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This volume discusses different approaches towards understanding the brain plasticity, aging, and regeneration of the nervous system in model organisms. Chapters guide the reader through an overview of model organisms, setup of an aquatic system, Tol2-mediated transgenesis, CRISPR/Cas9-mediated gene knock-out and knock-in, Cas13d-mediated gene knockdown, tissue clearing and expansion methods for brain and spinal cord imaging, Histological techniques, chromatin Immunoprecipitation Sequencing (ChIP-Seq) analysis, single-cell RNA sequencing analysis, BrdU/EdU incorporation assays, brain and spinal cord injury in African killifish, and emerging technologies and future perspectives in African killifish research. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory.
Authoritative and cutting-edge, Emerging Model Organisms aims to be a useful practical guide to researches to help further their study in this field.
List of contents
Establish a fishroom for African turquoise killifish Nothobranchius furzeri.- Gene editing and transgenic reporter assays in African killifish.- Spinal cord injury in African killifish.- Whole brain clearing and immunolabelling in the African killifish N. furzeri.- Studying Mating Behaviors in Nothobranchius furzeri.- A chromatin immunoprecipitation (ChIP) protocol for African turquoise killifish.- EdU labeling of proliferating cells in the African killifish Nothobranchius furzeri.- Immunofluorescence staining on cryosections with high autofluorescence in African killifish.- Rapid visualization of gene expression using chromogenic RNA in situ hybridization in African killifish.- Profiling gene expression in African turquoise killifish Nothobranchius furzeri embryos with RNA fluorescence in situ hybridization chain reaction (HCR).- Cell dissociation of tissues for single-cell analysis in African killifish.- Cryopreservation of the Mexican Tetra: Astyanax mexicanus.- Whole-mount multicolor fluorescent labeling by in situ hybridization in Astyanax mexicanus embryos and larvae.- Application of CRISPR-Cas9 for functional analysis in A. mexicanus.- Immunohistochemistry and in vivo neural imaging in A. mexicanus.- Sleep behavior analysis in Astyanax mexicanus.- A new method to analyze non-visual based social-like interactions in asocial cavefish.- A generalist's guide to quantifying swimming behavior within traditional and non-traditional teleost models.- Recording acoustic behaviour in Astyanax mexicanus fish:acquisition, decryption, interpretation.- Generation of mini-brains from hiPSCs.- Generating Human Pluripotent Stem Cell-Derived AssemBloids(TM) to Model Interneuron Migration and Immune Cell Interactions.- Multiplexed quantitative proteomics analysis of developing human brain organoids.
