Newly Developed Mg2 –Selective Fluorescent Probe Enables Visualization of Mg2 Dynamics in MitochondriaReportar como inadecuado




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Mg2+ plays important roles in numerous cellular functions. Mitochondria take part in intracellular Mg2+ regulation and the Mg2+ concentration in mitochondria affects the synthesis of ATP. However, there are few methods to observe Mg2+ in mitochondria in intact cells. Here, we have developed a novel Mg2+–selective fluorescent probe, KMG-301, that is functional in mitochondria. This probe changes its fluorescence properties solely depending on the Mg2+ concentration in mitochondria under physiologically normal conditions. Simultaneous measurements using this probe together with a probe for cytosolic Mg2+, KMG-104, enabled us to compare the dynamics of Mg2+ in the cytosol and in mitochondria. With this method, carbonyl cyanide p-trifluoromethoxy phenylhydrazone FCCP–induced Mg2+ mobilization from mitochondria to the cytosol was visualized. Although a FCCP–induced decrease in the Mg2+ 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 Mg2+ 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 Mg2+ concentration in mitochondria was observed in response to MPP+ in differentiated PC12 cells. These results indicate that KMG-301 is useful for investigating Mg2+ 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.



Autor: Yutaka Shindo, Tomohiko Fujii, Hirokazu Komatsu, Daniel Citterio, Kohji Hotta, Koji Suzuki, Kotaro Oka

Fuente: http://plos.srce.hr/



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