title: Single cell electroporation for longitudinal imaging of synaptic structure and function in the adult mouse neocortex in vivo.
abstract: Longitudinal imaging studies of neuronal structures in vivo have revealed rich dynamics in dendritic spines and axonal boutons. Spines and boutons are considered to be proxies for synapses. This implies that synapses display similar dynamics. However, spines and boutons do not always bear synapses, some may contain more than one, and dendritic shaft synapses have no clear structural proxies. In addition, synaptic strength is not always accurately revealed by just the size of these structures. Structural and functional dynamics of synapses could be studied more reliably using fluorescent synaptic proteins as markers for size and function. These proteins are often large and possibly interfere with circuit development, which renders them less suitable for conventional transfection or transgenesis methods such as viral vectors, in utero electroporation, and germline transgenesis. Single cell electroporation (SCE) has been shown to be a potential alternative for transfection of recombinant fluorescent proteins in adult cortical neurons. Here we provide proof of principle for the use of SCE to express and subsequently image fluorescently tagged synaptic proteins over days to weeks in vivo.
authors.affiliation: Department of Basic Neurosciences and The Center for Neuroscience, Centre Médical Universitaire (CMU), University of Geneva Geneva, Switzerland.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Pagès
authors.affiliation: Department of Basic Neurosciences and The Center for Neuroscience, Centre Médical Universitaire (CMU), University of Geneva Geneva, Switzerland.
authors.forename: Michele
authors.initials: M
authors.lastname: Cane
authors.affiliation: Department of Basic Neurosciences and The Center for Neuroscience, Centre Médical Universitaire (CMU), University of Geneva Geneva, Switzerland.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Randall
authors.affiliation: Itopie Informatique, Société Coopérative Geneva, Switzerland.
authors.forename: Luca
authors.initials: L
authors.lastname: Capello
authors.affiliation: Department of Basic Neurosciences and The Center for Neuroscience, Centre Médical Universitaire (CMU), University of Geneva Geneva, Switzerland.
authors.forename: Anthony
authors.initials: A
authors.lastname: Holtmaat
keywords: Single cell electroporation; calcium imaging; dendritic spine; in vivo; long-term imaging