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當前位置 > 首頁 > 技術文章 > 單個神經元O2消耗量、細胞內Ca2+濃度和線粒體膜電位的同時記錄

單個神經元O2消耗量、細胞內Ca2+濃度和線粒體膜電位的同時記錄

瀏覽次數:3213 發布日期:2009-6-17  來源:本站 僅供參考,謝絕轉載,否則責任自負
    非損傷微測技術用于神經毒性機制的研究
    谷氨酸毒性研究:單個神經元O2消耗量、細胞內Ca2+濃度和線粒體膜電位的同時記錄
    點擊查看大圖

    神經元是可興奮性細胞,在突觸活動及產生動作電位后,神經元要產生大量的ATP驅動離子泵以提高胞內的Na+和Ca2+水平。谷氨酸(Glu)是重要的神經遞質,負責快速突觸傳遞及突觸傳遞強度的長期變化,并參與認知和記憶等過程;但如果過度激活谷氨酸受體,谷氨酸會導致離子平衡破壞、細胞死亡等毒性反應。

    本文為明確谷氨酸神經毒性的機制,將非損傷微測技術與激光共聚焦技術結合,以大鼠幼崽大腦皮層的神經元為被測樣品,用非損傷微測技術(a self-referencing O2 electrode)檢測單個神經元O2消耗量(即O2內流),而用激光共聚焦技術檢測其胞內Ca2+濃度和線粒體膜電位。


    點擊查看大圖
    單個神經元細胞在Glu作用下O2消耗量、胞內Ca2+濃度
    和線粒體膜電位的變化過程

    研究發現,在谷氨酸作用下,細胞內Ca2+濃度上升,隨后O2消耗量增加,這期間線粒體膜電位也發生相應改變。該結論直接證實了下述谷氨酸毒性機理模型:谷氨酸受體被激活后能引起Ca2+內流,導致細胞內ATP損耗。

    將非損傷微測技術與激光共聚焦等技術相結合,檢測生物樣品內部和外部離子分子或其他信息的變化情況,已經成為揭示生命過程機理機制的重要手段。

    關鍵詞:非損傷微測技術(a self-referencing O2 electrode), 氧氣消耗量(oxygen consumption), 谷氨酸(glutamate);
    參考文獻:Gleichmann M, et al. J Neurochem, 2009,109: 644-655
    全文下載:http://www.xuyue.net/xylt/viewthread.php?tid=535&extra=page%3D1

    Abstract:
    In order to determine the sequence of cellular processes in glutamate toxicity, we simultaneously recorded O2 consumption,cytosolic Ca2+ concentration ([Ca2+]i), and mitochondrial membrane potential (mDw) in single cortical neurons. Oxygen consumption was measured using an amperometric selfreferencing platinum electrode adjacent to neurons in which [Ca2+]i and mDw were monitored with Fluo-4 and TMRE+,respectively, using a spinning disk laser confocal microscope.Excitotoxic doses of glutamate caused an elevation of [Ca2+]i followed seconds afterwards by an increase in O2 consumption which reached a maximum level within 1–5 min. A modest increase in mDw occurred during this time period, and then, shortly before maximalO2 consumption was reached, themDw, as indicated by TMRE+ fluorescence, dissipated. Maximal O2 consumption lasted up to 5 min and then declined together with mDw and ATP levels, while [Ca2+]i further increased. mDw and [Ca2+]i returned to baseline levels when neurons were treated
    with an NMDA receptor antagonist shortly after the [Ca2+]i increased. Our unprecedented spatial and time resolution revealed that this sequence of events is identical in all neurons, albeit with considerable variability in magnitude and kinetics of changes in O2 consumption, [Ca2+]i, and mDw. The data obtained using this new method are consistent with a model where Ca2+ influx causes ATP depletion, despite maximal mitochondrial respiration, minutes after glutamate receptor activation.
    Keywords: excitotoxicity, glutamate, oxygen consumption.

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