Single-molecule analysis of myocyte differentiation reveals bimodal lineage commitment.

Published

Journal Article

Cell differentiation is the foundation for tissue development and regeneration, disease modeling, and cell-based therapies. Although the differentiation of cell populations has been extensively studied in many systems, much less is known about the distribution of decision making of single cells within these populations. To characterize the differentiation of single skeletal muscle cells, we used single-molecule mRNA fluorescence in situ hybridization (smFISH) to precisely quantify the expression levels of the master myogenic regulatory factors MyoD and myogenin in individual myoblasts. We identified distinct cell states characterized by the number of myogenin transcripts expressed by a cell, with myoblasts stochastically transitioning to a myogenin-high state during differentiation. We also used MyoD overexpression to force the transdifferentiation of C3H10T1/2 cells into an induced myoblast phenotype. These reprogrammed cells revealed the presence of a critical threshold of MyoD expression required to initiate myogenin expression. These results provide quantitative single-molecule data to support the model of switch-like cell decision making and lineage specification.

Full Text

Duke Authors

Cited Authors

  • Gibson, TM; Gersbach, CA

Published Date

  • June 2015

Published In

Volume / Issue

  • 7 / 6

Start / End Page

  • 663 - 671

PubMed ID

  • 25953198

Pubmed Central ID

  • 25953198

Electronic International Standard Serial Number (EISSN)

  • 1757-9708

International Standard Serial Number (ISSN)

  • 1757-9694

Digital Object Identifier (DOI)

  • 10.1039/c5ib00057b

Language

  • eng