title: Viral gene transfer to developing mouse salivary glands.
abstract: Branching morphogenesis is essential for the formation of salivary glands, kidneys, lungs, and many other organs during development, but the mechanisms underlying this process are not adequately understood. Microarray and other gene expression methods have been powerful approaches for identifying candidate genes that potentially regulate branching morphogenesis. However, functional validation of the proposed roles for these genes has been severely hampered by the absence of efficient techniques to genetically manipulate cells within embryonic organs. Using ex vivo cultured embryonic mouse submandibular glands (SMGs) as models to study branching morphogenesis, we have identified new vectors for viral gene transfer with high efficiency and cell-type specificity to developing SMGs. We screened adenovirus, lentivirus, and 11 types of adeno-associated viruses (AAV) for their ability to transduce embryonic day 12 or 13 SMGs. We identified two AAV types, AAV2 and bovine AAV (BAAV), that are selective in targeting expression differentially to SMG epithelial and mesenchymal cell populations, respectively. Transduction of SMG epithelia with self-complementary (sc) AAV2 expressing fibroblast growth factor 7 (Fgf7) supported gland survival and enhanced SMG branching morphogenesis. Our findings represent, to our knowledge, the first successful selective gene targeting to epithelial vs. mesenchymal cells in an organ undergoing branching morphogenesis.
authors.affiliation: Laboratory of Cell and Developmental Biology, National Institute of Dental and Craniofacial Research, National Institutes of Health, Building 30, Room 426, 30 Convent Drive, MSC 4370, Bethesda, MD 20892-4370, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Hsu
authors.forename: G
authors.initials: G
authors.lastname: Di Pasquale
authors.forename: J S
authors.initials: JS
authors.lastname: Harunaga
authors.forename: T
authors.initials: T
authors.lastname: Onodera
authors.forename: M P
authors.initials: MP
authors.lastname: Hoffman
authors.forename: J A
authors.initials: JA
authors.lastname: Chiorini
authors.forename: K M
authors.initials: KM
authors.lastname: Yamada
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics; D004847:Epithelial Cells / Q000502:physiology; D005240:Feasibility Studies; D051523:Fibroblast Growth Factor 7 / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D049408:Luminescent Agents; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D009024:Morphogenesis / Q000235:genetics; D009924:Organ Culture Techniques; D010957:Plasmids / Q000235:genetics; D012469:Salivary Glands / Q000196:embryology*; D014022:Tissue Survival / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods; D014162:Transfection
chemical_list: C496454:Fgf7 protein, mouse; D049408:Luminescent Agents; D051523:Fibroblast Growth Factor 7; D049452:Green Fluorescent Proteins
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