Zebrafish as a model for drug screening

Zebrafish have become one of the most powerful and versatile models of vertebrate biology.

The Birth of a New Model Organism

As early as the 1930s, the development of the small teleost fish species. Danio rerio, commonly known as zebrafish, was studied

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The forward genetic screens generated vast collections of mutant lines, providing numerous insights into early embryonic development, organogenesis, neural development and behaviour.
More recently, in 2000, the first small-molecule screen in zebrafish was published.1

Peterson RT, Link BA, Dowling JE, Schreiber SL, Small molecule developmental screens reveal the logic and timing of vertebrate development, Proc Natl Acad Sci U S A, 2000;97:129659.

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The suitability of the model

Because of the tremendous opportunities they provide to analyse embryonic development. The zebrafish genome was recently sequenced by the Wellcome Trust Sanger Institute. This will aid in the interpretation of the extensive banks of genetic mutants that have been created and phenotyped.
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The features
1. Zebrafish are small in size (up to 3cm). 2.The embryonic development is rapid. 3.The body plan is laid out within 24 hours after fertilisation. 4.The optical clarity of the embryo enables in vivo observation of organogenesis, in particular with expression of fluorescent proteins. 5.External fertilisation and development together with high fecundity, 6.The cost of maintenance for fish is relatively low (less than 1% of the cost of mice). 7.Low cost and ability to maintain large numbers of animals in a small space, multiple genetic and molecular tools have been established.

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Drug screening strategies

The identification of new drugs can be divided into two approaches: Organism-based and target-based discovery., in which a whole organism is treated with a compound, has been the prevailing method of research. For example, in 1775 William Withering observed the beneficial effects of foxglove on a patient suffering from heart failure, and years later isolated digitalis, which is still an important therapy for heart failure.

More recently, target based discovery has become the method of choice in drug discovery. With this approach, the molecular causes of a disease are identified first and compounds that specifically interfere with these causes are then isolated using high-throughput screening (HTS). The zebrafish model may prove useful for both approaches.
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Your model of choice for screening

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The screening
the use of zebrafish to assess the action of compounds in a 96-well plate based. Each well containing up to five hatchlings. Various types of assay can be carried out. For example, potential drugs can be screened for their ability to correct a defect reflecting a human disease state (phenotypic rescue).

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assays have been developed to identify potential undesired clinical effects of a drug, from interference with heart function to visual disturbance.

Zebrafish hatchlings in 96well plate for high throughput drug testing

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The screening process

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Using fish embryos to screen for compounds that interfere with signaling pathways involved in early development : pathways known to play roles in a variety of disease processes. The compound "dorsomorphin" and demonstrated that it blocked BMP (bone morphogenetic protein) signaling, which has been implicated in anemia, inflammatory responses and bone-related disorders.

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 In examining dorsomorphin further, the investigators found that it had other "offtarget" effects -- it also blocked the VEGF (vascular endothelial growth factor) receptor and disrupted zebrafish blood vessel development, a process called angiogenesis.

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"Off-target effects contribute to side effects and limit the therapeutic potential of small molecule signaling inhibitors,"

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Role in environmental pollutant Screening Changes in gene expression patterns in zebrafish embryos resulting from exposure to environmental toxins can identify the individual toxins in the work place & environment.

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Zebrafish embryos were exposed to eleven common pollutants, including cadmium, mercury and arsenic. Scientists monitored changes in gene-expression profiles in order to predict the chemical that the embryo had been exposed to; the results were clear-cut for 10 out of 11 toxins

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Toxicity & safety evaluation

Zebrafish models have increasingly been used to assess drug toxicity and safety, largely because they are transparent, genetically tractable, and have organ systems that are very similar to those of humans. Zebrafish embryos are permeable to drugs.

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Zebrafish Swim Into Drug Development

ScienceDaily (Jan. 25, 2010) By combining the tools of medicinal chemistry and zebrafish biology, a team of Vanderbilt investigators has identified compounds that may offer therapeutic leads for bone-related diseases and cancer.

"For all their differences, humans and zebrafish aren't that dissimilar

"For every zebrafish gene we isolate, there is a related gene in humans."

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In Structureactivity relationships study

The zebrafish as a system is highly amenable to the study of structureactivity relationships (SARs). During the above screen for compounds that perturb the cell cycle.

One advantage of performing SAR studies in zebrafish is that they couple the analysis of binding affinity and ADME/toxicity

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In toxicology study
FDA points to technological deficits in toxicology as one of the primary causes of this pipeline problem, noting that in many cases, the approaches of the last century are still being used to assess this centurys candidates. New animal models are needed to test the safety of novel drug candidates, and the FDA report that an estimated 10% improvement in predicting failures before clinical trials would save US $100 million per drug in development costs. In addition to outdated technologies, toxicology frequently suffers by being divorced from the drug discovery process
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In Toxicology
The zebrafish is rapidly gaining acceptance as a promising animal model for toxicology. The ability to efficiently assess the toxicity of a large number of compounds enables whole libraries to be prescreened for potential toxicity

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In developmental biology..
How the brain and central nervous system develop. It helps that zebrafish embryos grow from just a single cell to having a forebrain, hindbrain, spinal column and eye within 24 hours. It also helps that the embryos are transparent and develop outside their mothers' bodies -- and can thus been seen under a microscope at every step of their development.
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Use for psychiatric disorder

Zebrafish are popular model systems in many areas of biomedical research. The first discovery of a zebrafish mutant with an apparent psychiatric disorder. When faced repeatedly with a stressful situation -- isolation from others -- the mutant fish stop swimming and hide in the corner of the tank for many minutes.
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