{"database":"biostudies-arrayexpress","file_versions":[],"scores":null,"additional":{"submitter":["Laurent Gilquin"],"organism":["Drosophila melanogaster"],"software":["Cell Ranger V5.0.0"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/E-MTAB-16520"],"description":["This study investigates how spatial and temporal signaling cues orchestrate the specification of mesodermal progenitors into distinct leg muscle lineages in Drosophila. Using single-cell RNA sequencing across developmental stages, we show that mesodermal precursors initially segregate into central and peripheral populations, defined by specific transcription factor codes such as Lim1, Bi, and Sox100B in central cells, and Hth and Unpg in peripheral cells which prefigure distal and proximal muscle fates, respectively. By computationally and genetically tracking central mesodermal precursors, we demonstrate that these early populations progressively subdivide into distinct muscle lineages, each characterized by unique combinations of transcription factors required for proper muscle development. We further show that the morphogens Wingless (Wg) and Decapentaplegic (Dpp) act in a spatiotemporal manner to control the stepwise evolution of these lineages. By integrating single-cell transcriptomics with spatial mapping and functional genetic analyses, this study establishes that epithelial-derived morphogens coordinate not only epithelial tissue patterning but also the synchronized specification of myoblasts with epithelial patterning, enabling the emergence of diverse leg muscles."],"repository":["biostudies-arrayexpress"],"sample_protocol":["Sample Collection - A FACS sorting was performed to isolate GFP+ cells.","Library Construction - Chromium Single Cell 3' Reagent Kit version v3.1; Single index","Nucleic Acid Extraction - None - Included in nucleic acid library construction protocol.","Sequencing - Illumina NextSeq500 sequencer, Paired-end high sequencing run (28 bp R1, 132 bp R2).","Library Construction - Chromium Single Cell 3' Reagent Kit version v3.1; Dual index (size index 1: 8, size index 2: 10)","Sample Treatment - MPs/myoblasts were labeled with a membrane GFP under the control of mef2-Gal4 transgene.","Sample Collection - Cells were dissociated using a protocol adapted from Harzer et al (doi: 10.1038/nprot.2013.062).","Growth Protocol - Crosses were performed on day 1. An egg collection of 3 hours was made the next day. The flies were raised at 25°C.","Sample Treatment - MPs/myoblasts were labeled with a membrane GFP under the control of 24B-Gal4 enhancer trap transgene."],"figure_sub":["Organization","MINSEQE Score","Assays and Data","Processed Data","MAGE-TAB Files"],"data_protocol":["Sequence Alignment - Reads were aligned with the Cell Ranger v5 pipeline using the Drosophila melanogaster genome (BDGP6.54 release 115) modified to include the GFP.","Data Transformation - Data were analyzed with Seurat toolkit (v4.1.1), using sctransform (v0.3.4) normalization method.","Data Transformation - Data were analyzed with Scanpy toolkit (v1.9.1), using bbknn (v1.5.1) integration method."],"omics_type":["Metabolomics","Unknown","Transcriptomics","Genomics","Proteomics"],"instrument_platform":["BD FACS Aria II","NextSeq 500"],"pubmed_abstract":["<h4>SUMMARY</h4>  Skeletal muscles display remarkable morphological diversity, but the developmental mechanisms specifying distinct muscle morphology remain poorly understood. Using Drosophila leg muscles as a model, we uncover how naïve mesodermal precursors progressively acquire lineage-restricted identities through a stepwise specification program guided by epithelial morphogens. Initially, multipotent mesodermal precursors become spatially and transcriptionally restricted into two broad lineages - proximal and distal - under the combined influence of Wg/Wnt1 and Dpp/BMP signals from the overlying epithelium. By tracing mesodermal precursors that eventually give rise to distal leg muscles, we reveal a second sequence of fate bifurcations that generate distinct muscle subtypes prior to myoblast fusion, as well as a separate lineage producing neuronal lamella cells. Focusing on a single muscle lineage, we show that Wg and Dpp act again during a second phase to control the spatial and temporal deployment of specific transcription factors, ultimately specifying a unique muscle identity. These findings demonstrate that epithelial morphogens not only pattern the epithelium but also orchestrate muscle diversity by promoting stepwise mesodermal specification. Mesodermal precursors translate morphogen signals over time and space to activate distinct transcriptional programs that operate in parallel with the general program of myogenesis, enabling the emergence of distinct muscle and non- muscle lineages whose unique identities underpin their specialized functions."],"study_type":["RNA-seq of coding RNA from single cells"],"species":["Drosophila melanogaster"],"pubmed_title":["Spatio-temporal control of myoblast identity drives muscle diversity in the Drosophila leg"],"pubmed_authors":["Camille Guillermin","Guillermin, C., Tribollet, V., Bouchet, M., Laurencon, A.,  Zhou, D.,  Sarnataro, S., Gilquin, L., Stevant, I., Marcy, G., Texeraud, E., Ghavi-helm, Y., Gillet, B., Hughes, S., Vonau, S., Enriquez, J.","Laurent Gilquin","Jonathan Enriquez"],"additional_accession":[]},"is_claimable":false,"name":"Single-cell RNA-Seq of leg disc mesodermal precursors labeled with GFP at five developmental stages","description":"This study investigates how spatial and temporal signaling cues orchestrate the specification of mesodermal progenitors into distinct leg muscle lineages in Drosophila. Using single-cell RNA sequencing across developmental stages, we show that mesodermal precursors initially segregate into central and peripheral populations, defined by specific transcription factor codes such as Lim1, Bi, and Sox100B in central cells, and Hth and Unpg in peripheral cells which prefigure distal and proximal muscle fates, respectively. By computationally and genetically tracking central mesodermal precursors, we demonstrate that these early populations progressively subdivide into distinct muscle lineages, each characterized by unique combinations of transcription factors required for proper muscle development. We further show that the morphogens Wingless (Wg) and Decapentaplegic (Dpp) act in a spatiotemporal manner to control the stepwise evolution of these lineages. By integrating single-cell transcriptomics with spatial mapping and functional genetic analyses, this study establishes that epithelial-derived morphogens coordinate not only epithelial tissue patterning but also the synchronized specification of myoblasts with epithelial patterning, enabling the emergence of diverse leg muscles.","dates":{"release":"2026-03-17T00:00:00Z","modification":"2026-03-17T10:18:19.257Z","creation":"2026-01-14T11:02:53.428Z"},"accession":"E-MTAB-16520","cross_references":{"ENA":["ERP187563"],"EFO":["EFO_0002944","EFO_0004170","EFO_0003789","EFO_0005684","EFO_0004917","EFO_0005518","EFO_0003816","EFO_0004184","EFO_0003969"],"doi":["10.1101/2025.09.10.675298"]}}