＜Symposium I＞A new imaging method of glucose transport in live cells

フォーマット:

論文

責任表示:

Yamada, Katsuya ; Yamamoto, Toshihiro ; Watanabe, Seiji ; Nishiuchi, Yuji ; Teshima, Tadashi ; Matsuoka, Hideaki ; Suga, Sechiko

言語:

英語

出版情報:

弘前大学大学院医学研究科・弘前医学会, 2010-07-08

著者名:

Yamada, Katsuya

Yamamoto, Toshihiro

Watanabe, Seiji

Nishiuchi, Yuji

Teshima, Tadashi

Matsuoka, Hideaki

Suga, Sechiko

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掲載情報:

弘前医学

ISSN:

0439-1721      

巻:

61

通号:

Supplement

開始ページ:

S23

終了ページ:

S25

バージョン:

publisher

概要:

Glucose transport activity in mammalian cells has been monitored by radiolabeled tracers such as [14C] 2-deoxy-D-glucose. However, due to their limited spatial and temporal resolution, measuring glucose uptake in single, living cells was di … ffi cult. We have developed a fl uorescent D-glucose derivative, 2-[N（- 7-nitrobenz-2-oxa-1,3-diazol-4-yl）amino]-2-deoxy-D-glucose （2-NBDG）, that allows a more sensitive measurement of glucose uptake in live cells. Indeed, the uptake of 2-NBDG takes place through glucose transporters （GLUTs） in a concentration-, time-, and temperature-dependent manner. Kinetic analysis revealed that apparent Km values for the uptake were similar to those reported by D-glucose and the nonmetabolizable glucose analogue, 3-O-methyl-D-glucose, found in pancreatic islet cells. This method can be combined with Ca2+ imaging and subsequent immunocytochemical identifi cation of cells. So far, 2-NBDG has been used for monitoring glucose uptake into a wide variety of mammalian cells including astrocytes and neurons. However, it has been pointed out that 2-NBDG method requires very accurate procedures, since the fl uorescence intensity is an arbitrary measure. In addition, temporal changes in health of cells in question should be verifi ed throughout the experiment by an independent way such as using patch clamp. To overcome the diffi culty, we synthesized transporter- unrecognizable glucose analogues as control for 2-NBDG uptake. These include 2-[N（- 7-Nitrobenz-2-oxa-1,3-diazol-4-yl）amino]-2-deoxy-L-glucose [2-NBDLG], the antipode of 2-NBDG. A combined use of transporter-recognizable（ D-isomer） and unrecognizable（ L-isomer） fl uorescent analogues allows us to investigate not only net stereospecifi c transport of glucose in live cells but also ligand-transporter interactions and non-transportermediated glucose movement. 続きを見る

URL:

http://hdl.handle.net/10129/3666