Probing Mitochondrial Permeability Transition Pore Activity in Nucleated Cells and Platelets by High-Throughput Screening Assays Suggests Involvement of Protein Phosphatase 2B in Mitochondrial Dynamics
| Autor: | Mao Xiang Chen, Matilde Caivano, Sumitra Ramachandran, Emma Ward, Sandy Toh, Mahmood Ahmed, A. Richard Rutter, Kirsten L. Searle, Yannick M. L. Lacroix |
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| Rok vydání: | 2018 |
| Předmět: |
Blood Platelets
animal diseases Cell Mitochondria Liver Mitochondrial Dynamics Mitochondrial Membrane Transport Proteins Tacrolimus Flow cytometry Green fluorescent protein chemistry.chemical_compound Nucleated cell Drug Discovery medicine Animals Humans Cyclophilin medicine.diagnostic_test Mitochondrial Permeability Transition Pore Chemistry Calcineurin MPTP High-Throughput Screening Assays Rats nervous system diseases Cell biology HEK293 Cells medicine.anatomical_structure nervous system Mitochondrial permeability transition pore cardiovascular system Molecular Medicine |
| Zdroj: | ASSAY and Drug Development Technologies. 16:445-455 |
| ISSN: | 1557-8127 1540-658X |
| DOI: | 10.1089/adt.2018.872 |
| Popis: | Mitochondrial permeability transition pore (mPTP) formation is well documented in isolated mitochondria. However, convincing detection of mPTP in whole cells remains elusive. In this study, we describe a high-throughput assay for Ca2+-activated mPTP opening in platelets using HyperCyt flow cytometry. In addition, we demonstrate that in several nucleated cells, using multiple approaches, the detection of cyclophilin D-dependent mPTP opening is highly challenging. Results with the mitochondrial-targeted Ca2+-sensing green fluorescent protein (mito-Case12) suggest the involvement of protein phosphatase 2B (PP2B; calcineurin) in regulating mitochondrial dynamics. Our results highlight the danger of relying on cyclosporine A alone as a pharmacological tool, and the need for comprehensive studies of mPTP in the cell. |
| Databáze: | OpenAIRE |
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