マウスに設置した頭窓を通して脳虚血モデルを作成した。蛍光ラベルした赤血球を全身投与後, 局所脳血流を測定しつつ, 共焦点顕微鏡にて脳皮質微小循環・毛細血管の動的変化をリアルタイムでin vivo ビデオモニターし, 一ヶ月以上にわたり反復して画像解析した。また, 適宜, 脳微小循環速度を解析した。さらに微小血管, アストロサイトを生体染色し, 脳虚血後の微小血管, 神経細胞, アストロサイトの形態変化を同時に二光子顕微鏡によって反復生体観察した。その後, 脳組織の結果と比較した。その結果, 脳虚血時脳微小循環調節における神経血管カップリングの詳細を観察し, その重要性を明らかにした。
A focal cerebral ischemia murine model with a cranial window was prepared. After systemic administration of fluorescently labelled red blood cells, the dynamic changes of the brain cortex microcirculation and the capillaries were repeatedly video-monitored in vivo for up to one month by means of confocal microscopy and analyzed in real time, while local blood flow in the brain was measured. The velocity of the brain microcirculation was also analyzed. In addition, vital staining of microvessels and astrocytes was performed, and the morphological changes of microvessels, nerve cells, and astrocytes after cerebral ischemia were simultaneously and repeatedly observed in vivo with a two-photon microscope. The results obtained were compared with ex-vivo results. This work has uncovered details of neurovascular coupling during blood flow regulation in the brain under cerebral ischemia was observed, and established its importance.
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