Neurochemical Research

, Volume 24, Issue 3, pp 351–358

Distinct Glycolysis Inhibitors Determine Retinal Cell Sensitivity to Glutamate-Mediated Injury

  • Ana Cristina Rego
  • Filipe M. Areias
  • Maria Sancha Santos
  • Catarina R. Oliveira
Article

DOI: 10.1023/A:1020977331372

Cite this article as:
Rego, A.C., Areias, F.M., Santos, M.S. et al. Neurochem Res (1999) 24: 351. doi:10.1023/A:1020977331372

Abstract

In this study, we analyzed how distinct glycolysis inhibitors influenced the redox status of retinal cells, used as a neuronal model. Three different approaches were used to inhibit glycolysis: the cells were submitted to iodoacetic acid (IAA), an inhibitor of glyceraldehyde 3-phosphate dehydrogenase, to 2-deoxy-glucose (DG) in glucose-free medium, which was used as a substitute of glucose, or in the absence of glucose. The redox status of the cells was evaluated by determining the reduction of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide). By the analysis of dose-response curves of MTT reduction, IAA showed values of IC50 = 7.02 × 10−5 M, whereas DG showed values of IC50 = 7.42 × 10−4 M. Upon 30 min-incubation, glucose deprivation, per se, did not significantly affect MTT reduction. We also evaluated the reduction of MTT as an indicator of cell injury by exposing the cells to 100 μM glutamate during the decrement of glycolysis function. In the presence of glutamate, for 2 h, there was a decrease in MTT reduction, which was potentiated in the presence of DG (10-20% decrease), in the presence of IAA (about 30% decrease) or in glucose-free medium (about 30% decrease). Major changes observed by the MTT assay, upon exposure to glutamate, indicative of changes in the redox status of retinal cells, were concomitant with variations in intracellular ATP. Under glucose deprivation, endogenous ATP decreased significantly from 38.9 ± 4.4 to 13.3 ± 0.7 nmol/mg protein after exposure to 100 μM glutamate. The results support a different vulnerability of retinal cells after being exposed to distinct forms of glycolysis inhibition.

ATP production glutamate glycolysis MTT reduction retinal cell injury 

Copyright information

© Plenum Publishing Corporation 1999

Authors and Affiliations

  • Ana Cristina Rego
    • 1
  • Filipe M. Areias
    • 1
  • Maria Sancha Santos
    • 1
  • Catarina R. Oliveira
    • 1
  1. 1.Center for Neurosciences of Coimbra and Faculty of MedicineUniversity of CoimbraCoimbraPortugal