title: Single-cell detection of microRNAs in developing vertebrate embryos after acute administration of a dual-fluorescence reporter/sensor plasmid.
abstract: The detection of microRNAs (miRNAs) at single-cell resolution is important for studying the role of these posttranscriptional regulators. Here, we use a dual-fluorescent green fluorescent protein (GFP)-reporter/monomeric red fluorescent protein (mRFP)-sensor (DFRS) plasmid, injected into zebrafish blastomeres or electroporated into defined tissues of mouse embryos in utero or ex utero, to monitor the dynamics of specific miRNAs in individual live cells. This approach reveals, for example, that in the developing mouse central nervous system, miR-124a is expressed not only in postmitotic neurons but also in neuronal progenitor cells. Collectively, our results demonstrate that acute administration of DFRS plasmids offers an alternative to previous in situ hybridization and transgenic approaches and allows the monitoring of miRNA appearance and disappearance in defined cell lineages during vertebrate development.
authors.affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
authors.forename: Davide
authors.initials: D
authors.lastname: De Pietri Tonelli
authors.forename: Federico
authors.initials: F
authors.lastname: Calegari
authors.forename: Ji-Feng
authors.initials: JF
authors.lastname: Fei
authors.forename: Tadashi
authors.initials: T
authors.lastname: Nomura
authors.forename: Noriko
authors.initials: N
authors.lastname: Osumi
authors.forename: Carl-Philipp
authors.initials: CP
authors.lastname: Heisenberg
authors.forename: Wieland B
authors.initials: WB
authors.lastname: Huttner
mesh_terms: D000818:Animals; D019070:Cell Lineage; D004622:Embryo, Mammalian / Q000737:chemistry*; D004625:Embryo, Nonmammalian; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D051379:Mice; D035683:MicroRNAs / Q000032:analysis*; D018485:Muscle Fibers, Skeletal / Q000737:chemistry; D018482:Muscle, Skeletal / Q000737:chemistry; D009474:Neurons / Q000737:chemistry; D010957:Plasmids; D013234:Stem Cells / Q000737:chemistry; D015027:Zebrafish / Q000196:embryology; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D035683:MicroRNAs; D000097573:Red Fluorescent Protein