P2X7 receptors at adult neural progenitor cells of the mouse subventricular zone
P2X7 receptors at adult neural progenitor cells of the mouse subventricular zone
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DOI：
10.1016/j.neuropharm.2013.05.017
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发表时间：
2013-10-01
影响因子：
4.7
通讯作者：
Rubini, Patrizia
中科院分区：
文献类型：
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作者：
Messemer, Nanette;Kunert, Christin;Rubini, Patrizia
神经发生需要新形成的祖细胞的增殖和随后的过剩细胞的死亡之间的平衡。RT-PCR和免疫细胞化学显示，P2 X7受体的mRNA和免疫反应性的培养的神经前体细胞（NPC）从成年小鼠脑室下区（SVZ）的制备的存在。全细胞膜片钳记录显示，在低Ca ~（2+）和零Mg ~（2+）浓度下，对ATP和原型P2 X7受体激动剂二苯甲酰-ATP（Bz-ATP）的内向电流反应显著增强。Bz-ATP诱导的电流在0 mV附近极性反转;在P2 X7（-/-）小鼠制备的NPC中，Bz-ATP未能引起膜电流。一般性P2 X/P2 Y受体拮抗剂PPADS和P2 X7选择性拮抗剂Brilliant Blue G和A-438079强烈抑制Bz-ATP的作用。长期应用Bz-ATP诱导的初始电流，缓慢增加到稳态响应。结合YO-PRO摄取的测定，这些实验表明受体通道的扩张和/或染料摄取途径的募集。通过Fura-2的Ca 2+成像显示，在Mg 2+缺乏的浴介质中，Bz-ATP引起[Ca 2 +]（i）完全取决于外部Ca 2+的存在的瞬变。MIT试验表明Bz-ATP处理导致细胞活力呈浓度依赖性降低。相应地，Bz-ATP导致活性caspase 3免疫反应性增加，表明P2 X7控制的细胞凋亡。在急性SVZ转基因Tg（巢蛋白/EGFP）小鼠脑切片，膜片钳记录确定P2 X7受体在NPC的药理学特性相同，他们培养的同行。我们认为，在NPC的凋亡/坏死的P2 X7受体可能是特别相关的病理条件下，导致ATP释放增加，从而可以抵消随后的过度细胞增殖。(C)2013爱思唯尔有限公司保留所有权利。
Neurogenesis requires the balance between the proliferation of newly formed progenitor cells and subsequent death of surplus cells. RT-PCR and immunocytochemistry demonstrated the presence of P2X7 receptor mRNA and immunoreactivity in cultured neural progenitor cells (NPCs) prepared from the adult mouse subventricular zone (SVZ). Whole-cell patch-clamp recordings showed a marked potentiation of the inward current responses both to ATP and the prototypic P2X7 receptor agonist dibenzoyl-ATP (Bz-ATP) at low Ca2+ and zero Mg2+ concentrations in the bath medium. The Bz-ATP-induced currents reversed their polarity near 0 mV; in NPCs prepared from P2X7(-/-) mice, Bz-ATP failed to elicit membrane currents. The general P2X/P2Y receptor antagonist PPADS and the P2X7 selective antagonists Brilliant Blue G and A-438079 strongly depressed the effect of Bz-ATP. Long-lasting application of Bz-ATP induced an initial current, which slowly increased to a steady-state response. In combination with the determination of YO-PRO uptake, these experiments suggest the dilation of a receptor-channel and/or the recruitment of a dye-uptake pathway. Ca2+-imaging by means of Fura-2 revealed that in a Mg2+-deficient bath medium Bz-ATP causes [Ca2+](i) transients fully depending on the presence of external Ca2+. The MIT test indicated a concentration-dependent decrease in cell viability by Bz-ATP treatment. Correspondingly, Bz-ATP led to an increase in active caspase 3 immunoreactivity, indicating a P2X7-controlled apoptosis. In acute SVZ brain slices of transgenic Tg(nestin/EGFP) mice, patch-clamp recordings identified P2X7 receptors at NPCs with pharmacological properties identical to those of their cultured counterparts. We suggest that the apoptotic/necrotic P2X7 receptors at NPCs may be of particular relevance during pathological conditions which lead to increased ATP release and thus could counterbalance the ensuing excessive cell proliferation. (C) 2013 Elsevier Ltd. All rights reserved.