Glioma-associated microglia and macrophages/monocytes display distinct electrophysiological properties and do not communicate via gap junctions.
Glioma-associated microglia and macrophages/monocytes display distinct electrophysiological properties and do not communicate via gap junctions.
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DOI：
10.1016/j.neulet.2014.09.035
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发表时间：
2014-11-07
影响因子：
2.5
通讯作者：
Kettenmann H
中科院分区：
文献类型：
--
作者：
Richter N;Wendt S;Georgieva PB;Hambardzumyan D;Nolte C;Kettenmann H
脑内小胶质细胞和外周巨噬细胞/单核细胞均浸润到胶质瘤中并促进胶质瘤生长。在本研究中，我们分析了耦合和膜电流在胶质瘤相关的小胶质细胞和巨噬细胞/单核细胞，并比较了控制和刺伤相关的小胶质细胞。使用Cx 3cr 1GFP/wt Ccr 2 RFP/wt敲入小鼠系，我们区分了在接种GL 261胶质瘤细胞后14-16天制备的急性脑切片中的胶质瘤相关小胶质细胞和巨噬细胞/单核细胞的膜电流。与其他疾病模型或细胞培养物相比，小胶质细胞的电流谱显示内向整流电流，这让人想起中间激活状态。巨噬细胞/单核细胞表现出更高的比外向电导和显着较低的电容，表明一个较小的膜面积比小胶质细胞。作为对照，我们还记录了来自对照小胶质细胞和刺伤相关小胶质细胞的电流。由于有小胶质细胞体外偶联的报道，我们将生物胞素注射到这些细胞中，并在固定切片后分析细胞偶联，并用Cy 3-缀合的链霉亲和素处理生物胞素标记。无论是控制小胶质细胞，神经胶质瘤相关的小胶质细胞和巨噬细胞/单核细胞，也没有刺伤相关的小胶质细胞显示出任何迹象的耦合。此外，进行qRT-PCR显示，在分离和分选的胶质瘤相关小胶质细胞和巨噬细胞/单核细胞上检测不到连接蛋白43，表明这些细胞不是偶联网络的一部分。
Both brain-resident microglia and peripheral macrophages/monocytes infiltrate into glioma and promote glioma growth. In the present study we analyzed coupling and membrane currents in glioma-associated microglia and macrophages/monocytes and compared this to control and stab wound-associated microglia. Using the Cx3cr1GFP/wt Ccr2RFP/wt knock-in mouse line, we distinguished membrane currents of glioma-associated microglia and macrophages/monocytes in acute brain slices prepared 14-16 days after inoculation of GL261 glioma cells. The current profile of microglia showed inward rectifying currents reminiscent of an intermediate activation state when compared to other disease models or cell culture. Macrophages/monocytes showed a higher specific outward conductance and a significantly lower capacitance indicative of a smaller membrane area than microglia. As controls, we also recorded currents from control microglia and stab wound-associated microglia. Since there are reports of microglial coupling in vitro, we injected biocytin into these cells and analyzed for cell coupling after fixing the slices and processed for biocytin labeling with Cy3- conjugated-Streptavidin. Neither control microglia nor glioma-associated microglia and macrophages/monocytes nor stab wound-associated microglia showed any sign of coupling. Moreover, performing qRT-PCR revealed that no connexin43 was detectable on isolated and sorted glioma-associated microglia and macrophages/monocytes, indicating that these cells are not part of a coupled network.
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