Neurogenesis plays a role in many physiological (memory formation) and pathological (stroke, depression) processes. However the mechanisms of postnatal stem cell proliferation and neurogenesis are still poorly understood. We characterized early neurogenesis in vitro in rat organotypic hippocampal slice cultures. Proliferation was assessed by bromodeoxyuridine incorporation, neurogenesis by bromodeoxyuridine-double labeling with doublecortin or beta-III tubulin. We showed for the first time that in addition to the dentate gyrus organotypic hippocampal slice cultures include a second neurogenic zone: the posterior periventricle, which is a part of the lateral ventricle wall. This structure lining the stratum oriens contained Nestin+ precursors. We could identify morphological and functional differences between dentate gyrus and posterior periventricle precursor populations. Our data demonstrate that basic fibroblast growth factor treatment induced a fast but short-lasting neurogenic response in the dentate gyrus while the posterior periventricle showed a more pronounced and long lasting neurogenic effect of basic fibroblast growth factor. Thus two neurogenic zones with different neurogenic properties were identified in organotypic hippocampal slice cultures.