title: Using injectoporation to deliver genes to mechanosensory hair cells.
abstract: Mechanosensation, the transduction of mechanical force into electrochemical signals, allows organisms to detect touch and sound, to register movement and gravity, and to sense changes in cell volume and shape. The hair cells of the mammalian inner ear are the mechanosensors for the detection of sound and head movement. The analysis of gene function in hair cells has been hampered by the lack of an efficient gene transfer method. Here we describe a method termed injectoporation that combines tissue microinjection with electroporation to express cDNAs and shRNAs in mouse cochlear hair cells. Injectoporation allows for gene transfer into dozens of hair cells, and it is compatible with the analysis of hair cell function using imaging approaches and electrophysiology. Tissue dissection and injectoporation can be carried out within a few hours, and the tissue can be cultured for days for subsequent functional analyses.
authors.affiliation: 1] Department of Molecular and Cellular Neuroscience, The Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, California, USA. [2].
authors.forename: Wei
authors.initials: W
authors.lastname: Xiong
authors.affiliation: Department of Molecular and Cellular Neuroscience, The Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wagner
authors.affiliation: Department of Molecular and Cellular Neuroscience, The Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Linxuan
authors.initials: L
authors.lastname: Yan
authors.affiliation: 1] Department of Molecular and Cellular Neuroscience, The Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, California, USA. [2].
authors.forename: Nicolas
authors.initials: N
authors.lastname: Grillet
authors.affiliation: Department of Molecular and Cellular Neuroscience, The Dorris Neuroscience Center, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Müller
mesh_terms: D000818:Animals; D002118:Calcium / Q000032:analysis / Q000378:metabolism; D004210:Dissection / Q000379:methods; D004594:Electrophysiology / Q000379:methods; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D018014:Gene Transfer Techniques / Q000295:instrumentation; D006198:Hair Cells, Auditory / Q000502:physiology*; D007091:Image Processing, Computer-Assisted; D008810:Mice, Inbred C57BL; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D008855:Microscopy, Electron, Scanning; D018408:Patch-Clamp Techniques
chemical_list: D002118:Calcium