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We used recombination\u2010independent doxycycline\u2010inducible myonucleus\u2010specific fluorescent labelling, tissue RNA\u2010sequencing, myonuclear DNA methylation analysis, multi\u2010omic integration, and single myonucleus RNA\u2010sequencing (smnRNA\u2010seq) to define the molecular characteristics of adult (6\u20138 month) and aged (24 month) murine skeletal muscle after acute mechanical overload (MOV). In adult and aged MOV muscles, we found that: 1) similarities in the transcriptional response to loading\u2014specifically in metabolism genes \u2013 were partly explained by a post\u2010transcriptional microRNA\u2010mediated mechanism that we corroborated using an inducible muscle fiber\u2010specific\n                    <jats:italic>miR\u20101<\/jats:italic>\n                    knockout model, 2) differences in age\u2010dependent transcriptional responses were linked to the magnitude and location of differential DNA methylation in resident myonuclei, specifically around genes such as\n                    <jats:italic>Myc<\/jats:italic>\n                    ,\n                    <jats:italic>Runx1<\/jats:italic>\n                    ,\n                    <jats:italic>Mybph<\/jats:italic>\n                    ,\n                    <jats:italic>Ankrd1<\/jats:italic>\n                    , collagen (\n                    <jats:italic>Col<\/jats:italic>\n                    ) genes, and minichromosome maintenance (\n                    <jats:italic>Mcm<\/jats:italic>\n                    ) genes, 3) adult and aged resident myonuclear transcriptomes had differing enrichment for innervation\u2010related transcripts as well as unique transcriptional profiles in an\n                    <jats:italic>Atf3+<\/jats:italic>\n                    \u201csarcomere assembly\u201d population after MOV, and 4) cellular deconvolution analysis and smnRNA\u2010seq\u00a0supports a role for neuromuscular junction regulation in age\u2010specific hypertrophic adaptation. These data are a roadmap for uncovering molecular targets to enhance aged muscle adaptability.\n                  <\/jats:p>","DOI":"10.1002\/advs.202521633","type":"journal-article","created":{"date-parts":[[2026,2,18]],"date-time":"2026-02-18T06:59:38Z","timestamp":1771397978000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["The Age\u2010Dependent Resident Myonuclear Multi\u2010Omic Response to an Acute Skeletal Muscle Hypertrophic Stimulus in Mice"],"prefix":"10.1002","volume":"13","author":[{"given":"Pieter J.","family":"Koopmans","sequence":"first","affiliation":[{"name":"Cell and Molecular Biology Graduate Program University of Arkansas  Fayetteville Arkansas USA"},{"name":"Molecular Muscle Mass Regulation (M3R) Laboratory, Department Health, Human Performance, &amp; Recreation University of Arkansas  Fayetteville Arkansas 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of Arkansas  Fayetteville Arkansas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas P.","family":"Greene","sequence":"additional","affiliation":[{"name":"Cachexia Research Laboratory, Department Health, Human Performance, &amp; Recreation University of Arkansas  Fayetteville Arkansas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John J.","family":"McCarthy","sequence":"additional","affiliation":[{"name":"Department of Physiology College of Medicine University of Kentucky  Lexington Kentucky USA"},{"name":"Center For Muscle Biology College of Health Sciences University of Kentucky  Lexington Kentucky USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed","family":"Ismaeel","sequence":"additional","affiliation":[{"name":"Department of Anatomy, Physiology, and Pharmacology, College of Medicine Auburn University  Auburn Alabama 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