Document Detail


Implications of glucose transporter protein type 1 (GLUT1)-haplodeficiency in embryonic stem cells for their survival in response to hypoxic stress.
MedLine Citation:
PMID:  14578187     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Glucose transporter protein type 1 (GLUT1) is a major glucose transporter of the fertilized egg and preimplantation embryo. Haploinsufficiency for GLUT1 causes the GLUT1 deficiency syndrome in humans, however the embryo appears unaffected. Therefore, here we produced heterozygous GLUT1 knockout murine embryonic stem cells (GT1+/-) to study the role of GLUT1 deficiency in their growth, glucose metabolism, and survival in response to hypoxic stress. GT1(-/-) cells were determined to be nonviable. Both the GLUT1 and GLUT3 high-affinity, facilitative glucose transporters were expressed in GT1(+/+) and GT1(+/-) embryonic stem cells. GT1(+/-) demonstrated 49 +/- 4% reduction of GLUT1 mRNA. This induced a posttranscriptional, GLUT1 compensatory response resulting in 24 +/- 4% reduction of GLUT1 protein. GLUT3 was unchanged. GLUT8 and GLUT12 were also expressed and unchanged in GT1(+/-). Stimulation of glycolysis by azide inhibition of oxidative phosphorylation was impaired by 44% in GT1(+/-), with impaired up-regulation of GLUT1 protein. Hypoxia for up to 4 hours led to 201% more apoptosis in GT1(+/-) than in GT1(+/+) controls. Caspase-3 activity was 76% higher in GT1(+/-) versus GT1(+/+) at 2 hours. Heterozygous knockout of GLUT1 led to a partial GLUT1 compensatory response protecting nonstressed cells. However, inhibition of oxidative phosphorylation and hypoxia both exposed their increased susceptibility to these stresses.
Authors:
Charles Heilig; Frank Brosius; Brian Siu; Luis Concepcion; Richard Mortensen; Kathleen Heilig; Min Zhu; Richard Weldon; Guimei Wu; David Conner
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The American journal of pathology     Volume:  163     ISSN:  0002-9440     ISO Abbreviation:  Am. J. Pathol.     Publication Date:  2003 Nov 
Date Detail:
Created Date:  2003-10-27     Completed Date:  2003-12-05     Revised Date:  2013-06-09    
Medline Journal Info:
Nlm Unique ID:  0370502     Medline TA:  Am J Pathol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1873-85     Citation Subset:  AIM; IM    
Affiliation:
Division of Nephrology, Johns Hopkins University School of Medicine, 720 Rutland Avenue, Ross 947, Baltimore, MD 21205, USA. cheilig@jhmi.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis / genetics
Blotting, Northern
Caspase 3
Caspases / metabolism
Cell Hypoxia
Cell Line
Embryo, Mammalian
Enzyme Inhibitors / pharmacology
Glucose Transporter Type 1
Haplotypes
Heterozygote
In Situ Nick-End Labeling
Mice
Mice, Knockout
Monosaccharide Transport Proteins / deficiency*,  genetics
Oxidative Phosphorylation / drug effects
Sodium Azide / pharmacology
Stem Cells / pathology*,  physiology*
Grant Support
ID/Acronym/Agency:
R01 DK 54507/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Enzyme Inhibitors; 0/Glucose Transporter Type 1; 0/Monosaccharide Transport Proteins; 0/Slc2a1 protein, mouse; 26628-22-8/Sodium Azide; EC 3.4.22.-/Casp3 protein, mouse; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases
Comments/Corrections

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