Methods to detect mitochondrial function.
The bioenergetic function of mitochondria can be investigated in intact cells by a variety of methods. A simple biochemical method to compare mitochondrial oxidative phosphorylation with glycolytic ATP synthesis takes advantage of the Pasteur effect, since the amount of lactate produced under basal conditions is an indication of glycolytic ATP, while the Delta-lactate (the difference between excess lactate produced after inhibition of respiration and basal lactate) represents ATP produced anaerobically in order to compensate for decreased oxidative phosphorylation after respiratory chain inhibition. The system has been validated in a series of cells, including human platelets and lymphocytes and lines cultured in vitro. Measurement of KCN-sensitive oxygen consumption by the cells and its sensitivity to uncouplers can be good supplementary indication of mitochondrial phosphorylative capacity.
Merlo-Pich
Milena
M
Dipartimento di Biochimica 'G. Moruzzi', University of Bologna, Via Irnerio 48, Bologna 40126, Italy.
Deleonardi
Giulia
G
Biondi
Annalisa
A
Lenaz
Giorgio
G
D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000368:Aged; D001669:Biochemical Phenomena; D001671:Biochemistry; D001792:Blood Platelets / Q000378:metabolism*; D002478:Cells, Cultured; D006019:Glycolysis; D006801:Humans; D019344:Lactic Acid / Q000378:metabolism; D008214:Lymphocytes / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D010101:Oxygen Consumption / Q000502:physiology; D013053:Spectrophotometry / Q000379:methods
D019344:Lactic Acid; D000255:Adenosine Triphosphate