| Project/Area Number |
02454481
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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 |
Physical pharmacy
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| Research Institution | Nagoya City University |
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Principal Investigator |
NAKANISHI Mamoru Nagoya City Univ. Professor Fac. Pharm. Sci., 薬学部, 教授 (90090472)
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| Co-Investigator(Kenkyū-buntansha) |
TORIGOE Chikako Nagoya City Univ. Instructor Fac. Pharm. Sci., 薬学部, 助手 (70237163)
INAGAKI Kenji Nagoya City Univ. Lecture Fac. Pharm. Sci., 薬学部, 講師 (00080193)
広瀬 順三 福山大学, 工学部, 助教授 (70080215)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1991: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1990: ¥3,500,000 (Direct Cost: ¥3,500,000)
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| Keywords | Calcium Ion / Image Analysis / Second Messenger / Immune Responses / Confocal Microscope / B cell / T cell / Basophile / 蛍光画像処理法 / カルシウムシグナル / 抗原レセプタ- / 脱感作 / プロテインキナ-ゼC / 共焦点レ-ザ顕微鏡 |
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| Research Abstract |
A confocal fluorescence microscope with an argon ion laser (488 nm) and a He-Cd laser (325 nm) , was used to study spatial heterogeneity of the calcium signals in rat basophilic leukemia cells (RBL-2H3 cells). After the stimulation of antigen (DNP_7 - BSA), fluo-3 fluorescence intensities increased in individual RBL-2H3 cells with different lag times. Timedependent profiles of the fluo-3 fluorescence intensities resembled closely the patterns of the sequential fluorescence ratio images of fura-2, which were used to measure [Ca^<2+>]_i in individual RBL-2H3 cells by a conventional fluorescence microscope. The present results by the confocal fluorescence microscope showed spatial heterogeneities of fluo-3 fluorescence intensities, suggesting the exsstence of the spatial heterogeneity of[Ca^<2+>]_i in RBL-2H3 cells. That is, the results showed that calcium signals first occurred transiently at pseudopods in RBL-2H3 cells and then the signals transferred to the central parts of the cells. In addition, from the fluorescence images of co-loaded Hoechst 33342 (DNA-specific probe) which were excited by a He-Cd laser, it was found that the fluorescence images of the nucleus were quite similar to those of the calcium signals mentioned above. This suggested that the receptor-mediated calcium signals were transferred not only to the cytoplasm but also to the nucleus.
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