Evaluation of M1-microglial activation by neurotoxic metals using optimized organotypic cerebral slice cultures
Evaluation of M1-microglial activation by neurotoxic metals using optimized organotypic cerebral slice cultures
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DOI：
10.2131/jts.44.471
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发表时间：
2019-01-01
影响因子：
2
通讯作者：
Hwang, Gi-Wook
中科院分区：
文献类型：
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作者：
Hoshi, Takayuki;Toyama, Takashi;Hwang, Gi-Wook
M1-小胶质细胞（神经毒性小胶质细胞）调节神经元发育和细胞死亡，并参与大脑中的许多病理。尽管器官型脑片培养被广泛用于研究神经元和小胶质细胞之间的串扰，但对小鼠大脑皮层切片中小胶质细胞的性质知之甚少。在这里，我们的目的是优化小鼠大脑切片文化，反映小胶质细胞的功能和评估的影响，神经毒性金属对M1-小胶质细胞的激活。从出生后第7天（P）小鼠制备的脑切片中的大多数小胶质细胞与成年小鼠脑中的成熟小胶质细胞相似，但是从P2小鼠制备的脑切片中的小胶质细胞是不成熟的，这是常规的制备条件。P7小鼠脑片中M1-小胶质细胞标志物（CD 16和CD 32）和炎性细胞因子（肿瘤坏死因子-α和白细胞介素-1 β）的表达程度高于P2小鼠脑片。这些结果表明，M1-小胶质细胞的活化可以更准确地评估在P7小鼠的脑切片比在P2小鼠。因此，我们接下来使用P7小鼠的脑切片检查了各种神经毒性金属对M1-小胶质细胞活化的影响，发现甲基汞刺激了M1-小胶质细胞的活化，但亚砷酸盐、铅和三丁基锡没有诱导这种活化。总之，本研究中使用的优化小鼠脑片培养物可以是一种有用的工具，用于研究在功能成熟的小胶质细胞存在下各种化学物质对中枢神经系统的影响。
M1-microglia (neurotoxic microglia) regulate neuronal development and cell death and are involved in many pathologies in the brain. Although organotypic brain slice cultures are widely used to study the crosstalk between neurons and microglia, little is known about the properties of microglia in the mouse cerebral cortex slices. Here, we aimed to optimize the mouse cerebral slice cultures that reflect microglial functions and evaluate the effects of neurotoxic metals on M1-microglial activation. Most microglia in the cerebral slices prepared from postnatal day (P) 7 mice were similar to mature microglia in adult mice brains, but those in the slices prepared from P2 mice were immature, which is a conventional preparation condition. The degree of expression of M1-microglial markers (CD16 and CD32) and inflammatory cytokines (tumor necrosis factor-alpha and interleukin-1 beta) by lipopolysaccharide, a representative microglia activator, in the cerebral slices of P7 mice were higher than that in the slices of P2 mice. These results indicate that M1-microglial activation can be evaluated more accurately in the cerebral slices of P7 mice than in those of P2 mice. Therefore, we next examined the effects of various neurotoxic metals on M1-microglial activation using the cerebral slices of P7 mice and found that methylmercury stimulated the activation to M1-microglia, but arsenite, lead, and tributyltin did not induce such activation. Altogether, the optimized mouse cerebral slice cultures used in this study can be a helpful tool to study the influence of various chemicals on the central nervous system in the presence of functionally mature microglia.