气候变化领域集成服务门户(CCPortal): Single-cell visualization and quantification of trace metals in Chlamydomonas lysosome-related organelles

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DOI	10.1073/pnas.2026811118
	Single-cell visualization and quantification of trace metals in Chlamydomonas lysosome-related organelles
	Schmollinger S.; Chen S.; Strenkert D.; Hui C.; Ralle M.; Merchant S.S.
发表日期	2021
ISSN	00278424
卷号	118 期号:16
英文摘要	The acidocalcisome is an acidic organelle in the cytosol of eukaryotes, defined by its low pH and high calcium and polyphosphate content. It is visualized as an electron-dense object by transmission electron microscopy (TEM) or described with mass spectrometry (MS)-based imaging techniques or multimodal X-ray fluorescence microscopy (XFM) based on its unique elemental composition. Compared with MS-based imaging techniques, XFM offers the additional advantage of absolute quantification of trace metal content, since sectioning of the cell is not required and metabolic states can be preserved rapidly by either vitrification or chemical fixation. We employed XFM in Chlamydomonas reinhardtii to determine single-cell and organelle trace metal quotas within algal cells in situations of trace metal overaccumulation (Fe and Cu). We found up to 70% of the cellular Cu and 80% of Fe sequestered in acidocalcisomes in these conditions and identified two distinct populations of acidocalcisomes, defined by their unique trace elemental makeup. We utilized the vtc1 mutant, defective in polyphosphate synthesis and failing to accumulate Ca, to show that Fe sequestration is not dependent on either. Finally, quantitation of the Fe and Cu contents of individual cells and compartments via XFM, over a range of cellular metal quotas created by nutritional and genetic perturbations, indicated excellent correlation with bulk data from corresponding cell cultures, establishing a framework to distinguish the nutritional status of single cells. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Heavy metal detoxification; Trace metal sequestration; Vacuole VTC XRF
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/179793
作者单位	California Institute for Quantitative Biosciences, University of California, Berkeley, CA 94720, United States; Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, United States; Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095, United States; Institute for Genomics and Proteomics, University of California, Los Angeles, CA 90095, United States; Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439, United States; Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, CA 94550, United States; Molecular and Medical Genetics Department, Oregon Health and Sciences University, Portland, OR 97239, United States; Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, United States
推荐引用方式 GB/T 7714	Schmollinger S.,Chen S.,Strenkert D.,et al. Single-cell visualization and quantification of trace metals in Chlamydomonas lysosome-related organelles[J],2021,118(16).
APA	Schmollinger S.,Chen S.,Strenkert D.,Hui C.,Ralle M.,&Merchant S.S..(2021).Single-cell visualization and quantification of trace metals in Chlamydomonas lysosome-related organelles.Proceedings of the National Academy of Sciences of the United States of America,118(16).
MLA	Schmollinger S.,et al."Single-cell visualization and quantification of trace metals in Chlamydomonas lysosome-related organelles".Proceedings of the National Academy of Sciences of the United States of America 118.16(2021).