Combining BrdU-Labeling to Detection of Neuronal Markers to Monitor Adult Neurogenesis in Hydra

Article Article scientifique Article Combining BrdU-Labeling to Detection of Neuronal Markers to Monitor Adult Neurogenesis in Hydra eng 2020 author Buzgariu Wanda Christa 642187 author Curchod Marie-Laure 22802 author Perruchoud Chrystelle 676031 author Galliot Brigitte 0000-0001-7596-8284 29979 3-24 10.1007/978-1-4939-9732-9_1 31552646 1064-3745 https://link.springer.com/protocol/10.1007/978-1-4939-9732-9_1 The nervous system is produced and maintained in adult Hydra through the continuous production of nerve cells and mechanosensory cells (nematocytes or cnidocytes). De novo neurogenesis occurs slowly in intact animals that replace their dying nerve cells, at a faster rate in animals regenerating their head as a complete apical nervous system is built in few days. To dissect the molecular mechanisms that underlie these properties, a precise monitoring of the markers of neurogenesis and nematogenesis is required. Here we describe the conditions for an efficient BrdU-labeling coupled to an immunodetection of neuronal markers, either regulators of neurogenesis, here the homeoprotein prdl-a, or neuropeptides such as RFamide or Hym-355. This method can be performed on whole-mount animals as well as on macerated tissues when cells retain their morphology. Moreover, when antibodies are not available, BrdU-labeling can be combined with the analysis of gene expression by whole-mount in situ hybridization. This co-immunodetection procedure is well adapted to visualize and quantify the dynamics of de novo neurogenesis. Upon continuous BrdU labeling, the repeated measurements of BrdU-labeling indexes in specific cellular populations provide a precise monitoring of nematogenesis as well as neurogenesis, in homeostatic or developmental conditions. Hydra nervous system Interstitial stem cells Neurogenesis Nematogenesis In situ Hybridization Immunofluorescence Hydroxyurea BrdU Prdl-a Hym-355 RFamide Dewey 590 Swiss National Science Foundation Regulation of stem cell plasticity in Hydra 31003A_149630 CH Project funding Swiss National Science Foundation grant 31003_169930 CH 44 Département de génétique et évolution 162 false 821 Groupe Galliot Methods in Molecular Biology 2047 6d00c6a1-bbbb-4f4b-bb59-568aeae4f231 Article (Accepted version) Public 2020_Buzgariu_Methods_Mol_Biol_author_format(3).pdf 3156143 application/pdf Restricted 2020-10-06 openAccess Faculté des sciences / Section de biologie / Département de génétique et évolution

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Combining BrdU-Labeling to Detection of Neuronal Markers to Monitor Adult Neurogenesis in Hydra	Methods in Molecular Biology			2020	510	235

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Curchod, Marie-Laure