Source:http://linkedlifedata.com/resource/pubmed/id/19100620
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
2009-3-9
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pubmed:abstractText |
Mitochondrial Ca(2+) activates many processes, from mitochondrial metabolism to opening of the permeability transition pore (PTP) and apoptosis. However, there is considerable controversy regarding the free mitochondrial [Ca(2+)] ([Ca(2+)](M)) levels that can be attained during cell activation or even in mitochondrial preparations. Studies using fluorescent dyes (rhod-2 or similar), have reported that phosphate precipitation precludes [Ca(2+)](M) from increasing above 2-3 microM. Instead, using low-Ca(2+)-affinity aequorin probes, we have measured [Ca(2+)](M) values more than two orders of magnitude higher. We confirm here these values by making a direct in situ calibration of mitochondrial aequorin, and we show that a prolonged increase in [Ca(2+)](M) to levels of 0.5-1mM was actually observed at any phosphate concentration (0-10mM) during continuous perfusion of 3.5-100 microM Ca(2+)-buffers. In spite of this high and maintained (>10 min) [Ca(2+)](M), mitochondria retained functionality and the [Ca(2+)](M) drop induced by a protonophore was fully reversible. In addition, this high [Ca(2+)](M) did not induce PTP opening unless additional activators (phenyl arsine oxide, PAO) were present. PAO induced a rapid, concentration-dependent and irreversible drop in [Ca(2+)](M). In conclusion [Ca(2+)](M) levels of 0.5-1mM can be reached and maintained for prolonged periods (>10 min) in phosphate-containing medium, and massive opening of PTP requires additional pore activators.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Aequorin,
http://linkedlifedata.com/resource/pubmed/chemical/Arsenicals,
http://linkedlifedata.com/resource/pubmed/chemical/Buffers,
http://linkedlifedata.com/resource/pubmed/chemical/Calcium,
http://linkedlifedata.com/resource/pubmed/chemical/Carbonyl Cyanide...,
http://linkedlifedata.com/resource/pubmed/chemical/Mutant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphates,
http://linkedlifedata.com/resource/pubmed/chemical/oxophenylarsine
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pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
1532-1991
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pubmed:author | |
pubmed:issnType |
Electronic
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pubmed:volume |
45
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
243-50
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pubmed:meshHeading |
pubmed-meshheading:19100620-Aequorin,
pubmed-meshheading:19100620-Animals,
pubmed-meshheading:19100620-Arsenicals,
pubmed-meshheading:19100620-Buffers,
pubmed-meshheading:19100620-Calcium,
pubmed-meshheading:19100620-Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone,
pubmed-meshheading:19100620-Cattle,
pubmed-meshheading:19100620-Cells, Cultured,
pubmed-meshheading:19100620-Chromaffin Cells,
pubmed-meshheading:19100620-HeLa Cells,
pubmed-meshheading:19100620-Humans,
pubmed-meshheading:19100620-Membrane Potential, Mitochondrial,
pubmed-meshheading:19100620-Mitochondria,
pubmed-meshheading:19100620-Mutant Proteins,
pubmed-meshheading:19100620-Perfusion,
pubmed-meshheading:19100620-Permeability,
pubmed-meshheading:19100620-Phosphates
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pubmed:year |
2009
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pubmed:articleTitle |
Mitochondrial free [Ca2+] levels and the permeability transition.
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pubmed:affiliation |
Instituto de Biología y Genética Molecular (IBGM), Departamento de Bioquímica y Biología Molecular y Fisiología, Universidad de Valladolid and Consejo Superior de Investigaciones Científicas (CSIC), Ramón y Cajal, 7, E-47005 Valladolid, Spain.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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