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oai:uvadoc.uva.es:10324/59852021-06-23T09:48:09Zcom_10324_1133com_10324_931com_10324_894col_10324_1209

Vay, Laura 2c53b216-f48d-4528-b236-82b247556625 500 Hernández San Miguel, Esther 7debb415-c184-4201-b951-b4b4648840b5 500 Domínguez Lobatón, María Carmen 70e473344399afbe 500 0000-0001-9574-3082 Moreno Díaz-Calderón, Alfredo a94b8921ed763ed9 500 0000-0002-7444-5198 Montero Zoccola, María Teresa 0ea84f9ae27b5cd1 500 0000-0001-7702-6653 Álvarez Martín, Javier cff9feef39aa55ff 500 0000-0003-0636-5521 2014-09-16T11:07:28Z 2014-09-16T11:07:28Z 2009 Cell Calcium, 2009, vol. 45, p. 243-250 0143-4160 http://uvadoc.uva.es/handle/10324/5985 10.1016/j.ceca.2008.10.007 243 250 Cell Calcium 45 Producción Científica Mitochondrial Ca2+ 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 [Ca2+] ([Ca2+]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 [Ca2+]M from increasing above 2–3 M. Instead, using low-Ca2+-affinity aequorin probes, we have measured [Ca2+]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 [Ca2+]M to levels of 0.5–1mM was actually observed at any phosphate concentration (0–10mM) during continuous perfusion of 3.5–100 MCa2+-buffers. In spite of this high and maintained (>10 min) [Ca2+]M, mitochondria retained functionality and the [Ca2+]M drop induced by a protonophore was fully reversible. In addition, this high [Ca2+]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 [Ca2+]M. In conclusion [Ca2+]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. application/pdf eng Elsevier Ltd. info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/4.0/ Attribution-NonCommercial-NoDerivatives 4.0 International Calcio - Metabolismo Mitochondrial free [Ca2+] levels and the permeability transition info:eu-repo/semantics/article SI CC-LICENSE license_url license_url text/plain 49 https://uvadoc.uva.es/bitstream/10324/5985/2/license_url 4afdbb8c545fd630ea7db775da747b2f MD5 2 license_text license_text text/html; charset=utf-8 22117 https://uvadoc.uva.es/bitstream/10324/5985/3/license_text dd6580d2d5007383f0e67b904850adc9 MD5 3 license_rdf license_rdf application/rdf+xml; charset=utf-8 23253 https://uvadoc.uva.es/bitstream/10324/5985/4/license_rdf cd76e7886171c964e259dcf5e912e299 MD5 4 LICENSE license.txt license.txt text/plain 4250 https://uvadoc.uva.es/bitstream/10324/5985/5/license.txt 909e634ba52becf192e4e9b4bcde7863 MD5 5 ORIGINAL ALVAREZ 23- Mitochondrial free [Ca2+] levels.pdf ALVAREZ 23- Mitochondrial free [Ca2+] levels.pdf PD-113 application/pdf 647152 https://uvadoc.uva.es/bitstream/10324/5985/1/ALVAREZ%2023-%20Mitochondrial%20free%20%5bCa2%2b%5d%20levels.pdf a19c27a71009ad371231708131888aca MD5 1 THUMBNAIL ALVAREZ 23- Mitochondrial free [Ca2+] levels.pdf.jpg ALVAREZ 23- Mitochondrial free [Ca2+] levels.pdf.jpg IM Thumbnail image/jpeg 2160 https://uvadoc.uva.es/bitstream/10324/5985/6/ALVAREZ%2023-%20Mitochondrial%20free%20%5bCa2%2b%5d%20levels.pdf.jpg 3187fa575bb34e176ac6cc403c7a65b7 MD5 6 10324/5985 oai:uvadoc.uva.es:10324/5985 2021-06-23 11:48:09.245 UVaDOC repositorio@uva.es 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