Lineage tracing of nuclei in skeletal myofibers uncovers distinct transcripts and interplay between myonuclear populations

• Published in: Nature communications Vol. 15; no. 1; pp. 9372 - 19
• Main Authors: Sun, Chengyi, Swoboda, Casey O., Morales, Fabian Montecino, Calvo, Cristofer, Petrany, Michael J., Parameswaran, Sreeja, VonHandorf, Andrew, Weirauch, Matthew T., Lepper, Christoph, Millay, Douglas P.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30.10.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 38; 38/91; 45; 631/136/2060/2068; 631/136/83; 631/1647/514/1949; 631/80/642/1363; 64/60; Animals; Cell Differentiation - genetics; Cell fusion; Cell Lineage - genetics; Cell Nucleus - genetics; Cell Nucleus - metabolism; Gene expression; Gene fusion; Gene regulation; Gene sequencing; Humanities and Social Sciences; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; multidisciplinary; Muscle Development - genetics; Muscle Fibers, Skeletal - cytology; Muscle Fibers, Skeletal - metabolism; Muscle, Skeletal - cytology; Muscle, Skeletal - growth & development; Muscle, Skeletal - metabolism; Muscles; Musculoskeletal system; Nuclear fusion; Nuclei; Nuclei (cytology); Populations; Science; Science (multidisciplinary); Skeletal muscle; Stem cells; Stimuli; Tracing; Transcription
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-53510-z

Multinucleated skeletal muscle cells need to acquire additional nuclei through fusion with activated skeletal muscle stem cells when responding to both developmental and adaptive growth stimuli. A fundamental question in skeletal muscle biology has been the reason underlying this need for new nuclei in cells that already harbor hundreds of nuclei. Here we utilize nuclear RNA-sequencing approaches and develop a lineage tracing strategy capable of defining the transcriptional state of recently fused nuclei and distinguishing this state from that of pre-existing nuclei. Our findings reveal the presence of conserved markers of newly fused nuclei both during development and after a hypertrophic stimulus in the adult. However, newly fused nuclei also exhibit divergent gene expression that is determined by the myogenic environment to which they fuse. Moreover, accrual of new nuclei through fusion is required for nuclei already resident in adult myofibers to mount a normal transcriptional response to a load-inducing stimulus. We propose a model of mutual regulation in the control of skeletal muscle development and adaptations, where newly fused and pre-existing myonuclear populations influence each other to maintain optimal functional growth. Skeletal muscle cells contain hundreds of nuclei, but can also add new nuclei in response to various stimuli. Here, the authors perform lineage tracing on the newly fused nuclei showing that these exhibit unique transcriptional states depending on the stimulus.