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首页> 外文期刊>Cellular Signalling >Functional decreases in P2X(7) receptors are associated with retinoic acid-induced neuronal differentiation of Neuro-2a neuroblastoma cells
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Functional decreases in P2X(7) receptors are associated with retinoic acid-induced neuronal differentiation of Neuro-2a neuroblastoma cells

机译:P2X(7)受体的功能下降与视黄酸诱导的Neuro-2a神经母细胞瘤细胞的神经元分化有关。

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Neuro-2a (N2a) cells are derived from spontaneous neuroblastoma of mouse and capable to differentiate into neuronal-like cells. Recently, P2X(7) receptor has been shown to sustain growth of human neuroblastoma cells but its role during neuronal differentiation remains unexamined. We characterized the role of P2X(7) receptors in the retinoic acid (RA)-differentiated N2a cells. RA induced N2a cells differentiation into neurite bearing and neuronal specific proteins, microtubule-associated protein 2 (MAP2) and neuronal specific nuclear protein (NeuN), expressing neuronal-like cells. Interestingly, the RA-induced neuronal differentiation was associated with decreases in the expression and function of P2X(7) receptors. Functional inhibition of P2X(7) receptors by P2X(7) receptor selective antagonists, 5'-triphosphate, periodate-oxidized 2',3'-dialdehyde ATP (oATP), brilliant blue G (BBG) or A438079 induced neurite outgrowth. In addition, RA and oATP treatment stimulated the expression of neuron-specific class III beta-tubulin (TuJ1), and knockdown of P2X(7) receptor expression by siRNA induced neurite outgrowth. To elucidate the possible mechanism, we found the levels of basal intracellular Ca2+ concentrations ([Ca2+](i)) were decreased in either RA- or oATP-differentiated or P2X(7) receptor knockdown N2a cells. Simply cultured N2a cells in low Ca2+ medium induced a 2-fold increase in neurite length. Treatment of N2a cells with ATP hydrolase apyrase and the P2X(7) receptors selective antagonist oATP or BBG decreased cell viability and cell number. Nevertheless, oATP but not BBG decreased cell proliferation and cell cycle progression. These results suggest for the first time that decreases in expression/function of P2X(7) receptors are involved in neuronal differentiation. We provide additional evidence shown that the ATP release-activated P2X(7) receptor is important in maintaining cell survival of N2a neuroblastoma cells.
机译:Neuro-2a(N2a)细胞源自小鼠自发性神经母细胞瘤,能够分化为神经元样细胞。最近,已显示P2X(7)受体可维持人类神经母细胞瘤细胞的生长,但其在神经元分化过程中的作用仍未经检查。我们表征了视黄酸(RA)分化的N2a细胞中的P2X(7)受体的作用。 RA诱导N2a细胞分化为神经突样和神经元特异性蛋白,微管相关蛋白2(MAP2)和神经元特异性核蛋白(NeuN),表达神经元样细胞。有趣的是,RA诱导的神经元分化与P2X(7)受体的表达和功能下降有关。由P2X(7)受体选择性拮抗剂,5'-三磷酸,高碘酸盐氧化的2',3'-二醛ATP(oATP),亮蓝G(BBG)或A438079诱导的神经突向外生长抑制P2X(7)受体。此外,RA和oATP处理可刺激神经元特异性III类β-微管蛋白(TuJ1)的表达,并通过siRNA诱导的神经突增生抑制P2X(7)受体的表达。为了阐明可能的机制,我们发现在RA或oATP分化的或P2X(7)受体敲低的N2a细胞中,基础细胞内Ca2 +浓度([Ca2 +](i))的水平降低。在低Ca2 +培养基中简单培养的N2a细胞诱导神经突长度增加2倍。用ATP水解酶腺苷三磷酸酶和P2X(7)受体选择性拮抗剂oATP或BBG处理N2a细胞会降低细胞活力和细胞数量。但是,oATP而不是BBG会降低细胞增殖和细胞周期进程。这些结果首次表明,P2X(7)受体的表达/功能下降与神经元分化有关。我们提供的其他证据表明,ATP释放激活的P2X(7)受体在维持N2a神经母细胞瘤细胞的细胞存活中很重要。

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