Shibuki Katsuei, Hishida Ryuichi, Murakami Hiroatsu, Kudoh Masaharu, Kawaguchi Tadashi, Watanabe Masatoshi, Watanabe Shunsuke, Kouuchi Takeshi, Tanaka Ryuichi
Departments of Neurophysiology, Niigata University, Asahi-machi, Niigata 951-8585, Japan.
J Physiol. 2003 Jun 15;549(Pt 3):919-27. doi: 10.1113/jphysiol.2003.040709. Epub 2003 May 2.
We used autofluorescence of mitochondrial flavoproteins to image cortical neural activity in the rat. Green autofluorescence in blue light was examined in slices obtained from rat cerebral cortex. About half of the basal autofluorescence was modulated by the presence or absence of O2 or glucose in the medium. Repetitive electrical stimulation at 20 Hz for 1 s produced a localized fluorescence increase in the slices. The amplitude of the increase was 27 +/- 2 % (mean +/- S.D., n = 35). Tetrodotoxin or diphenyleneiodonium, an inhibitor of flavoproteins, blocked the autofluorescence responses. The autofluorescence responses were not observed in slices perfused with calcium-, glucose- or O2-free medium. In the primary somatosensory cortex of rats anaesthetized with urethane (1.5 g kg-1, I.P.), an activity-dependent increase in autofluorescence of 20 +/- 4 % (n = 6) was observed after electrical cortical stimulation at 100 Hz for 1 s, and an increase of 2.6 +/- 0.5 % (n = 33) after vibratory skin stimulation at 50 Hz for 1 s applied to the plantar hindpaw. These responses were large enough to allow visualization of the neural activity without having to average a number of trials. The distribution of the fluorescence responses after electrical or vibratory skin stimulation was comparable to that of the cortical field potentials in the same rats. The fluorescence responses were followed by an increase in arterial blood flow. The former were resistant to an inhibitor of nitric oxide synthase, while the latter was inhibited. Thus, activity-dependent changes in the autofluorescence of flavoproteins are useful for functional brain imaging in vivo.
我们利用线粒体黄素蛋白的自发荧光对大鼠的皮质神经活动进行成像。在取自大鼠大脑皮质的切片中检测了蓝光下的绿色自发荧光。约一半的基础自发荧光受到培养基中氧气或葡萄糖存在与否的调节。以20Hz重复电刺激1秒可使切片中的荧光局部增加。增加的幅度为27±2%(平均值±标准差,n = 35)。河豚毒素或黄素蛋白抑制剂二苯碘鎓可阻断自发荧光反应。在灌注无钙、无葡萄糖或无氧培养基的切片中未观察到自发荧光反应。在用氨基甲酸乙酯(1.5 g kg-1,腹腔注射)麻醉的大鼠初级体感皮质中,以100Hz电刺激皮质1秒后,观察到自发荧光的活性依赖性增加为20±4%(n = 6),对后足底进行50Hz振动皮肤刺激1秒后,增加了2.6±0.5%(n = 33)。这些反应足够大,无需对多次试验进行平均就能可视化神经活动。电刺激或振动皮肤刺激后荧光反应的分布与同一只大鼠皮质场电位的分布相当。荧光反应后动脉血流量增加。前者对一氧化氮合酶抑制剂有抗性,而后者受到抑制。因此,黄素蛋白自发荧光的活性依赖性变化可用于体内功能性脑成像。