Analysis of polarized transport mechanisms in salivary duct cells.

唾液管细胞极化运输机制的分析。

• 批准号: 23792129
• 负责人: SHITARA Akiko
• 金额: $ 2.66万
• 依托单位: Health Sciences University of Hokkaido
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23792129/
• 关键词: 極性輸送; 唾液腺; イメージング; アデノウイルスベクター; Halo tag; 小胞輸送

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.

Na ~+/K ~+ ATP酶是唾液腺转运至基底侧膜的标志蛋白之一。我们构建了表达Na+/K+ ATP酶的融合Halo标签的质粒载体，并将其转染到HeLa细胞中，利用封闭配体和荧光配体分析了新合成的Halo标签与Na+/K+ ATP酶融合后的转运动力学，发现大部分Na+/K+ ATP酶不通过内体转运。我们制备了表达Halo-tag Na ~+/K ~+ ATP酶的腺病毒，并将其转染到下颌下腺中，通过免疫染色证实分子在下颌下腺的基底外侧膜上表达。消化酶消化法分散转染的下颌下腺，荧光配体染色可见活细胞质膜特异性染色。

项目成果

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

使用共价标签分析唾液腺细胞中的囊泡运输

• 发表时间: 2012
• 作者: 設楽彰子;荒川俊哉;田隈泰信
• 通讯作者: 田隈泰信

ゴルジ体のリボン構造形成におけるVAMP4の役割

VAMP4在高尔基带结构形成中的作用

• 发表时间: 2011
• 作者: 設楽彰子;荒川俊哉;田隈泰信;Arakawa T,Okayama M,Mizoguchi I,Shitara A,Takuma T;Shitara A,Okayama M,Arakawa T,Mizoguchi T,Takuma T;Takuma T,Shitara A,Okayama M,Arakawa T,Mizoguchi T;Shitara A,Arakawa T,Okayama M,Mizoguchi T,Takuma T
• 通讯作者: Shitara A,Arakawa T,Okayama M,Mizoguchi T,Takuma T

Expression of functional Stim1-mKO1 in rat submandibular acinar cells by retrograde ductal injection of an adenoviral vector

• DOI: 10.1016/j.archoralbio.2011.06.001
• 发表时间: 2011-11-01
• 期刊: ARCHIVES OF ORAL BIOLOGY
• 影响因子: 3
• 作者: Morita, Takao;Tanimura, Akihiko;Tojyo, Yosuke
• 通讯作者: Tojyo, Yosuke

Expression of functional Stim1 in rat submandibular acinar cells by retrograde ductal injection of adenovirus

逆行导管注射腺病毒在大鼠颌下腺泡细胞中表达功能性Stim1

• 发表时间: 2011
• 作者: 設楽彰子;荒川俊哉;田隈泰信;Arakawa T,Okayama M,Mizoguchi I,Shitara A,Takuma T;Shitara A,Okayama M,Arakawa T,Mizoguchi T,Takuma T;Takuma T,Shitara A,Okayama M,Arakawa T,Mizoguchi T;Shitara A,Arakawa T,Okayama M,Mizoguchi T,Takuma T;Morita T,Tanimura A,Shitara A,Suzuki Y,Nezu A,Takuma T,Tojyo Y
• 通讯作者: Morita T,Tanimura A,Shitara A,Suzuki Y,Nezu A,Takuma T,Tojyo Y

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

机械应力诱导人牙周膜细胞的基因表达

• 发表时间: 2011
• 作者: 設楽彰子;荒川俊哉;田隈泰信;Arakawa T,Okayama M,Mizoguchi I,Shitara A,Takuma T
• 通讯作者: Arakawa T,Okayama M,Mizoguchi I,Shitara A,Takuma T