{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T13:47:16Z","timestamp":1790948836349,"version":"4.1.0"},"reference-count":47,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,3,3]],"date-time":"2025-03-03T00:00:00Z","timestamp":1740960000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)"},{"name":"Open Access Publishing Fund of Philipps-Universit\u00e4t Marburg"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>MicroRNAs function as post-transcriptional regulators in gene expression and control a broad range of biological processes in metazoans. The formation of multinucleated muscles is essential for locomotion, growth, and muscle repair. microRNAs have also emerged as important regulators for muscle development and function. In order to identify new microRNAs required for muscle formation, we have performed a large microRNA overexpression screen. We screened for defects during embryonic and adult muscle formation. Here, we describe the identification of mir-276a as a regulator for muscle migration during testis formation. The mir-276a overexpression phenotype in testis muscles resembles the loss-of-function phenotype of heartless. A GFP sensor assay reveals that the 3\u2032UTR of heartless is a target of mir-276a. Furthermore, we found that mir-276a is essential for the proper development of indirect flight muscles and describe a method for determining the number of nuclei for each of the six longitudinal muscle fibers (DLMs), which are part of the indirect flight muscles.<\/jats:p>","DOI":"10.3390\/cells14050368","type":"journal-article","created":{"date-parts":[[2025,3,3]],"date-time":"2025-03-03T09:04:49Z","timestamp":1740992689000},"page":"368","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["mir-276a Is Required for Muscle Development in Drosophila and Regulates the FGF Receptor Heartless During the Migration of Nascent Myotubes in the Testis"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2931-7717","authenticated-orcid":false,"given":"Mathieu","family":"Preu\u00dfner","sequence":"first","affiliation":[{"name":"Department of Biology, Developmental Biology, Philipps University Marburg, Karl-von-Frisch Str. 8, 35037 Marburg, Germany"},{"name":"Department of Developmental Biology of Vertebrates, Institute of Cell Biology and Neuroscience, Goethe University Frankfurt am Main, 60438 Frankfurt am Main, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maik","family":"Bischoff","sequence":"additional","affiliation":[{"name":"Department of Biology, Developmental Biology, Philipps University Marburg, Karl-von-Frisch Str. 8, 35037 Marburg, Germany"},{"name":"Department of Biology, University of North Carolina at Chapel Hill, CB#3280, Chapel Hill, NC 27599-3280, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3081-477X","authenticated-orcid":false,"given":"Susanne Filiz","family":"\u00d6nel","sequence":"additional","affiliation":[{"name":"Department of Biology, Developmental Biology, Philipps University Marburg, Karl-von-Frisch Str. 8, 35037 Marburg, Germany"},{"name":"Department of Biology, Molecular Embryology, Philipps University Marburg, Karl-von-Frisch Str. 8, 35037 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1481","DOI":"10.1126\/science.272.5267.1481","article-title":"Twist: A myogenic switch in Drosophila","volume":"272","author":"Baylies","year":"1996","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2373","DOI":"10.1101\/gad.9.19.2373","article-title":"Lethal of scute, a proneural gene, participates in the specification of muscle progenitors during Drosophila embryogenesis","volume":"9","author":"Carmena","year":"1995","journal-title":"Genes Dev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1242\/dev.112.3.755","article-title":"The embryonic origin of imaginal discs in Drosophila","volume":"112","author":"Bate","year":"1991","journal-title":"Development"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.semcdb.2020.02.009","article-title":"Muscle development: A view from adult myogenesis in Drosophila","volume":"104","author":"Laurichesse","year":"2020","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/S0092-8674(00)00024-6","article-title":"Drosophila dumbfounded: A myoblast attractant essential for fusion","volume":"102","author":"Coutts","year":"2000","journal-title":"Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4229","DOI":"10.1242\/dev.128.21.4229","article-title":"rst and its paralogue kirre act redundantly during embryonic muscle development in Drosophila","volume":"128","author":"Bonengel","year":"2001","journal-title":"Development"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4489","DOI":"10.1242\/dev.128.22.4489","article-title":"Drosophila Lame duck, a novel member of the Gli superfamily, acts as a key regulator of myogenesis by controlling fusion competent myoblast development","volume":"128","author":"Duan","year":"2001","journal-title":"Development"},{"key":"ref_8","first-page":"133","article-title":"Minc function in Drosophila myogenesis","volume":"129","author":"Coutts","year":"2002","journal-title":"Development"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Rout, P., Preu\u00dfner, M., and \u00d6nel, S.F. (2022). Drosophila melanogaster: A Model System to Study Distinct Genetic Programs in Myoblast Fusion. Cells, 11.","DOI":"10.3390\/cells11030321"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Boukhatmi, H. (2021). Drosophila, an Integrative Model to Study the Features of Muscle Stem Cells in Development and Regeneration. Cells, 10.","DOI":"10.3390\/cells10082112"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/S0092-8674(02)00744-4","article-title":"Sex-specific deployment of FGF signaling in Drosophila recruits mesodermal cells into the male genital imaginal disc","volume":"109","author":"Ahmad","year":"2002","journal-title":"Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1016\/j.ydbio.2012.07.022","article-title":"Multinucleated smooth muscles and mononucleated as well as multinucleated striated muscles develop during establishment of the male reproductive organs of Drosophila melanogaster","volume":"370","author":"Kuckwa","year":"2012","journal-title":"Dev. Biol."},{"key":"ref_13","first-page":"329","article-title":"A new level of plasticity: Drosophila smooth-like testes muscles compensate failure of myoblast fusion","volume":"143","author":"Kuckwa","year":"2016","journal-title":"Development"},{"key":"ref_14","first-page":"1876","article-title":"Myotube migration to cover and shape the testis of Drosophila depends on Heartless, Cadherin\/Catenin, and myosin II","volume":"6","author":"Fritzen","year":"2017","journal-title":"Biol. Open"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1038\/s41467-020-20362-2","article-title":"Filopodia-based contact stimulation of cell migration drives tissue morphogenesis","volume":"12","author":"Bischoff","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Bischoff, M.C., Norton, J.E., Munguia, E.A., Clark, S.E., Gurley, N.J., Korankye, R., Gyabaah, E.A., Encarnacion, T., Serody, C.J., and Jones, C.D. (2025). A large reverse-genetic screen identifies numerous regulators of testis nascent myotube collective cell migration and collective organ sculpting. Mol. Biol. Cell, 36.","DOI":"10.1091\/mbc.E24-10-0456"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1016\/j.semcdb.2017.11.017","article-title":"Drosophila adult muscle development and regeneration","volume":"72","author":"Gunage","year":"2017","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1135","DOI":"10.1083\/jcb.141.5.1135","article-title":"Patterning Muscles Using Organizers: Larval Muscle Templates and Adult Myoblasts Actively Interact to Pattern the Dorsal Longitudinal Flight Muscles of Drosophila","volume":"141","author":"Roy","year":"1998","journal-title":"J. Cell Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1361","DOI":"10.1242\/dev.125.8.1361","article-title":"Twist and Notch negatively regulate adult muscle differentiation in Drosophila","volume":"125","author":"Anant","year":"1998","journal-title":"Development"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/j.ydbio.2006.03.022","article-title":"Notch pathway repression by vestigial is required to promote indirect flight muscle differentiation in Drosophila melanogaster","volume":"295","author":"Bernard","year":"2006","journal-title":"Dev. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e03126","DOI":"10.7554\/eLife.03126","article-title":"Identification of a new stem cell population that generates Drosophila flight muscles","volume":"3","author":"Gunage","year":"2014","journal-title":"eLife"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ydbio.2020.05.009","article-title":"FGF signaling promotes myoblast proliferation through activation of wingless signaling","volume":"464","author":"Vishal","year":"2020","journal-title":"Dev. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4040","DOI":"10.1242\/dev.077495","article-title":"Bidirectional Notch activation represses fusion competence in swarming adult Drosophila myoblasts","volume":"139","author":"Gildor","year":"2012","journal-title":"Development"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.cell.2018.03.006","article-title":"Metazoan MicroRNAs","volume":"173","author":"Bartel","year":"2018","journal-title":"Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2007","DOI":"10.1002\/jcp.25854","article-title":"Interplay between miRNAs and human diseases","volume":"233","author":"Paul","year":"2018","journal-title":"J. Cell. Physiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"730","DOI":"10.1101\/gad.9.6.730","article-title":"Drosophila MEF2, a transcription factor that is essential for myogenesis","volume":"9","author":"Bour","year":"1995","journal-title":"Genes. Dev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1126\/science.7839146","article-title":"Requirement of MADS domain transcription factor D-MEF2 for muscle formation in Drosophila","volume":"267","author":"Lilly","year":"1995","journal-title":"Science"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1006\/dbio.1996.9987","article-title":"Wingless signaling induces Nautilus expression in the ventral mesoderm of the Drosophila embryo","volume":"176","author":"Ranganayakulu","year":"1996","journal-title":"Dev. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"40171","DOI":"10.1074\/jbc.M110.139337","article-title":"WAVE forms hetero- and homo-oligomeric complexes at integrin junctions in Drosophila visualized by bimolecular fluorescence complementation","volume":"285","author":"Gohl","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1387\/ijdb.8735928","article-title":"Muscle development and attachment to the epidermis is accompanied by expression of beta 3 and beta 1 tubulin isotypes, respectively","volume":"40","author":"Buttgereit","year":"1996","journal-title":"Int. J. Dev. Biol."},{"key":"ref_31","first-page":"e2438","article-title":"Paraffin-Embedded and Frozen Sections of Drosophila Adult Muscles","volume":"46","author":"Kucherenko","year":"2010","journal-title":"J. Vis. Exp."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.ymeth.2014.02.037","article-title":"A guide to study Drosophila muscle biology","volume":"68","author":"Weitkunat","year":"2014","journal-title":"Methods"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1543","DOI":"10.1534\/genetics.112.145383","article-title":"Functional characterization of Drosophila microRNAs by a novel in vivo library","volume":"192","author":"Schertel","year":"2012","journal-title":"Genetics"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2179","DOI":"10.1126\/science.286.5447.2179","article-title":"Microarray analysis of Drosophila development during metamorphosis","volume":"286","author":"White","year":"1999","journal-title":"Science"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1242\/dev.113.1.67","article-title":"Development of the indirect flight muscles of Drosophila","volume":"113","author":"Fernandes","year":"1991","journal-title":"Development"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1016\/j.ydbio.2005.09.007","article-title":"Transcription of Myocyte enhancer factor-2 in adult Drosophila myoblasts is induced by the steroid hormone ecdysone","volume":"288","author":"Lovato","year":"2005","journal-title":"Dev. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1038\/s41576-023-00611-y","article-title":"microRNAs in action: Biogenesis, function and regulation","volume":"24","author":"Shang","year":"2023","journal-title":"Nat. Rev. Genet."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Panni, S., Lovering, R.C., Porras, P., and Orchard, S. (2020). Non-coding RNA regulatory networks. Biochim. Biophys. Acta Gene Regul. Mech., 1863.","DOI":"10.1016\/j.bbagrm.2019.194417"},{"key":"ref_39","first-page":"E2965","article-title":"mir-276a strengthens Drosophila circadian rhythms by regulating timeless expression","volume":"113","author":"Chen","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","first-page":"e21222","article-title":"Regulatory mechanism of daily sleep by miR-276a","volume":"35","author":"Zhang","year":"2021","journal-title":"FASEB J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"5821","DOI":"10.1523\/JNEUROSCI.4004-12.2013","article-title":"MicroRNA-276a functions in ellipsoid body and mushroom body neurons for naive and conditioned olfactory avoidance in Drosophila","volume":"33","author":"Li","year":"2013","journal-title":"J. Neurosci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"jkac239","DOI":"10.1093\/g3journal\/jkac239","article-title":"Drosophila FMRP controls miR-276-mediated regulation of nejire mRNA for space-filling dendrite development","volume":"12","author":"Li","year":"2022","journal-title":"G3 Genes|Genomes|Genetics"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1038\/nsmb.1409","article-title":"The regulatory activity of microRNA* species has substantial influence on microRNA and 3\u2032 UTR evolution","volume":"15","author":"Okamura","year":"2008","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"4519","DOI":"10.1096\/fj.201701185RR","article-title":"Ecdysone-responsive microRNA-252-5p controls the cell cycle by targeting Abi in Drosophila","volume":"32","author":"Lim","year":"2018","journal-title":"FASEB J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1111\/imb.12872","article-title":"Ecdysone-induced microRNA miR-276a-3p controls developmental growth by targeting the insulin-like receptor in Drosophila","volume":"32","author":"Lee","year":"2023","journal-title":"Insect Mol. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1438","DOI":"10.1016\/j.cub.2024.02.056","article-title":"Inter-organ steroid hormone signaling promotes myoblast fusion via direct transcriptional regulation of a single key effector gene","volume":"34","author":"Ruan","year":"2024","journal-title":"Curr. Biol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1006\/nbdi.2002.0510","article-title":"Novel features of dFMR1, the Drosophila orthologue of the fragile X mental retardation protein","volume":"11","author":"Schenck","year":"2002","journal-title":"Neurobiol. Dis."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/5\/368\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:46:04Z","timestamp":1760028364000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/5\/368"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,3]]},"references-count":47,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["cells14050368"],"URL":"https:\/\/doi.org\/10.3390\/cells14050368","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,3]]}}}