TMB-8 and thapsigargin modulate purine release from dissociated primary cultures of rat brain astrocytes.

P Ballerini, R Ciccarelli, P Di Iorio, P Giuliani, D Francano, G Fanò, F Caciagli
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Abstract

In our previous studies, the evoked purine outflow from rat brain cultured astrocytes was reported to be Na+ independent and K+ and [Ca2+]e partially dependent. Thus, the eventual [Ca2+]i influence on purine astrocyte release was investigated in an attempt to better characterize the ionic requirements of this mechanism in cells which serve many complex and still partly unknown functions within the CNS. TMB-8 and Thapsigargin (drugs described as able to inhibit and increase the ion efflux from its internal stores respectively) and BAPTA/AM (able to chelate the cytoplasmic free Ca2+), were used. TMB-8 and BAPTA/AM decreased, whereas Thapsigargin enhanced glial purine outflow. These findings suggest a significant [Ca2+]i dependence of the electrically evoked purine efflux from cultured astrocytes even though further investigations using fluorescent probes are needed.

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TMB-8和thapsignargin调节大鼠脑星形胶质细胞分离原代培养嘌呤的释放。
在我们之前的研究中,大鼠脑培养星形胶质细胞诱发的嘌呤流出是Na+独立的,K+和[Ca2+]e部分依赖。因此,研究了[Ca2+]i对嘌呤星形胶质细胞释放的最终影响,试图更好地表征这种机制在细胞中的离子需求,这种机制在中枢神经系统内服务于许多复杂且仍部分未知的功能。使用TMB-8和Thapsigargin(分别能够抑制和增加其内部储存的离子外排的药物)和BAPTA/AM(能够螯合胞质游离Ca2+)。TMB-8和BAPTA/AM减少,而Thapsigargin增加了胶质嘌呤流出。这些发现表明,尽管需要使用荧光探针进行进一步的研究,但培养星形胶质细胞电诱发嘌呤外排的[Ca2+]i依赖性显著。
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