Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants.
Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants.
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DOI：
10.1371/journal.pone.0135238
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Cuadros MA
中科院分区：
文献类型：
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作者：
Ferrer-Martín RM;Martín-Oliva D;Sierra-Martín A;Carrasco MC;Martín-Estebané M;Calvente R;Martín-Guerrero SM;Marín-Teva JL;Navascués J;Cuadros MA
小胶质细胞在神经退行性变中的作用仍然存在争议。我们研究了小胶质细胞在视网膜中是否具有神经毒性或神经保护功能。在体外用米诺环素处理来自10日龄小鼠的视网膜外植体以抑制小胶质细胞活化，用LPS处理以增加小胶质细胞活化，或用负载氯膦酸盐的脂质体（Lip-Clo）处理以消耗小胶质细胞。用流式细胞术检测视网膜细胞的存活率，用TUNEL法检测死亡细胞的分布。采用流式细胞术和免疫细胞化学方法分析小胶质细胞的免疫表型、形态学特征及其分布。用米诺环素处理视网膜外植体减少了小胶质细胞的活化，同时显著降低了细胞活力，增加了TUNEL标记细胞的存在。这种处理还阻止了小胶质细胞向细胞死亡最多的外核层迁移。LPS处理增加了小胶质细胞活化，但对细胞活力或小胶质细胞分布没有影响。最后，用Lip-Clo去除部分小胶质细胞降低了视网膜外植体中的细胞活力，显示出与米诺环素类似的效果。因此，当去除小胶质细胞或抑制其活化时，体外培养的视网膜外植体中的细胞活力降低，表明小胶质细胞在该系统中具有神经营养作用。
The role of microglia during neurodegeneration remains controversial. We investigated whether microglial cells have a neurotoxic or neuroprotective function in the retina. Retinal explants from 10-day-old mice were treated in vitro with minocycline to inhibit microglial activation, with LPS to increase microglial activation, or with liposomes loaded with clodronate (Lip-Clo) to deplete microglial cells. Flow cytometry was used to assess the viability of retinal cells in the explants and the TUNEL method to show the distribution of dead cells. The immunophenotypic and morphological features of microglia and their distribution were analyzed with flow cytometry and immunocytochemistry. Treatment of retinal explants with minocycline reduced microglial activation and simultaneously significantly decreased cell viability and increased the presence of TUNEL-labeled cell profiles. This treatment also prevented the migration of microglial cells towards the outer nuclear layer, where cell death was most abundant. The LPS treatment increased microglial activation but had no effect on cell viability or microglial distribution. Finally, partial microglial removal with Lip-Clo diminished the cell viability in the retinal explants, showing a similar effect to that of minocycline. Hence, cell viability is diminished in retinal explants cultured in vitro when microglial cells are removed or their activation is inhibited, indicating a neurotrophic role for microglia in this system.