The restriction of calcium influx in metaphase and post-metaphase stages of cell division revealed by imaging secondary ion mass spectrometry (SIMS).
The restriction of calcium influx in metaphase and post-metaphase stages of cell division revealed by imaging secondary ion mass spectrometry (SIMS).
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DOI：
10.1111/jmi.13182
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发表时间：
2023-05
影响因子：
2
通讯作者：
Chandra, Subhash
中科院分区：
文献类型：
--
作者：
Chandra, Subhash
关键词：
采用离子显微镜的二次离子质谱（SIMS）同位素成像技术观察单个肾上皮LLC-PK1细胞的钙内流。研究中使用的CAMECA IMS-3f SIMS仪器能够以500纳米的空间分辨率产生单细胞的同位素图像。由于仪器的高真空要求，细胞采用冻裂法低温制备，冷冻的冻干细胞用于SIMS分析。通过将细胞暴露于细胞外缓冲液中的44Ca稳定同位素中10分钟，直接成像钙的内流。测量了质量为44的44Ca内流和质量为40 （40Ca）的内源性钙在同一细胞中的分布。间期细胞与分裂细胞的直接比较表明，钙的内流在细胞分裂的中期和后期受到限制。这种限制在细胞分裂后期解除。与间期细胞相比，在钙内流限制下，10分钟内44Ca的净内流约为一半。在钙内流限制下，有丝分裂染色体与细胞质间的44Ca浓度相同。这些观察结果表明，细胞分裂时，内质网（ER）钙摄取受到钙内流限制。
A secondary ion mass spectrometry (SIMS) based isotopic imaging technique of ion microscopy was used for observing calcium influx in single renal epithelial LLC-PK1 cells. The CAMECA IMS-3f SIMS instrument, used in the study, is capable of producing isotopic images of single cells at 500 nm spatial resolution. Due to the high vacuum requirements of the instrument the cells were prepared cryogenically with a freeze-fracture method and frozen freeze-dried cells were used for SIMS analysis. The influx of calcium was imaged directly by exposure of cells to 44Ca stable isotope in the extracellular buffer for 10 min. The 44Ca influx was measured at mass 44 and the distribution of endogenous calcium at mass 40 (40Ca) in the same cell. A direct comparison of interphase cells to cells undergoing division revealed that calcium influx is restricted in metaphase and post-metaphase stages of cell division. This restriction is lifted in late cytokinesis. The net influx of 44Ca in 10 min. was approximately half under calcium influx restriction in comparison to interphase cells. Under calcium influx restriction the 44Ca concentration was the same between the mitotic chromosome and the cytoplasm. These observations indicate that the endoplasmic reticulum (ER) calcium uptake is compromised under calcium influx restriction in cells undergoing division.
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影响因子：
7.4
作者：
AUSSERER, WA;LING, YC;MORRISON, GH
通讯作者：
MORRISON, GH
影响因子：
2
作者：
CHANDRA, S;MORRISON, GH;WOLCOTT, CC
通讯作者：
WOLCOTT, CC
影响因子：
7.4
作者：
Chandra, S;Smith, DR;Morrison, GH
通讯作者：
Morrison, GH
DOI：
10.1046/j.1365-2818.2001.00944.x
发表时间：
2001-11-01
期刊：
JOURNAL OF MICROSCOPY-OXFORD
影响因子：
--
作者：
Chandra, S
通讯作者：
Chandra, S
DOI：
10.1073/pnas.1821399116
发表时间：
2019-05-21
影响因子：
11.1
作者：
Yu, Fang;Hubrack, Satanay Z.;Machaca, Khaled
通讯作者：
Machaca, Khaled