Abstract
BackgroundThe 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.ResultsThe 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.ConclusionWhenever 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.
Highlights
The morphopathological picture of a subset of equine myopathies is compatible with a primary mitochondrial disease, but functional confirmation in vivo is still pending
Epidemiological studies aimed at delineating the precise environmental conditions that increase the risk of developing atypical myopathy would gain from a standardized laboratory test that lends itself to mass screening
A standardized analytic procedure is proposed along with a critical appraisal of the crucial questions of reproducibility, effect of time elapsed between blood sampling and laboratory processing and reference values
Summary
The morphopathological picture of a subset of equine myopathies is compatible with a primary mitochondrial disease, but functional confirmation in vivo is still pending. Equine atypical myopathy affects horses and ponies kept on pasture and not exercised either prior to or at the time of the first clinical signs. The morphopathological picture of the disease suggested a mitochondrial disorder the characteristics of which are compatible with those of equine toxic myopathies due to plant, bacterial or fungal toxins [4,5,6,7,8,9]. Morphologic clues of mitochondrial dysfunction in these equine diseases have accumulated, a functional confirmation is still pending. Epidemiological studies aimed at delineating the precise environmental conditions that increase the risk of developing atypical myopathy would gain from a standardized laboratory test that lends itself to mass screening. Reliable methods for the sensitive determination of a possible alteration of mitochondrial function in equine tissues are desirable
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