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利用活细胞中超分辨率形态相关细胞器识别技术对多个细胞器中的锌进行同步追踪。

Simultaneous Zn tracking in multiple organelles using super-resolution morphology-correlated organelle identification in living cells.

机构信息

State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering, Nanjing University, 210023, Nanjing, China.

Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH, 45267, USA.

出版信息

Nat Commun. 2021 Jan 4;12(1):109. doi: 10.1038/s41467-020-20309-7.

DOI:10.1038/s41467-020-20309-7
PMID:33397937
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7782730/
Abstract

Zn plays important roles in metabolism and signaling regulation. Subcellular Zn compartmentalization is essential for organelle functions and cell biology, but there is currently no method to determine Zn signaling relationships among more than two different organelles with one probe. Here, we report simultaneous Zn tracking in multiple organelles (Zn-STIMO), a method that uses structured illumination microscopy (SIM) and a single Zn fluorescent probe, allowing super-resolution morphology-correlated organelle identification in living cells. To guarantee SIM imaging quality for organelle identification, we develop a new turn-on Zn fluorescent probe, NapBu-BPEA, by regulating the lipophilicity of naphthalimide-derived Zn probes to make it accumulate in multiple organelles except the nucleus. Zn-STIMO with this probe shows that CCCP-induced mitophagy in HeLa cells is associated with labile Zn enhancement. Therefore, direct organelle identification supported by SIM imaging makes Zn-STIMO a reliable method to determine labile Zn dynamics in various organelles with one probe. Finally, SIM imaging of pluripotent stem cell-derived organoids with NapBu-BPEA demonstrates the potential of super-resolution morphology-correlated organelle identification to track biospecies and events in specific organelles within organoids.

摘要

Zn 在新陈代谢和信号调节中起着重要作用。细胞内 Zn 区室化对于细胞器功能和细胞生物学至关重要，但目前还没有一种方法可以用一个探针来确定超过两个不同细胞器之间的 Zn 信号关系。在这里，我们报告了一种在多个细胞器中同时追踪 Zn（Zn-STIMO）的方法，该方法使用结构光照明显微镜（SIM）和单个 Zn 荧光探针，允许在活细胞中进行超分辨率形态相关的细胞器识别。为了保证细胞器识别的 SIM 成像质量，我们通过调节萘酰亚胺衍生的 Zn 探针的亲脂性来开发了一种新的 Zn 荧光探针 NapBu-BPEA，使它除了细胞核之外还可以积累在多个细胞器中。使用这种探针的 Zn-STIMO 表明，HeLa 细胞中 CCCP 诱导的线粒体自噬与不稳定 Zn 的增强有关。因此，直接通过 SIM 成像进行细胞器识别使 Zn-STIMO 成为一种用一个探针确定各种细胞器中不稳定 Zn 动力学的可靠方法。最后，使用 NapBu-BPEA 对多能干细胞衍生的类器官进行 SIM 成像，证明了超分辨率形态相关细胞器识别在追踪类器官内特定细胞器中的生物物种和事件的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/630fef2bce2d/41467_2020_20309_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/f231c34d5676/41467_2020_20309_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/d3f3b343a6ab/41467_2020_20309_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/1031f35ace4e/41467_2020_20309_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/62825be9d6d0/41467_2020_20309_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/0a8e621508a0/41467_2020_20309_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/5c67d871c7a5/41467_2020_20309_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/916817522033/41467_2020_20309_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/630fef2bce2d/41467_2020_20309_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/f231c34d5676/41467_2020_20309_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/d3f3b343a6ab/41467_2020_20309_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/1031f35ace4e/41467_2020_20309_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/62825be9d6d0/41467_2020_20309_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/0a8e621508a0/41467_2020_20309_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/5c67d871c7a5/41467_2020_20309_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/916817522033/41467_2020_20309_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6937/7782730/630fef2bce2d/41467_2020_20309_Fig8_HTML.jpg

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