Ratiometric-pericam-mt, a novel tool to evaluate intramitochondrial zinc.
Zn(2+) can enter mitochondria and promote a plethora of physiological and patho-physiological effects. The issue of measuring changes in intramitochondrial levels is therefore critical. Past studies have employed fluorescent Zn(2+) indicators, like Rhod-2 and RhodZin-3, however, the use of these probes is impaired by their extramitochondrial sequestration. In this study, we show that the ratiometric mitochondria-targeted pericam, RPmt, can be employed to detect changes of intramitochondrial free Zn(2+) ([Zn(2+)](m)) levels. Using RPmt in neuronal and non neuronal cell lines we demonstrate that mitochondria can take up the cation mobilized from the cytosolic pool of protein-bound Zn(2+) and that mitochondrial Zn(2+) sequestration is largely mediated by the activity of the Ca(2+) uniporter.
Department of Basic and Applied Medical Science, Center for Excellence on Aging, University 'G. d'Annunzio', Chieti, Italy.
Teresa
T
Caporale
Domenico
D
Ciavardelli
Carmine
C
Di Ilio
Paola
P
Lanuti
Denise
D
Drago
Stefano L
SL
Sensi
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002118:Calcium / Q000032:analysis; D002460:Cell Line; D003600:Cytosol / Q000737:chemistry*; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D008164:Luminescent Proteins / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000737:chemistry*; D009474:Neurons; D014162:Transfection; D015032:Zinc / Q000032:analysis*
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D015032:Zinc; D002118:Calcium