Fischer H G, Eder C, Hadding U, Heinemann U
Institut für Medizinische Mikrobiologie und Virologie, Heinrich-Heine-Universität, Düsseldorf, Germany.
Neuroscience. 1995 Jan;64(1):183-91. doi: 10.1016/0306-4522(94)00398-o.
Microglia were enriched in brain cell cultures from newborn mice as a result of supplementation with the growth factors macrophage colony-stimulating factor or granulocyte/macrophage colony-stimulating factor. When separately administered these two cytokines promote the outgrowth of loosely adherent cells with similar morphology which stained positive for CD11b and nonspecific esterase. Microglial cells isolated from both types of culture were electrophysiologically characterized using the whole cell configuration of the patch-clamp technique. Different resting membrane potentials were measured. In response to hyperpolarizing and depolarizing voltage commands 68 of 91 macrophage colony-stimulating factor-cultured microglial cells exhibited only an inward rectifying potassium current. By contrast, an outward potassium current was observed on 71 of 95 granulocyte/macrophage colony-stimulating factor-grown cells. Parallel testing of their capability for antigen presentation proved the activated functional state of these microglial cells. They induce antigen-specific T cell response without prior stimulus. In comparison, cells developed with macrophage colony-stimulating factor failed to present antigen. In such resting microglia a short-term treatment with granulocyte/macrophage colony-stimulating factor or interferon-gamma provoked a strong appearance of outward potassium currents, however, only the interferon-gamma-trigger resulted in efficient antigen presentation. The differential induction of both functional parameters suggests the detection of outward potassium currents to provide an electrophysiological activation marker of microglia which is subjected to cytokine regulation but not compellingly linked to antigen presentation.
通过补充生长因子巨噬细胞集落刺激因子或粒细胞/巨噬细胞集落刺激因子，小胶质细胞在新生小鼠的脑细胞培养物中得到富集。单独给予这两种细胞因子时，它们会促进形态相似的松散贴壁细胞的生长，这些细胞对CD11b和非特异性酯酶呈阳性染色。使用膜片钳技术的全细胞配置对从这两种培养物中分离出的小胶质细胞进行电生理特性分析。测量到了不同的静息膜电位。在对超极化和去极化电压指令的反应中，91个巨噬细胞集落刺激因子培养的小胶质细胞中有68个仅表现出内向整流钾电流。相比之下，在95个粒细胞/巨噬细胞集落刺激因子培养的细胞中有71个观察到外向钾电流。对它们的抗原呈递能力进行的平行测试证明了这些小胶质细胞的激活功能状态。它们无需预先刺激就能诱导抗原特异性T细胞反应。相比之下，用巨噬细胞集落刺激因子培养的细胞未能呈递抗原。在这种静息小胶质细胞中，用粒细胞/巨噬细胞集落刺激因子或干扰素-γ进行短期处理会引发外向钾电流的强烈出现，然而，只有干扰素-γ触发的结果导致有效的抗原呈递。这两个功能参数的差异诱导表明，检测外向钾电流可提供一种小胶质细胞的电生理激活标志物，其受细胞因子调节，但与抗原呈递没有必然联系。
