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Effects of anthopleurin-Q on the intracellular free Ca2+ concentration in cultured rat cortical neurons

机译：花青素-Q对大鼠皮质神经元细胞内游离钙离子浓度的影响

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摘要

The present study was designed to investigate the mechanism underlying the intracellular free Ca2+ concentration ([Ca2+]i) modulated by Anthopleurin-Q (AP-Q), a sea anemone toxin, using whole-cell patch clamp and fluorescence digital imaging techniques. Results indicated that the overall Ca2+ concentration could be augmented in presence of AP-Q. The increase of [Ca2+]i induced by AP-Q was eliminated in Na+-free solution, Ca2+-free solution or in presence of TTX. However, the Ca2+ increase induced by AP-Q could not be influenced by cyclopiazonic acid (CPA), a specific inhibitor of the endoplasmic reticulum Ca2+-ATPase pump. We furthermore demonstrated that voltage-gated calcium channels (VGCCs) blocker verapamil, or inhibitor of the reverse operation Na+-Ca2+ exchanger NiCl2 attenuated AP-Q-induced [Ca2+]i elevation. Furthermore, the inactivation process of Na+ current (I Na) was significantly delayed with slightly change of its amplitude by AP-Q. These findings demonstrated that neuron voltage-gated Na+ channels are also targets of AP-Q. Overall, the present results suggested that AP-Q induced calcium influx via Na+-dependent activation of voltage-gated sodium channels (VGSCs), VGCCs and reverse operation of the Na+/Ca 2+exchanger.
机译：本研究旨在使用全细胞膜片钳和荧光数字成像技术研究海藻毒素Anthopleurin-Q（AP-Q）调节细胞内游离Ca2 +浓度（[Ca2 +] i）的潜在机制。结果表明，在AP-Q存在下，总的Ca2 +浓度可以增加。在无Na +溶液，无Ca2 +溶液或TTX存在下，消除了AP-Q诱导的[Ca2 +] i的增加。但是，AP-Q诱导的Ca2 +增加不受内质网Ca2 + -ATPase泵的特异性抑制剂环吡嗪酸（CPA）的影响。我们进一步证明，电压门控钙通道（VGCC）阻断剂维拉帕米或逆向操作Na + -Ca2 +交换剂NiCl2的抑制剂可减弱AP-Q诱导的[Ca2 +] i升高。此外，Na +电流（I Na）的失活过程因AP-Q幅度的轻微变化而明显延迟。这些发现表明，神经元电压门控的Na +通道也是AP-Q的靶标。总体而言，目前的结果表明，AP-Q通过依赖Na +的电压门控钠通道（VGSC），VGCC和Na + / Ca 2+交换器的反向运行来诱导钙内流。

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