Excitotoxic cell death of ES derived neural lineage cells

描述：(摘自摘要)创伤性脊髓的治疗方法 目前伤势有限。本节提出的实验， 项目I是新的计划项目申请的一部分。此计划项目 有探索治疗潜力的总体长期目标 胚胎干细胞来源的神经系细胞移植 进入了受伤的脊髓。项目I的具体目标是 测定ES细胞对兴奋性毒性和氧化损伤的易感性 在体外，并确定遗传和药理学方法来阻断 这个漏洞可以用来最大化ES细胞在 移植。 项目I的第一部分将使用全细胞膜片钳测试ESNLC 录音和Fura-2染料显微荧光法验证许多神经细胞 表达功能性NMDA、AMPA/海人藻酸、γ-氨基丁酸(GABA)和 甘氨酸受体，并确定I组mGluRs的患病率。 随后，项目的主要部分将检验Es的假设 神经元和少突胶质细胞对兴奋性毒性的易感性 (由添加外源性兴奋性毒素或暴露于 氧气-葡萄糖缺乏)，这种脆弱性可以减弱 使用NMDA或AMPA/海人藻酸受体阻滞剂、抗氧化剂或I组mGluR 封锁接近了。它还将检验接触NT-3的假设 会增强ES神经细胞对兴奋性毒性的易感性。如果这是 将致力于确定这种神经营养因子介导的 损伤增强是由trks介导的，并可被抗氧化剂减弱。 接近了。这些体外实验的结果将在 项目III和项目IV中进行的活体实验的设计，其中 受脊髓挫伤的啮齿动物将接受ESNLC 移植。

DESCRIPTION: (From abstract) Therapeutic approaches to traumatic spinal cord injury are currently limited. Experiments proposed in the present section, Project I, are part of a new Program Project application. This Program Project has the overall long-term goal of exploring the therapeutic potential of transplanting embryonic stem (ES) cell-derived neural lineage cells (ESNLCs) into the traumatically-injured spinal cord. The specific goal of Project I is to determine the vulnerability of ES cells to excitotoxic and oxidative insults in vitro, and to identify genetic and pharmacological approaches to blocking this vulnerability that can be used to maximize ES cell survival after transplantation. The first part of Project I will test ESNLCs with whole-cell patch clamp recordings and fura-2 dye microfluorimetry to verify that many neuronal cells express functional NMDA, AMPA/kainate, gamma-amino-butyric acid (GABA) and glycine receptors, and to determine the prevalence of Group I mGluRs. Subsequently, the main portion of the Project will test the hypothesis that ES neurons and oligodendrocytes develop vulnerability to excitotoxicity in vitro (induced either by addition of exogenous excitotoxins, or exposure to oxygen-glucose deprivation), and that this vulnerability can be attenuated using NMDA or AMPA/kainate receptor blockade, anti-oxidant, or Group I mGluR blockade approaches. It will also test the hypothesis that exposure to NT-3 will enhance ES neuronal cell vulnerability to excitotoxicity. If this is the case, effort will be directed to determining whether this neurotrophin-mediated injury enhancement is mediated by trks, and can be attenuated by anti-oxidant approaches. The results of these in vitro experiments will be exploited in the design of the in vivo experiments carried out in Projects III and IV, where rodents subjected to spinal cord contusion injury will receive ESNLC transplants.