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Pharmacological study on intracerebral GABA mechanism

脑内GABA作用机制的药理学研究

基本信息

批准号：
62570095
负责人：
SATO Katsuhiko
金额：
$ 1.15万
依托单位：
Tokyo Medical 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：
1987
资助国家：
日本
起止时间：
1987 至 1989
项目状态：
已结题

项目摘要

1. GABA immunohistochemistry. (1) Study of intraspinal GABA neuron The GABA neuron in the rat spinalcord issued yellowishgreen FITC-specific fluorescence. Specific fluorescence showing the localization of CABA-positive hells and GABA-positive fibers was observed in the spinal anterior horn and posterior horn. To compare the distribution of intraspinal GABA with that of a synthetic enzyme, we prepared specimens by an indirect fluorescent antibody technique using a GAD antibody, and studied for the distribution state on the same level. As a result, we found that the GAD distribution agreed with GABA localization, but the fluorescent intensity and distribution density were less than those of the specific fluorescence of GABA. Accordingly, we believe that using a GABA antibody directly is more accurate in histochemical ; searches for GABA-positive cells and positive fibers. (2) Study of intracerebral GABA neuron. We have studied immunohistochemically the nigra-striatal GABA pathway in the … More rat, and compared it with GAD localization, and compared it with GAD localization. As a result, GABA-positive cells and positive fibers issued green fluorescence in the stratum. Study of GAD-positive cells and positive fibers on the same level revealed that their distribution almost agreed with GABA distribution. GABA- positive fibers issued green fluorescence diffusedly at substantia nigra zoma compacta. The distribution of GAD-positive cells on the same level agreed with GABA distribution.2. Biochemical analysis of GABA content in brain tissues. After pretreatment, the rat brain tissue sample to which a reaction reagent had been added was injected into 10-25ul HPLC. The peak width may become too large if the injection amount exceeds 25ul, with about 20-25 ng at 254 nm 0.01 AUF. The brain tissue sample showed a value of 18 ng/25 ul by a standard curve, and the retention time was 3.40 minutes. The total amount was 400 ul, which meant 288 ng/400ul. Accordingly the content in the sample was 2.88 ug/ml. In analysis of intracerebral GABA by HPLC, in this experiment a reliable and possible value appears to be a concentration of about 2 ug/ml. However, a study is still being made on it. Less

1. GABA免疫组织化学。(1)脊髓内GABA神经元的研究大鼠脊髓内GABA神经元发出黄绿色的fitc特异性荧光。在脊髓前角和后角观察到特异性荧光显示gaba阳性的hell和gaba阳性纤维的定位。为了比较GABA与合成酶在椎管内的分布，我们采用GAD抗体间接荧光抗体技术制备标本，研究其在同一水平上的分布状态。结果，我们发现GAD的分布与GABA的定位一致，但荧光强度和分布密度小于GABA的特异性荧光。因此，我们认为直接使用GABA抗体在组织化学上更准确；寻找gaba阳性细胞和阳性纤维。(2)脑内GABA神经元的研究。我们用免疫组织化学方法研究了…More大鼠黑质纹状体GABA通路，并将其与GAD定位进行了比较，并与GAD定位进行了比较。结果，gaba阳性细胞和阳性纤维在地层中发出绿色荧光。在同一水平上对gad阳性细胞和阳性纤维的研究表明，它们的分布与GABA的分布基本一致。GABA阳性纤维在黑质致密斑处弥漫性发出绿色荧光。gad阳性细胞在同一水平上的分布与GABA的分布一致。脑组织GABA含量的生化分析。预处理后，加入反应试剂的大鼠脑组织样品注射到10-25ul HPLC中。当进样量超过25ul时，峰宽可能过大，在254 nm 0.01 AUF时，峰宽约为20-25 ng。脑组织样品的标准曲线值为18 ng/25 ul，保留时间为3.40 min。总量为400ul，即288 ng/400ul。因此，样品中的含量为2.88 ug/ml。在高效液相色谱法分析脑内GABA时，本实验中可靠且可能的值似乎是2 ug/ml左右的浓度。然而，一项研究仍在进行中。少