Comparative effects of the herbicide dinitro-o-cresol on mitochondrial bioenergetics

The herbicide dinitro-o-cresol (DNOC) was evaluated for its effects on bioenergetic activities of potato tuber mitochondria to elucidate its mechanism of action and to compare its toxicological properties with those of the chemically related uncoupler dinitrophenol (DNP). DNOC acts as a typical uncoupler, similarly to the classical uncouplers DNP and carbonylcyanide p-trifluoromethoxyphenyl-hydrazone (FCCP). Low concentrations of DNOC (< 100 micromolar) maximally stimulate succinate-supported respiration of plant mitochondria, with simultaneous collapse of trans-membrane electrical potential, more efficiently than DNP. The herbicide makes the plant mitochondrial membrane more permeable to protons, acting as a protonophore even in non-energized mitochondria. High concentrations of DNOC (> 100 micromolar) act also more efficiently than DNP simultaneously as a protonophore and inhibitor of respiration, especially when respiration is supported by substrates that are transported to the matrix. The efficiency of DNOC is decreased with increase of mitochondrial protein, BSA and exogenous orthophosphate. Although similar effects were observed for animal and plant mitochondria, rat-liver mitochondrial respiration was more sensitive to DNOC then plant mitochondria. Furthermore, in the presence of DNOC, liver mitochondria exhibited a higher state 3 respiratory coupling level than potato tuber mitochondria, as a result of a considerable stimulation (60%) of state 3 respiration. In conclusion, DNOC is a more potent mitochondrial uncoupler and respiratory chain inhibitor than DNP, although their chemical structures are very similar. Apparently, the additional methyl group of DNOC increases its efficiency as an uncoupler and as an inhibitor, as compared to DNP. Plant mitochondria were shown to be as useful as animal mitochondria in evaluating the toxicity of these xenobiotics.