Neurogenesis in hippocampal slice cultures.
A major challenge in studying neurogenesis in the adult brain is gaining access to neural stem cells for experimental manipulation. We developed an approach utilizing mouse hippocampal organotypic cultures to characterize neurogenesis under controlled conditions. After 2 weeks in culture, double immunostaining using the mitotic marker BrdU and cell type-specific markers revealed persistent proliferation of various cell types. The birth of new neurons was restricted to a third subgranular germinal zone as shown by analysis of the expression pattern of the proneural transcription factor neurogenin-2 and colocalization of BrdU with neuronal phenotypic markers. The regional distribution of newly born neurons closely resembled that observed in vivo in the adult hippocampus. Furthermore, neurogenesis was increased by chronic application of epidermal growth factor (EGF) and abolished by adding serum to the culture medium. Our study therefore establishes the hippocampal slice culture as a promising ex vivo model for investigating neurogenesis.
Raineteau
Olivier
O
Brain Research Institute, University of Zurich, CH-8057 Zurich, Switzerland. rainet@hifo.unizh.ch
Rietschin
Lotty
L
Gradwohl
Gérard
G
Guillemot
François
F
Gähwiler
Beat H
BH
D000818:Animals; D000831:Animals, Newborn; D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D001798:Blood Proteins / Q000494:pharmacology; D001973:Bromodeoxyuridine; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D004815:Epidermal Growth Factor / Q000378:metabolism / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009924:Organ Culture Techniques / Q000379:methods; D010641:Phenotype; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D001798:Blood Proteins; D001973:Bromodeoxyuridine; D004815:Epidermal Growth Factor; D009419:Nerve Tissue Proteins; C102938:Neurog2 protein, mouse