成年大鼠脑细胞内钙的测定

Measurement of Free Intracellular Ca 2+ in Isolarted Brain Cells from Adult Rats

目的：探讨成年大鼠脑细胞分离及其细胞内钙（［Ｃａ２＋］ｉ）的测定。方法：低浓度胰蛋白酶消化及机械吹打等方法分离脑细胞，以Ｆｕｒａ－２／ＡＭ为Ｃａ２＋－敏感的荧光指示剂，应用荧光分光光度计测定［Ｃａ２＋］ｉ。结果：分离的脑细胞存活率在９５％以上，在细胞外Ｃａ２＋１ｍｍｏｌ／Ｌ的情况下［Ｃａ２＋］ｉ为（２７０±３５）ｎｍｏｌ／Ｌ，高钾使［Ｃａ２＋］ｉ急剧增加。结论：本方法分离成年大鼠脑细胞简单、可靠，且能准确反映［Ｃａ２＋］ｉ的变化。

Objective:We explored the dissociation method for the brain cells of adult rat and the measurement of free intracellular calcium([Ca 2+ ]i).Methods:Digesting with low concentration of trypsin and gently triturating methods were used to dissociate brain cells.[Ca 2+ ]i was measured by fluorescence spectrometer with fura 2/AM as Ca 2+ sensitive fluorescent indicator.Results:The cellular viability rate was over 95%,the resting [Ca 2+ ]i level was (270±35)nmol/L on the condition that the extracellular concentration of Ca 2+ was 1 mmol/L,high concentration of K + made [Ca 2+ ]i was significantly increased.Conclusions:The method used to dissociate brain cells of adult rat was simple and reliable and could accurately reflect the change of [Ca 2+ ]i.