1989 Fiscal Year Final Research Report Summary
Pharmacological study on intracerebral GABA mechanism
| Project/Area Number |
62570095
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General pharmacology
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| Research Institution | Tokyo Medical University |
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Principal Investigator |
SATO Katsuhiko Tokyo Med. Coll., Med. fac., Ass. Prof., 医学部, 助教授 (00133372)
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| Project Period (FY) |
1987 – 1989
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| Keywords | GABA neuron / GAD neuron / immunohistochemistry / indirect fluorescent antibody technique / high-performance liquid chromatography / fluorescent histochemistry |
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| Research Abstract |
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
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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
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