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Trypanocidal action of bisphosphonium salts through a mitochondrial target
A study headed by Abdulsalam A.M. Alkhaldi of the University of Glasgow delineates the trypanocidal action of bisphosphonium salts through a mitochondrial target in bloodstream-form (BF) Trypanosoma brucei (T. brucei). After being informed that lipophilic bisphosphonium salts exhibited antiprotozoal activity by targeting mitochondrial succinate dehydrogenase complexes (SDH) in the closely related Leishmania genus, Alkhaldi's team tested whether a similar phenomenon would occur in T. brucei.
Alkhaldi's team narrowed their area of focus to the mitochondria following observations that bisphosphonium compounds CD38 and AHI-9 inhibited trypanosomes from replicating mitochondrial kinetoplast DNA and/or starting kinetoplast division. In BF T. brucei treated with CD38 and AHI-9, three factors were noted for evaluation of changes in proper mitochondrial function. A dose-dependent reduction of mitochondrial membrane potential and ATP levels, accompanied by a rise in intracellular calcium consistent with mitochondrial depolarization were recognized via fluorescence flow cytometry using the rhodamine indicator TMRE, an ATP bioluminescence assay kit, and fluorescent microplate reading using a Screen Quest™ Fluo-8 Calcium Kit. Eventually the researchers pinpointed the source of mitochondrial impairment to be from bisphosphonium inhibition of F0F1 ATPase instead of SDH as previously suspected.
In this project, Alkhadhi's team was able to confirm mitochondrial dysfunction in BF T. brucei by detecting the activity of fluorophores such as calcium-sensitive Fluo-8. Fluo-8, the active dye of the Screen Quest™ Fluo-8 Calcium Kit, is a novel fluorescent indicator optimized for unparalleled brilliance and sensitivity in its signal and robust cell loading capability. The Screen Quest™ Fluo-8 Kit provides all necessary reagents and protocols for easy application of Fluo-8 to any experiment requiring accurate quantification or tracking of intracellular calcium concentration or response.
Alkhaldi's team narrowed their area of focus to the mitochondria following observations that bisphosphonium compounds CD38 and AHI-9 inhibited trypanosomes from replicating mitochondrial kinetoplast DNA and/or starting kinetoplast division. In BF T. brucei treated with CD38 and AHI-9, three factors were noted for evaluation of changes in proper mitochondrial function. A dose-dependent reduction of mitochondrial membrane potential and ATP levels, accompanied by a rise in intracellular calcium consistent with mitochondrial depolarization were recognized via fluorescence flow cytometry using the rhodamine indicator TMRE, an ATP bioluminescence assay kit, and fluorescent microplate reading using a Screen Quest™ Fluo-8 Calcium Kit. Eventually the researchers pinpointed the source of mitochondrial impairment to be from bisphosphonium inhibition of F0F1 ATPase instead of SDH as previously suspected.
In this project, Alkhadhi's team was able to confirm mitochondrial dysfunction in BF T. brucei by detecting the activity of fluorophores such as calcium-sensitive Fluo-8. Fluo-8, the active dye of the Screen Quest™ Fluo-8 Calcium Kit, is a novel fluorescent indicator optimized for unparalleled brilliance and sensitivity in its signal and robust cell loading capability. The Screen Quest™ Fluo-8 Kit provides all necessary reagents and protocols for easy application of Fluo-8 to any experiment requiring accurate quantification or tracking of intracellular calcium concentration or response.
References
- Abdulsalam A.M. Alkhaldi, Jan Martinek , Brian Panicucci, Christophe Dardonville, Alena Zíkova, Harry P. de Koning. Trypanocidal action of bisphosphonium salts through a mitochondrial target in bloodstream form Trypanosoma brucei. International Journal for Parasitology: Drugs and Drug Resistance 6 (2016) 23e34.
- Screen Quest™ Fluo-8 Medium Removal Calcium Assay Kit. AAT Bioquest, n.d. Web. 14 July 2016
Original created on December 22, 2016, last updated on December 22, 2016
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