Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells
<p>Abstract</p> <p>Background</p> <p>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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2007-09-01
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| Series: | BMC Veterinary Research |
| Online Access: | http://www.biomedcentral.com/1746-6148/3/25 |
| _version_ | 1818752740189274112 |
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| author | Coignoul Freddy Baise Etienne Sarlet Michaël Fett Thomas Cassart Dominique Desmecht Daniel |
| author_facet | Coignoul Freddy Baise Etienne Sarlet Michaël Fett Thomas Cassart Dominique Desmecht Daniel |
| author_sort | Coignoul Freddy |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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.</p> <p>Conclusion</p> <p>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.</p> |
| first_indexed | 2024-12-18T04:56:16Z |
| format | Article |
| id | doaj.art-0f88019a61fc498ea2407cff839ee6c7 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-12-18T04:56:16Z |
| publishDate | 2007-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-0f88019a61fc498ea2407cff839ee6c72022-12-21T21:20:15ZengBMCBMC Veterinary Research1746-61482007-09-01312510.1186/1746-6148-3-25Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cellsCoignoul FreddyBaise EtienneSarlet MichaëlFett ThomasCassart DominiqueDesmecht Daniel<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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.</p> <p>Conclusion</p> <p>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.</p>http://www.biomedcentral.com/1746-6148/3/25 |
| spellingShingle | Coignoul Freddy Baise Etienne Sarlet Michaël Fett Thomas Cassart Dominique Desmecht Daniel Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells BMC Veterinary Research |
| title | Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells |
| title_full | Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells |
| title_fullStr | Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells |
| title_full_unstemmed | Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells |
| title_short | Flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells |
| title_sort | flow cytometric probing of mitochondrial function in equine peripheral blood mononuclear cells |
| url | http://www.biomedcentral.com/1746-6148/3/25 |
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