Systems neuroscienceIdentification and characterization of two neurogenic zones in interface organotypic hippocampal slice cultures
Section snippets
OHC
Organotypic hippocampal cultures were prepared according to the standard interface method (Stoppini et al., 1991). Hippocampal slices were prepared from postnatal days 7–9 (P7-9) Wistar rats (Harlan Winkelmann GmbH, Borchen, Germany). After decapitation, the brain was removed, hippocampi were dissected and transversely sliced into a 350μm thickness on a McIlwain tissue chopper (The Mickle Laboratory Engineering Co., Guildford, UK). The slices were then carefully separated with two pairs of fine
Identification and in situ distribution of CNS cell types in OHC
OHC were prepared from P7-9 Wistar rats and cultivated for two weeks before experiments were performed. For the general experimental setup see Fig. 1A. To characterize the three-dimensional structure of OHC, different cell types were detected by immunohistochemistry in unsectioned (“whole mounts”) and cross-sectioned 14 DIV slices. Astrocytes (GFAP+), precursor cells (Nestin+), migrating neuroblasts (DCX+), young and mature neurons (β-III tubulin+), mature neurons (NeuN+) and microglia (IB4+)
Discussion
The aim of this work was to investigate the early neurogenesis in vitro in interface OHC. Hippocampal slices were prepared from P7-9 rats and maintained in vitro for 14 d before the proliferating cells were labeled with BrdU.
The initial trauma during preparation and subsequent cultivation stimulated astrocytes to proliferate and form a glial barrier ensheathing the slice (GE). The presence of activated astrocytes expressing Nestin and activated microglia indicated a protective function of the
Acknowledgments
Work was supported by graduate program “Biologische Grundlagen von Erkrankungen des Nervensystems,” Medizinische Fakultät, Otto-von Guericke-Universität, Magdeburg and European Grant QLK3-CT-2001-00407.
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Neurogenesis: A process ontogenically linked to brain cavities and their content, CSF
2020, Seminars in Cell and Developmental BiologyCitation Excerpt :Neural precursors in the adult brain dentate gyrus are a heterogeneous population, which includes developmental radial glia cells (Xu et al. 2015; Berg et al. 2018). The hippocampal neurogenic niche preserves its ability to respond to CSF [86] and consequently to respond to embryonic stimulus like FGF2 [87]. The original neural precursor population close to the ventricular system named Sub-ependymal zone (SEZ) remains in the adult with intense replicating activity and maintains pluripotentiality, being considered the primitive neurogenic niche incapable of incorporating new precursors to the dentate gyrus, meanwhile in the SGZ, precursors display reduced mitogenic activity and are considered Neuronal restricted precursors.
Evidence for the contribution of BDNF-TrkB signal strength in neurogenesis: An organotypic study
2015, Neuroscience LettersCitation Excerpt :In the present study, we used long-term hippocampal organotypic cultures as an ex vivo system for the study of neurogenesis. Application of hippocampal organotypic slice cultures for the story of neurogenesis is relatively rare [14,15,17–21]. The pattern of maturation of newly born neurons in the OHSC is comparable to the one described in vivo [14].
Enhanced neurogenesis in organotypic cultures of rat hippocampus after transient subfield-selective excitotoxic insult induced by domoic acid
2012, NeuroscienceCitation Excerpt :The decrease in the percentage of damaged and dead cells is presumably because of removal as evidenced by microglial activation, and was paralleled by a significantly increased incorporation of the proliferation marker BrdU into the SGZ of the dentate gyrus and the CA1 subfield, a result that shows clearly that moderate excitotoxicity in OHSC stimulates a robust and sustained proliferative response within the injured hippocampal subfields. At least four possible sources of progenitor cells in the hippocampus have been described: (1) ongoing proliferation of neuronal precursor cells takes place up to adulthood in the SGZ of the dentate gyrus (Kaplan and Hinds, 1977; Cameron et al., 1993), and new neurons can be generated within the SGZ and even migrate to CA1 and CA3 regions (Daval et al., 2004); (2) the CA1 itself has been shown as an origin of dividing cells (Nakatomi et al., 2002); (3) cells migrating from the periventricular zone near the hippocampal formation have been reported to contribute to replacement of neurons of the CA1 region (Daval et al., 2004; Chechneva et al., 2005), and finally, (4) proliferating astroglia and microglia are known to incorporate BrdU during their S-phase. Our data do not support proliferation of astroglia as the explanation for enhanced incorporation of BrdU, although DOM neurotoxicity has been reported to induce astrogliosis in vivo in rodents after acute exposure (Appel et al., 1997; Ananth et al., 2003) and rapid changes in GFAP staining in the CNS have been observed after excitotoxic lesions (Eriksdotter-Nilsson et al., 1987).
Synaptic integration of newly generated neurons in rat dissociated hippocampal cultures
2011, Molecular and Cellular NeuroscienceCitation Excerpt :The continued occurrence of neurogenesis in vitro in hippocampal slice cultures (Kamada et al., 2004; Raineteau et al., 2004) offers advantages for studying the regulation of neurogenesis by various drugs or growth factors. Using this approach it has been shown that serum (Raineteau et al., 2004) and pilocarpine-induced seizure activity (Poulsen et al., 2005) inhibit neurogenesis whereas growth factors (Raineteau et al., 2004; Chechneva et al., 2005; Poulsen et al., 2005) and glutamate receptor antagonists (Poulsen et al., 2005) increase the generation of neurons by slice cultures. The dendrites and axons of newborn neurons in slice cultures have also been examined in detail along with mEPSCs and mIPSCs, showing that new neurons can integrate and mature normally in vitro (Raineteau et al., 2006).