Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate.
Imaging mitochondrial flux in single cells with a FRET sensor for pyruvate.
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DOI：
10.1371/journal.pone.0085780
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Barros LF
中科院分区：
文献类型：
--
作者：
San Martín A;Ceballo S;Baeza-Lehnert F;Lerchundi R;Valdebenito R;Contreras-Baeza Y;Alegría K;Barros LF
线粒体通量目前可以在低分辨率下获得。在这里，我们介绍了一种用于丙酮酸的基因编码的FRET传感器，以及在高时间分辨率下定量测量完整个体细胞中丙酮酸运输、丙酮酸产生和线粒体丙酮酸消耗的方法。在HEK293细胞、神经元和星形胶质细胞中，线粒体丙酮酸摄取在生理水平上是饱和的，这表明代谢率是由细胞器的固有属性决定的，而不是底物的可获得性。该传感器的潜力在神经元中得到了进一步的证明，在神经元中，发现线粒体流量在短暂的theta爆发引发的钙瞬变的几秒钟内增加了300%，而血糖水平保持不变。相反，星形细胞线粒体对细胞外ATP引起的类似的钙瞬变不敏感。我们预计，丙酮酸传感器提供的改进的分辨率将对对线粒体功能感兴趣的基础和应用研究人员具有实际意义。
Mitochondrial flux is currently accessible at low resolution. Here we introduce a genetically-encoded FRET sensor for pyruvate, and methods for quantitative measurement of pyruvate transport, pyruvate production and mitochondrial pyruvate consumption in intact individual cells at high temporal resolution. In HEK293 cells, neurons and astrocytes, mitochondrial pyruvate uptake was saturated at physiological levels, showing that the metabolic rate is determined by intrinsic properties of the organelle and not by substrate availability. The potential of the sensor was further demonstrated in neurons, where mitochondrial flux was found to rise by 300% within seconds of a calcium transient triggered by a short theta burst, while glucose levels remained unaltered. In contrast, astrocytic mitochondria were insensitive to a similar calcium transient elicited by extracellular ATP. We expect the improved resolution provided by the pyruvate sensor will be of practical interest for basic and applied researchers interested in mitochondrial function.
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影响因子：
7.4
作者：
Gerencser, Akos A.;Neilson, Andy;Choi, Sung W.;Edman, Ursula;Yadava, Nagendra;Oh, Richard J.;Ferrick, David A.;Nicholls, David G.;Brand, Martin D.
通讯作者：
Brand, Martin D.
影响因子：
4.1
作者：
Dimmer, KS;Friedrich, B;Bröer, S
通讯作者：
Bröer, S
影响因子：
5.3
作者：
Ruminot, Ivan;Gutierrez, Robin;Felipe Barros, L.
通讯作者：
Felipe Barros, L.
DOI：
10.1042/bj20091515
发表时间：
2010-01-15
期刊：
The Biochemical journal
影响因子：
--
作者：
Ovens MJ;Davies AJ;Wilson MC;Murray CM;Halestrap AP
通讯作者：
Halestrap AP
影响因子：
64.8
作者：
Lee, Youngjin;Morrison, Brett M.;Li, Yun;Lengacher, Sylvain;Farah, Mohamed H.;Hoffman, Paul N.;Liu, Yiting;Tsingalia, Akivaga;Jin, Lin;Zhang, Ping-Wu;Pellerin, Luc;Magistretti, Pierre J.;Rothstein, Jeffrey D.
通讯作者：
Rothstein, Jeffrey D.