title: Polyethylenimine-mediated expression of transgenes in the acinar cells of rats salivary glands in vivo.
abstract: Non viral-mediated transfection of plasmid DNA provides a fast and reliable way to express various transgenes in selected cell populations in live animals. Here, we show an improvement of a previously published method that is based on injecting plasmid DNA into the ductal system of the salivary glands in live rats. Specifically, using complexes between plasmid DNA and polyethyleneimine (PEI) we show that the expression of the transgenes is directed selectively to the salivary acinar cells. PEI does not affect the ability of cells to undergo regulated exocytosis, which was one of the main drawbacks of the previous methods. Moreover PEI does not affect the proper localization and targeting of transfected proteins, as shown for the apical plasma membrane water channel aquaporin 5 (AQP5). Overall, this approach, coupled with the use of intravital microscopy, permits to conduct localization and functional studies under physiological conditions, in a rapid, reliable, and affordable fashion.
authors.affiliation: Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health Bethesda, MD, USA.
authors.forename: Monika
authors.initials: M
authors.lastname: Sramkova
authors.affiliation: Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health Bethesda, MD, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Parente
authors.affiliation: Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health Bethesda, MD, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Wigand
authors.affiliation: Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health Bethesda, MD, USA.
authors.forename: Myo-Pale'
authors.initials: MP
authors.lastname: Aye
authors.affiliation: Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health Bethesda, MD, USA.
authors.forename: Akiko
authors.initials: A
authors.lastname: Shitara
authors.affiliation: Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health Bethesda, MD, USA.
authors.forename: Roberto
authors.initials: R
authors.lastname: Weigert
keywords: acinar cells; aquaporin 5; in vivo transfection; intravital microscopy; non-viral gene delivery; polyethyleneimine; salivary glands