Gomes P, Chevalier J, Boesmans W, Roosen L, van den Abbeel V, Neunlist M, Tack J, Vanden Berghe P
Center for Gastroenterological Research, Katholieke Universiteit Leuven, Leuven, Belgium.
Neurogastroenterol Motil. 2009 Aug;21(8):870-e62. doi: 10.1111/j.1365-2982.2009.01302.x. Epub 2009 Mar 26.
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 Ca(2+)-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 Ca(2+)-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 ADPbetas. 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 ADPbetas. 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.
神经胶质细胞与神经元之间动态相互作用的重要性在中枢神经系统和肠神经系统中日益受到认可。然而，除了其保护作用外，关于肠神经胶质细胞相互作用的了解甚少。本研究旨在利用光学技术在小鼠培养模型中研究神经胶质细胞间的通讯。将完整的胚胎（E13）肠道进行酶解，接种在盖玻片上，并用免疫组织化学和Ca(2+)成像技术进行研究。假定的祖细胞样细胞（同时表达PGP9.5和S-100）在大约5天内分化为表达典型细胞特异性标志物的神经胶质细胞或神经元。神经胶质细胞与神经元的比例可通过特定补充剂（N2、G5）进行调控。神经元和神经胶质细胞通过对去极化（高K(+)）或溶血磷脂酸的Ca(2+)反应以及典型标志物的表达在功能上得以鉴定。神经元对乙酰胆碱、二甲基苯基哌嗪、5-羟色胺、三磷酸腺苷和电刺激有反应，而神经胶质细胞对三磷酸腺苷和β-ADP有反应。MRS2179对神经胶质细胞反应的抑制表明P2Y1受体参与其中。神经元刺激也会引起延迟的神经胶质细胞反应，苏拉明和催化核苷酸分解的外源性腺苷三磷酸双磷酸酶可使其减弱。相反，ecto-ATPase抑制剂ARL-67156可增强神经胶质细胞反应。在这种小鼠肠共同培养体系中，使用光学技术可轻松监测功能性神经胶质细胞和神经元。神经胶质细胞可被三磷酸腺苷或β-ADP直接激活。神经元细胞的激活（二甲基苯基哌嗪、K(+)）会在神经胶质细胞中引起继发性反应，可通过调节三磷酸腺苷和二磷酸腺苷的分解来进行调节。这有力地支持了肠神经元与神经胶质细胞之间旁分泌嘌呤能通讯的参与。
