2000 Fiscal Year Annual Research Report

外分泌における水電解質輸送と開口分泌の協調機構

Research Project

Project/Area Number	10044334
Research Institution	Okazaki National Research Institutes
Principal Investigator	村上政隆岡崎国立共同研究機構, 生理学研究所, 助教授 (10104275)
Co-Investigator(Kenkyū-buntansha)	吉垣純子日本大学, 松戸歯学部, 助手 (40256904) 瀬川彰久北里大学, 医学部, 講師 (50154638) 杉谷博士日本大学, 松戸歯学部, 助教授 (20050114) 吉村啓一北海道大学, 歯学部, 助教授 (30000938) 瀬尾芳輝京都府立医科大学, 医学部, 講師 (90179317)
Keywords	水分泌 / 水電解質輸送 / 蛋白分泌 / 開口放出 / 唾液腺 / 輸送体 / チャンネル / 細胞内情報伝達
Research Abstract	本研究は、大量の水分泌と開口分泌を観察できる唾液腺に対象を限定、小胞輸送により運ばれた分泌顆粒が細胞膜に融合し開口放出を起こすプロセスについて分子生理的、形態学的に水電解質輸送と開口分泌を計測、水輸送・開口分泌両システム間の協調機構を検討することを目的とし実施した。平成12年度以下の実績が得られた。 A.分泌経過チーム:開口分泌(粘液分泌)のモデルとして、ラット顎下線の血管潅流・単離細胞標本を作製し、カルバコール(ムスカリン受容体刺激)イソプロテレノール(βアドレナリン性受容体刺激)について水分泌・ムチン分泌・酸素消費の時間経過を測定、平成10-12年度で明らかにならなかったβアドレナリン性受容体刺激による水分泌と酸素消費の増強効果が低濃度カルバコール刺激で見い出され、従来の結果と合せ増強効果が細胞エネルギー供給系の能力により制限されることを示唆した。 B.形態チームは、光学・透過電子・走査電子顕微鏡を用い、細胞間分泌細管IC(管腔)と周辺微細構造を観察した。ムスカリン受容体刺激ではIC周辺のrERの拡張、mitochondoria電子密度増加、まばらな開口放出像、microvilli減少、ICの初期一過性拡大が観察された。βアドレナリン性受容体刺激により、ICに多くの開口分泌がみられ、その周辺にさらに分泌顆粒が開口分泌する像が観察された。またmicrovilliは消失した。IC周辺のrERの拡張mitochondoria電子密度増加も同時に観測された。分泌活動時の、rERの拡張は細胞内Ca代謝と、mitochondoria電子密度増加はエネルギー代謝増加と関連づけられる。 C.開口放出関連蛋白チームは、免疫組織化学法により耳下腺ではVAMP2が分泌顆粒膜に、syntaxin4とSNAP23が管腔膜に発現するが、顎下腺・舌下腺ではsyntazxin1Aが腺房細胞に局在することを見い出した。粘液分泌と漿液分泌で開口分泌の分子機序が異なる可能性が示された。 D.分泌物質の運動性グループは、標識デキストランの腺房上皮通過を測定結果を解析し、ラット顎下腺ではムスカリン受容体刺激により直径5Åの傍細胞分泌経路が開通すると推定した。本年度、開口分泌と水輸送の協調機構として、1)エネルギー供給の制限があること2)水輸送関連蛋白の移送と蛋白分泌の共通制御の可能性、3)傍細胞経路の調節の存在、が明らかになった。

Research Products
(38 results)

All Other

All Publications (38 results)

[Publications] Murakami M: "Relationship between amylase and fluid secretion in the iso1ated perfused whole parotid gland of the rat."Eur J Morphology. 38(4). 243-247 (2000)
[Publications] Murakami M: "Fluid and amylase secretion by perfused parotid gland : physio-morphological approach."J Korean Med Sci. 15. S38-39 (2000)
[Publications] Yoshimura K: "Carbachol-induced [Ca^<2+>]_i increase,but not activation of protein kinase C, stimulates exocytosis in rat parotid acini."Journal of Physiology. 522. 403-416 (2000)
[Publications] Seo Y: "Anisotropic and restricted diffusion of water in sciatic nerve : A 2H double-quantum-filtered NMR study."Magn.Reson.Med. 42. 461-466 (1999)
[Publications] Seo Y: "1H double-quantum-filtered MR imaging as a new tool for assessment of healing of the ruptured Achilles tendon."Magn.Reson.Med.. 42. 884-889 (1999)
[Publications] Tsoref T: "Slice-selective proton double quantum filtered MRI of joint connective tissues."J.Magn.Reson.Imag.. 11. 336-341 (2000)
[Publications] Takamiya H.: "Characteristics of proton NMR T2 relaxation of water in the normal and regenerating tendon."Jpn.J.Physiol.. 50(6). 569-576 (2000)
[Publications] Sogami,M: "Application of the transition state theory to water transport across cell membrane."Biochi.Biophys.Acta. (In press). (2001)
[Publications] Kusaka,Y: "Intradiscal solid phase displacement as a determinant of the centripetal fluid shift in the loaded intervertebral disc."Spine. 26(In press). (2001)
[Publications] Navon,G: "Multiquantum filters and order in tissues."NMR in Biomedicine. 14(In press). (2001)
[Publications] Kukimoto M: "Crystallization and preliminary X-ray difraction analysis of the extracellular domain of the cell surface antigen CD38 complexed with gnaglioside."J.Biochem.. 127. 181-184 (2000)
[Publications] Sawada M: "Ribose 1,5-bisphosphate is a putative regulator of fructose 6-phosphate/fructose 1,6-bisphosphate cycle in liver."Int.J.Biochem.Cell Biol.. 32. 447-454 (2000)
[Publications] Ozaki I: "Ribose l,5-bisphosphate inhibits fructose-1,6-bisphosphatase in rat kidney cortex."Comp.Biochem.Physiol.. 125B. 97-102 (2000)
[Publications] Nakao S: "Bradykinin potentiates prostaglandin E2 release in the human gingival fibroblasts pretreated with interleukin-1B via Ca2+ mobilization."Eur.J.Pharmacol.. 385. 247-253 (2000)
[Publications] Nakao S: "Activation of NFkB is necessary for IL-1B-induced cyclooxygenese-2(COX-2) expression in human gingival fibroblasts."Mol.Cell.Biochem.. 209. 113-118 (2000)
[Publications] Hara-Yokoyama M: "Complex gnagliosides as cell surface inhibitors for the ecto-NAD+ glycohydrolase of CD38."Biochemistry. 40. 888-895 (2001)
[Publications] Fujita-Yoshigaki,J.: "Simulation of Regulated Exocytosis of Amylase from Salivary Parotid Acinar Cells by a Consecutive Reaction Model Comprising Two Sequential First-order Reactions."J.Theor.Biol.. 204. 165-177 (2000)
[Publications] Segawa A: "Cell biology of human salivary secretion."Eur J Mophology. 38. 237-241 (2000)
[Publications] Riva A: "A high resolution SEM study of human minor salivary glands."Eur J Mophology. 38. 219-226 (2000)
[Publications] Ambu R: "Scanning electron microscopy of the interior of cells in Hurthle cell tumors."Ultrastructural Pathology. 24. 211-219 (2000)
[Publications] Curry MR: "Single water channels of Aquaporin-1 do not obey the Kedem-Katchalsky equations"J.Membrane Biol. (In press). (2001)
[Publications] Gerelsaikhan,T.: "Transmembrane topology of the secretory Na-K-2Cl cotransporter studied by in vitro translation."J Biol Chem. 275. 40471-40477 (2000)
[Publications] Moore-Hoon,M.L: "Molecular characterization of the cation-chloride cotransporter family."Eur.J.Morphology. 36. S137-141 (1998)
[Publications] Evans,R.L: "New insights into the upregulation and function of the salivary Na^+-K^+-2Cl^- cotransporter."Eur.J.Morphology. 36. S142-146 (1998)
[Publications] Moore-Hoon,M.L: "Increased expression of the secretory Na^+-K^+-2Cl^- cotransporter with differentiation of a human intestinal cell line."Biochem.Biophys.Res.Comm.. 244. 15-19 (1998)
[Publications] Evans,R.L: "Evidence for a physiological role of NH_4^+ transport on the secretory Na^+-K^+-2Cl^- cotransporter."Biochem.Biophys.Res.Comm.. 245. 301-306 (1998)
[Publications] Moore-Hoon,M.L: "Molecular and topological characterization of the rat parotid Na^+-K^+-2Cl^- cotransporter."Biochim.Biophys.Acta. 1373. 261-269 (1998)
[Publications] Turner,R.J.: "Ion transport and signalling in human labial glands."Arch.Oral Biol.. 44. S15-19 (1999)
[Publications] Kurihara,K: "Characterization of the phosphorylation event resulting in upregulation of the salivary Na+-K+-2Cl-cotransporter."Biochemistry. 39. 3718-3724 (2000)
[Publications] Evans,R.L.: "Severe impairment of salivation in Na^+/K^+/2Cl^- cotransporter (NKCC1)-deficient mice."J.Biol.Chem.. 275. 26720-26726 (2000)
[Publications] Gerelsaikhan,T.: "Membrane topology and function of the secretory Na^+-K^+-2Cl^- cotransporter (NKCC1)."J.Kor.Acad.Med.. 15. S3-4 (2000)
[Publications] Tanimura,A.: "Evidence that type I, II and III inositol 1,4,5-trisphosphate receptors can occur as integral plasma membrane proteins."J.Biol.Chem.. 275. 27488-27493 (2000)
[Publications] Shimono,M.: "Connexins in salivary glands"European J Morphology. 38. 257-261 (2000)
[Publications] Hashimoto,S: "Tight junctions in the rat parotid gland"European J Morphology. 38. 263-267 (2000)
[Publications] Shimono,M: "Connexins in the developing salivary glands"European J Morphology. 36. S112-117 (2000)
[Publications] Benes C: "Alterations in PKCδ phosphorlation and activity in rat parotid acinar cells by receptor-dependent and receptor-independent stimuli."Am J Physiology. (In press). (2001)
[Publications] Whittembury G: "The Kidney : Physiology & Pathophysiology.Eds : Seldin SG & Giebisch G.Philadelphia, Lippincot, Williams & Wilkins"Coupled transport of water and solutes across epithelia. (2000)
[Publications] Hill AE: "Int.Revs,Cytology."Paracellular fluid transport by epithelia.(In press). (2001)

2000 Fiscal Year Annual Research Report

外分泌における水電解質輸送と開口分泌の協調機構

Principal Investigator

村上 政隆 岡崎国立共同研究機構, 生理学研究所, 助教授 (10104275)

Research Products

[Publications] Murakami M: "Relationship between amylase and fluid secretion in the iso1ated perfused whole parotid gland of the rat."Eur J Morphology. 38(4). 243-247 (2000)

[Publications] Murakami M: "Fluid and amylase secretion by perfused parotid gland : physio-morphological approach."J Korean Med Sci. 15. S38-39 (2000)

[Publications] Yoshimura K: "Carbachol-induced [Ca^<2+>]_i increase,but not activation of protein kinase C, stimulates exocytosis in rat parotid acini."Journal of Physiology. 522. 403-416 (2000)

[Publications] Seo Y: "Anisotropic and restricted diffusion of water in sciatic nerve : A 2H double-quantum-filtered NMR study."Magn.Reson.Med. 42. 461-466 (1999)

[Publications] Seo Y: "1H double-quantum-filtered MR imaging as a new tool for assessment of healing of the ruptured Achilles tendon."Magn.Reson.Med.. 42. 884-889 (1999)

[Publications] Tsoref T: "Slice-selective proton double quantum filtered MRI of joint connective tissues."J.Magn.Reson.Imag.. 11. 336-341 (2000)

[Publications] Takamiya H.: "Characteristics of proton NMR T2 relaxation of water in the normal and regenerating tendon."Jpn.J.Physiol.. 50(6). 569-576 (2000)

[Publications] Sogami,M: "Application of the transition state theory to water transport across cell membrane."Biochi.Biophys.Acta. (In press). (2001)

[Publications] Kusaka,Y: "Intradiscal solid phase displacement as a determinant of the centripetal fluid shift in the loaded intervertebral disc."Spine. 26(In press). (2001)

[Publications] Navon,G: "Multiquantum filters and order in tissues."NMR in Biomedicine. 14(In press). (2001)

[Publications] Kukimoto M: "Crystallization and preliminary X-ray difraction analysis of the extracellular domain of the cell surface antigen CD38 complexed with gnaglioside."J.Biochem.. 127. 181-184 (2000)

[Publications] Sawada M: "Ribose 1,5-bisphosphate is a putative regulator of fructose 6-phosphate/fructose 1,6-bisphosphate cycle in liver."Int.J.Biochem.Cell Biol.. 32. 447-454 (2000)

[Publications] Ozaki I: "Ribose l,5-bisphosphate inhibits fructose-1,6-bisphosphatase in rat kidney cortex."Comp.Biochem.Physiol.. 125B. 97-102 (2000)

[Publications] Nakao S: "Bradykinin potentiates prostaglandin E2 release in the human gingival fibroblasts pretreated with interleukin-1B via Ca2+ mobilization."Eur.J.Pharmacol.. 385. 247-253 (2000)

[Publications] Nakao S: "Activation of NFkB is necessary for IL-1B-induced cyclooxygenese-2(COX-2) expression in human gingival fibroblasts."Mol.Cell.Biochem.. 209. 113-118 (2000)

[Publications] Hara-Yokoyama M: "Complex gnagliosides as cell surface inhibitors for the ecto-NAD+ glycohydrolase of CD38."Biochemistry. 40. 888-895 (2001)

[Publications] Fujita-Yoshigaki,J.: "Simulation of Regulated Exocytosis of Amylase from Salivary Parotid Acinar Cells by a Consecutive Reaction Model Comprising Two Sequential First-order Reactions."J.Theor.Biol.. 204. 165-177 (2000)

[Publications] Segawa A: "Cell biology of human salivary secretion."Eur J Mophology. 38. 237-241 (2000)

[Publications] Riva A: "A high resolution SEM study of human minor salivary glands."Eur J Mophology. 38. 219-226 (2000)

[Publications] Ambu R: "Scanning electron microscopy of the interior of cells in Hurthle cell tumors."Ultrastructural Pathology. 24. 211-219 (2000)

[Publications] Curry MR: "Single water channels of Aquaporin-1 do not obey the Kedem-Katchalsky equations"J.Membrane Biol. (In press). (2001)

[Publications] Gerelsaikhan,T.: "Transmembrane topology of the secretory Na-K-2Cl cotransporter studied by in vitro translation."J Biol Chem. 275. 40471-40477 (2000)

[Publications] Moore-Hoon,M.L: "Molecular characterization of the cation-chloride cotransporter family."Eur.J.Morphology. 36. S137-141 (1998)

[Publications] Evans,R.L: "New insights into the upregulation and function of the salivary Na^+-K^+-2Cl^- cotransporter."Eur.J.Morphology. 36. S142-146 (1998)

[Publications] Moore-Hoon,M.L: "Increased expression of the secretory Na^+-K^+-2Cl^- cotransporter with differentiation of a human intestinal cell line."Biochem.Biophys.Res.Comm.. 244. 15-19 (1998)

[Publications] Evans,R.L: "Evidence for a physiological role of NH_4^+ transport on the secretory Na^+-K^+-2Cl^- cotransporter."Biochem.Biophys.Res.Comm.. 245. 301-306 (1998)

[Publications] Moore-Hoon,M.L: "Molecular and topological characterization of the rat parotid Na^+-K^+-2Cl^- cotransporter."Biochim.Biophys.Acta. 1373. 261-269 (1998)

[Publications] Turner,R.J.: "Ion transport and signalling in human labial glands."Arch.Oral Biol.. 44. S15-19 (1999)

[Publications] Kurihara,K: "Characterization of the phosphorylation event resulting in upregulation of the salivary Na+-K+-2Cl-cotransporter."Biochemistry. 39. 3718-3724 (2000)

[Publications] Evans,R.L.: "Severe impairment of salivation in Na^+/K^+/2Cl^- cotransporter (NKCC1)-deficient mice."J.Biol.Chem.. 275. 26720-26726 (2000)

[Publications] Gerelsaikhan,T.: "Membrane topology and function of the secretory Na^+-K^+-2Cl^- cotransporter (NKCC1)."J.Kor.Acad.Med.. 15. S3-4 (2000)

[Publications] Tanimura,A.: "Evidence that type I, II and III inositol 1,4,5-trisphosphate receptors can occur as integral plasma membrane proteins."J.Biol.Chem.. 275. 27488-27493 (2000)

[Publications] Shimono,M.: "Connexins in salivary glands"European J Morphology. 38. 257-261 (2000)

[Publications] Hashimoto,S: "Tight junctions in the rat parotid gland"European J Morphology. 38. 263-267 (2000)

[Publications] Shimono,M: "Connexins in the developing salivary glands"European J Morphology. 36. S112-117 (2000)

[Publications] Benes C: "Alterations in PKCδ phosphorlation and activity in rat parotid acinar cells by receptor-dependent and receptor-independent stimuli."Am J Physiology. (In press). (2001)

[Publications] Whittembury G: "The Kidney : Physiology & Pathophysiology.Eds : Seldin SG & Giebisch G.Philadelphia, Lippincot, Williams & Wilkins"Coupled transport of water and solutes across epithelia. (2000)

[Publications] Hill AE: "Int.Revs,Cytology."Paracellular fluid transport by epithelia.(In press). (2001)

村上政隆岡崎国立共同研究機構, 生理学研究所, 助教授 (10104275)