Morphological analysis of intrinsic circuit of the cerebral cortex

大脑皮层内在回路的形态学分析

• 批准号: 05680658
• 负责人: KANEKO Takeshi
• 金额: $ 1.34万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05680658/
• 关键词: Cerebral Cortex; Intrinsic Circuit; Glutaminase; Glutamatergic Neuron; GAB Aergic Neuron; Intracellular recording; Staining; Immunohis to chemistry; Slice Method; カルシウム結合蛋白質; サブスタンスP受容体; シナプス

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

为了以自下而上的方式理解大脑皮层，我们必须阐明大脑皮层中内在电路的细节。然而，自从早期的高尔基体浸渍研究以来，在这一研究领域没有取得重大进展。电子显微镜研究揭示了微电路，如突触，但不适合描绘大脑皮层的电路结构的轮廓。本研究采用细胞内记录、生物胞素注射和免疫细胞化学相结合的方法，利用脑片研究大脑皮层内回路的结构。首先，在皮层切片中记录膜特性和突触反应。注射生物胞素后，将切片固定，切片并进行免疫细胞化学处理。采用三重免疫荧光法对切片进行转氨酶、小清蛋白、钙结合蛋白或天冬氨酸转氨酶染色。黑猩猩 ...更多信息 用与荧光染料缀合的抗生物素蛋白标记的Led神经元由此研究它们是谷氨酸能的还是GAB能的。谷氨酰胺酶是谷氨酸递质的合成酶，存在于锥体神经元中，但不存在于大脑皮层的含GABA的神经元中（T.Kaneko and N.Mizuno，Neuroscience，vol.61，pp.839- 849，1994）。在荧光显微镜下观察后，用DAB/镍通过ABC法使生物胞素注射的细胞显色为蓝黑色。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的那些，通过PAP方法用TAPM/对甲酚在同一切片中进一步免疫染色为红色。用照相机重建法研究了细胞内标记细胞的轴突与化学分类神经元之间的连接。以这种方式，我们可以揭示电和化学分类的神经元如何与化学分类的神经元连接（Kaneko等人，Comp. Neurol.第345卷，第161 - 171页，第172 -184页）。我们现在已经发现并研究了大脑皮层第VI层中的两种类型的锥体细胞，即精氨酸酶阳性和精氨酸酶阴性的锥体细胞。少

项目成果

Kaneko T,Shigemoto R,Nakanishi S and Mizuno N: "Mor-phological and chemical characteristics of substance P receptor-immunoreactive neurons in the rat neocortex" Neuroscience. vol.60 (no.1) May. 199-211 (1994)

Kaneko T、Shigemoto R、Nakanishi S 和 Mizuno N：“大鼠新皮质中 P 物质受体免疫反应性神经元的形态和化学特征”神经科学。

金子武嗣: "Morphological and chemical characteristics of substance P receptor-immunoreactive neurons in the neocortex" Neuroscience. 60. 199-211 (1994)

Takeshi Kaneko：“新皮质中 P 物质受体免疫反应性神经元的形态和化学特征”《神经科学》60. 199-211 (1994)。

Kaneko T,Caria MA and Asanuma H: "Information pro-cessing within the motor cortex. I.Responses of morpho-logically identified motor cortical cells to stimulation of the somatosensory cortex" The Journal of Compar-ative Neurology. vol.345 (no.2) July. 161-

Kaneko T、Caria MA 和 Asanuma H：“运动皮层内的信息处理。I.形态学识别的运动皮层细胞对体感皮层刺激的反应”《比较神经学杂志》。

金子武嗣: "Glutamate-synthesizing enzymes in GABAergic neurons of the neocortex:a double immunofluorescence study in the rat" Neuroscience. 61. 839-849 (1994)

Takeshi Kaneko：“新皮质 GABA 能神经元中的谷氨酸合成酶：大鼠的双重免疫荧光研究”《神经科学》61. 839-849 (1994)。

Kaneko T,Minami M,Satoh M,and Mizuno N: "Im-munocytochemical localization of mu-opioid receptor in the rat caudate-putamen" Neuroscience Letters. vol.184 (no.3) Jan.149-152 (1995)

Kaneko T、Minami M、Satoh M 和 Mizuno N：“大鼠尾壳核中 mu-阿片受体的免疫细胞化学定位”神经科学快报。