Germline Development in the Zebrafish

This volume details a wide range of methods, ranging from beginner through advanced, used to

further study zebrafish and fish germline.. Chapter guide readers through cultivating and manipulating germ cells, imaging of germline processes and the molecular analysis of their, protein, and RNA. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Germline Development in the Zebrafish: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Separation of Oocyte and Follicle Layer for Gene Expression Analysis in Zebrafish.- The Xenopus Oocyte as an Expression System for Functional Analyses of Fish Aquaporins.- Computer-Assisted Sperm Analysis to Test Environmental Toxicants.- Cryopreservation and Transplantation of Spermatogonial Stem Cells.- Masculinization of Zebrafish through Partial Depletion of Primordial Germ Cells by Injecting Diluted Morpholino Oligonucleotides into Embryos.- Chemical Genetics: Manipulating the Germline with Small Molecules.- In vitro Induction of Teleost PGCs.- Applying Rho Pathway Inhibitors to Investigate Germ Plasm Localization.- Cryopreservation of Pooled Sperm Samples.- Quantifying Tissue Tension in the Granulosa Layer after Laser Surgery.- In-vivo and Ex-vivo CT Imaging of Zebrafish Gonads.- Live and Time-lapse Imaging of Early Oogenesis and Meiotic Chromosomal Dynamics in Cultured Juvenile Zebrafish Ovaries.- Detection of the Polar Body after Fertilization.- Labelling the Micropylar Cell in Zebrafish Whole-mount and Cryo-sectioned Follicles.- Visualisation of Transcriptional Activity in Early Zebrafish Primordial Germ Cells.- Experimental Methodologies to Visualise Early Cell Biology in Zebrafish Embryos.- Observation of Medaka Larval Gonads by Immunohistochemistry and Confocal Laser Microscopy.- Methods for Visualization of RNA and Cytoskeletal Elements In The Early Zebrafish Embryo.- Glyoxal Fixation as an Alternative for Zebrafish Embryo Immunostaining.- Histological Analysis of Gonads in Zebrafish.- Manipulating and Visualizing the Germline with Transgenic Lines.- Liquid Chromatography and Tandem Mass Spectrometry in Label Free Protein Quantification of Zebrafish (Danio rerio) Eggs.- In vivo Protein Lifetime Measurements across Multiple Organs in the Zebrafish.- In vivo Imaging of Protein Interactions in the Germ Plasm with Bimolecular Fluorescent Complementation.- The Zygotic Genome Activation: Critical Prelude to the Most Important Time of Your Life.- Identification of RNA-binding Protein Landscapes Across Zebrafish Embryonic Transcriptome via iCLIP Approach.- Tethered Function Assay to Study RNA Regulatory Proteins in Zebrafish Embryos.- -Massively Parallel Analysis of Regulatory RNA Sequences.- Exploring Translational Control of Maternal mRNAs in Zebrafish.