| Monitoring mitochondrial [Ca(2+)] dynamics with rhod-2, ratiometric pericam and aequorin. | |
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MedLine Citation:
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PMID: 20667591 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The dynamics of mitochondrial [Ca(2+)] ([Ca(2+)](M)) plays a key role in a variety of cellular processes. The most important methods available to monitor [Ca(2+)](M) are fluorescent dyes such as rhod-2 and specifically targeted proteins such as aequorin and pericam. However, significant discrepancies, both quantitative and qualitative, exist in the literature between the results obtained with different methods. We have made here a systematic comparison of the response of several fluorescent dyes, rhod-2 and rhod-FF, and two Ca(2+)-sensitive proteins, aequorin and pericam. Our results show that measurements obtained with aequorin and pericam are consistent in terms of dynamic Ca(2+) changes. Instead, fluorescent dyes failed to follow Ca(2+) changes adequately, especially during repetitive stimulation. In particular, measures obtained with rhod-2 or rhod-FF evidenced the previously reported Ca(2+)-dependent inhibition of mitochondrial Ca(2+) uptake, but data obtained with aequorin or pericam under the same conditions did not. The reason for the loss of response of fluorescent dyes is unclear. Loading with these dyes produced changes in mitochondrial morphology and membrane potential, which were small and reversible at low concentrations (1-2 microM), but produced large and prolonged damage at higher concentrations. In addition, cells loaded with low concentrations of rhod-2 suffered large changes in mitochondrial morphology after light excitation. Our results suggest that [Ca(2+)](M) data obtained with these dyes should be taken with care. |
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Authors:
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Rosalba I Fonteriz; Sergio de la Fuente; Alfredo Moreno; Carmen D Lobatón; Mayte Montero; Javier Alvarez |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-07-27 |
Journal Detail:
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Title: Cell calcium Volume: 48 ISSN: 1532-1991 ISO Abbreviation: Cell Calcium Publication Date: 2010 Jul |
Date Detail:
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Created Date: 2010-08-27 Completed Date: 2010-12-01 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8006226 Medline TA: Cell Calcium Country: Netherlands |
Other Details:
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Languages: eng Pagination: 61-9 Citation Subset: IM |
Copyright Information:
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Copyright 2010 Elsevier Ltd. All rights reserved. |
Affiliation:
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Instituto de Biología y Genética Molecular (IBGM), Departamento de Bioquímica y Biología Molecular y Fisiología, Facultad de Medicina, Universidad de Valladolid and Consejo Superior de Investigaciones Científicas (CSIC), Ramón y Cajal, 7, E-47005 Valladolid, Spain. |
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| MeSH Terms | |
Descriptor/Qualifier:
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Aequorin
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diagnostic use* Calcium / metabolism* Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / pharmacology Fluorescent Dyes / diagnostic use* Green Fluorescent Proteins / metabolism Hela Cells Heterocyclic Compounds, 3-Ring / diagnostic use Humans Microscopy, Confocal Mitochondria / drug effects, metabolism* Mitochondrial Proteins / diagnostic use* |
| Chemical | |
Reg. No./Substance:
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0/Fluorescent Dyes; 0/Heterocyclic Compounds, 3-Ring; 0/Mitochondrial Proteins; 132523-91-2/rhod-2; 147336-22-9/Green Fluorescent Proteins; 370-86-5/Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; 50934-79-7/Aequorin; 7440-70-2/Calcium |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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