Imaging of oxygen gradients in giant umbrella cells: an ex vivo PLIM study.
Imaging of oxygen gradients in giant umbrella cells: an ex vivo PLIM study.
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巨型伞状细胞中氧梯度的成像：一项离体 PLIM 研究。
DOI：
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
D. Papkovsky
中科院分区：
文献类型：
--
作者：
Alexander V. Zhdanov;A. Golubeva;I. Okkelman;J. Cryan;D. Papkovsky
O2在有氧代谢和细胞和组织功能的调节中起着关键作用。组织中O2水平的局部差异和波动已被很好地记录下来；然而，O2微梯度的生理意义，特别是在亚细胞水平上，仍然知之甚少。利用穿透细胞的O2探针铂-葡萄糖和共聚焦荧光显微镜，我们观察了位于膀胱上皮表层的单个巨型(>；100-μm)伞状细胞中O2的分布。我们优化了对小鼠膀胱内表面进行体内磷光染色以及随后对切除的活组织进行体外分析的条件。影像实验显示呼吸伞状细胞内有明显的(≤85μM)和异质性的脱氧，整个细胞的径向O2梯度高达40μM，或∼0.6μM/μm。深度脱氧区(5-15μM O2)与富含极化线粒体的区域相对应。线粒体呼吸的药理激活降低了伞状细胞的氧合和O2梯度，而抗霉素A的抑制消除了这些梯度，并导致组织逐渐恢复到环境水平。O2分布的详细三维地图可能用于细胞内依赖O2的酶反应和下游过程的建模，如低氧诱导因子信号转导。利用共聚焦成像对细胞内和组织内O2梯度进行进一步的体外和体内研究，可以揭示膀胱和其他组织中调控O2依赖(病理)生理过程的分子机制。
O2 plays a pivotal role in aerobic metabolism and regulation of cell and tissue function. Local differences and fluctuations in tissue O2 levels are well documented; however, the physiological significance of O2 microgradients, particularly at the subcellular level, remains poorly understood. Using the cell-penetrating phosphorescent O2 probe Pt-Glc and confocal fluorescence microscopy, we visualized O2 distribution in individual giant (>100-μm) umbrella cells located superficially in the urinary bladder epithelium. We optimized conditions for in vivo phosphorescent staining of the inner surface of the mouse bladder and subsequent ex vivo analysis of excised live tissue. Imaging experiments revealed significant (≤85 μM) and heterogeneous deoxygenation within respiring umbrella cells, with radial O2 gradients of up to 40 μM across the cell, or ∼0.6 μM/μm. Deeply deoxygenated (5-15 μM O2) regions were seen to correspond to the areas enriched with polarized mitochondria. Pharmacological activation of mitochondrial respiration decreased oxygenation and O2 gradients in umbrella cells, while inhibition with antimycin A dissipated the gradients and caused gradual reoxygenation of the tissue to ambient levels. Detailed three-dimensional maps of O2 distribution potentially can be used for the modeling of intracellular O2-dependent enzymatic reactions and downstream processes, such as hypoxia-inducible factor signaling. Further ex vivo and in vivo studies on intracellular and tissue O2 gradients using confocal imaging can shed light on the molecular mechanisms regulating O2-dependent (patho)physiological processes in the bladder and other tissues.
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影响因子：
7.4
作者：
Kelley, Eric E.;Khoo, Nicholas K. H.;Hundley, Nicholas J.;Malik, Umair Z.;Freeman, Bruce A.;Tarpey, Margaret M.
通讯作者：
Tarpey, Margaret M.
含肌红蛋白的骨骼肌纤维中细胞内氧梯度的模型研究。
影响因子：
3.4
作者：
Federspiel,WJ
通讯作者：
Federspiel,WJ
影响因子：
4.1
作者：
Moore, Marc;Moore, Ruben;McFetridge, Peter S.
通讯作者：
McFetridge, Peter S.
影响因子：
4.8
作者：
Wang, Zhen J.;Joe, Bonnie N.;Coakley, Fergus V.;Zaharchuk, Greg;Busse, Reed;Yeh, Benjamin M.
通讯作者：
Yeh, Benjamin M.
DOI：
10.1002/cphc.200800296
发表时间：
2008-08-25
期刊：
Chemphyschem : a European journal of chemical physics and physical chemistry
影响因子：
--
作者：
Finikova OS;Lebedev AY;Aprelev A;Troxler T;Gao F;Garnacho C;Muro S;Hochstrasser RM;Vinogradov SA
通讯作者：
Vinogradov SA