Main description:

This volume provides key updates on several methods from the first edition as well as including new novel techniques to address the most recent technological developments and their applications in retinal research Chapters guide readers through gene identification approaches, detailed protocols to generate functional retinal pigment epithelium cells, mouse retina and other animal models, fundus imaging and angiography, and cell-based treatment approaches. 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 tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Retinal Degeneration: Methods and Protocols, Second Edition aims to ensure successful scientific work in the further study of this vital field.

Contents:

Part I: Molecular Genetics and Tools

1 Identification and Analysis of Genes Associated with Inherited Retinal Diseases

Mubeen Khan, Zeinab Fadaie, Stephanie S. Cornelis, Frans P.M. Cremers, and Susanne Roosing

2 eQTL Analysis in Retinal Research

Felix Grassmann

3 Testing for known Retinal Degeneration Mutants in Mouse Strains

Khalid Rashid, Katharina Dannhausen, and Thomas Langmann

4 CRISPR/Cas9 Gene Editing In Vitro and in Retinal Cells In Vivo

Daniela Benati, Valeria Marigo, and Alessandra Recchia

5 Monitoring Surface Reactions by Combined Western Blot-ELISA Analysis

Yuchen Lin, Sarah Irmscher, and Christine Skerka

Part II: Cell/Tissue Culture Models

6 Generation of Functional Retinal Pigment Epithelium from Human Induced Pluripotent Stem Cells

Caroline Brandl

7 Advanced Analysis of Photoreceptor Outer Segment Phagocytosis by RPE Cells in Culture

Francesca Mazzoni, Yingyu Mao, and Silvia C. Finnemann

8 Porcine RPE/Choroidal Explant Cultures

Alexa Klettner and Yoko Miura

9 The Mouse Retinal Organoid Trisection Recipe: Efficient Generation of 3D Retinal Tissue from Mouse Embryonic Stem Cells

Manuela Voelkner, Thomas Kurth, and Mike O. Karl

10 Cell Death Analysis in Retinal Cultures

Sarah L Roche, Ana M Ruiz-Lopez, and Thomas G Cotter

11 Fate Mapping In Vivo to Distinguish Bona Fide Microglia Versus Recruited Monocyte-derived Macrophages in Retinal Disease

Nancy J Reyes, Rose Mathew, and Daniel R Saban

Part III: Animal Models

12 Light Damage Models of Retinal Degeneration

Christian Grimm and Charlotte E. Reme

13 Induction and Readout of Oxygen-Iinduced Retinopathy

Raffael Liegl, Claudia Priglinger, and Andreas Ohlmann

14 Generation and Analysis of Xenopus laevis Models of Retinal Degeneration using CRISPR/Cas9

Joanna M. Feehan, Paloma Stanar, Beatrice M. Tam, Colette Chiu, and Orson L. Moritz

15 Gene Knockdown in Zebrafish (Danio rerio) as a Tool to Model Photoreceptor Diseases

Holger Dill and Utz Fischer

16 Drosophila melanogaster - A Valuable Genetic Model Organism to Elucidate the Biology of Retinitis Pigmentosa

Malte Lehmann, Elisabeth Knust, and Sarita Hebbar

Part IV: Retinal Imaging

17 Retinal Fundus Imaging in Mouse Models of Retinal Diseases

Anne F. Alex, Maged Alnawaiseh, Peter Heiduschka, and Nicole Eter

18 Phenotyping of Mouse Models with OCT

G. Alex Ochakovski and M. Dominik Fischer

19 Cell-specific Markers for the Identification of Retinal Cells and Subcellular Organelles by Immunofluorescence Microscopy

Laurie L. Molday, Christiana L. Cheng, and Robert S. Molday

20 Immuno-TEM/STEM in Retinal Research

Sanae Sakami and Krzysztof Palczewski

21 Noninvasive Two-photon Microscopy Imaging of Mouse Retina and Retinal Pigment Epithelium

Grazyna Palczewska, Timothy S. Kern, and Krzysztof Palczewski

Part V: Therapies

22 Cell-based Therapy for Retinal Disease: The New Frontier

Marco Zarbin

23 In Vitro Evaluation of AAV Vectors for Retinal Gene Therapy

Johanna Wagner, Christian Schoen, Elvir Becirovic, Martin Biel, and Stylianos Michalakis

24 Nanoparticles Targeting Retinal and Choroidal Capillaries In Vivo

Alexandra Haunberger and Achim Goepferich

25 Optimized Subretinal Injection Technique for Gene Therapy Approaches

Regine Muhlfriedel, Stylianos Michalakis, Marina Garcia Garrido, Vithiyanjali Sothilingam, Christian Schoen, Martin Biel, and Mathias W. Seeliger