Multi-isotope imaging mass spectrometry quantifies stem cell division and metabolism.
Multi-isotope imaging mass spectrometry quantifies stem cell division and metabolism.
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DOI：
10.1038/nature10734
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发表时间：
2012-01-15
期刊：
影响因子：
64.8
通讯作者：
Lechene, Claude P.
中科院分区：
文献类型：
--
作者：
Steinhauser, Matthew L.;Bailey, Andrew P.;Senyo, Samuel E.;Guillermier, Christelle;Perlstein, Todd S.;Gould, Alex P.;Lee, Richard T.;Lechene, Claude P.
具有稳定同位素标记的质谱学在发现生物的动态状态方面一直是开创性的，但仅限于大宗组织或细胞。我们开发了多同位素成像质谱仪(MIMS)，使我们能够以亚微米分辨率观察和测量稳定的同位素掺入。在这里，我们将MIMS应用于不同的生物，包括果蝇、老鼠和人类。我们测试了“永生链假说”，该假说预测，在不对称干细胞分裂期间，含有较老模板DNA的染色体被分离到注定将保留干细胞的子代，从而确保一生的遗传稳定性。用15N-胸腺嘧啶核苷标记小鼠从孕期到出生后8周，追踪4wk后，小肠隐窝细胞分裂未发现15N标记残留。在给予15N-胸腺嘧啶核苷脉冲追踪的成年小鼠中，我们发现增殖的隐窝细胞稀释了符合随机链分离的标记。我们通过对果蝇脂肪转化和翻译到人类造血系统的原理验证研究，证明了MIMS的广泛应用。这些研究表明，MIMS提供了使用其他技术无法获得的稳定同位素标记的高分辨率定量，并且广泛适用于生物和医学研究。
Mass spectrometry with stable isotope labels has been seminal in discovering the dynamic state of living matter but is limited to bulk tissues or cells. We developed multi-isotope imaging mass spectrometry (MIMS) that allowed us to view and measure stable isotope incorporation with sub-micron resolution. Here we apply MIMS to diverse organisms, including Drosophila, mice, and humans. We test the “immortal strand hypothesis,” which predicts that during asymmetric stem cell division chromosomes containing older template DNA are segregated to the daughter destined to remain a stem cell, thus insuring lifetime genetic stability. After labeling mice with 15N-thymidine from gestation through post-natal week 8, we find no 15N label retention by dividing small intestinal crypt cells after 4wk chase. In adult mice administered 15N-thymidine pulse-chase, we find that proliferating crypt cells dilute label consistent with random strand segregation. We demonstrate the broad utility of MIMS with proof-of-principle studies of lipid turnover in Drosophila and translation to the human hematopoietic system. These studies show that MIMS provides high-resolution quantitation of stable isotope labels that cannot be obtained using other techniques and that is broadly applicable to biological and medical research.
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影响因子：
56.9
作者：
Schoenheimer, R;Rittenberg, D
通讯作者：
Rittenberg, D
影响因子：
5.2
作者：
Sotiropoulou, Panagiota A.;Candi, Aurelie;Blanpain, Cedric
通讯作者：
Blanpain, Cedric
DOI：
10.1073/pnas.1013004108
发表时间：
2011-01-04
影响因子：
11.1
作者：
Montgomery, Robert K.;Carlone, Diana L.;Breault, David T.
通讯作者：
Breault, David T.
影响因子：
30.8
作者：
Sangiorgi, Eugenio;Capecchi, Mario R.
通讯作者：
Capecchi, Mario R.
影响因子：
11.4
作者：
Schepers, Arnout G.;Vries, Robert;Clevers, Hans
通讯作者：
Clevers, Hans