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Molecular mechanisms of salivary exocytosis-endocytosis coupling

Research Project

Project/Area Number 09671866
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Morphological basic dentistry
Research InstitutionKitasato University

Principal Investigator

SEGAWA Akihisa  Kitasato Univ. School of Medicine, Assistant Professor, 医学部, 講師 (50154638)

Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsSalivary glands / exocytosis / endocytosis / calcium / IPィイD23ィエD2 receptor
Research Abstract

This research project is aimed to clarify the molecular mechanisms involved in exocytosis-endocytosis coupling in salivary glands by morphological/functional/biochemical approaches. This year, analyses were performed on whole gland perfusion system, calcium signaling and parotid glands in l3-galactosidase knock-out mice.
1) Whole gland perfusion: Isolated whole parotid glands of rats were perfused arterially with isoproterenol (β-agonist) or carbachol (muscarinic agonist), and secreted saliva and tissues were collected sequentially; β-stimulation evoked exocytosis following which the fused granule membranes were removed one by one. Muscarinic stimulation inhibited this removal process. Since muscarinic stimulation causes fluid secretion but β-stimulation does not, it is suggested that membrane retrieval process plays significant roles in regulation of fluid secretion.
2) Calcium signaling: High speed cofocal microscopy of living parotid glands showed the synchronized response in acini and the non-synchronized response in ducts. Gap junction was detected between acinar cells but not duct cells. Acini are likely to function as a syncitium.
3) β-galactosidase knock-out mice: β-galactosidase is involved in the lysosomal degradation of membrane components. Parotid acinar cells in mice deficient in this enzyme revealed vacuolar profiles in the cytoplasm. Electron microscopic observation demonstrated that the vacuoles appeared in the RER region. Unexpectedly, this enzyme is suggested to play roles in the biosynthetic processes rather than the degradation.

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (32 results)

All Other

All Publications (32 results)

  • [Publications] A. Segawa: "Measurement of secretion in confocal microscopy"Methods in Endymology. 307. 328-340 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Nowroozi, S Kim, A Segawa et al: "Hih Lenals of GM_1Gangliosidase in the panotid gland"Otoloryngologic Clinics of North America. 32. 779-791 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Takamura, S. Yamashita, A. Segawa: "Hillisecord analyses of Ca^<2+> initiation sites evolsed muscarinic stimulation in exocine alimar cells"Biochem Biophys. Res. Comm.. 259. 656-660 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K. Yoshimura, M. Murakami, A. Segawa: "Carbachol induced Ca increase, but not activation of protein kinase c, stimulates exocytsis in rat parotid acim"J. Physiol.(London). 522. 403-416 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Segawa, F. Loffredo et al: "Cell Biology of human salivary secretion"European Journal of Morphology. (in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Murakami, K. Yoshimura, A. Segawa et al: "Relationship between anuylase and fluid secretion in the isolated parfiosel abobe pawtid gland of the rat"European Journal of Morpholoy. (in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A Segawa: "Measurement of secretion in confocal microscopy."Methods in Enzymology. Vol 307"Confocal Microscopy" (PM Conn ed) Academic Press. 328-340 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H Takemura, S Yamashina, A Segawa: "Millisecond analysis of Ca2+ initiation sites evoked by muscarinic receptor stimulation in exocrine acinar cells"Biochemical Biophysical Research Communications. 259. 656-660 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N Nowroozi, S Kim, A Segawa, H Warita, D Rice, PC Denny, JH Zernik: "High levels of GM1 ganglioside and the GM1-galactosidase in the partoid gland - a new model for secretory mechanisms of the partoid gland"Otolaryngologic Clinics of North America. 32. 779-791 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K Yoshimura, M Murakami, A Segawa: "Carbachol induced Ca increase, but not activation of protein kinase C, stimulates exocytosis in rat parotid acini"Joumal of physiology (London). 522. 403-416 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A Segawa, F Loffredo, R Puxeddu, S Yamashina, F Testa Riva, A Riva: "Cell biology of human saliva secretion"European Journal of Morphology. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M Murakami, K Yoshimura, A Segawa, F Loffredo, A Riva: "Relationship between amylase and fluid secretion in the isolated perfused whole parotid of the rat"European Journal of Morphology. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A.Segawa: "Measurement of secretion in confocal microscopy"Methodo in Engymology. 307. 328-340 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N.Nowrozi,Skim,A Segawa et al.: "High levels of GM_1 -gangliosidase in the parotid gland"Otolauynagologi Clinis of North America. 32. 779-791 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] H.Takemura,S Yamashina,A.Segawa: "Millisecond analysis of Ca^<2+> initiation evoked by muocarinic stinulation in exocine acimar cells"Biochem.Biophys.Res.Comm.. 259. 656-660 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] K.Yoshimura,M.Murakami,A.Segawa: "Carbachol indnced Ca increar, but not activation of protein kinase C, stinulates exocytsies in rat parotid acim"J.Physiol.(London). (in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] A.Segawa,F Lottredo et al: "Cell Biology of human saliray secretion"European Journal of Mophology. (in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] M.Murakami,K.Yoshimura,A.Segawa et al: "Relation letumn analysis and"European Journal of Mophology. (in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] A.Riva et al: "Human salivary gland parenchymal cell seen by SEM from the cytoplasmic side using a new osmiusn mausation method" European Journal of Morphology. 36. 35-40 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Segawa et al: "Cytoskelctal regulation of human salivary secution studied by high resolution electron micsoscopy and enfocal laser microscopy" European Journal of Morphology. 36. 41-45 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Segawa et al: "Exocytosis in human salivay glands visualized by high-resolution scanning elechon microciopy" Cell Tissue Res.291. 325-336 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Segawa: "Exocytosis-removal cycle of single secretory gramulis visualized by confocal laser microscopy" Bioimges. 5. 153-155 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Miki-Yamamoto Hino et al: "Apical vesicles bearing inositol 1,4,5-hiphosphate receptors in the Ca^<2+> initiation site of ductol epithelium of submanclibulor gland" Journal of Cell Biology. 141. 135-142 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Segawa: "Heasurement of secretion" Methods in Enzymology. (in press).

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Segawa: "Recent Advances in Microscopy of Cells,Tissues and Organo" Antonio Delfino Editore(Rome), 363-366 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Segawa et al: "Glandular Mechonismo of Saliroy Secretion" Karger(Basel), 89-100 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Akihisa Segawa et al: "Exciytosis in human salivary glands visualized by high scaoluhion scanning electron microscopy" Cell and Tissue Research. (in press). (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Alessandio Riva et al: "The paresnchy mal cells of the human salivary glands seen at SEM from the cytoplasmic side by a new osumium macechin metho" European Journal of Horphology. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Akihisa Segawa et al: "Cytosheletol reyulation of human salivary sicution studied by high resolution olictionsmicusciyuy and comfocal lason mirmy." European Journal of Morphology. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Miki Yamamoto-Hino et al: "Apical vesicles bearing inositol 1,4,5-Tirphosphate receptors in the Ca^<2+> initiation site of ductal epthelium of sulimonclibilar gland" Journal of Cell Biology. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Akihisa Segawa: "Receut Advances in Microscopy of Cells,Tissues and Orgons" Malpighi Academy (Rome) (in press), (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Akihisa Segawa: "Glandular Mechanisms of Salivary Secretion" Karger (Basel) (in press), (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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