Atmospheric oxygen tension slows myoblast proliferation via mitochondrial activation.
Atmospheric oxygen tension slows myoblast proliferation via mitochondrial activation.
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DOI：
10.1371/journal.pone.0043853
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Partridge TA
中科院分区：
文献类型：
--
作者：
Duguez S;Duddy WJ;Gnocchi V;Bowe J;Dadgar S;Partridge TA
线粒体活性抑制增殖，并且是成肌细胞分化所必需的。成肌细胞增殖也受到标准组织培养中使用的20%氧气水平的抑制。我们假设，线粒体活性将是更大的高氧（20%O2）相对于更多的生理氧气（5%O2）。在5%和20%氧气下培养来自分离的肌纤维和条件永生化的H-2K成肌细胞的小鼠原代成肌细胞。通过细胞计数、EdU标记和CFSE稀释测定的增殖在20%氧气下较慢。原代成肌细胞中MyoD的表达在20%氧气下延迟，但肌原性（通过融合指数测量）略高。基于FACS的线粒体活性指标测量和ATP水平的光度测量显示，线粒体在20%氧气下表现出更大的膜电位和更高水平的活性氧（ROS），伴随着细胞内ATP的升高。线粒体质量不受影响。低浓度的CCCP（一种呼吸链解偶联剂）和寡霉素A（一种ATP合酶抑制剂）均增加了成肌细胞增殖的速率。ROS被研究为线粒体逆行信号传导的潜在机制，但是通过N-乙酰半胱氨酸（NAC）或α-苯基-N-叔丁基硝酮（PBN）清除ROS水平并不能挽救高氧条件下细胞分裂的抑制速率，这表明存在其他途径。在20%的氧气下，来自老年小鼠的原代成肌细胞的增殖比来自年轻成年小鼠的增殖慢，但在5%的氧气下没有差异。这些结果暗示线粒体调节作为成肌细胞对氧张力反应的机制解释。5%氧气对老年小鼠成肌细胞增殖率的拯救表明，在20%氧气的标准组织培养中，卫星细胞增殖与年龄相关的下降是一个主要的人为因素。它支持了这样一种观点，即这些与年龄有关的变化至少部分是由环境因素引起的，而不是卫星细胞固有的特征。
Mitochondrial activity inhibits proliferation and is required for differentiation of myoblasts. Myoblast proliferation is also inhibited by the ∼20% oxygen level used in standard tissue culture. We hypothesize that mitochondrial activity would be greater at hyperoxia (20% O2) relative to more physiological oxygen (5% O2). Murine primary myoblasts from isolated myofibres and conditionally immortalized H-2K myoblasts were cultured at 5% and 20% oxygen. Proliferation, assayed by cell counts, EdU labeling, and CFSE dilution, was slower at 20% oxygen. Expression of MyoD in primary myoblasts was delayed at 20% oxygen, but myogenicity, as measured by fusion index, was slightly higher. FACS-based measurement of mitochondrial activity indicators and luminometric measurement of ATP levels revealed that mitochondria exhibited greater membrane potential and higher levels of Reactive Oxygen Species (ROS) at 20% oxygen with concomitant elevation of intracellular ATP. Mitochondrial mass was unaffected. Low concentrations of CCCP, a respiratory chain uncoupler, and Oligomycin A, an ATP synthase inhibitor, each increased the rate of myoblast proliferation. ROS were investigated as a potential mechanism of mitochondrial retrograde signaling, but scavenging of ROS levels by N-acetyl-cysteine (NAC) or α-Phenyl-N-tert-butylnitrone (PBN) did not rescue the suppressed rate of cell division in hyperoxic conditions, suggesting other pathways. Primary myoblasts from older mice showed a slower proliferation than those from younger adult mice at 20% oxygen but no difference at 5% oxygen. These results implicate mitochondrial regulation as a mechanistic explanation for myoblast response to oxygen tension. The rescue of proliferation rate in myoblasts of aged mice by 5% oxygen suggests a major artefactual component to age-related decline of satellite cell proliferation in standard tissue culture at 20% oxygen. It lends weight to the idea that these age-related changes result at least in part from environmental factors rather than characteristics intrinsic to the satellite cell.
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影响因子：
13.1
作者：
Finley LW;Haigis MC
通讯作者：
Haigis MC
影响因子：
3.3
作者：
Barani, AE;Durieux, AC;Freyssenet, D
通讯作者：
Freyssenet, D
DOI：
10.1073/pnas.88.12.5096
发表时间：
1991-06-01
影响因子：
11.1
作者：
JAT, PS;NOBLE, MD;KIOUSSIS, D
通讯作者：
KIOUSSIS, D
影响因子：
3.7
作者：
Duguez, S;Sabido, O;Freyssenet, D
通讯作者：
Freyssenet, D
影响因子：
5.2
作者：
Collins, Charlotte A.;Zammit, Peter S.;Partridge, Terence A.
通讯作者：
Partridge, Terence A.