Retinal Development: Methods and Protocols

ISBN-13: 9781617798474 / Angielski / Twarda / 2012 / 367 str.

Shu-Zhen Wang

Opis

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.

Spis treści

Part I: Manipulation of Gene Expression in vivo

 1.  Conditional Control of Gene Expression in the Mouse Retina

Qian Ding and Lin Gan

 2. Generation Transgenic Xenopus using Restriction Enzyme-Mediated Intergration

Mohammad Haeri and Barry E. Knox

3. In vivo Functional Analysis of Transcription Factor--response Element Interaction using Transgenic Xenopus laevis

Heithem M. El-Hodiri, Yi Pan, and Lisa E. Kelly

4. Subretinal Delivery and Electroporation in Pigmented and Nonpigmented Adult Mouse Eyes

John M. Nickerson, Penny Goodman, Micah A. Chrenek, Christiana J. Johnson, Lennart Berglin, T. Michael Redmond, Jeffrey H. Boatright

5. In Ovo Eye Electroporation

Teri L. Belecky-Adams, Scott R. Hudson, and Sarika Tiwari

6. Targeted Microinjection of Synthetic mRNAs to Alter Retina Gene Expression in Xenopus Embryos

 Sally A. Moody

 Also see Chapter 8.

Part II: Tracing Cell Fate

7. Testing Retina Fate Commitment in Xenopus by Blastomere Deletion, Transplantation and Explant Culture

 Sally A. Moody

 8. Application of Cre-loxP Recombination for Lineage Tracing of Adult Zebrafish Retinal Stem Cells

Rajesh Ramachandran, Aaron Reifler, Jin Wan, and Daniel Goldman

 9. Fate Tracing of neurogenin2-expressing Cells in the Mouse Retina using CreER^TM - LacZ

Wenxin Ma and Shu-Zhen Wang

Part III: In vitro Systems

10. In Vitro Explant Culture and Related Protocols for the Study of Mouse Retinal Development

Kangxin Jin and Mengqing Xiang

11. In vitro Biochemical Assays to Monitor Rhodopsin Function

Joshua Sammons and Alecia K. Gross

12. Transfection of Primary Embryonic Chicken Retinal Cells using Cationic Lipid

Yi-Wen Hsieh and Xian-Jie Yang

13. Production of High Titer RCAS Retrovirus

 Run-Tao Yan and Shu-Zhen Wang

14. Chick RPE Transdifferentiation Assay for Proneural Activities

Shu-Zhen Wang and Run-Tao Yan

Part IV: Regeneration/Stem Cells

15. Studying the Generation of Regenerated Retinal Neuron from Müller glia in the Mouse Eye

Mike O. Karl and Thomas A. Reh

16. Production and Transplantation of Retinal Cells from Human and Mouse Embryonic Stem Cells

Anna La Torre, Deepak A Lamba, Anu Jayabalu, Thomas A Reh

17. Light-induced Photoreceptor Degeneration in the Retina of the Zebrafish

Scott Taylor, Jing Chen, Jing Luo and Peter Hitchcock

18. Microarray-based Gene Profiling Analysis of Müller Glia-derived Retinal Stem Cells in Light-damaged Retinas from Adult Zebrafish

Zhao Qin and Pamela A. Raymond

Also see Chapter 8.

 Part V: Function/Imaging

 19: Measuring Rodent Electroretinograms to Assess Retinal Function

Molly E. Clark and Timothy W. Kraft

 20. Functional Imaging of Retinal Photoreceptors and Inner Neurons using Stimulus-evoked Intrinsic Optical Signals

Xin-Cheng Yao and Yi-Chao Li

Part VI: Emerging Methodologies

21. Use of Laser Capture Microdissection for Analysis of Retinal mRNA/miRNA Expression and DNA Methylation

Laszlo Hackler Jr, Tomohiro Masuda, Verity F. Oliver, Shannath L. Merbs,  and Donald J.Zack

22. Revealing Looping Organization of Mammalian Photoreceptor Genes Using Chromosome Conformation Capture (3C) Assays

Guang-Hua Peng and Shiming Chen

23. Retinal Transcriptome Profiling by Directional Next Generation Sequencing Using 100 ng of Total RNA

Matthew John Brooks, Harsha Karur Rajasimha, and Anand Swaroop

24. Exome Sequencing: Capture and Sequencing of All Human Coding Regions for Disease Gene Discovery

Rinki Ratna Priya, Harsha Karur Rajasimha, Matthew Brooks, and Anand Swaroop

 25. Determination of Post-translational Modifications of Photoreceptor Differentiation Factor NRL: Focus on SUMOylation

Jerome E. Roger, Jacob Nellissery and Anand Swaroop