查和表征在我们的核心设施中发现的选定的新基因 通过与目标1-4相关的在慢性低氧过程中调节的基因芯片分析。 在这项拨款申请中提出的实验，目的是阐明 神经营养受体与“血管”受体的复杂动态相互作用 出生后发育过程中中枢神经系统神经元和内皮细胞的信号转导及其意义 在慢性亚致死性缺氧期间的中断。在下一个赠款期间积累的知识 将促进新的合理的治疗方法和药物的发展 在这些细胞类型的调节过程中，它们将更加具体和选择性 早产儿出生后脑发育的关键期。

DESCRIPTION (provided by applicant): Our hypothesis is that there is a complex, dynamic interaction between neurotrophic receptor and "vascular" receptor signaling in neurons and endothelia in the CNS during postnatal development and it is disrupted during chronic sublethal hypoxia. During this grant period, using a variety of methodologies, we propose to: 1. Characterize the expression of neurotrophic (NT) receptors (BDNF receptors) and VEGF & angiopoietin receptors (VEGFR1 & 2, Tie2) on neurons and endothelia in the CNS (cortex) in vivo and in vitro in normoxic and hypoxic conditions. 2. Characterize the interactions and the signaling pathways involved following NT (trkB & p75NGF) and VEGF (VEGFR1 & R2) receptor engagement in neurons in vitro in normoxic and hypoxic conditions. Specifically, we will determine: the effects of VEGFR signaling upon NT receptor expression and signaling in cerebral neurons; the effects of NT signaling upon VEGF receptor expression and signaling in cerebral neurons; and the effects of sequential NT and VEGF (and VEGF and NT) receptor engagement upon neuronal proliferation, survival and differentiation. 3. Characterize the interactions and the signaling pathways involved following NT (trkB & p75NGF) and VEGF (VEGFR1 & R2) receptor engagement in cerebral endothelial cells in vitro in normoxic and hypoxic conditions. Specifically, we will determine; the effects of VEGFR signaling upon NT receptor (trkB & p75NGF) and ligand (BDNF) expression in cerebral microvascular endothelial cells; the effects of NT signaling (via BDNF) upon VEGF receptor expression and signaling in cerebral microvascular endothelial cells; and the effects of sequential NT and VEGF (and VEGF and NT) receptor engagement upon endothelial proliferation, survival and differentiation. 4. Determine NT and VE--GF receptor and ligand expression patterns and interactions in cocultures of neurons & cerebral microvascular endothelial (RBE4) cells; RBE4 cells & astrocytes; and neurons, RBE4 cells and astrocytes in vitro in normoxic and hypoxic conditions. 5. Investigate and characterize selected novel genes identified in our core facility by cDNA array analyses that are modulated during chronic hypoxia and pertinent to Aims 1 - 4. The experiments proposed in this grant application have as their goal the elucidation of the complex, dynamic interactions between neurotrophic receptor and "vascular" receptor signaling in neurons and endothelia in the CNS during postnatal development and their disruptions during chronic sublethal hypoxia. Knowledge accrued during the next grant period will facilitate the development of new and novel rational therapeutic approaches and agents which will be more specific and selective in their modulation of these cell types during the critical period of postnatal brain development in the premature newborn.