| Project/Area Number |
07554071
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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 | 展開研究 |
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| Research Field |
動物生理・代謝
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
TAKAHATA Masakazu Hokkaido Univ., Grad.Sch.of Sci., Pro., 大学院・理学研究科, 教授 (10111147)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Etsuro Hokkaido Univ., Grad.Sch.of Sci., Assoc.Pro., 大学院・理学研究科, 助教授 (80203131)
NAGAYAMA Toshiki Hokkaido Univ., Grad.Sch.of Sci., Assoc.Pro., 大学院・理学研究科, 助教授 (80218031)
SUZUKI Noriyo Hokkaido Univ., Grad.Sch.of Sci., Assoc.Pro., 大学院・理学研究科, 助教授 (10001851)
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| Project Period (FY) |
1995 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1996: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | invertebrate / crayfish / optical recording system / dendrite / synaptic activity / calcium fluorescence indicator / image processing / central neuron / 蛍光細胞内染色 / LDS細胞 / 画像解析 / 入力シナプス |
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| Research Abstract |
An optical recording system that can be used in electrophysiological experiments using the whole animal preparation of invertebrates has been developed to visualize the change in intracellular calcium concentration associated with the synaptic activity. The system consists of an epifluorescence device for exciting Fura-2 that is iontophoretically injected into a cell, a dissecting microscope with the working distance of 187mm, a Peltier-cooled CCD video camera, and a personal computer with newly developed programs for image analysis. To test the performance of this recording system, nerve cells in the terminal abdominal ganglion of crayfish were loaded with the dye and observed from the dorsal side with the CCD exposure time of 16sec. The whole cell structure having the soma, dendrites and axon could be visualized in situ together with the electrode tip under the dissecting microscope by either 380nm or 340nm excitation. When the sensory nerve bundle was stimulated repeatedly at 1Hz during the exposure time, a decrease in the fluorescence of the dye to excitation at 380nm was detected in a specific dendritic region of the cell. However, any significant change in the ratio of fluorescence by 380nm to that by 340nm excitation during synaptic activation could not be detected probably due to the relatively weak strength of the latter. It is concluded that although the sensitivity and time resolution are not enough to quantify the dynamics of intracellular calcium ions, the presently developed system can be used to localize the functional synaptic region within a cell by visualizing the change in intracellular calcium concentration due to synaptic activity together with the whole cell structure.
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