Conditional labeling of newborn granule cells to visualize their integration into established circuits in hippocampal slice cultures.
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.
Raineteau
Olivier
O
Brain Research Institute, University of Zurich, Zurich, Switzerland. olr21@cam.ac.uk
Hugel
Sylvain
S
Ozen
Ilknur
I
Rietschin
Lotty
L
Sigrist
Markus
M
Arber
Silvia
S
Gähwiler
Beat H
BH
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
D051792:Basic Helix-Loop-Helix Proteins; D001973:Bromodeoxyuridine; D049452:Green Fluorescent Proteins; D009419:Nerve Tissue Proteins; D013629:Tamoxifen; C102938:Neurog2 protein, mouse