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BMC veterinary research2007; 3; 25; doi: 10.1186/1746-6148-3-25

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

Abstract: 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. Results: 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. Conclusions: 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.
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Publication Date: 2007-09-28 PubMed ID: 17903245PubMed Central: PMC2045081DOI: 10.1186/1746-6148-3-25Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research presents a standardized flow cytometric procedure to analyze mitochondrial function in horse peripheral blood mononuclear cells using a dye known as JC-1. The researchers investigate various aspects like the effects of time passed between blood sampling and laboratory processing, and the influence of age and sex on the results. They suggest this test could assist in the study and prevention of horse diseases attributed to mitochondrial dysfunction.

Research Methodology and Key Findings

This research involved developing and testing a standardized flow cytometric procedure to analyze mitochondrial function in the peripheral blood mononuclear cells of horses. The key aspects of the study include:

  • Use of the cationic dye JC-1: The researchers used a specific dye that accumulates in the mitochondria depending on its potential, changing its fluorescence from green to orange. This enables the optical measurement of mitochondrial membrane potential through the orange/green fluorescence intensity ratio.
  • Technical considerations: The study aimed to critically appraise crucial questions relating to technical aspects of the procedure, such as its reproducibility, the impact of time elapsed between drawing samples and processing in the lab, and the establishment of reference values.
  • Stable results: The findings showed that the orange and green fluorescence values associated with JC-1, along with their ratio, remained stable over time. They were not influenced by the age or sex of the horse and were highly sensitive to intoxication with a mitochondrial potential dissipator.
  • Time restrictions for samples: The study notably found that the values remained stable, provided that no more than five hours elapsed between taking the blood sample and processing it in the lab. This gave a specific time window within which samples must be tested using this procedure.

Conclusion and Implications

Based on the study’s findings, the researchers suggested this flow cytometric procedure as a reliable method to quantitatively measure mitochondrial function in horses. They provided specific instructions for sample collection and handling, proposing that blood samples should be kept at room temperature in sodium citrate tubes. Importantly, they must be processed within a maximum of five hours after collection.

The development of this test could potentially aid in the confirmation and study of equine myopathies currently associated with mitochondrial dysfunction. Furthermore, it could also play a key role in the prevention of these conditions, leading to better health outcomes for horses.

Cite This Article

APA
Cassart D, Fett T, Sarlet M, Baise E, Coignoul F, Desmecht D. (2007). Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells. BMC Vet Res, 3, 25. https://doi.org/10.1186/1746-6148-3-25

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 3
Pages: 25

Researcher Affiliations

Cassart, Dominique
  • Department of Pathology, Faculty of Veterinary Medicine, University of Liege, Sart Tilman B43, B-4000 Liège, Belgium. Dominique.Cassart@ulg.ac.be
Fett, Thomas
    Sarlet, Michaël
      Baise, Etienne
        Coignoul, Freddy
          Desmecht, Daniel

            MeSH Terms

            • Animals
            • Benzimidazoles / chemistry
            • Carbocyanines / chemistry
            • Female
            • Flow Cytometry / methods
            • Flow Cytometry / veterinary
            • Fluorescent Dyes / chemistry
            • Horses / blood
            • Horses / physiology
            • Leukocytes, Mononuclear / physiology
            • Male
            • Membrane Potential, Mitochondrial / physiology
            • Mitochondria / physiology
            • Reference Values
            • Reproducibility of Results

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            Citations

            This article has been cited 3 times.
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