Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells

• Published in: BMC veterinary research Vol. 3; no. 1; p. 25
• Main Authors: Cassart, Dominique, Fett, Thomas, Sarlet, Michaël, Baise, Etienne, Coignoul, Freddy, Desmecht, Daniel
• Format: Journal Article Web Resource
• Language: English
• Published: England BioMed Central Ltd 28-09-2007; BioMed Central; BMC
• Subjects: Analysis and chemistry; Animals; Benzimidazoles - chemistry; Blood; Carbocyanines - chemistry; Creatine kinase; Development and progression; Diagnosis; Female; Flow Cytometry - methods; Flow Cytometry - veterinary; Fluorescent Dyes - chemistry; Health aspects; Horses - blood; Horses - physiology; Leukocytes, Mononuclear - physiology; Life sciences; Male; Membrane Potential, Mitochondrial - physiology; Mitochondria; Mitochondria - physiology; Muscle diseases; Médecine vétérinaire & santé animale; Physiological aspects; Properties; Reference Values; Reproducibility of Results; Risk factors; Sciences du vivant; Veterinary medicine & animal health; Belgium
• ISSN: 1746-6148; 1746-6148
• DOI: 10.1186/1746-6148-3-25

The morphopathological picture of a subset of equine myopathies is compatible with a primary mitochondrial disease, but functional confirmation in vivo is still pending. The cationic dye JC-1 exhibits potential-dependent accumulation in mitochondria that is detectable by a fluorescence shift from green to orange. As a consequence, mitochondrial membrane potential can be optically measured by the orange/green fluorescence intensity ratio. A flow cytometric standardized analytic procedure of the mitochondrial function of equine peripheral blood mononuclear cells is proposed along with a critical appraisal of the crucial questions of technical aspects, reproducibility, effect of time elapsed between blood sampling and laboratory processing and reference values.
The JC-1-associated fluorescence orange and green values and their ratio were proved to be stable over time, independent of age and sex and hypersensitive to intoxication with a mitochondrial potential dissipator. Unless time elapsed between blood sampling and laboratory processing does not exceed 5 hours, the values retrieved remain stable. Reference values for clinically normal horses are given.
Whenever a quantitative measurement of mitochondrial function in a horse is desired, blood samples should be taken in sodium citrate tubes and kept at room temperature for a maximum of 5 hours before the laboratory procedure detailed here is started. The hope is that this new test may help in confirming, studying and preventing equine myopathies that are currently imputed to mitochondrial dysfunction.