title: Conditional labeling of newborn granule cells to visualize their integration into established circuits in hippocampal slice cultures.
abstract: The dentate gyrus continues to produce new granule cells throughout life. Understanding the mechanisms underlying their integration into the pre-existing hippocampal circuitry is of crucial importance. In the present study, we developed an approach allowing visual tracking of newborn granule cells in hippocampal organotypic slices. By crossing neurogenin 2 (Ngn2-CreER) with Cre-reporter mice expressing YFP or GFP reporter genes, it was possible to observe living cells after treating slice cultures with 4-hydroxytamoxifen to induce Cre recombinase activation. Colocalization of GFP with the mitotic marker BrdU demonstrated that the GFP-expressing granule cells were born in vitro. They mature and integrate normally into the hippocampal circuitry, as shown using morphological and electrophysiological techniques. This ex vivo approach therefore offers a highly accessible model to study the effects of long-term treatments on maturation and integration of newborn granule cells.
authors.affiliation: Brain Research Institute, University of Zurich, Zurich, Switzerland. olr21@cam.ac.uk
authors.forename: Olivier
authors.initials: O
authors.lastname: Raineteau
authors.forename: Sylvain
authors.initials: S
authors.lastname: Hugel
authors.forename: Ilknur
authors.initials: I
authors.lastname: Ozen
authors.forename: Lotty
authors.initials: L
authors.lastname: Rietschin
authors.forename: Markus
authors.initials: M
authors.lastname: Sigrist
authors.forename: Silvia
authors.initials: S
authors.lastname: Arber
authors.forename: Beat H
authors.initials: BH
authors.lastname: Gähwiler
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D001973:Bromodeoxyuridine / Q000378:metabolism; D002454:Cell Differentiation; D049108:Cell Enlargement; D049109:Cell Proliferation; D018891:Dentate Gyrus / Q000166:cytology*; D015870:Gene Expression / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D051379:Mice; D008822:Mice, Transgenic; D009415:Nerve Net / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000235:genetics; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000379:methods; D013629:Tamoxifen / Q000494:pharmacology
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D001973:Bromodeoxyuridine; D049452:Green Fluorescent Proteins; D009419:Nerve Tissue Proteins; D013629:Tamoxifen; C102938:Neurog2 protein, mouse