Mouse lines with photo-activatable mitochondria to study mitochondrial dynamics.
Mouse lines with photo-activatable mitochondria to study mitochondrial dynamics.
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DOI：
10.1002/dvg.22050
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发表时间：
2012-11
期刊：
影响因子：
1.5
通讯作者：
Chan, David C.
中科院分区：
文献类型：
--
作者：
Pham, Anh H.;McCaffery, J. Michael;Chan, David C.
许多病理状态涉及线粒体融合、分裂或转运的失调。这些动态事件通常在细胞系中进行研究，因为在组织中追踪线粒体具有挑战性。为了研究组织和疾病模型中的线粒体动力学，我们产生了两个具有光活化线粒体（PhAM）的小鼠品系。在PhAMfloxed系中，光可转化荧光蛋白Dendra 2（mito-Dendra 2）的细胞定位形式与上游loxP侧翼终止信号一起沿着靶向遍在表达的Rosa 26基因座。Cre在PhAMfloxed细胞中的表达导致明亮的mito-Dendra 2荧光，而对线粒体形态没有不利影响。当与Cre驱动程序杂交时，PhAMfloxed系在特定细胞类型中表达mito-Dendra 2，即使在具有高细胞密度的组织中也可以追踪线粒体。在第二条线（PhAMexcised）中，mito-Dendra 2的表达是普遍存在的，允许在广泛的活组织和固定组织中分析线粒体。通过使用光转换技术，我们直接测量了培养细胞以及骨骼肌等组织中的线粒体融合事件。这些小鼠品系有助于在广泛的原代细胞和组织中分析线粒体动力学，并可用于检查发育过渡和疾病状态中的线粒体。
Many pathological states involve dysregulation of mitochondrial fusion, fission, or transport. These dynamic events are usually studied in cells lines because of the challenges in tracking mitochondria in tissues. To investigate mitochondrial dynamics in tissues and disease models, we generated two mouse lines with photo-activatable mitochondria (PhAM). In the PhAMfloxed line, a mitochondrially localized version of the photo-convertible fluorescent protein Dendra2 (mito-Dendra2) is targeted to the ubiquitously expressed Rosa26 locus, along with an upstream loxP-flanked termination signal. Expression of Cre in PhAMfloxed cells results in bright mito-Dendra2 fluorescence without adverse effects on mitochondrial morphology. When crossed with Cre drivers, the PhAMfloxed line expresses mito-Dendra2 in specific cell types, allowing mitochondria to be tracked even in tissues that have high cell density. In a second line (PhAMexcised), the expression of mito-Dendra2 is ubiquitous, allowing mitochondria to be analyzed in a wide range of live and fixed tissues. By using photo-conversion techniques, we directly measured mitochondrial fusion events in cultured cells as well as tissues such as skeletal muscle. These mouse lines facilitate analysis of mitochondrial dynamics in a wide spectrum of primary cells and tissues, and can be used to examine mitochondria in developmental transitions and disease states.
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DOI：
10.1523/jneurosci.1233-11.2012
发表时间：
2012-01-04
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Magrané J;Sahawneh MA;Przedborski S;Estévez ÁG;Manfredi G
通讯作者：
Manfredi G
影响因子：
3.5
作者：
Chen H;Chan DC
通讯作者：
Chan DC
DOI：
10.1083/jcb.200903065
发表时间：
2009-09-21
期刊：
The Journal of cell biology
影响因子：
--
作者：
Wakabayashi J;Zhang Z;Wakabayashi N;Tamura Y;Fukaya M;Kensler TW;Iijima M;Sesaki H
通讯作者：
Sesaki H
影响因子：
64.5
作者：
Chen H;Vermulst M;Wang YE;Chomyn A;Prolla TA;McCaffery JM;Chan DC
通讯作者：
Chan DC
影响因子：
--
作者：
CARDULLO, RA;BALTZ, JM
通讯作者：
BALTZ, JM