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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn2+ with genetically encoded sensors
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Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn2+ with genetically encoded sensors

机译：使用遗传编码传感器测量稳态和动态内质网和高尔基体Zn2 +

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Zn~(2+) plays essential roles in biology, and cells have adopted exquisite mechanisms for regulating steady-state Zn~(2+) levels. Although much is known about total Zn~(2+) in cells, very little is known about its subcellular distribution. Yet defining the location of Zn~(2+) and how it changes with signaling events is essential for elucidating how cells regulate this essential ion. Here we create fluorescent sensors genetically targeted to the endoplasmic reticulum (ER) and Golgi to monitor steady-state Zn~(2+) levels as well as flux of Zn~(2+) into and out of these organelles. These studies reveal that ER and Golgi contain a concentration of free Zn~(2+) that is 100 times lower than the cytosol. Both organelles take up Zn~(2+) when cytosolic levels are elevated, suggesting that the ER and Golgi can sequester elevated cytosolic Zn~(2+) and thus have the potential to play a role in influencing Zn~(2+) toxicity. ER Zn~(2+) homeostasis is perturbed by small molecule antagonists of Ca~(2+) homeostasis and ER Zn~(2+) is released upon elevation of cytosolic Ca~(2+) pointing to potential exchange of these two ions across the ER. This study provides direct evidence that Ca~(2+)signaling can influence Zn~(2+) homeostasis and vice versa, that Zn~(2+) dynamics may modulate Ca~(2+)signaling.
机译：Zn〜（2+）在生物学中起着至关重要的作用，细胞采用了精妙的机制来调节稳态Zn〜（2+）的水平。尽管人们对细胞中总Zn〜（2+）的了解很多，但对其亚细胞分布的了解却很少。然而，定义Zn〜（2+）的位置及其随信号事件的变化方式对于阐明细胞如何调节该基本离子至关重要。在这里，我们创建了遗传上针对内质网（ER）和高尔基体的荧光传感器，以监测稳态Zn〜（2+）水平以及Zn〜（2+）流入和流出这些细胞器的流量。这些研究表明，ER和高尔基体中的游离Zn〜（2+）浓度比胞质溶胶低100倍。当细胞质水平升高时，两个细胞器都吸收Zn〜（2+），这表明ER和高尔基体可以螯合升高的细胞质Zn〜（2+），因此有可能在影响Zn〜（2+）毒性中发挥作用。 ER Zn〜（2+）稳态受到Ca〜（2+）稳态的小分子拮抗剂的干扰，并且当细胞质Ca〜（2+）升高时释放ER Zn〜（2+），表明这两个离子的潜在交换在急诊室。这项研究提供了直接的证据，表明Ca〜（2+）信号可以影响Zn〜（2+）的稳态，反之亦然，Zn〜（2+）动力学可以调节Ca〜（2+）信号。

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