Why zebrafish screening?
Recently, zebrafish-based screening has gained favor as an alternative to mammalian screening due to cost, throughput and reduced ethical concerns. Zebrafish are a useful model for drug development because of their high biological similarity to humans. Studies in ontogeny and organogenesis have shown that their primary organ systems are very similar to humans, and the synteny between zebrafish and Homo sapiens is as high as 70-80%.
Zebrafish are beneficial for screening assays because of their high fecundity, transparency of embryos for viewing organ structures, and ease of gene manipulation. Their small size allows embryos to be placed in microtiter plates and treated with compounds. Phenotypes can then be measured on a high content screening system.
The ImageXpress® High-Content Screening System provides optimal flexibility for acquiring high quality images with a large field of view. MetaXpress® High Content Image Acquisition and Analysis Software enables image analysis for a wide range of applications with simple workflows, and in combination with AcuityXpress High Content Informatics Software for data mining, this end-to-end solution dramatically improves throughput for zebrafish-based in vivo screening.
Zebrafish-based in vivo high content screening applications
There has been a multitude of human disease models developed in zebrafish such as:
- Metabolic syndrome: obesity (visceral fat), dyslipidemia, fatty liver, glucose tolerance disorder
- Xenotransplantation of human cancer cells: tumor angiogenesis, distant metastasis
- Circulatory disease: cardiac failure, druginduced arrhythmia
- Central nervous system disorder: deafness, visual disturbance, smell disorder, epilepsy, developmental disorder, sleep arousal disorder, myodystrophy (ALS)
Several examples of how highcontent imaging assays can be used for investigating these disorders are shown here.
Modeling tumor metastasis after xenotransplantation of human cancer cells1
Monitoring inhibition of angiogenesis
Cardiac function analysis
Gene knockdown quantitation2
Targeted image acquisition of a specific area of the well or zebrafish
For objects that may not be in the same area of every well, targeted imaging can be used to pinpoint the object of interest using low magnification and then return to those coordinates to acquire a higher magnification image.
Zebrafish embryos are a valuable vertebrate model for translational research. Using the ImageXpress High-Content Screening Systems to acquire images of zebrafish in focus from head to tail with a large field of view and high-speed z-stacking provides the ability to measure phenotypes characteristic of disease or toxicity. High-throughput in vivo imaging of zebrafish in combination with MetaXpress High-Content Analysis software empowers laboratories to increase their throughput dramatically and screen thousands of compounds in just a few days. Set up and run automated imaging screens to monitor inhibition of angiogenesis, quantitate gene knockdown, and measure ototoxicity and neurotoxicity with this end-to-end solution.
1. Zhang, B., et al., Quantitative phenotyping-based in vivo chemical screening in a zebrafish model of leukemia stem cell xenotransplantation, PLoS One, 2014 Jan 15; 9(1).
2. Umemoto, N., et al., Fluorescent-based methods for gene knockdown and functional cardiac imaging in zebrafish, Mol Biotechnol, 2013 Oct; 55(2): 131-42.
3. Kanungo, J., et al., In vivo imaging and quantitative analysis of changes in axon length using transgenic zebrafish embryos. Neurotoxicol Teratol, 2011 Nov-Dec; 33(6): 618-23.
4. Diekmann, H., et al., Characterization of optic nerve regeneration using transgenic zebrafish. Front Cell Neurosci, 2015 April 9; 9:118.
5. Huan, H., et al., High-throughput screening for bioactive molecules using primary cell culture of transgenic zebrafish embryos. Cell Rep, 2012 Sep 27; 2(3):695-704.