RRC ID 68641
Author Shindo Y, Fujii T, Komatsu H, Citterio D, Hotta K, Suzuki K, Oka K.
Title Newly developed Mg2+-selective fluorescent probe enables visualization of Mg2+ dynamics in mitochondria.
Journal PLoS One
Abstract Mg(2+) plays important roles in numerous cellular functions. Mitochondria take part in intracellular Mg(2+) regulation and the Mg(2+) concentration in mitochondria affects the synthesis of ATP. However, there are few methods to observe Mg(2+) in mitochondria in intact cells. Here, we have developed a novel Mg(2+)-selective fluorescent probe, KMG-301, that is functional in mitochondria. This probe changes its fluorescence properties solely depending on the Mg(2+) concentration in mitochondria under physiologically normal conditions. Simultaneous measurements using this probe together with a probe for cytosolic Mg(2+), KMG-104, enabled us to compare the dynamics of Mg(2+) in the cytosol and in mitochondria. With this method, carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP)-induced Mg(2+) mobilization from mitochondria to the cytosol was visualized. Although a FCCP-induced decrease in the Mg(2+) concentration in mitochondria and an increase in the cytosol were observed both in differentiated PC12 cells and in hippocampal neurons, the time-courses of concentration changes varied with cell type. Moreover, the relationship between mitochondrial Mg(2+) and Parkinson's disease was analyzed in a cellular model of Parkinson's disease by using the 1-methyl-4-phenylpyridinium ion (MPP(+)). A gradual decrease in the Mg(2+) concentration in mitochondria was observed in response to MPP(+) in differentiated PC12 cells. These results indicate that KMG-301 is useful for investigating Mg(2+) dynamics in mitochondria. All animal procedures to obtain neurons from Wistar rats were approved by the ethical committee of Keio University (permit number is 09106-(1)).
Volume 6(8)
Pages e23684
Published 2011-1-1
DOI 10.1371/journal.pone.0023684
PII PONE-D-11-06469
PMID 21858208
PMC PMC3156752
MeSH 1-Methyl-4-phenylpyridinium / pharmacology Animals Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / pharmacology Cation Transport Proteins / genetics Cation Transport Proteins / metabolism Cells, Cultured Cytosol / chemistry Cytosol / metabolism Fluorescent Dyes / chemical synthesis Fluorescent Dyes / chemistry* Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Hippocampus / cytology Ion Transport / drug effects Magnesium / chemistry Magnesium / metabolism* Microscopy, Confocal Mitochondria / chemistry Mitochondria / drug effects Mitochondria / metabolism* Mitochondrial Proteins / genetics Mitochondrial Proteins / metabolism Molecular Dynamics Simulation* Molecular Structure Neurons / chemistry Neurons / drug effects Neurons / metabolism PC12 Cells Proton Ionophores / pharmacology RNA, Small Interfering / genetics Rats Rats, Wistar Spectrometry, Fluorescence Transfection
IF 2.74
Human and Animal Cells PC-12(RCB0009)