| Project/Area Number |
61440025
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
General physiology
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| Research Institution | Hokkaido University |
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Principal Investigator |
KANNO Tomio Hokkaido University, Faculty of Veterinary Medicine, Professor, 獣医学部, 教授 (50009982)
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| Co-Investigator(Kenkyū-buntansha) |
SAITO Toshiyuki Hokkaido University, Faculty of Veterinary Medicine, Instructor, 獣医学部, 助手 (10162215)
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| Project Period (FY) |
1986 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥24,100,000 (Direct Cost: ¥24,100,000)
Fiscal Year 1989: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1988: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1987: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1986: ¥12,500,000 (Direct Cost: ¥12,500,000)
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| Keywords | Intracellular concentration of calcium ion / Pancreatic acinar cell / Cholecystokinin / Stimulus-secretion coupling / Secretory response of exocrine gland / Nomarski apparatus / Fura-2 / Rough-surface endoplasmic reticulum / 刺激ー放出連関 / furaー2 / チトクローム / 酸化還元位 / CCK-8 / 膵消化酵素放出反応 / 膵液流量上昇反応 / 栄養効果 / 細胞内のCaイオン / (Na^+、K^+)ATPase / CCKー8 / 下垂体神経葉 / CNa^+,K^+)ATPase / ミトコンドリア / チトクロームa(【a_3】),bおよびC+【C_1】 / 酸化還元反応 / 消化酵素放出反応 / 肥大膵 |
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| Research Abstract |
Microspectrofluorimetry with fura-2 was utilized to monitor cytosolic concentration of calcium ion, [Ca^<2+>]_i, in single cells of the isolated pancreatic acini during continuous stimulation with C-terminal octapeptide of cholecystokinin (CCK-8). Prior to the microspectrofluorimtry, the same cells were examined under the Nomarski differential interference contrast optical system. Regional morphological differences could be resolved, insofar as, the apical half of an acinar cell contained a large number of zymogen granules (ZG region), whereas the basal half appeared clear with few granulation (region of endoplasmic reticulum; ER region). When acinar cells were continuously stimulated with 30pM CCK-8, which is known to cause sustained increase in amylase release from pancreatic acini, oscillation of [Ca^<2+>]_i was usually observed in the ER region, whereas a single transient increase in [Ca^<2+>]_i was usually detected in the ZG region. The oscillatory change in [Ca^<2+>]_i consisted of three components: 1) an initial transient increase flowed by 2) a gradual decline, upon which 30 pM CCK-8 during perfusion with a Ca^<2+>-free solution, the initial transient increase in [Ca^<2+>]_i remained but the secondary oscillations were abolished. A single transient increase in [Ca^<2+>]_i without any subsequent oscillation was observed when the acinar cells were continuously stimulated with 100pM CCK-8. Secretory responses of acinar cells to 30pM CCK-8 were not interfered with fura-2 AM (10mum), but these to 100 pM CCK-8 were significantly inhibited with fura-2 AM. These results show that the CCK-8-induced oscillatory changes in [Ca^<2+>]_i may play a significant role in the stimulus-secretion coupling in the pancreatic acinar cell.
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