Department of Cell and Developmental Biology, Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel.
Department of Cell and Developmental Biology, Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel; Center for Research on Pain, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel.
Neurosci Lett. 2020 Sep 14;735:135212. doi: 10.1016/j.neulet.2020.135212. Epub 2020 Jun 25.
In light of the general shift from rats to mice as the leading rodent model in neuroscience research we used c-Fos expression as a tool to survey brain regions in the mouse in which neural activity differs between the states of wakefulness and pentobarbital-induced general anesthesia. The aim was to complement prior surveys carried out in rats. In addition to a broad qualitative review, 28 specific regions of interest (ROIs) were evaluated quantitatively. Nearly all ROIs in the cerebral cortex showed suppressed activity during anesthesia. Subcortically, however, some ROIs showed suppression, some showed little change, and some showed increased activity. The overall picture was similar to the rat. Special attention was devoted to ROIs significantly activated during anesthesia, as such loci might actively drive the transition to anesthetic unconsciousness rather than responding passively to inhbitory agents distributed globally (the "wet blanket" hypothesis). Twelve such "anesthesia-on" ROIs were identified: the paraventricular hypothalamic nucleus, supraoptic nucleus, tuberomamillary nucleus, lateral habenular nucleus, dentate gyrus, nucleus raphe pallidus, central amygdaloid nucleus, perifornical lateral hypothalamus, ventro-lateral preoptic area, lateral septum, paraventricular thalamic nucleus and zona incerta. The same primary anti-FOS antibody was used in all mice, but two alternative reporter systems were employed: ABC-diaminobenzidine and the currently more popular AlexaFluor488. Fluorescence tagging revealed far fewer FOS-immunoreactive neurons, sounding an alert that the reporter system chosen can have major effects on results obtained.
鉴于在神经科学研究中，老鼠已普遍取代大鼠成为主要的啮齿类动物模型，我们使用 c-Fos 表达作为一种工具来检测小鼠在清醒状态和戊巴比妥诱导的全身麻醉状态下神经活动不同的脑区。目的是补充先前在大鼠中进行的调查。除了广泛的定性综述外，还对 28 个特定的感兴趣区域 (ROI) 进行了定量评估。在麻醉期间，几乎所有大脑皮层的 ROI 都显示出活性受到抑制。然而，在皮质下区域，一些 ROI 显示出抑制，一些显示出变化不大，还有一些显示出活性增加。总体情况与大鼠相似。特别关注在麻醉期间明显激活的 ROI，因为这些区域可能主动驱动向麻醉无意识的转变，而不是被动响应分布在全身的抑制性药物（“湿毯子”假说）。确定了 12 个这样的“麻醉开启”ROI：室旁下丘脑核、视上核、结节乳头核、外侧缰核、齿状回、苍白球 Raphe 核、杏仁中央核、peri 外侧下丘脑、腹外侧视前区、外侧隔核、室旁丘脑核和未定带。所有小鼠都使用相同的抗 FOS 抗体，但使用了两种替代报告系统：ABC-二氨基联苯胺和目前更流行的 AlexaFluor488。荧光标记显示出的 FOS 免疫反应性神经元要少得多，这表明所选择的报告系统可以对获得的结果产生重大影响。
