In vivo imaging of activated microglia in a mouse model of focal cerebral ischemia by two-photon microscopy

通过双光子显微镜对小鼠局灶性脑缺血模型中活化的小胶质细胞进行体内成像

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作者:Seoyeon Bok, Taejun Wang, Chan-Ju Lee, Seong-Uk Jeon, Young-Eun Kim, Jeongwoo Kim, Beom-Ju Hong, Calvin Jinse Yoon, Sungjee Kim, Seung-Hoon Lee, Hak Jae Kim, Il Han Kim, Ki Hean Kim, G-One Ahn

Abstract

Microglia are brain resident macrophages rapidly responding to various stimuli to exert appropriate inflammatory responses. Although they have recently been exploited as an attractive candidate for imaging neuroinflammation, it is still difficult to visualize them at the cellular and molecular levels. Here we imaged activated microglia by establishing intracranial window chamber (ICW) in a mouse model of focal cerebral ischemia by using two-photon microscopy (TPM), in vivo. Intravenous injection of fluorescent antibodies allowed us to detect significantly elevated levels of Iba-1 and CD68 positive activated microglia in the ipsilateral compared to the contralateral side of the infarct. We further observed that indomethacin, a non-steroidal anti-inflammatory drug significantly attenuated CD68-positive microglial activation in ICW, which was further confirmed by qRT-PCR biochemical analyses. In conclusion, we believe that in vivo TPM imaging of ICW would be a useful tool to screen for therapeutic interventions lowering microglial activation hence neuroinflammation.

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