January 1997
Volume 38, Issue 1
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Articles  |   January 1997
Glucose dependence of glycolysis, hexose monophosphate shunt activity, energy status, and the polyol pathway in retinas isolated from normal (nondiabetic) rats.
Author Affiliations
  • B S Winkler
    Eye Research Institute, Oakland University, Rochester, MI 48309-4401, USA.
  • M J Arnold
    Eye Research Institute, Oakland University, Rochester, MI 48309-4401, USA.
  • M A Brassell
    Eye Research Institute, Oakland University, Rochester, MI 48309-4401, USA.
  • D R Sliter
    Eye Research Institute, Oakland University, Rochester, MI 48309-4401, USA.
Investigative Ophthalmology & Visual Science January 1997, Vol.38, 62-71. doi:
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      B S Winkler, M J Arnold, M A Brassell, D R Sliter; Glucose dependence of glycolysis, hexose monophosphate shunt activity, energy status, and the polyol pathway in retinas isolated from normal (nondiabetic) rats.. Invest. Ophthalmol. Vis. Sci. 1997;38(1):62-71.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: To measure glucose-dependent metabolic activities and selected parameters of the polyol pathway in retinas isolated from normal rats to test the hypothesis recently proposed by Van den Enden et al that incubation of whole retinas for 2 hours with elevated concentrations of glucose results in activation of the polyol pathway, which is the cause of a redox imbalance, as measured by an increase in the retinal cytosolic lactate-pyruvate ratio and a diabetic-like state. METHODS: Retinas obtained from nondiabetic rats and separated from other ocular tissues were incubated for several hours in incubation medium containing glucose at concentrations ranging from 5 to 30 mM. Measurements were made under aerobic and anaerobic conditions of lactic acid production, retinal adenosine triphosphate (ATP), lactic acid content, the hexose monophosphate shunt pathway, aldose reductase activity, and levels of sorbitol and galactitol. Morphology was examined by light microscopy at the end of the incubations. RESULTS: Incubation of isolated rat retinas with 20 mM glucose increased lactic acid production by approximately 25% in comparison to the rate observed in 5mM glucose under aerobic and anaerobic conditions. The content of ATP and lactate in the retinas after a 2-hour incubation in the presence of oxygen and 20 mM glucose was equal to the amounts found in fresh tissues, whereas these metabolites declined, respectively, by 25% and 45% when 5 mM glucose was used. The activity of the hexose monophosphate shunt pathway in isolated rat retinas was not increased increased significantly when the concentration of glucose was raised from 5 to 30 mM. Aldose reductase activity and polyols were below our limits of detection, 0.5 nmol/minute.mg protein and 3.5 nmol/retina, respectively, under all conditions tested. The morphologic appearance of the retina was similar in the presence of normal and high concentrations of glucose. CONCLUSIONS: These results show that incubation of isolated rat retinas, obtained from nondiabetic rats, with elevated concentrations of glucose for 2 hours leads to increases in glycolysis and a higher tissue content of lactic acid and ATP in comparison to values obtained with 5 mM glucose. However, the magnitude of the glucose-dependent increase in the retinal level of lactate in the current study and in that of Van den Enden et al is six to seven times greater than the calculated flux of glucose through the polyol pathway. These results, therefore, do not support the hypothesis of Van den Enden et al. Rather, it is suggested that supranormal concentrations of glucose yield more lactate and ATP in a whole retina because they optimize the supply of this essential nutrient to cells throughout the tissue by overcoming diffusional limitations that result when the retina is separated from its normal choroidal and intraretinal blood supplies.

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