| Project/Area Number |
01470129
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
製造化学・食品
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| Research Institution | Kyoto University |
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Principal Investigator |
SASAKI Ryuzo Kyoto University, Agriculture, Professor, 農学部, 教授 (60077378)
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| Co-Investigator(Kenkyū-buntansha) |
IKURA Koji Kyoto University, Agriculture, Assistant, 農学部, 助手 (00101246)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1990: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | Erythropoiesis / Erythropoietin / Kidney / Dietary protein / Monoclonal antibody / 単クロ-ン抗体 / 赤血球前駆細胞 / 腎細胞 / 造血因子 / 食餌蛋白質 |
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| Research Abstract |
Erythrocytes have a limited life span ; mammals have to produce erythrocytes for their whole life. Recruitment of matured erythrocytes is achieved by proliferation and differentiation of undifferentiated hematopoietic precursors. Erythropoietin (EPO), a glycoprotein produced by kidney, is the major physiological stimulator of these processes. When protein-deprived died was given to rats, the rate of erythropoiesis is impaired. We have shown that this impairment is due to abrupt cessation of EPO synthesis that appears to occur immediately after deprivation of protein in the diet. The purpose of this project is to elucidate the mechanism by which the EPO synthesis is under control of protein intake.
(1) To establish the method of EPO measurement with high sensitivity, high accuracy, and feasibility, monoclonal antibodies that recognize different epitopes of EPO have been prepared. The established method, a sandwich-type enzyme-linked immunoassay, measures EPO as low as 5 mU/ml within several hours. (2) The Kidney cells responsible for EPO production have been unclear. By the use of the monoclonal antibody, we stained the kidney of rat in which EPO production is induced by bleeding. The microscopic observation of the stained cells suggests that the peritubular endothelial cells are most likely candidate for production of EPO. (3) To develop the in vitro culture system of the kidney cells producing EPO, the kidney cortex of the anemic rat was disrupted by collagenase to yield single cell suspension. The EPO production was detected in the supernatant of the cultured cells. Concentration, purification, and identification of the cells producing EPO have being attempted so that we can test effect of the sera from the rats to which either protein-deprived or protein-supplied diet is given on EPO production of the cultured cells.
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