RT info:eu-repo/semantics/article
T1 Monitoring mitochondrial [Ca2+] dynamics with rhod-2, ratiometric pericam and aequorin
A1 Fonteriz García, Rosalba Inés
A1 Fuente Pérez, Sergio de la
A1 Moreno Díaz-Calderón, Alfredo
A1 Domínguez Lobatón, María Carmen
A1 Montero Zoccola, María Teresa
A1 Álvarez Martín, Javier
K1 Calcio - Metabolismo
AB The dynamics of mitochondrial [Ca2+] ([Ca2+]M) plays a key role in a variety of cellular processes. Themost important methods available to monitor [Ca2+]M are fluorescent dyes such as rhod-2 and specificallytargeted proteins such as aequorin and pericam. However, significant discrepancies, both quantitativeand qualitative, exist in the literature between the results obtained with different methods. We havemade here a systematic comparison of the response of several fluorescent dyes, rhod-2 and rhod-FF,and two Ca2+-sensitive proteins, aequorin and pericam. Our results show that measurements obtainedwith aequorin and pericam are consistent in terms of dynamic Ca2+ changes. Instead, fluorescent dyesfailed to follow Ca2+ changes adequately, especially during repetitive stimulation. In particular, measuresobtained with rhod-2 or rhod-FF evidenced the previously reported Ca2+-dependent inhibition ofmitochondrial Ca2+ uptake, but data obtained with aequorin or pericam under the same conditions didnot. The reason for the loss of response of fluorescent dyes is unclear. Loading with these dyes producedchanges in mitochondrial morphology and membrane potential, which were small and reversible at lowconcentrations (1–2 M), but produced large and prolonged damage at higher concentrations. In addition,cells loaded with low concentrations of rhod-2 suffered large changes in mitochondrial morphologyafter light excitation. Our results suggest that [Ca2+]M data obtained with these dyes should be taken withcare.
PB Elsevier Ltd.
SN 0143-4160
YR 2010
FD 2010
LK http://uvadoc.uva.es/handle/10324/5959
UL http://uvadoc.uva.es/handle/10324/5959
LA eng
NO Cell Calcium, 2010, vol. 48, p. 61-69
NO Producción Científica
DS UVaDOC
RD 24-nov-2024