Ultrastructural studies on the intracellular trafficking of membranes and HCl-secreting proteins in gastric parietal cells by high-pressure freezing.

• Project/Area Number: 23590238
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General anatomy (including Histology/Embryology)
• Research Institution: University of Miyazaki
• Principal Investigator: SAWAGUCHI Akira 宮崎大学, 医学部, 教授 (30336292)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2011: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 高圧凍結技法 / 胃粘膜 / 胃酸分泌 / 壁細胞 / 超微形態 / 細胞膜動態 / 免疫組織化学 / 蛋白局在 / TRIM50蛋白

Research Abstract

We investigated the intracellular trafficking of membranes and HCl-screting proteins in rat gastric parietal cells by taking advantages of the ex vivo experimental model, named rat isolated gastric mucosa, and the high-pressure freezing followed by freeze substitution cryo-techniques.
As results, we obtained a new insight concerning the intracellular trafficking of membrane fusion proteins, such as syntaxin3 and Hip1r, and ion-transporter channel proteins, such as KCNQ1, between the resting state and the active acid-secreting state of the parietal cell by the application of immuno-histochemistry. In addition, we revealed a unique change in functional morphology in the gastric parietal cells of TRIM50 knock-out mice at the electron microscopical level.

Research Products

• [Journal Article] 樹脂包埋補助ツールを用いたピンポイント電顕試料作製法 (2013)
  • Author(s): 髙橋伸育、豊嶋典世、澤口朗
  • Journal Title: 顕微鏡 Volume: 48(2) Pages: 113-117
• [Journal Article] 超微形態と生体物質の保存に優れた加圧凍結技法 (2013)
  • Author(s): 澤口朗
  • Journal Title: Plant Morphology Volume: 25 Pages: 7-10
  • NAID: 130004691678
• [Journal Article] TRIM50 regulates vesicular trafficking for acid secretion in gastric parietal cells (2012)
  • Author(s): Nishi, M., Aoyama, F., Kisa, F., Zhu, H., Sun, M., Lin, P., Ohta, H., Van, B., Yamamoto, S., Kakizawa, S., Sakai, H., Ma, J., Sawaguchi, A., Takeshima, H
  • Journal Title: J of Biol Chem Volume: 287(40) Pages: 33523-32
  • Peer Reviewed
• [Journal Article] TRIM50 regulates vesicular trafficking for acid secretion in gastric parietal cells. (2012)
  • Author(s): Nishi, M., Aoyama, F., Kisa, F., Zhu, H., Sun, M., Lin, P., Ohta, H., Van, B., Yamamoto, S., Kakizawa, S., Sakai, H., Ma, J., Sawaguchi, A., Takeshima, H.
  • Journal Title: Journal of Biological Chemistry Volume: 287 Pages: 33523-33532
  • Peer Reviewed
• [Journal Article] Functional transformation of gastric parietal cells and intracellular trafficking of ion channels/transporters in the apical canalicular membrane associated with acid secretion (2011)
  • Author(s): Aoyama, F., Sawaguchi, A
  • Journal Title: Biol Pharm Bull Volume: 34 Pages: 813-816
  • NAID: 130000738054
• [Journal Article] Functional transformation of gastric parietal cells and intracellular trafficking of ion channels/transporters in the apical canalicular membrane associated with acid secretion. (2011)
  • Author(s): Aoyama, F., Sawaguchi, A.
  • Journal Title: Biol Pharm Bull Volume: 34 Pages: 813-816
  • NAID: 130000738054
• [Presentation] 胃酸分泌後壁細胞のプロトンポンプ回収機構におけるオートファジーの関与 (2013)
  • Author(s): 澤口朗, 青山典世, 高橋伸育
  • Organizer: 第45回日本臨床分子形態学会総会・学術集会
  • Place of Presentation: 福岡
  • Year and Date: 2013-09-14
• [Presentation] 胃酸分泌後壁細胞のプロトンポンプ回収機構におけるオートファジーの関与 (2013)
  • Author(s): 澤口 朗, 青山典世, 高橋伸育
  • Organizer: 第45回臨床分子形態学会総会・学術集会
  • Place of Presentation: 福岡県福岡市
• [Presentation] ラット単離胃粘膜モデルで見い出されたオートファジーを介する胃小窩壁細胞剥離現象の凍結技法を応用した機能形態解析 (2012)
  • Author(s): 豊嶋典世、髙橋伸育、井手惣幸、澤口朗
  • Organizer: 第68回日本解剖学会九州支部学術集会
  • Place of Presentation: 久留米
  • Year and Date: 2012-10-13
• [Presentation] 加圧凍結技法の基礎と応用の歴史 (2012)
  • Author(s): 澤口朗
  • Organizer: 日本植物学会第76回大会
  • Place of Presentation: 姫路
  • Year and Date: 2012-09-15
• [Presentation] Autophagic degeneration and subsequent exfoliation of gastric pit-parietal cells into the gastric lumen associated with a stimulation of acid-secretion : an in vitro study observed in isolated rat gastric mucosa model (2012)
  • Author(s): Aoyama, F., Takahashi, N., Ide, S., Sawaguchi, A
  • Organizer: 14th International Congress of Histochemistry and Cytochemistry
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-08-26
• [Presentation] 加圧凍結技法が広げる次世代電顕解析の可能性 (2012)
  • Author(s): 澤口朗、豊嶋典世、髙橋伸育
  • Organizer: 日本顕微鏡学会第68回学術講演会
  • Place of Presentation: つくば
  • Year and Date: 2012-05-14
• [Presentation] 凍結技法と単離胃粘膜モデルを応用した胃酸分泌関連疾患の病態解析 (2012)
  • Author(s): 豊嶋典世、髙橋伸育、井手惣幸、澤口朗
  • Organizer: 第117回日本解剖学会全国学術集会
  • Place of Presentation: 山梨
  • Year and Date: 2012-03-27
• [Presentation] A secretagouge-induced microtubule dynamics facilitate HK-ATPase vesicle transport in gastric parietal cell. (2012)
  • Author(s): Natarajan, P., Sawaguchi, A., Okamoto, T., Forte, J.G.
  • Organizer: The American Society for Cell Biology 52th Annual Meeting
  • Place of Presentation: San Francisco, USA
• [Presentation] 高圧凍結技法を応用した胃酸分泌関連蛋白の局在解析 (2011)
  • Author(s): 澤口朗、豊嶋典世、高橋伸育
  • Organizer: 第52回日本組織細胞化学会総会・学術集会
  • Place of Presentation: 金沢
  • Year and Date: 2011-09-24
• [Presentation] 高圧凍結技法を応用した胃酸分泌関連蛋白の局在解析 (2011)
  • Author(s): 澤口 朗、豊嶋典世、高橋伸育
  • Organizer: 第52回日本組織細胞化学会総会・学術集会
  • Place of Presentation: 石川県金沢市