Single-molecule transcript counting of stem-cell markers in the mouse intestine
Author(s)Itzkovitz, Shaul Shalev; Lyubimova, Anna; Blat, Irene C.; Maynard, Mindy; van Es, Johan; Lees, Jacqueline; Jacks, Tyler E.; Clevers, Hans; van Oudenaarden, Alexander; ... Show more Show less
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Determining the molecular identities of adult stem cells requires technologies for sensitive transcript detection in tissues. In mouse intestinal crypts, lineage-tracing studies indicated that different genes uniquely mark spatially distinct stem-cell populations, residing either at crypt bases or at position +4, but a detailed analysis of their spatial co-expression has not been feasible. Here we apply three-colour single-molecule fluorescent in situ hybridization to study a comprehensive panel of intestinal stem-cell markers during homeostasis, ageing and regeneration. We find that the expression of all markers overlaps at crypt-base cells. This co-expression includes Lgr5, Bmi1 and mTert, genes previously suggested to mark distinct stem cells. Strikingly, Dcamkl1 tuft cells, distributed throughout the crypt axis, co-express Lgr5 and other stem-cell markers that are otherwise confined to crypt bases. We also detect significant changes in the expression of some of the markers following irradiation, indicating their potential role in the regeneration process. Our approach can enable the sensitive detection of putative stem cells in other tissues and in tumours, guiding complementary functional studies to evaluate their stem-cell properties.
available in PMC 2012 July 1.
DepartmentMassachusetts Institute of Technology. Materials Processing Center; David H. Koch Institute for Integrative Cancer Research at MIT; Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of Physics
Nature Cell Biology
Nature Publishing Group
Itzkovitz, Shalev et al. “Single-molecule Transcript Counting of Stem-cell Markers in the Mouse Intestine.” Nature Cell Biology 14.1 (2011): 106–114. Web.
Author's final manuscript