Work

Single-molecule analysis of myocyte differentiation reveals bimodal lineage commitment

Year: 2015

Type: article

Abstract: 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.

Source: Integrative Biology

DOI: 10.1039/c5ib00057b

Authors: Tyler M. Gibson, Charles A. Gersbach

Institutions: Duke University, Duke Medical Center

Language: English

Topic: Muscle Physiology and Disorders

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Institutes of Health, Directorate for Biological Sciences, National Institute of General Medical Sciences

Awards: University Training Program in Biomolecular and Tissue Engineering, Engineering Morphogenetic Factors for Enhanced Genetic Reprogramming, Duke Center for Systems Biology, G5622705143