ATP-dependent paracrine communication between enteric neurons and glia in a primary cell culture derived from embryonic mice
ATP-dependent paracrine communication between enteric neurons and glia in a primary cell culture derived from embryonic mice
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DOI：
10.1111/j.1365-2982.2009.01302.x
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发表时间：
2009-08-01
影响因子：
3.5
通讯作者：
Vanden Berghe, P.
中科院分区：
文献类型：
--
作者：
Gomes, P.;Chevalier, J.;Vanden Berghe, P.
无论是在中枢神经系统还是肠神经系统，神经胶质细胞和神经元之间的动态相互作用的重要性越来越被认识到。然而，除了它们的保护作用外，人们对肠道神经胶质细胞的相互作用知之甚少。目的是利用光学技术研究小鼠培养模型中的神经胶质细胞间通讯。完整的胚胎（E13）内脏被酶解离，播种在盖上，用免疫组织化学和Ca2+成像研究。假定的祖细胞样细胞（表达PGP9.5和S-100）在大约5天内分化为表达典型细胞特异性标记的胶质细胞或神经元。神经胶质-神经元比例可以通过特定的补充来控制（N2, G5）。神经元和胶质细胞通过它们对去极化（高K+）或溶血磷脂酸的Ca2+反应和典型标记物的表达在功能上进行了鉴定。神经元对ACh、DMPP、5-HT、ATP和电刺激有反应，而胶质细胞对ATP和ADP β s有反应。MRS2179抑制胶质细胞的反应表明P2Y1受体参与其中。神经元刺激也引起延迟的神经胶质反应，苏拉明和催化核苷酸分解的外源性酵素酶可以减少这种反应。相反，ARL-67156（一种外链atp酶抑制剂）增强了胶质细胞的反应。在这种小鼠肠道共培养中，使用光学技术可以很容易地监测功能性胶质细胞和神经元。神经细胞可以被ATP或ADP β s直接激活。神经元细胞的激活（DMPP, K+）引起神经细胞的继发性反应，这可以通过调节ATP和ADP的分解来调节。这有力地支持了肠神经元和神经胶质之间的旁分泌嘌呤能通讯的参与。
P>The importance of dynamic interactions between glia and neurons is increasingly recognized, both in the central and enteric nervous system. However, apart from their protective role, little is known about enteric neuro-glia interaction. The aim was to investigate neuro-glia intercellular communication in a mouse culture model using optical techniques. Complete embryonic (E13) guts were enzymatically dissociated, seeded on coverslips and studied with immunohistochemistry and Ca2+-imaging. Putative progenitor-like cells (expressing both PGP9.5 and S-100) differentiated over approximately 5 days into glia or neurons expressing typical cell-specific markers. The glia-neuron ratio could be manipulated by specific supplements (N2, G5). Neurons and glia were functionally identified both by their Ca2+-response to either depolarization (high K+) or lysophosphatidic acid and by the expression of typical markers. Neurons responded to ACh, DMPP, 5-HT, ATP and electrical stimulation, while glia responded to ATP and ADP beta s. Inhibition of glial responses by MRS2179 suggests involvement of P2Y1 receptors. Neuronal stimulation also caused delayed glial responses, which were reduced by suramin and by exogenous apyrases that catalyse nucleotide breakdown. Conversely, glial responses were enhanced by ARL-67156, an ecto-ATPase inhibitor. In this mouse enteric co-culture, functional glia and neurons can be easily monitored using optical techniques. Glial cells can be activated directly by ATP or ADP beta s. Activation of neuronal cells (DMPP, K+) causes secondary responses in glial cells, which can be modulated by tuning ATP and ADP breakdown. This strongly supports the involvement of paracrine purinergic communication between enteric neurons and glia.