β-Amyloid-mediated inhibition of redox activity (MTT reduction) is not an indicator of astroglial degeneration

Abstract

The reduction in 3-[4,5-dimethylthiazol]-2,5-diphenyltetrazolium bromide (MTT) to a coloured formazan compound by cultured cells has been extensively used as an in vitro model for understanding neurobiological mechanisms involved in amyloid beta-protein (A beta-mediated cell death. In primary cultures of astrocytes, very low concentrations of aggregated A beta 1-4C, but not A beta 4C-1, produced a significant inhibition in the reduction of the dye MTT. This inhibitory response was rapid and persisted as long as A beta 1-4C was present in the culture medium. Such a severe reduction in cell redox activity for days failed to cause death of astroglial cells, measured in terms of trypan blue uptake and lactate dehydrogenase release. Interleukin-1 beta (IL-1 beta), which is known to attenuate excitotoxic neurodegeneration, had no effect on A beta 1-4C-induced inhibition of MTT reduction. These results suggest that even though inhibition of MTT reduction represents an early indicator of the A beta 1-4C mediated cell injury, without other corroborating evidence, it should not be used as a measure of cell death.