Atomic Force Microscopy on Living Cells: Aldosterone–Induced Localized Cell Swelling
Atomic Force Microscopy on Living Cells: Aldosterone–Induced Localized Cell Swelling
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活细胞原子力显微镜：醛固酮诱导的局部细胞肿胀
DOI：
10.1159/000025869
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发表时间：
2008
影响因子：
2.8
通讯作者：
H. Oberleithner
中科院分区：
文献类型：
--
作者：
S. Schneider;P. Pagel;J. Storck;Y. Yano;B. Sumpio;J. Geibel;H. Oberleithner
用于对活细胞的表面进行三维成像。仪器的分辨率主要受活细胞的弹性和柔软性的限制。然而，由于扫描大小和时间分辨率的原因，有可能获得1-100 nm范围内的分辨率。此外，无论细胞形态如何，都可以测量活细胞的体积。我们应用原子力显微镜在静止和应变条件下获得单个培养的牛主动脉和人脐带内皮细胞的形态信息，并同时测量细胞体积对醛固酮的响应。越来越多的证据表明，类固醇激素，如醛固酮、孕酮、维生素D或蜕皮类固醇，对细胞内电解质、肾脏电解质排泄和细胞体积也有快速的(范围从几秒到几分钟)非基因组影响[1-6]。从生理学的角度来看，激素诱导的细胞体积变化最近被认为是调节细胞代谢的关键信号[7-10]。细胞体积的类似变化已被证明影响内皮细胞的新陈代谢[11-14]。内皮细胞紧密地包裹在血管的管腔表面，在调节血管张力、血管重塑以及在人类动脉硬化和高血压的发病机制中发挥着重要作用。已有研究表明，内皮细胞肿胀可能会干扰细胞间的相互作用，导致跨内皮细胞通透性增加，这是动脉硬化发生的前驱机制[15，16]。在这项研究中，我们测试了血管内皮细胞在紧张状态下对醛固酮的反应是否会改变其体积。这样的变化已经在库尔特计数器测量的人类白细胞中显示出来。与或多或少呈球形并生活在悬浮液中的白细胞不同，内皮细胞表现出复杂的形态并附着在底物上。因此，只有使用更复杂的技术，才能测量生理条件下内皮细胞的离散细胞体积变化。利用原子力显微镜，我们可以精确地测量单个活体内皮细胞的绝对细胞体积，占总细胞体积的B1%。在加入醛固酮前，模拟动脉血压的机械应激内皮细胞体积为1,827b1，2fl(n=16)。激素作用5min后，细胞体积增加28%。25分钟后，尽管培养液中持续存在醛固酮，细胞体积仍恢复正常。质膜Na+/H+交换的阻断剂阿米洛利可阻止醛固酮引起的初始容量增加。此外，通过使用仪器附带的软件，我们能够减去在醛固酮刺激前后获得的单个EC的三维图像。剩下的是以三维方式可视化的净体积变化(图2)。1)。我们可以定位肾脏基础研究中的肿胀或收缩
for imaging the surface of living cells in three dimensions. The resolution of the instrument is mainly limited by the elasticity and softness of living cells. However, it is possible to achieve a resolution in the range of 1–100 nm due to scan size and time resolution. Moreover, it is possible to measure the volume of the living cells regardless of cell morphology. We applied AFM to obtain morphological information on individual cultured endothelial cells (ECs) of bovine aorta and human umbilical cord under stationary and strain conditions and to simultaneously measure changes in cell volume in response to aldosterone. There is increasing evidence that steroid hormones like aldosterone, progesterone, vitamin D or ecdysteroids also have rapid (range seconds to minutes), nongenomic effects on intracellular electrolytes, renal electrolyte excretion and cell volume [1–6]. From the physiological point of view, hormoneinduced changes in cell volume have recently been discussed as the crucial signal in regulating cell metabolism [7–10]. Similar changes in cell volume have been shown to affect metabolism in ECs [11–14]. ECs tightly coat the luminal surface of blood vessels and play an important role in the regulation of vascular tone, vascular remodeling, and in the pathogenesis of arteriosclerosis and hypertension in humans. It has been shown that swelling of ECs may disturb cell-to-cell interactions resulting in an increase in transendothelial permeability, a precuror mechanism in the development of arteriosclerosis [15, 16]. In this study we tested whether ECs under tension change their volume in response to aldosterone. Such changes were already shown in human leukocytes measured by Coulter counter. In contrast to leukocytes which are more or less spherical and live in suspension, ECs exhibit a complex morphology and adhere to a substrate. Thus, measurements of discrete cell volume changes in ECs under physiological conditions are only feasible with more sophisticated techniques. By using AFM we could precisely measure the absolute cell volume, B1% of total cell volume, of individual living ECs. Before addition of aldosterone the cell volume of mechanically stressed ECs mimicking arterial blood pressure was 1,827 B1 2 fl (n = 16). Cell volume was found to increase by 28% 5 min after hormone exposure. 25 min later cell volume was back to normal despite the continuous presence of aldosterone in the medium. Amiloride, a blocker of the plasma membrane Na+/H+ exchanger, prevented the initial aldosterone-induced volume increase. Moreover, by using the software coming with the instrument, we were able to subtract the three-dimensional images of an individual EC obtained before and after stimulation with aldosterone. What remains is t e net volume change visualized in a three-dimensional manner (fig. 1). We could localize the swelling or shrinkage Basic Renal Research
DOI：
10.1152/ajpcell.1996.270.4.c990
发表时间：
1996-04
期刊：
The American journal of physiology
影响因子：
--
作者：
T. J. Wiese;J. Dunlap;C. Conner;J. Grzybowski;W. Lowe;M. Yorek
通讯作者：
T. J. Wiese;J. Dunlap;C. Conner;J. Grzybowski;W. Lowe;M. Yorek
失血性休克期间骨骼肌毛细血管的管腔变窄和内皮细胞肿胀。
DOI：
--
发表时间：
1989
期刊：
Circulatory shock
影响因子：
--
作者：
Mazzoni,MC;Borgstrom,P;Intaglietta,M;Arfors,KE
通讯作者：
Arfors,KE