| Project/Area Number |
06557045
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
KANAIDE Hideo KYUSHU UNIVERSITY,FACULTY OF MEDICINE,PROFESSOR, 医学部, 教授 (80038851)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIMURA Junji KYUSHU UNIVERSITY,FACULTY OF MEDICINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (90237727)
KOBAYASHI Sei KYUSHU UNIVERSITY,FACULTY OF MEDICINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (80225515)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 1996: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1995: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1994: ¥8,000,000 (Direct Cost: ¥8,000,000)
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| Keywords | Vascular endothelial cells / vascular smooth muscle cells / measurements in vivo / cytosolic calcium concentration / fura-2 / フラ-ス |
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| Research Abstract |
1. THE DEVELOPMENT OF A NEW SYSTEM FOR MONITORING THE FUNCTIONS OF THE VASCULAR ENDOTHELIAL AND SMOOTH MUSCLE CELLS : The system we have developed has following characteristics ; (1) Using a grating and a chopper mirror driven by a stepping motor, it became possible to alternately (10 Hz) illuminate the tissue sample with 340 and 380 nm excitation lights for the measurement of intracellular calcium concentration, [Ca] i. (2) the chopper system is controlled through a personal computer by investigator designed software. (3) Fluorescence images are obtained using a chopper mirror and a SIT camera. A display flame is 640x400 dots and a 8 bit flame. (4) Fluorescence ratio (F340/F380) images can be obtained at 5 Hz. (5) It is possible to simultaneously record changes in the [Ca] i levels at 30 points in an image on the TV monitor. (6) Low cost for making the system.
2. NEW FINDINGS OBTAINED USING THIS SYSTEM,FRONT-SURFACE FLUORIMETRY AND TECHNIQUES IN MOLECULAR BIOLOGY : (1) Using the newly developed technique, the simultaneous fluorimetry of fura-2 and diaminonaphthalene, we found the [Ca] i-NO production relationships in the endothelial cells in situ. (2) Using front-sueface fluorimetry, reverse transcription polymerase chain reaction, and the renal and coronary arteries, we found that both adrenomedullin and endothelin might act as autocrine/paracrione signal transmitters in these tissues.
3. HUMANA PRESS (NJ : U.S.A) asked us to write a chapter "Measurement of [Ca] i in muscle strips using front-surface fluorimetry" of a book on "Calcium signaling protocols (edited by DG Lambert)", a part of the Methods in Molecular Biology series. It is in press.
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