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Multi-isotope imaging mass spectrometry quantifies stem cell division and metabolism

机译：多同位素成像质谱法可量化干细胞分裂和代谢

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摘要

Mass spectrometry with stable isotope labels has been seminal in discovering the dynamic state of living matter1'2, 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 submicrometre resolution3'4. 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 DN A are segregated to the daughter destined to remain a stem cell, thus insuring lifetime genetic stability. After labelling mice with ~(15)N-thymidine from gestation until post-natal week 8, we find no ~(15)N label retention by dividing small intestinal crypt cells after a four-week chase. In adult mice administered ~(15)N-thymidine pulse-chase, we find that proliferating crypt cells dilute the ~(15)N 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 haematopoietic system. These studies show that MIMS provides high-resolution quantification of stable isotope labels that cannot be obtained using other techniques and that is broadly applicable to biological and medical research.
机译：具有稳定同位素标记的质谱技术在发现生物物质1'2的动态状态方面具有开创性意义，但仅限于大块组织或细胞。我们开发了多同位素成像质谱（MIMS），使我们能够查看和测量稳定的同位素，并具有亚微米分辨率3'4。在这里，我们将MIMS应用于包括果蝇，小鼠和人类在内的多种生物。我们测试了“永生链假说”，该假说预言了在不对称干细胞分裂过程中，包含较旧模板DN A的染色体将被分离到预定保留干细胞的子代，从而确保了终生遗传稳定性。从妊娠至出生后第8周，用〜（15）N-胸腺嘧啶核苷标记小鼠后，我们在追踪4周后将小肠隐窝细胞分开，发现没有〜（15）N标记保留。在成年小鼠施用〜（15）N-胸苷脉冲追踪中，我们发现增殖的隐窝细胞稀释了〜（15）N标记，与随机链分离相符。我们用果蝇脂质转换和向人类造血系统翻译的原理研究证明了MIMS的广泛用途。这些研究表明，MIMS可提供稳定同位素标记的高分辨率定量，这些同位素标记无法使用其他技术获得，并且广泛适用于生物学和医学研究。

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  • 来源
    《Nature》 |2012年第7382期|p.516-519|共4页
  • 作者

    Matthew L. Steinhauser; Andrew P. Bailey; Samuel E. Senyo; Christelle Guillermier; Todd S. Perlstein; Alex P. Gould; Richard T. Lee; Claude P. Lechene;

  • 作者单位

    Department of Medicine, Division of Cardiovascular Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.,Harvard Medical School, Boston, Massachusetts 02115, USA.;

    Division of Physiology and Metabolism, Medical Research Council National Institute for Medical Research, Mill Hill, London NW7 1AA, UK.;

    Department of Medicine, Division of Cardiovascular Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.,Harvard Medical School, Boston, Massachusetts 02115, USA.;

    Harvard Medical School, Boston, Massachusetts 02115, USA.,National Resource for Imaging Mass Spectroscopy, 65 Landsdowne St., Cambridge, Massachusetts 02139, USA.,Department of Medicine, Division of Genetics, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.;

    Department of Medicine, Division of Cardiovascular Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.,Harvard Medical School, Boston, Massachusetts 02115, USA.;

    Division of Physiology and Metabolism, Medical Research Council National Institute for Medical Research, Mill Hill, London NW7 1AA, UK.;

    Department of Medicine, Division of Cardiovascular Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.,Harvard Medical School, Boston, Massachusetts 02115, USA.,Harvard Stem Cell Institute,Cambridge, Massachusetts 02138, USA;

    Harvard Medical School, Boston, Massachusetts 02115, USA.,National Resource for Imaging Mass Spectroscopy, 65 Landsdowne St., Cambridge, Massachusetts 02139, USA.,Department of Medicine, Division of Genetics, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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