Dynamic imaging of somatosensory cortical activity in the rat visualized by flavoprotein autofluorescence
Dynamic imaging of somatosensory cortical activity in the rat visualized by flavoprotein autofluorescence
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DOI：
10.1113/jphysiol.2003.040709
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发表时间：
2003-06-15
影响因子：
5.5
通讯作者：
Tanaka, R
中科院分区：
文献类型：
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作者：
Shibuki, K;Hishida, R;Tanaka, R
我们使用线粒体黄素蛋白的自发荧光来成像大鼠的皮质神经活动。在从大鼠大脑皮层获得的切片中检查蓝光下的绿色自发荧光。大约一半的基础自发荧光是通过培养基中是否存在 0 或葡萄糖来调节的。 20 Hz 持续 1 秒的重复电刺激会在切片中产生局部荧光增加。增加幅度为 27 +/- 2 %（平均值 +/- S.D.，n = 35）。河豚毒素或二亚苯基碘鎓（一种黄素蛋白抑制剂）可阻断自发荧光反应。在灌注不含钙、葡萄糖或 O-2 的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）麻醉的大鼠的初级体感皮层中，在 100 Hz 的电皮层刺激 1 s 后，观察到活动依赖性自发荧光增加 20 +/- 4 % (n = 6)，在 50 Hz 的振动皮肤刺激 1 s 后观察到自发荧光增加 2.6 +/- 0.5 % (n = 33)。 1 秒应用于足底后爪。这些反应足够大，可以使神经活动可视化，而无需对多次试验进行平均。电或振动皮肤刺激后荧光反应的分布与同一大鼠的皮质场电位的分布相当。荧光反应之后是动脉血流量的增加。前者对一氧化氮合酶抑制剂具有抗性，而后者则受到抑制。因此，黄素蛋白自发荧光的活动依赖性变化对于体内功能性脑成像是有用的。
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 0, 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 O-2-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.