| Project/Area Number |
09660331
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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 |
Basic veterinary science/Basic zootechnical science
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| Research Institution | Azabu University |
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Principal Investigator |
FUJISE Hiroshi Azabu University, School of Veterinary Medicine, Professor, 獣医学部, 教授 (40106232)
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| Co-Investigator(Kenkyū-buntansha) |
OCHIAI Hideharu Azabu University, Institute of Biosciences, Lecturer, 生物科学総合研究所, 助手 (20247307)
IKEDA Teruo Azabu University, School of Veterinary Medicine, Associate Professor, 獣医学部, 助教授 (60151297)
SHIROTA Kinji Azabu University, Institute of Biosciences, Professor, 生物科学総合研究所, 教授 (70147974)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥2,700,000 (Direct Cost: ¥2,700,000)
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| Keywords | Dog / potassium / Amino acids / Red blood cells / Erythroblasts / Gene / K-Cl cotransporter / Water channel / K-Cl共輸送 / K-CI共輸送 |
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| Research Abstract |
Progenitor of red blood cells were induced from mononuclear cells separated by Ficoll-Conray from peripheral blood in dog with hereditary high K and low amino acid transport. Firstly the mononuclear cells were incubated in IMDM medium with FCS and phytohemaglutinin-stimulated leukocyte conditioned medium, then they were cultured in the IMDM medium with erythropoietin. These cells were positive with benzidine, glycophorin and hemoglobin, thus it was confirmed that these cells differentiated to erythroblasts.
cDNA cloning of K-Cl cotransporter (KCCI) , aquapolin 1 (AQP1) and Na-dependent glutamate transporter (GLAST) was performed, and full length cDNAs of KCC1 and AQP1 were cloned, and fragment cDNA of GLAST was cloned from the erythroid progenitor and kidney cells. These cDNAs were highly identical to those of other animals reported.
Xenopus oocytes injected with cRNA of AQP1 permeated water more than 3-fold compare to the oocytes injected with only water. Therefore, it was strongly suggested that the AQP1 act for water permeation in the dog red blood cells and kidney cells. HEK293 transfected with KCC1 are investigating for ion transport. In future, mutant gene constructed by site directed mutagenesis will be transfected to the cells to investigate cooperation of AQP1 and KCC1 for volume regulation.
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