Functional activity of permeability transition pore in energized and deenergized rat liver mitochondria
Permeability transition pore (mPTP) opening was studied under energized and deenergized conditions in rat liver mitochondria, and the effect of membrane depolarization on mPTP activity was evaluated. To assess mPTP activity, cyclosporine-sensitive swelling and cyclosporine sensitive Ca2+ efflux from...
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| Format: | Article |
| Language: | English |
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National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry
2020-12-01
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| Series: | The Ukrainian Biochemical Journal |
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| Online Access: | http://ukrbiochemjournal.org/wp-content/uploads/2020/12/Akopova_6_20.pdf |
| _version_ | 1797668828775186432 |
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| author | O. V. Akopova L. I. Kolchinskaya V. I. Nosar |
| author_facet | O. V. Akopova L. I. Kolchinskaya V. I. Nosar |
| author_sort | O. V. Akopova |
| collection | DOAJ |
| description | Permeability transition pore (mPTP) opening was studied under energized and deenergized conditions in rat liver mitochondria, and the effect of membrane depolarization on mPTP activity was evaluated. To assess mPTP activity, cyclosporine-sensitive swelling and cyclosporine sensitive Ca2+ efflux from mitochondria was studied using light absorbance techniques. In energized mitochondria, mPTP opening in sub-conductance states, at [Ca2+] ≤ Ka, contributed positively to the rate of respiration, without affecting ΔΨm. Threshold Ca2+ concentrations were found, which excess resulted in fast mitochondrial depolarization upon mPTP opening. An estimate of mPTP activity by cyclosporine-sensitive Ca2+ transport under energized and deenergized conditions have shown that membrane depolarization by protonophore CCCP essentially increased initial rate (V0), at simultaneous decrease of the half-time (t1/2) of Ca2+ efflux, which indicated mPTP activation, as compared to energized mitochondria. However, only partial release of Ca2+ via mPTP upon membrane depolarization was observed. With the use of selective blockers of Ca2+ uniporter and mPTP, ruthenium red (RR) and cyclosporine A (CsA), partial contribution of Ca2+ uniporter and mPTP in Ca2+ transport was found. “Titration” of Ca2+ transport by adding RR at different times from the onset of depolarization showed that depolarization dramatically reduced “life span” of mPTP as compared to energized mitochondria, which agreed with the kinetic characteristics of CsA-sensitive Ca2+ transport after the abolition of ΔΨm. Ca2+ added from the outer side of mitochondrial membrane produced dual effect on mPTP activity: activation at the onset of depolarization, but consequent promotion of mPTP closure. Based on the experiments, it was concluded that mitochondrial energization was required for prolonged mPTP functioning in sub-conductance states, whereas membrane depolarization promoted the transition of mPTP to inactive state during calcium release from mitochondria. |
| first_indexed | 2024-03-11T20:34:09Z |
| format | Article |
| id | doaj.art-3e5fdd1ef9b94dacb5e7a804affdf26d |
| institution | Directory Open Access Journal |
| issn | 2409-4943 2413-5003 |
| language | English |
| last_indexed | 2024-03-11T20:34:09Z |
| publishDate | 2020-12-01 |
| publisher | National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry |
| record_format | Article |
| series | The Ukrainian Biochemical Journal |
| spelling | doaj.art-3e5fdd1ef9b94dacb5e7a804affdf26d2023-10-02T07:19:18ZengNational Academy of Sciences of Ukraine, Palladin Institute of BiochemistryThe Ukrainian Biochemical Journal2409-49432413-50032020-12-01926637610.15407/ubj92.06.063Functional activity of permeability transition pore in energized and deenergized rat liver mitochondriaO. V. Akopova0https://orcid.org/0000-0002-0928-5087L. I. Kolchinskaya1V. I. Nosar2Bogomoletz Institute of Physiology, National Academy of Sciences of Ukraine, KyivBogomoletz Institute of Physiology, National Academy of Sciences of Ukraine, KyivBogomoletz Institute of Physiology, National Academy of Sciences of Ukraine, KyivPermeability transition pore (mPTP) opening was studied under energized and deenergized conditions in rat liver mitochondria, and the effect of membrane depolarization on mPTP activity was evaluated. To assess mPTP activity, cyclosporine-sensitive swelling and cyclosporine sensitive Ca2+ efflux from mitochondria was studied using light absorbance techniques. In energized mitochondria, mPTP opening in sub-conductance states, at [Ca2+] ≤ Ka, contributed positively to the rate of respiration, without affecting ΔΨm. Threshold Ca2+ concentrations were found, which excess resulted in fast mitochondrial depolarization upon mPTP opening. An estimate of mPTP activity by cyclosporine-sensitive Ca2+ transport under energized and deenergized conditions have shown that membrane depolarization by protonophore CCCP essentially increased initial rate (V0), at simultaneous decrease of the half-time (t1/2) of Ca2+ efflux, which indicated mPTP activation, as compared to energized mitochondria. However, only partial release of Ca2+ via mPTP upon membrane depolarization was observed. With the use of selective blockers of Ca2+ uniporter and mPTP, ruthenium red (RR) and cyclosporine A (CsA), partial contribution of Ca2+ uniporter and mPTP in Ca2+ transport was found. “Titration” of Ca2+ transport by adding RR at different times from the onset of depolarization showed that depolarization dramatically reduced “life span” of mPTP as compared to energized mitochondria, which agreed with the kinetic characteristics of CsA-sensitive Ca2+ transport after the abolition of ΔΨm. Ca2+ added from the outer side of mitochondrial membrane produced dual effect on mPTP activity: activation at the onset of depolarization, but consequent promotion of mPTP closure. Based on the experiments, it was concluded that mitochondrial energization was required for prolonged mPTP functioning in sub-conductance states, whereas membrane depolarization promoted the transition of mPTP to inactive state during calcium release from mitochondria.http://ukrbiochemjournal.org/wp-content/uploads/2020/12/Akopova_6_20.pdfrat liver mitochondriacalciumpermeability transition poreca2+ transportmembrane potential |
| spellingShingle | O. V. Akopova L. I. Kolchinskaya V. I. Nosar Functional activity of permeability transition pore in energized and deenergized rat liver mitochondria The Ukrainian Biochemical Journal rat liver mitochondria calcium permeability transition pore ca2+ transport membrane potential |
| title | Functional activity of permeability transition pore in energized and deenergized rat liver mitochondria |
| title_full | Functional activity of permeability transition pore in energized and deenergized rat liver mitochondria |
| title_fullStr | Functional activity of permeability transition pore in energized and deenergized rat liver mitochondria |
| title_full_unstemmed | Functional activity of permeability transition pore in energized and deenergized rat liver mitochondria |
| title_short | Functional activity of permeability transition pore in energized and deenergized rat liver mitochondria |
| title_sort | functional activity of permeability transition pore in energized and deenergized rat liver mitochondria |
| topic | rat liver mitochondria calcium permeability transition pore ca2+ transport membrane potential |
| url | http://ukrbiochemjournal.org/wp-content/uploads/2020/12/Akopova_6_20.pdf |
| work_keys_str_mv | AT ovakopova functionalactivityofpermeabilitytransitionporeinenergizedanddeenergizedratlivermitochondria AT likolchinskaya functionalactivityofpermeabilitytransitionporeinenergizedanddeenergizedratlivermitochondria AT vinosar functionalactivityofpermeabilitytransitionporeinenergizedanddeenergizedratlivermitochondria |