Functional validation of adult hippocampal organotypic cultures as an in vitro model of brain injury
Functional validation of adult hippocampal organotypic cultures as an in vitro model of brain injury
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DOI：
10.1016/j.brainres.2003.12.009
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发表时间：
2004-03-19
期刊：
影响因子：
2.9
通讯作者：
Cho, SG
中科院分区：
文献类型：
--
作者：
Finley, M;Fairman, D;Cho, SG
为了确定海马锥体神经元在成熟器官型培养物中是否保留真实的功能特性，从年轻成年大鼠（P20-21）建立海马切片培养物。在体外培养7天，神经元层保持紧密的组织，而不是在围产期切片中经常观察到的锥体神经元的扩大重组。CA 3和CA 1锥体神经元响应于电流注入而激发动作电位，并表现出自发和诱发的突触电流，表明神经元功能完整且海马神经回路正常。我们还测试了切片培养对缺血性损伤的神经元敏感性。急性缺血模式导致选择性死亡的锥体神经元在CA 1区，这是防止治疗与NMDA拮抗剂，MK-801。在缺血期间检测到兴奋性和抑制性氨基酸神经递质的大量流出，与急性切片中显示的变化一致。总之，从年轻成年大鼠制备的海马器官型培养物在体外保持神经元结构和突触活性，并且可以与急性切片系统平行用于模拟成熟脑组织以检查缺血病理生理学和神经保护治疗。(C)2004 Elsevier B. V.保留所有权利。
To determine whether hippocampal pyramidal neurons retain authentic functional properties in mature organotypic culture, hippocampal slice cultures were established from young adult rats (P20-21). Cultures maintained 7 days in vitro retained tight organization of neuronal layers, as opposed to the widening restructure of pyramidal neurons often observed in perinatal slices. CA3 and CA1 pyramidal neurons fired action potentials in response to current injection and exhibited spontaneous and evoked synaptic currents, indicating intact neuronal function and normal hippocampal neural circuitry. We also tested neuronal sensitivity of slice cultures to ischemic injury. Acute ischemic paradigm resulted in selective death of pyramidal neurons in the CA1 region, which was prevented by treatment with an NMDA-antagonist, MK-801. Robust efflux of excitatory and inhibitory amino acid neurotransmitters was detected during ischemia, consistent with changes shown in acute slices. In summary, hippocampal organotypic cultures prepared from young adult rats maintained neuronal architecture and synaptic activity in vitro and can be used in parallel with an acute slice system to model mature brain tissue to examine ischemic pathophysiology and neuroprotective treatment. (C) 2004 Elsevier B.V. All rights reserved.