A high-throughput respirometric assay for mitochondrial biogenesis and toxicity.
A high-throughput respirometric assay for mitochondrial biogenesis and toxicity.
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DOI：
10.1016/j.ab.2010.04.040
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发表时间：
2010-09-01
影响因子：
2.9
通讯作者：
Schnellmann RG
中科院分区：
文献类型：
--
作者：
Beeson CC;Beeson GC;Schnellmann RG
Mitochondria are a common target of toxicity for drugs and other chemicals, and results in decreased aerobic metabolism and cell death. In contrast, mitochondrial biogenesis restores cell vitality and there is a need for new agents to induce biogenesis. Current cell-based models of mitochondrial biogenesis or toxicity are inadequate because cultured cell lines are highly glycolytic with minimal aerobic metabolism and altered mitochondrial physiology. In addition, there are no high-throughput, real-time assays that assess mitochondrial function. We adapted primary cultures of renal proximal tubular cells (RPTC) that exhibit in vivo levels of aerobic metabolism, are not glycolytic, and retain higher levels of differentiated functions and used the Seahorse Biosciences analyzer to measure mitochondrial function in real time in multi-well plates. Using uncoupled respiration as a marker of electron transport chain (ETC) integrity, the nephrotoxicants cisplatin, HgCl2 and gentamicin exhibited mitochondrial toxicity prior to decreases in basal respiration and cell death. Conversely, using FCCP-uncoupled respiration as a marker of maximal ETC activity, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), SRT1720, resveratrol, daidzein, and metformin produced mitochondrial biogenesis in RPTC. The merger of the RPTC model and multi-well respirometry results in a single high throughput assay to measure mitochondrial biogenesis and toxicity, and nephrotoxic potential.
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影响因子：
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作者：
Thukral, SK;Nordone, PJ;Afshari, CA
通讯作者：
Afshari, CA
影响因子：
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Gerencser, Akos A.;Neilson, Andy;Choi, Sung W.;Edman, Ursula;Yadava, Nagendra;Oh, Richard J.;Ferrick, David A.;Nicholls, David G.;Brand, Martin D.
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影响因子：
3.8
作者：
Soltoff, S P;Mandel, L J
通讯作者：
Mandel, L J