Document Detail

Glucose metabolism in freshly isolated Müller glial cells from a mammalian retina.
MedLine Citation:
PMID:  1377718     Owner:  NLM     Status:  MEDLINE    
Glucose metabolism was studied in isolated retinal Müller glial cells from the juvenile guinea pig. Cells, once enzymatically isolated and purified, were identified by morphological criteria, positive vimentin immunoreactivity, and histochemical staining for glycogen. Purified suspensions of Müller cells were obtained in quantities sufficient for biochemical analysis (approximately 2 x 10(5)/pair of retinas) and light microscopic autoradiography. In bicarbonate-buffered Ringer's medium containing 3H-2-deoxyglucose and no glucose, greater than or equal to 80% of the glucose analogue taken up intracellularly by Müller cells was phosphorylated to 3H-2-deoxyglucose-6-phosphate. In autoradiographs, this non-metabolized product provided visual evidence of glucose phosphorylation: the distribution of cell grains mirrored the morphology of individual Müller cells in situ. Exposure to the glycolytic inhibitor iodoacetate (500 microM) caused an 85% decrease in adenosine triphosphate (ATP) content; concomitantly, 3H-2-deoxyglucose-6-phosphate decreased by 90% and paralleled a dramatic decrease of cell labelling in autoradiographs, while levels of 3H-2-deoxyglucose did not change. In the continual absence of glucose, glycogen content decreased with time and this decrease was slowed by 36% in the presence of iodoacetate. This indicated that, in control conditions, glycosyl units from glycogen sustain cellular metabolism, and hence 3H-2-deoxyglucose phosphorylation. 3H-2-deoxyglucose-6-phosphate concentration was 43-fold less than that of ATP in the control conditions so that depletion of ATP during iodoacetic acid (IAA)-blocked glycolysis was not due to hexokinase activity. These results demonstrate that this preparation is adequate for quantitative studies of glucose metabolism at the cellular and molecular level in an important metabolic compartment of the mammalian retina.
C L Poitry-Yamate; M Tsacopoulos
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of comparative neurology     Volume:  320     ISSN:  0021-9967     ISO Abbreviation:  J. Comp. Neurol.     Publication Date:  1992 Jun 
Date Detail:
Created Date:  1992-08-06     Completed Date:  1992-08-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0406041     Medline TA:  J Comp Neurol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  257-66     Citation Subset:  IM    
Department of Oto-neuro-ophthalmology, University of Geneva Medical School, Switzerland.
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MeSH Terms
Adenosine Triphosphate / metabolism
Deoxyglucose / metabolism
Fluorescent Antibody Technique
Glucose / metabolism*
Glycogen / metabolism
Guinea Pigs
Indicators and Reagents
Iodoacetates / pharmacology
Neuroglia / metabolism*
Retina / cytology,  metabolism*
Staining and Labeling
Reg. No./Substance:
0/Indicators and Reagents; 0/Iodoacetates; 154-17-6/Deoxyglucose; 50-99-7/Glucose; 56-65-5/Adenosine Triphosphate; 9005-79-2/Glycogen

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