| Project/Area Number |
02557012
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General pharmacology
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| Research Institution | Kansai Medical University |
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Principal Investigator |
INAGAKI Chiyoko Kansai medical University Professor, 医学部, 教授 (60025640)
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| Co-Investigator(Kenkyū-buntansha) |
URIU Toshiko Kansai medical University Assistant, 医学部, 助手 (40203608)
HARA Mitsuyoshi Kansai medical University Assistant, 医学部, 助手 (50192282)
OTANI Hitomi Kansai medical University Assistant, 医学部, 助手 (40140272)
INOUE Masafumi Kansai medical University Lecturer, 医学部, 講師 (90203257)
OMORI Kyoko Kansai medical University Lecturer, 医学部, 講師 (90152256)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥10,400,000 (Direct Cost: ¥10,400,000)
Fiscal Year 1992: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1991: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥6,800,000 (Direct Cost: ¥6,800,000)
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| Keywords | Chloride ion / Cl^- / Cl^--sensitive probe / corneal endothelial cell / retinal pigmemt epithelial cell / hippocampal pyramidal cell / Cl^- pump / Na^+ / K^+ / Cl^- cotransporter / Cl^-感受性蛍光色素 / 細胞内Cl^-濃度 / エタクリン酸 / フロセミド / 海馬神経細胞 |
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| Research Abstract |
1. Cl^--sensitive probe: A Cl^--sensitive fluorescent dye,N-(6-methoxyquinolyl) acetoethyl ester (MQAE, purity > 99%) was synthesized by the method of Verkman.
2. Multi-channel window photometer:The fluorescence intensity of a MQAE-loaded cell was continuously recorded on an inverted microscope equipped with a night vision camera-based multi-channel window photometer, which was devised to measure the photo intensity at desired multiple windows in a microscopic view. Using this equipment, changes in the intracellular Cl^- concentration ([Cl^-]_i) were monitored to elucidate the mechanisms for its regulation.
3. Corneal endothelial cell:Stationary [Cl^-]_i of rabbit corneal endothelial cells was 30.1*1.6 mM. Efflux of Cl^- with alpha_2^-adrenoceptor stimulation,and an increase in Cl^- permeability of plasma membranes with beta-adrenoceptor stimulation were suggested.
4. Retinal pigment epithelial cell (RPE): Stationary [Cl^-]_i of rabbit RPE was approx. 90 mM. Atrial natriuretic peptide was demonstrated to accumulate through activation of Na^+/K^+/Cl^- co-transporter regulated via cGMP-dependent protein kinase.
5. Hippocampal pyramidal cell:Stationary [Cl^-]_i of rat hippocampal pyramidal cells was 6.63*1.56 mM. An ATP-driven Cl^- pump extrudes Cl^- and Na^+/K^+/Cl^- co-transporter accumulates the same anion.
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