Morphological analysis of intrinsic circuit of the cerebral cortex

为了自下而上地理解大脑皮层，我们必须阐明大脑皮层内在回路的细节。然而，自早期的高尔基浸渍研究以来，在这一研究领域没有取得重大进展。电镜研究显示了突触等微电路，但不适合描绘大脑皮层的电路结构轮廓。在这项研究中，我们旨在通过细胞内记录、生物细胞素注射和免疫细胞化学相结合的方法研究大脑皮层的内在电路结构。首先，在皮层切片上记录膜特性和突触反应。注射生物细胞素后，固定切片，切片，进行免疫细胞化学处理。用三重免疫荧光法对切片进行谷氨酰胺酶、小白蛋白、钙结合蛋白或天冬氨酸转氨酶染色。用亲和素与荧光染料结合标记的impa…More led神经元，研究其是谷氨酸能还是GAB能。谷氨酰胺酶是一种合成谷氨酸的递质酶，存在于锥体神经元中，而不存在于大脑皮层中含有谷氨酰胺的神经元中（T.Kaneko和N.Mizuno, Neuroscience, vol.61, pp.839-849,1994）。另一方面，小白蛋白、钙结合蛋白和转氨酶定位于大脑皮层的GAB能神经元。荧光显微镜下观察，注射生物细胞素的细胞用DAB/镍ABC法制成蓝黑色。化学分类的皮质神经元，如表达P物质受体或非磷酸化形式的神经丝蛋白，在同一切片上用PAP方法用TAPM/对甲酚进一步免疫染色成红色。利用透明摄像重建技术研究细胞内标记细胞与化学分类神经元之间的轴突连接。通过这种方式，我们可以揭示电学和化学分类的神经元是如何与化学分类的神经元相连的（Kaneko等人，Comp. Neurol.）。第345卷，第161-171,172-184页)。我们现在已经发现并正在研究大脑皮层第六层的两种锥体细胞，谷氨酰胺酶阳性和谷氨酰胺酶阴性。少

In order to understand the cerebral cortex in a bottom-up manner, we have to elucidate the detail of the intrinsic circuitry in the cerebral cortex. However, since earlier Golgi impregnation studies, there have been no significant advancement in this field of study. Electron microscopic studies revealed the microcircuitry such as synapses, but were not suitable for drawing the outline of the circuit architecture of the cerebral cortex. In this study, we aimed at investigating the structure of the intrinsic circuitry of the cerebral cortex by a combined method intracellular recording, biocytin injection and immunocytochemistry using brain slices. First, the membrane properties and synaptic responses were recorded in the cortical slices. After the injection of biocytin, the slices were fixed, cut into sections and processed for immunocytochemistry. The sections were stained for glutaminase, parvalbumin, calbindin or aspartate aminotransferase by triple immunofluorescence method. The impa … More led neurons, which were labeled with avidin conjugated with fluorescent dye, were thereby studied whether they were glutamatergic or GAB Aergic. Glutaminase is a synthesizing enzyme of transmitter glutamate, and present in pyramidal neurons, but not in GAB-containing neurons of the cerebral cortex ( T.Kaneko and N.Mizuno, Neuroscience, vol.61, pp.839-849,1994) .On the other hand, parvalbumin, calbindin and aminotransferdse are localized in GAB Aergic neurons in the cerebral cortex. After the observation under a fluorescence microscope, the biocytin-injected cells were developed blue black by the ABC method with DAB/nickel. Chemically classified cortical neurons such as those expressing substance P receptors or non-phosphorylated form of neurofilament proteins were further immunostained red in the same section by the PAP method with TAPM/p-cresol. The connection of the axons from the intracellularly labeled cells onto the chemically classified neurons were investigated by camera lucida reconstruction. In this way, we could reveal how the electrically and chemically classified neurons were connected with chemically classified neurons (Kaneko et al. , Comp. Neurol. vol.345, pp.161-171,172-184) . We have now found and been investigating the two types of pyramidal cells in the layr VI of the cerebral cortex, glutaminase-positive and glutaminase-negative ones. Less