1992 Fiscal Year Final Research Report Summary
Multi-point measurement of intracellular chloride concentrations.
| 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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| Keywords | Chloride ion / Cl^- / Cl^--sensitive probe / corneal endothelial cell / retinal pigmemt epithelial cell / hippocampal pyramidal cell / Cl^- pump / Na^+ / K^+ / Cl^- cotransporter |
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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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[Publications] Inoue,M.,Hara,M.,Zeng,X. -T.,Hirose,T.,Ohnishi,S.,Yasukura,T.,Uriu,T.,Minato,A. and Inagaki,C.: "An ATP-driven Cl^- pump regulates Cl^- concentrations in rat hippocampal neurons." Neurosci. Lett.134. 75-78 (1991)
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[Publications] Hara,M.,Inoue,M.,Yasukura,T.,Ohnishi,S.,Mikami,Y. and Inagaki,C.: "Uneven distribution of intracellular Cl^- in rat hippocampal neurons." Neurosci. Lett.143. 135-138 (1992)
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[Publications] Ohnishi,S.,Hara,M.,Inoue,M.,Yamashita,T.,Kumazawa,T.,Minato,A. and Inagaki,C.: "Delayed shortening and shrinkage of cochlear outer hair cells." Am. J. Physiol.263. C1088-C1095 (1992)
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