title: Stable, conditional, and muscle-fiber-specific expression of electroporated transgenes in chick limb muscle cells.
abstract: Limb muscle formation is spread out over time and, consequently, muscle cells are not easy to target at any particular stages. We aimed to design a technique to study gene function in the different steps of limb muscle formation. We have associated transposon-mediated gene transfer and a tetracycline-dependent activation method with forelimb somite electroporation. In addition, we have established a new vector combining a differentiated-muscle-cell-specific promoter and the transposon system, which allows stable gene expression in limb differentiated muscle cells and not in muscle progenitors. Using these methods, we observed that conditional Fgf4 expression in muscle cells at the onset of fetal muscle differentiation and specific Fgf4 expression in differentiated muscle cells alter muscle fiber formation in chick limbs. Limb somite electroporation with these set of vectors allowing stable, conditional, and differentiated-muscle-specific expression of transgenes opens new perspectives for investigating gene function at various steps of limb muscle formation.
authors.affiliation: CNRS, UMR7622, Biologie Moléculaire et Cellulaire du Développement, Université Pierre et Marie Curie, Paris, France.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Aline
authors.initials: A
authors.lastname: Bonnet
authors.forename: Marie Claire
authors.initials: MC
authors.lastname: Delfini
authors.forename: Koichi
authors.initials: K
authors.lastname: Kawakami
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Delphine
authors.initials: D
authors.lastname: Duprez
mesh_terms: D000818:Animals; D002642:Chick Embryo; D018274:Electroporation / Q000379:methods*; D005121:Extremities / Q000196:embryology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D009132:Muscles / Q000166:cytology* / Q000378:metabolism*; D019170:Somites / Q000166:cytology / Q000378:metabolism; D019076:Transgenes / Q000235:genetics*; D019895:Transposases / Q000235:genetics / Q000378:metabolism
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019895:Transposases