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Single-nucleus RNA-seq identifies transcriptional heterogeneity in multinucleated skeletal myofibers

By Michael J. Petrany, Casey O Swoboda, Chengyi Sun, Kashish Chetal, Xiaoting Chen, Matthew T Weirauch, Nathan Salomonis, Douglas P. Millay

Posted 15 Apr 2020
bioRxiv DOI: 10.1101/2020.04.14.041400 (published DOI: 10.1038/s41467-020-20063-w)

While the majority of cells contain a single nucleus, cell types such as trophoblasts, osteoclasts, and skeletal myofibers require multinucleation. One advantage of multinucleation can be the assignment of distinct functions to different nuclei, but comprehensive interrogation of transcriptional heterogeneity within multinucleated tissues has been challenging due to the presence of a shared cytoplasm. Here, we utilized single-nucleus RNA-sequencing (snRNA-seq) to determine the extent of transcriptional diversity within multinucleated skeletal myofibers. Nuclei from mouse skeletal muscle were profiled across the lifespan, which revealed the emergence of distinct myonuclear populations in postnatal development and their reactivation in aging muscle. Our datasets also provided a platform for discovery of novel genes associated with rare specialized regions of the muscle cell, including markers of the myotendinous junction and functionally validated factors expressed at the neuromuscular junction. These findings reveal that myonuclei within syncytial muscle fibers possess distinct transcriptional profiles that regulate muscle biology. ### Competing Interest Statement The authors have declared no competing interest.

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