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Article Dans Une Revue Journal of visualized experiments : JoVE Année : 2023

Studying Muscle Transcriptional Dynamics at Single-molecule Scales in Drosophila

Résumé

Skeletal muscles are large syncytia made up of many bundled myofibers that produce forces and enable body motion. Drosophila is a classical model to study muscle biology. The combination of both Drosophila genetics and advanced omics approaches led to the identification of key conserved molecules that regulate muscle morphogenesis and regeneration. However, the transcriptional dynamics of these molecules and the spatial distribution of their messenger RNA within the syncytia cannot be assessed by conventional methods. Here we optimized an existing singlemolecule RNA fluorescence in situ hybridization (smFISH) method to enable the detection and quantification of individual mRNA molecules within adult flight muscles and their muscle stem cells. As a proof of concept, we have analyzed the mRNA expression and distribution of two evolutionary conserved transcription factors, Mef2 and Zfh1/Zeb. We show that this method can efficiently detect and quantify single mRNA molecules for both transcripts in the muscle precursor cells, adult muscles, and muscle stem cells. © 2023 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.
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Dates et versions

hal-04260328 , version 1 (26-10-2023)

Identifiants

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Emma Leroux, Nourhene Ammar, Savannah Moinet, Thierry Pecot, Hadi Boukhatmi. Studying Muscle Transcriptional Dynamics at Single-molecule Scales in Drosophila. Journal of visualized experiments : JoVE, 2023, 199, ⟨10.3791/65713⟩. ⟨hal-04260328⟩
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