Brain mapping with L-proline microinjection on the cardiovascular action and antibody raising against proline synthetic enzyme
| Project/Area Number |
12670040
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General physiology
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
TAKEMOTO Yumi Faculty of Medicine, Physiology, Hiroshima University, Research associate, 医学部, 助手 (60188224)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | L-proline / amino acids / proline synthetic enzyme / cardiovascular system / antibody / medulla / rats / arterial blood pressure |
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| Research Abstract |
1. Brain mapping with L-proline microinjections : (1) Proline stimulation evoked a depressor and bradycardic changes as glutamate did, but through a central mechanism different from that of glutamate, in the nucleus tractus solitarii of anesthetized rats. (2) Proline injections into the caudal ventrolateral medulla induced a depressor action in the anesthetized rat.
2. Raising antibody against L-proline synthetic enzyme : (1) Immunization of peptide fragment antigen made by synthesis. On the basis of an amino acid sequence of human proline synthetic enzyme, peptide fragment parts with highly expected antigenic nature were selected. The peptide fragments synthesized and bound with rabbit serum albumin were immunized and serum from the immunized rabbits was stored. (2) Evaluation of antibody. Before the evaluation, a method for measurement of enzymatic activity of proline synthesis was established using the rat brains. Two methods with HPLC or TLC and with optical density of NADPH were tested. Economical choice was the method by NADPH. By this method, it was possible to detect enzymatic activity of the extract concentrated with an affinity chromatography from the rat brain. This was first evidence for the existence of proline synthetic enzyme in rat brain. Antibodies against the peptide fragments were surely raised, so that they are available to detect the enzyme with the fragment part on the surface after degeneration by the tissue preparation. Raised serum or partially purified antibody with peptide bound affinity chromatography had no inhibitory action on the enzymatic activity from rat brain, suggesting no involvement of peptide parts in the enzymatic activity.
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Report
(3 results)
Research Products
(7 results)
