title: Single-cell Microinjection for Cell Communication Analysis.
abstract: Gap junctions are intercellular channels that allow the communication of neighboring cells. This communication depends on the contribution of a hemichannel by each neighboring cell to form the gap junction. In mammalian cells, the hemichannel is formed by six connexins, monomers with four transmembrane domains and a C and N terminal within the cytoplasm. Gap junctions permit the exchange of ions, second messengers, and small metabolites. In addition, they have important roles in many forms of cellular communication within physiological processes such as synaptic transmission, heart contraction, cell growth and differentiation. We detail how to perform a single-cell microinjection of Lucifer Yellow to visualize cellular communication via gap-junctions in living cells. It is expected that in functional gap junctions, the dye will diffuse from the loaded cell to the connected cells. It is a very useful technique to study gap junctions since you can evaluate the diffusion of the fluorescence in real time. We discuss how to prepare the cells and the micropipette, how to use a micromanipulator and inject a low molecular weight fluorescent dye in an epithelial cell line.
authors.affiliation: Institute Oswaldo Cruz, Laboratory of Cellular Communication, Oswaldo Cruz Foundation.
authors.forename: Anael Viana Pinto
authors.initials: AV
authors.lastname: Alberto
authors.affiliation: Institute Oswaldo Cruz, Laboratory of Cellular Communication, Oswaldo Cruz Foundation.
authors.forename: André G
authors.initials: AG
authors.lastname: Bonavita
authors.affiliation: Institute Oswaldo Cruz, Laboratory of Cellular Communication, Oswaldo Cruz Foundation.
authors.forename: Antonio A
authors.initials: AA
authors.lastname: Fidalgo-Neto
authors.affiliation: Institute Oswaldo Cruz, Laboratory of Cellular Communication, Oswaldo Cruz Foundation.
authors.forename: Filipe
authors.initials: F
authors.lastname: Berçot
authors.affiliation: Institute Oswaldo Cruz, Laboratory of Cellular Communication, Oswaldo Cruz Foundation; alveslaa@gmail.com.
authors.forename: Luiz A
authors.initials: LA
authors.lastname: Alves
mesh_terms: D000818:Animals; D001692:Biological Transport; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005456:Fluorescent Dyes / Q000008:administration & dosage / Q000493:pharmacokinetics; D017629:Gap Junctions / Q000502:physiology; D007546:Isoquinolines / Q000008:administration & dosage* / Q000493:pharmacokinetics; D051379:Mice; D008845:Microinjections; D008856:Microscopy, Fluorescence; D023421:Models, Animal
chemical_list: D005456:Fluorescent Dyes; D007546:Isoquinolines; C017475:lucifer yellow