Robotic injection of zebrafish embryos for high-throughput screening in disease models

Spaink, Herman P., Cui, Chao, Wiweger, Malgorzata I., Jansen, Hans J., Veneman, Wouter J., Marin-Juez, Ruben, de Sonneville, Jan, Ordas, Anita, Torraca, V., van der Ent, Wietske, Leenders, William P., Meijer, Annemarie H., Snaar-Jagalska, B. Ewa and Dirks, Ron P. 2013. Robotic injection of zebrafish embryos for high-throughput screening in disease models. Methods. 62 (3), pp. 246-254. https://doi.org/10.1016/j.ymeth.2013.06.002

TitleRobotic injection of zebrafish embryos for high-throughput screening in disease models
TypeJournal article
AuthorsSpaink, Herman P., Cui, Chao, Wiweger, Malgorzata I., Jansen, Hans J., Veneman, Wouter J., Marin-Juez, Ruben, de Sonneville, Jan, Ordas, Anita, Torraca, V., van der Ent, Wietske, Leenders, William P., Meijer, Annemarie H., Snaar-Jagalska, B. Ewa and Dirks, Ron P.
Abstract

The increasing use of zebrafish larvae for biomedical research applications is resulting in versatile models for a variety of human diseases. These models exploit the optical transparency of zebrafish larvae and the availability of a large genetic tool box. Here we present detailed protocols for the robotic injection of zebrafish embryos at very high accuracy with a speed of up to 2000 embryos per hour. These protocols are benchmarked for several applications: (1) the injection of DNA for obtaining transgenic animals, (2) the injection of antisense morpholinos that can be used for gene knock-down, (3) the injection of microbes for studying infectious disease, and (4) the injection of human cancer cells as a model for tumor progression. We show examples of how the injected embryos can be screened at high-throughput level using fluorescence analysis. Our methods open up new avenues for the use of zebrafish larvae for large compound screens in the search for new medicines.

JournalMethods
Journal citation62 (3), pp. 246-254
ISSN1046-2023
Year2013
PublisherElsevier
Publisher's version
License
CC BY-NC-SA 3.0
File Access Level
Open (open metadata and files)
Digital Object Identifier (DOI)https://doi.org/10.1016/j.ymeth.2013.06.002
Publication dates
Published15 Aug 2013

Related outputs

Editorial: Zebrafish Models for Human Disease Studies
Zang, L., Torraca, V., Shimada, Y. and Nishimura, N. 2022. Editorial: Zebrafish Models for Human Disease Studies. Frontiers in Cell and Developmental Biology. 10 861941. https://doi.org/10.3389/fcell.2022.861941

Septins promote caspase activity and coordinate mitochondrial apoptosis
Hoan Van Ngo, Stevens Robertin, Dominik Brokatzky, Magdalena K. Bielecka, Damián Lobato‐Márquez, Vincenzo Torraca and Serge Mostowy 2022. Septins promote caspase activity and coordinate mitochondrial apoptosis. Cytoskeleton. Advanced online publication. https://doi.org/10.1002/cm.21696

Editorial: Nucleic Acid-Associated Inflammation
Laguette, N., Langevin, C., Olagnier, D., Torraca, V., Vanpouille-Box, C. and Verrier, E.R. 2021. Editorial: Nucleic Acid-Associated Inflammation. Frontiers in Immunology. 12. https://doi.org/10.3389/fimmu.2021.791580

Disruption of Cxcr3 chemotactic signaling alters lysosomal function and renders macrophages more microbicidal
Frida Sommer, Vincenzo Torraca, Yufei Xie, Aliede E. in ‘t Veld, Joost Willemse and Annemarie H. Meijer 2021. Disruption of Cxcr3 chemotactic signaling alters lysosomal function and renders macrophages more microbicidal. Cell Reports. 35 (2) 109000. https://doi.org/10.1016/j.celrep.2021.109000

Deficiency in the autophagy modulator Dram1 exacerbates pyroptotic cell death of Mycobacteria-infected macrophages
Rui Zhang, Monica Varela, Gabriel Forn-Cuní, Vincenzo Torraca, Michiel van der Vaart and Annemarie H. Meijer 2020. Deficiency in the autophagy modulator Dram1 exacerbates pyroptotic cell death of Mycobacteria-infected macrophages. Cell Death and Disease. 11 277. https://doi.org/10.1038/s41419-020-2477-1

Shigella sonnei
Vincenzo Torraca, Kathryn Holt and Serge Mostowy 2020. Shigella sonnei. Trends in Microbiology. 28 (8), pp. P696-697. https://doi.org/10.1016/j.tim.2020.02.011

Frontline Science: Antagonism between regular and atypical Cxcr3 receptors regulates macrophage migration during infection and injury in zebrafish
Sommer, Frida, Torraca, V., Kamel, Sarah M., Lombardi, Amber and Meijer, Annemarie H. 2020. Frontline Science: Antagonism between regular and atypical Cxcr3 receptors regulates macrophage migration during infection and injury in zebrafish. Journal of Leukocyte Biology. 107 (2), pp. 185-203. https://doi.org/10.1002/jlb.2hi0119-006r

Chemokine Receptors and Phagocyte Biology in Zebrafish
Sommer, Frida, Torraca, V. and Meijer, Annemarie H. 2020. Chemokine Receptors and Phagocyte Biology in Zebrafish. Frontiers in Immunology. 11 325. https://doi.org/10.3389/fimmu.2020.00325

Supplementary RNAseq dataset files
Frida Sommer, Torraca, V. and Annemarie H. Meijer 2020. Supplementary RNAseq dataset files. Zenodo. https://doi.org/10.5281/zenodo.3833847

Analysis tools to quantify dissemination of pathology in zebrafish larvae
David R. Stirling, Oniz Suleyman, Eliza Gil, Philip M. Elks, Vincenzo Torraca, Mahdad Noursadeghi and Gillian S. Tomlinson 2020. Analysis tools to quantify dissemination of pathology in zebrafish larvae. Scientific Reports. 10 3149. https://doi.org/10.1038/s41598-020-59932-1

In vivo biomolecular imaging of zebrafish embryos using confocal Raman spectroscopy
Håkon Høgset, Conor C. Horgan, James P. K. Armstrong, Mads S. Bergholt, Vincenzo Torraca, Qu Chen, Timothy J. Keane, Laurence Bugeon, Margaret J. Dallman, Serge Mostowy and Molly M. Stevens 2020. In vivo biomolecular imaging of zebrafish embryos using confocal Raman spectroscopy. Nature Communications . 11 6172. https://doi.org/10.1038/s41467-020-19827-1

Shigella sonnei O-Antigen Inhibits Internalization, Vacuole Escape, and Inflammasome Activation
Watson, Jayne L., Sanchez-Garrido, Julia, Goddard, Philippa J., Torraca, V., Mostowy, Serge, Shenoy, Avinash R. and Clements, Abigail 2019. Shigella sonnei O-Antigen Inhibits Internalization, Vacuole Escape, and Inflammasome Activation. mBio. 10 (6) e02654-19. https://doi.org/10.1128/mbio.02654-19

Shigella sonnei infection of zebrafish reveals that O-antigen mediates neutrophil tolerance and dysentery incidence
Vincenzo Torraca, Myrsini Kaforou, Jayne Watson, Gina M. Duggan, Hazel Guerrero-Gutierrez, Sina Krokowski, Michael Hollinshead, Thomas B. Clarke, Rafal J. Mostowy, Gillian S. Tomlinson, Vanessa Sancho-Shimizu, Abigail Clements and Serge Mostowy 2019. Shigella sonnei infection of zebrafish reveals that O-antigen mediates neutrophil tolerance and dysentery incidence. PLOS Pathogens. 15 (12) e1008006. https://doi.org/10.1371/journal.ppat.1008006

RNAseq Profiling of Leukocyte Populations in Zebrafish Larvae Reveals a cxcl11 Chemokine Gene as a Marker of Macrophage Polarization During Mycobacterial Infection
Rougeot, Julien, Torraca, V., Zakrzewska, Ania, Kanwal, Zakia, Jansen, Hans J., Sommer, Frida, Spaink, Herman P. and Meijer, Annemarie H. 2019. RNAseq Profiling of Leukocyte Populations in Zebrafish Larvae Reveals a cxcl11 Chemokine Gene as a Marker of Macrophage Polarization During Mycobacterial Infection. Frontiers in Immunology. 10 832. https://doi.org/10.3389/fimmu.2019.00832

CXCR4 signaling regulates metastatic onset by controlling neutrophil motility and response to malignant cells
Annemarie Meijer and Torraca, V. 2019. CXCR4 signaling regulates metastatic onset by controlling neutrophil motility and response to malignant cells. Scientific Reports. 9 2399. https://doi.org/10.1038/s41598-019-38643-2

Macrophages target Salmonella by Lc3-associated phagocytosis in a systemic infection model
Masud, Samrah, Prajsnar, Tomasz K., Torraca, V., Lamers, Gerda E. M., Benning, Marianne, Van der Vaart, Michiel and Meijer, Annemarie H. 2019. Macrophages target Salmonella by Lc3-associated phagocytosis in a systemic infection model. Autophagy. 15 (5), pp. 796-812. https://doi.org/10.1080/15548627.2019.1569297

Meeting report: Zebrafish Infection and Immunity 2019
Torraca, V., Gomes, Margarida C., Sarris, Milka and Mostowy, Serge 2019. Meeting report: Zebrafish Infection and Immunity 2019. Lab Animal. 48, pp. 284-287. https://doi.org/10.1038/s41684-019-0400-0

Zebrafish Infection: From Pathogenesis to Cell Biology
Vincenzo Torraca and Serge Mostowy 2018. Zebrafish Infection: From Pathogenesis to Cell Biology. Trends in Cell Biology. 28 (2), pp. P143-156. https://doi.org/10.1016/j.tcb.2017.10.002

Shigella-Induced Emergency Granulopoiesis Protects Zebrafish Larvae from Secondary Infection
Willis, Alexandra R., Torraca, V., Gomes, Margarida C., Shelley, Jennifer, Mazon-Moya, Maria, Filloux, Alain, Lo Celso, Cristina and Mostowy, Serge 2018. Shigella-Induced Emergency Granulopoiesis Protects Zebrafish Larvae from Secondary Infection. mBio. 9 (3) e00933-18. https://doi.org/10.1128/mbio.00933-18

Septins restrict inflammation and protect zebrafish larvae from Shigella infection
Maria J. Mazon-Moya, Alexandra R. Willis, Vincenzo Torraca, Laurent Boucontet, Avinash R. Shenoy, Emma Colucci-Guyon and Serge Mostowy 2017. Septins restrict inflammation and protect zebrafish larvae from Shigella infection. PLOS Pathogens. 13 (6) e1006467. https://doi.org/10.1371/journal.ppat.1006467

The inflammatory chemokine Cxcl18b exerts neutrophil-specific chemotaxis via the promiscuous chemokine receptor Cxcr2 in zebrafish
Torraca, V., Otto, Natasja A., Tavakoli-Tameh, Aidin and Meijer, Annemarie H. 2017. The inflammatory chemokine Cxcl18b exerts neutrophil-specific chemotaxis via the promiscuous chemokine receptor Cxcr2 in zebrafish. Developmental & Comparative Immunology. 67, pp. 57-65. https://doi.org/10.1016/j.dci.2016.10.014

The chemokine receptor CXCR4 promotes granuloma formation by sustaining a mycobacteria-induced angiogenesis programme
Annemarie Meijer and Torraca, V. 2017. The chemokine receptor CXCR4 promotes granuloma formation by sustaining a mycobacteria-induced angiogenesis programme. Scientific Reports. 7 45061. https://doi.org/10.1038/srep45061

Functional analysis reveals no transcriptional role for the glucocorticoid receptor beta-isoform in zebrafish
Chatzopoulou, Antonia, Schoonheim, Peter J., Torraca, V., Meijer, Annemarie H., Spaink, Herman P. and Schaaf, Marcel J.M. 2017. Functional analysis reveals no transcriptional role for the glucocorticoid receptor beta-isoform in zebrafish. Molecular and Cellular Endocrinology. 447, pp. 61-70. https://doi.org/10.1016/j.mce.2017.02.036

Modeling Infectious Diseases in the Context of a Developing Immune System
Masud, Samrah, Torraca, V. and Meijer, Annemarie H. 2017. Modeling Infectious Diseases in the Context of a Developing Immune System. Current Topics in Developmental Biology. 124, pp. 277-329. https://doi.org/10.1016/bs.ctdb.2016.10.006

Septins and Bacterial Infection
Torraca, V. and Mostowy, Serge 2016. Septins and Bacterial Infection. Frontiers in Cell and Developmental Biology. https://doi.org/10.3389/fcell.2016.00127

The CXCR3-CXCL11 signaling axis mediates macrophage recruitment and dissemination of mycobacterial infection
Torraca, V., Cui, Chao, Boland, Ralf, Bebelman, Jan-Paul, van der Sar, Astrid M., Smit, Martine J., Siderius, Marco, Spaink, Herman P. and Meijer, Annemarie H. 2015. The CXCR3-CXCL11 signaling axis mediates macrophage recruitment and dissemination of mycobacterial infection. Disease Models and Mechanisms. 8 (3), pp. 253-269. https://doi.org/10.1242/dmm.017756

Macrophage-pathogen interactions in infectious diseases: new therapeutic insights from the zebrafish host model
Torraca, V., Masud, S., Spaink, H.P. and Meijer, A.H. 2014. Macrophage-pathogen interactions in infectious diseases: new therapeutic insights from the zebrafish host model. Disease Models and Mechanisms. 7 (7), pp. 785-797. https://doi.org/10.1242/dmm.015594

Ultra-small graphene oxide functionalized with polyethylenimine (PEI) for very efficient gene delivery in cell and zebrafish embryos
Zhou, Xiang, Laroche, Fabrice, Lamers, Gerda E. M., Torraca, V., Voskamp, Patrick, Lu, Tao, Chu, Fuqiang, Spaink, Herman P., Abrahams, Jan Pieter and Liu, Zunfeng 2012. Ultra-small graphene oxide functionalized with polyethylenimine (PEI) for very efficient gene delivery in cell and zebrafish embryos. Nano Research. 5, pp. 703-709. https://doi.org/10.1007/s12274-012-0254-x

Permalink - https://westminsterresearch.westminster.ac.uk/item/vq5z5/robotic-injection-of-zebrafish-embryos-for-high-throughput-screening-in-disease-models


Share this

Usage statistics

22 total views
16 total downloads
These values cover views and downloads from WestminsterResearch and are for the period from September 2nd 2018, when this repository was created.