Retinal Development

This book details many of the protocols used for a wide range of experiments, including manipulating gene expression in vivo, retinal regeneration and stem cell-based replacement, and ERG (function) recording and non-invasive imaging.

In recent years, there have been major advances in the concepts and methodologies used in the study of retinal development at both cellular and molecular levels. These advanced methodologies have allowed and will continue to allow researchers to gain new insights into the molecular mechanisms underlying retinal development. In Retinal Development: Methods and Protocols, expert researchers in the field detail many of the protocols used for a wide range of experiments. These include protocols and techniques for manipulating gene expression in vivo, tracing cell fates with modernized classic blastomere manipulation in Xenopus and with Cre-based technique in mouse and in zebrafish, retinal regeneration and stem cell-based replacement, and ERG (function) recording and non-invasive imaging. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Retinal Development: Methods and Protocols provides methodologies crucial to the success of increasingly more complex and often challenging investigations in the fields of retinal development and other biological and biomedical research.

Details methodolgies that can be modified and applied to the studies of rential development Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Conditional Control of Gene Expression in the Mouse Retina.-Generation of Transgenic Xenopus Using Restriction Enzyme-Mediated Integration.-In vivo Functional Analysis of Transcription Factor--response Element Interaction using Transgenic Xenopus laevis.-Subretinal Delivery and Electroporation in Pigmented and Nonpigmented Adult Mouse Eyes.-In Ovo Eye Electroporation.-Targeted Microinjection of Synthetic mRNAs to Alter Retina Gene Expression in Xenopus Embryos.-Testing Retina Fate Commitment in Xenopus by Blastomere Deletion, Transplantation and Explant Culture.-Application of Cre-loxP Recombination for Lineage Tracing of Adult Zebrafish Retinal Stem Cells.-Fate Tracing of neurogenin2-expressing Cells in the Mouse Retina using CreERTM - LacZ.-In Vitro Explant Culture and Related Protocols for the Study of Mouse Retinal Development.-In vitro Biochemical Assays to Monitor Rhodopsin Function.-Transfection of Primary Embryonic Chicken Retinal Cells using Cationic Lipid.-Production of High Titer RCAS Retrovirus.-Chick RPE Transdifferentiation Assay for Proneural Activities.-Studying the Generation of Regenerated Retinal Neuron from Müller glia in the Mouse Eye.-Production and Transplantation of Retinal Cells from Human and Mouse Embryonic Stem Cells.-Light-induced Photoreceptor Degeneration in the Retina of the Zebrafish.-Microarray-based Gene Profiling Analysis of Müller Glia-derived Retinal Stem Cells in Light-damaged Retinas from Adult Zebrafish.-Measuring Rodent Electroretinograms to Assess Retinal Function.-Functional Imagingof Retinal Photoreceptors and Inner Neurons using Stimulus-evoked Intrinsic Optical Signals.-Use of Laser Capture Microdissection for Analysis of Retinal mRNA/miRNA Expression and DNA Methylation.-Revealing Looping Organization of Mammalian Photoreceptor Genes Using Chromosome Conformation Capture (3C) Assays.-Retinal Transcriptome Profiling by Directional Next Generation Sequencing Using 100 ng of Total RNA.-Exome Sequencing: Capture and Sequencing of All Human Coding Regions for Disease Gene Discovery.-Determination of Post-translational Modifications of Photoreceptor Differentiation Factor NRL: Focus on SUMOylation.