Main description:

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.

Contents:

Part I: African killifish (Nothobranchius furzeri)

1. Establish a fishroom for African turquoise killifish Nothobranchius furzeri

Michelle B. Vilensky and Chi-Kuo Hu

2. Gene editing and transgenic reporter assays in African killifish

Wei Wang

3. Spinal cord injury in African killifish

Yufei Lou and Wei Wang

4. Whole brain clearing and immunolabelling in the African killifish N. furzeri

Augusto Ortega Granillo, Fengyan Deng, Hannah Wilson, Yongfu Wang, and Alejandro Sanchez Alvarado

5. Studying Mating Behaviors in Nothobranchius furzeri

Bernadette D. Johnson and Adam G. Jones

6. A chromatin immunoprecipitation (ChIP) protocol for African turquoise killifish

Jiaxing Jin and Deqing Hu

7. EdU labeling of proliferating cells in the African killifish Nothobranchius furzeri

Augusto Ortega Granillo, Fengyan Deng, Yongfu Wang, Wei Wang, Alejandro Sanchez Alvarado

8. Immunofluorescence staining on cryosections with high autofluorescence in African killifish

Shiqi Liu and Wei Wang

9. Rapid visualization of gene expression using chromogenic RNA in situ hybridization in African killifish

Mengjun Liu, Shiqi Liu, and Wei Wang

10. Profiling gene expression in African turquoise killifish Nothobranchius furzeri embryos with RNA fluorescence in situ hybridization chain reaction (HCR)

Lu Jia and Chi-Kuo Hu

11. Cell dissociation of tissues for single-cell analysis in African killifish

Liping Zhang and Wei Wang

Part II: Mexican cavefish (Astyanax mexicanus)

12. Cryopreservation of the Mexican Tetra: Astyanax mexicanus

Diana P. Baumann and Andrew Ingalls

13. Whole-mount multicolor fluorescent labeling by in situ hybridization in Astyanax mexicanus embryos and larvae

Maryline Blin, Sylvie Retaux, and Jorge Torres-Paz

14. Application of CRISPR-Cas9 for functional analysis in A. mexicanus Itzel Sifuentes-Romero, Estephany Ferrufino, and Johanna E. Kowalko

15. Immunohistochemistry and in vivo neural imaging in A. mexicanus

Evan Lloyd, Robert A. Kozol, Erik R. Duboue, and Alex C. Keene

16. Sleep behavior analysis in Astyanax mexicanus

Pierce Hutton, Evan Lloyd, Madelyn Dotson, and Alex C. Keene

17. A new method to analyze non-visual based social-like interactions in asocial cavefish

Motoko Iwashita and Masato Yoshizawa

18. A generalist's guide to quantifying swimming behavior within traditional and non-traditional teleost models

Luke Olsen, Richard Alexander, Jaya Krishnan, and Nicolas Rohner

19. Recording acoustic behaviour in Astyanax mexicanus fish:

acquisition, decryption, interpretation

Carole Hyacinthe

Part III: Mini-brain

20. Generation of mini-brains from hiPSCs

Yan Wang, Dai Tsuchiya and Chongbei Zhao

21. Generating Human Pluripotent Stem Cell-Derived AssemBloids (TM) to Model Interneuron Migration and Immune Cell Interactions

Negin Imani Farahani, Jeanne Chan, Adam Anonuevo, Leon H. Chew, and Erin Knock

22. Multiplexed quantitative proteomics analysis of developing human brain organoids

Laurence Florens, Anita Saraf, Yan Hao, Ying Zhang, Yan Wang, Tari J. Parmely, Chongbei Zhao, and Michael P. Washburn