Details

Autor(en) / Beteiligte

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

Titel

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

Ist Teil von

• Nature (London), 2012-01, Vol.481 (7382), p.516-519

Ort / Verlag

London: Nature Publishing Group UK

Erscheinungsjahr

2012

Quelle

EBSCOhost Psychology and Behavioral Sciences Collection

Beschreibungen/Notizen

• Multi-isotope imaging mass spectrometry is used to investigate the ‘immortal strand hypothesis’, Drosophila lipid metabolism and human lymphopoiesis. Quantifying metabolism Multi-isotope imaging mass spectrometry (MIMS) is a new and generally applicable method for the study of DNA replication, lipid and protein turnover and cell fate in animals and humans. In a proof-of-principle study, MIMS was used to test the 'immortal strand hypothesis', which proposes that stem cells maintain a master genetic template that is protected from cancer-causing mutations. The hypothesis remains hotly debated, in part because of the difficulties involved in testing it experimentally. Stable isotope incorporation was viewed and measured by MIMS in mammalian intestinal cell division, Drosophila melanogaster lipid metabolism and human lymphopoiesis. Mass spectrometry with stable isotope labels has been seminal in discovering the dynamic state of living matter 1 , 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 resolution 3 , 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 DNA 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.