We have characterized the glucose-transport system in soleus muscle from female GLUT4-null mice to determine whether GLUT1, 3 or 5 account for insulin-stimulated glucose-transport activity. Insulin increased 2-deoxyglucose uptake 2.8- and 2.1-fold in soleus muscle from wild-type and GLUT4-null mice, respectively. Cytochalasin B, an inhibitor of GLUT1- and GLUT4-mediated glucose transport, inhibited insulin-stimulated 2-deoxyglucose uptake by > 95% in wild-type and GLUT4-null soleus muscle. Addition of 35 mM fructose to the incubation media was without effect on insulin-stimulated 3-O-methylglucose transport activity in soleus muscle from either genotype, whereas 35 mM glucose inhibited insulin-stimulated (20 nM) 3-O-methylglucose transport by 65% in wild-type and 99% in GLUT4-null mice. We utilized the 2-N-4-1-(1-azi-2,2,2-t r i f l u o r o e t h y l ) b e n z o y l - 1, 3 - b i s (D - m a n n o s e - 4 - y l o x y ) - 2 - p ro p y lamine (ATB-BMPA) exofacial photolabel to determine if increased cell-surface GLUT1 or GLUT4 content accounted for insulin-stimulated glucose transport in GLUT4-null muscle. In wild-type soleus muscle, cell-surface GLUT4 content was increased by 2.8-fold under insulin-stimulated conditions and this increase corresponded to the increase in 2-deoxyglucose uptake. No detectable cell-surface GLUT4 was observed in soleus muscle from female GLUT4-null mice under either basal or insulin-stimulated conditions. Basal cell-surface GLUT1 content was similar between wild-type and GLUT4-null mice, with no further increase noted in either genotype with insulin exposure. Neither GLUT3 nor GLUT5 appeared to account for insulin-stimulated glucose-transport activity in wild-type or GLUT4-null muscle. In conclusion, insulin-stimulated glucose-transport activity in female GLUT4-null soleus muscle is mediated by a facilitative transport process that is glucose- and cytochalasin B-inhibitable, but which is not labelled strongly by ATB-BMPA.
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September 1999
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Research Article|
August 24 1999
In vitro analysis of the glucose-transport system in GLUT4-null skeletal muscle
Jeffrey W. RYDER;
Jeffrey W. RYDER
*Department of Clinical Physiology, Karolinska Hospital, S-171 76, Stockholm, Sweden
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Yuichi KAWANO;
Yuichi KAWANO
*Department of Clinical Physiology, Karolinska Hospital, S-171 76, Stockholm, Sweden
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Alexander V. CHIBALIN;
Alexander V. CHIBALIN
*Department of Clinical Physiology, Karolinska Hospital, S-171 76, Stockholm, Sweden
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Jorge RINCÓN;
Jorge RINCÓN
*Department of Clinical Physiology, Karolinska Hospital, S-171 76, Stockholm, Sweden
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Tsu-Shuen TSAO;
Tsu-Shuen TSAO
†Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, 10461-1602, U.S.A.
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Antine E. STENBIT;
Antine E. STENBIT
†Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, 10461-1602, U.S.A.
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Terry COMBATSIARIS;
Terry COMBATSIARIS
†Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, 10461-1602, U.S.A.
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Jing YANG;
Jing YANG
‡Department of Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, U.K.
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Geoffrey D. HOLMAN;
Geoffrey D. HOLMAN
‡Department of Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, U.K.
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Maureen J. CHARRON;
Maureen J. CHARRON
†Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, 10461-1602, U.S.A.
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Juleen R. ZIERATH
Juleen R. ZIERATH
1
*Department of Clinical Physiology, Karolinska Hospital, S-171 76, Stockholm, Sweden
1To whom correspondence should be addressed (jrz@klinfys.ks.se).
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Publisher: Portland Press Ltd
Received:
May 20 1999
Accepted:
June 21 1999
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1999
1999
Biochem J (1999) 342 (2): 321–328.
Article history
Received:
May 20 1999
Accepted:
June 21 1999
Citation
Jeffrey W. RYDER, Yuichi KAWANO, Alexander V. CHIBALIN, Jorge RINCÓN, Tsu-Shuen TSAO, Antine E. STENBIT, Terry COMBATSIARIS, Jing YANG, Geoffrey D. HOLMAN, Maureen J. CHARRON, Juleen R. ZIERATH; In vitro analysis of the glucose-transport system in GLUT4-null skeletal muscle. Biochem J 1 September 1999; 342 (2): 321–328. doi: https://doi.org/10.1042/bj3420321
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