Analysis of polarized transport mechanisms in salivary duct cells.

2013 Fiscal Year Final Research Report

• Project/Area Number: 23792129
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Functional basic dentistry
• Research Institution: Health Sciences University of Hokkaido
• Principal Investigator: SHITARA Akiko 北海道医療大学, 歯学部, 助教 (30508718)
• Project Period (FY): 2011 – 2013
• Keywords: 極性輸送 / 唾液腺 / イメージング

Research Abstract

Na+/K+ ATPase is one of the marker protein which is transported to the basolateral membrane in salivary gland. We made the plasmid vector which express Na+/K+ ATPase fused with Halo tag and transfected it to the HeLa cells.
We analyzed the transport dynamics of newly synthesized Halo tag fusing with Na+/K+ ATPase by using blocking ligand and fluorescence ligand, and we have shown that most of Na+/K+ ATPase are not transported via endosome. We made adeno virus which express Halo-tag Na+/K+ ATPase and transfected it to the submandibular gland, and we confirmed that molecules are expressed on the basolateral membrane of submandibular gland by immunostaining. Transfected submandibular gland was dispersed by digestive enzyme and stained with fluorescent ligands, and we could observe the specific staining of plasma membrane in living cells.

Research Products

• [Journal Article] Gravity loading induces adenosine triphosphate release and phosphorylation of extracellular signal-regulated kinases in human periodontal ligament cells (2014)
  • Author(s): Ito M,Arakawa T,Okayama M,Shitara A,Mizoguchi I,Takuma T
  • Journal Title: J Investig Clin Dent Volume: (印刷中)
  • DOI: 10.1111/jicd.12049
  • Peer Reviewed
• [Journal Article] VAMP4 is required to maintain the ribbon structure of the Golgi apparatus (2013)
  • Author(s): Shitara A,Shibui T,Okayama M,Arakawa T,Mizoguchi I,Sakakura Y,Takuma T
  • Journal Title: Mol Cell Biochem. Volume: 380(1-2) Pages: 11-21
  • DOI: 10.1007/s11010-013-1652-4
  • Peer Reviewed
• [Journal Article] Isoproterenol stimulates transient SNAP23-VAMP2 interaction in rat parotid glands (2013)
  • Author(s): Takuma T,Shitara A,Arakawa T,Okayama M,Mizoguchi I,Tajima Y
  • Journal Title: FEBS Lett. Volume: 587(6) Pages: 583-589
  • DOI: 10.1016/j.febslet.2013.01.039
  • Peer Reviewed
• [Journal Article] SNARE proteins are not excessive for the formation of post-Golgi SNARE complexes in HeLa cells (2012)
  • Author(s): Okayama M,Shitara A,Arakawa T,Tajima Y,Mizoguchi I,Takuma T
  • Journal Title: Mol Cell Biochem. Volume: 366(1-2) Pages: 159-168
  • DOI: 10.1007/s11010-012-1293-z
  • Peer Reviewed
• [Journal Article] Expression of functional Stim1-mKO1 in rat submandibular acinar cells by retrograde ductal injection of an adenoviral vector (2011)
  • Author(s): Morita T,Tanimura A,Shitara A,Suzuki Y,Nezu A,Takuma T,Tojyo Y
  • Journal Title: Arch Oral Biol. Volume: 56(11) Pages: 1356-1365
  • DOI: 10.1016/j.archoralbio.2011.06.001
  • Peer Reviewed
• [Journal Article] Spontaneous oscillations in rat parotid ducts are associated with cell serlling (2011)
  • Author(s): Shitara A,Tojyo Y,Tanimura A
  • Journal Title: Journal of Oral Bioscience Volume: 53(4) Pages: 304-311
  • Peer Reviewed
• [Journal Article] Diversity and Spatio-Temporal Properties of Calcium Responces in Salivary Ducts (2011)
  • Author(s): Tanimura A,Shitara A,Tojyo Y
  • Journal Title: Journal of Oral Bioscience Volume: 53(1) Pages: 48-56
  • Peer Reviewed
• [Presentation] 共有結合性タグを用いた唾液腺細胞における小胞輸送の解析 (2012)
  • Author(s): Shitara A,Arakawa T,Takuma T
  • Organizer: The 53rd Annual Meeting of Japan Association for Oral Biology
  • Place of Presentation: Fukushima
  • Year and Date: 2012-09-14
• [Presentation] Expression of functional Stim1 in rat submandibular acinar cells by retrograde ductal injection of adenovirus (2011)
  • Author(s): Morita T,Tanimura A,Shitara A,Suzuki Y,Nezu A,Takuma T,Tojyo Y
  • Organizer: Gordon Research Conferences Salivary Glands & Exocrine Biology
  • Place of Presentation: Garveston,USA
  • Year and Date: 20110200
• [Presentation] ヒト歯根膜細胞のメカニカルストレスによる遺伝子発現 (2011)
  • Author(s): Arakawa T,Okayama M,Mizoguchi I,Shitara A,Takuma T
  • Organizer: The 53rd Annual Meeting of Japan Association for Oral Biology
  • Place of Presentation: Gifu
  • Year and Date: 2011-10-02
• [Presentation] ゴルジ体のリボン構造形成におけるVAMP4の重要性 (2011)
  • Author(s): Shitara A,Okayama M,Arakawa T,Mizoguchi T,Takuma T
  • Organizer: The 53rd Annual Meeting of Japan Association for Oral Biology
  • Place of Presentation: Gifu
  • Year and Date: 2011-10-01
• [Presentation] ラット顎下腺の調節性開口分泌に関わるSNAE複合体 (2011)
  • Author(s): Takuma T,Shitara A,Okayama M,Arakawa T,Mizoguchi T
  • Organizer: The 53rd Annual Meeting of Japan Association for Oral Biology
  • Place of Presentation: Gifu
  • Year and Date: 2011-10-01
• [Presentation] ゴルジ体のリボン構造形成におけるVAMP4の役割 (2011)
  • Author(s): Shitara A,Arakawa T,Okayama M,Mizoguchi T,Takuma T
  • Organizer: The 84th Annual Meeting of the Japanese Biochemical Society
  • Place of Presentation: Kyoto
  • Year and Date: 2011-09-23