1996 Fiscal Year Final Research Report Summary
Analysis of the intracellular second messenger pathway in the intestinal epithelial cell
| Project/Area Number |
08044331
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Field |
General physiology
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| Research Institution | University of Shizuoka |
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Principal Investigator |
KUWAHARA Atsukazu Univ. of Shizuoka, 大学院, 助教授 (60142890)
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| Co-Investigator(Kenkyū-buntansha) |
SUGIYA Hiroshi Nihon Univ. Sch., of Dentistry, 松戸歯学部, 助教授 (20050114)
MOTIZUKI Tohru Univ. of Shizuoka, 薬学部, 助教授 (00117780)
FUJIMIYA Mineko Shiga Univ. of Medical Sci, 医学部, 助教授 (10199359)
RECHKEMMER G. Inst., fur Ernaehrungsphysiologie, 部長
LABURTHE M. Inserm, Frace, 部長
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| Project Period (FY) |
1996 – 1997
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| Keywords | Guinea - pig / Distal colon / Chloride secretion / Cytosolic free calcium / Tachykinin / Crypt cell / Conforcal micorscopy / Ion transport |
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| Research Abstract |
In epithelial cells of the intestine , fluid secretion is driven by the accumulation of ClィイD1-ィエD1 in the cytosol by the action of the NaィイD1+ィエD1-KィイD1+ィエD1-2ClィイD1-ィエD1 co - transporter , the NaィイD1+ィエD1-HィイD1+ィエD1 exchanger which are in the basolateral membrane . Chloride ion then enter the lumen through channels in the apical membrane and this electrogenic anion efflux is balanced by KィイD1+ィエD1 effluex through KィイD1+ィエD1 channel in the basolateral and possibly the apical membrane . Various neurpeptides including substance P control the rate of secretion by increasing the intracellular CaィイD12+ィエD1 concentration which in turn activates the ClィイD1-ィエD1channels and the KィイD1+ィエD1 channels . Substance P has been reported to activate several intracellular signal transduction system . In despite of the physiological importance of chloride secretion and its regulation by substance P in distal colon , neither the colonic epithelium signaling mechanisms nor the specific receptor present on
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these cells have been characterized .
Thus , We have examined the effect of substance P on CaィイD12+ィエD1ィイD2IィエD2 of isolated crypt cells from guinea - pig colon by confocal laser scanning microscope . Substance P , neurokinin A and neurokinin B evoked a concentration - dependent in crease in [CaィイD12+ィエD1]ィイD2iィエD2 with a rank order of potencies of substance P > neurokinin A >> neurokinin B . The selective NK-1 receptor antagonist , FK888 reduced the responses to substance P and neurokini A . In extracellular CaィイD12+ィエD1 free condition , integrated pattern of substance P - induced increase in [CaィイD12+ィエD1]ィイD2iィエD2 were significantly decreased ; substance P - induced increase in [CaィイD12+ィエD1]ィイD2IiィエD2 decreased to 50 % of control . Thapsigargin also decreased the substance P - induced increase in [CaィイD12+ィエD1]ィイD2iィエD2 to 60 % of control . Combination of exracellular CaィイD12+ィエD1 free and thapsigargin pretreatment caused a greater decrease of substance P- induced increase in [CaィイD12+ィエD1]ィイD2iィエD2 . The results suggest that both extracellular CaィイD12+ィエD1 and intracellular thapsigargin - sensitive CaィイD12+ィエD1 and intracellular thapsigargin - sensitive CaィイD12+ィエD1 stores contribute to substance P - induced increase in [CaィイD12+ィエD1]ィイD2iィエD2 . Less
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[Publications] Yanaihara N., Mochizuki T,. Kuwahara A., Hoshino M., Kakuyama H., Iguchi K., Iwanaga T., Li, J., Futai Y., Kanno T., Yamabe K., and Yanaihara C.: "Endocrine and Gastrointestinal Action of Galanin"Annales of the New York Academy of Sciences Vol. 863. 129-142 (1998)
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