Zebrafish assays for drug toxicity screening

Zebrafish are vertebrate organisms that are of growing interest for preclinical drug discovery applications. Zebrafish embryos develop most of the major organ systems present in mammals, including the cardiovascular, nervous and digestive systems, in < 1 week. Additional characteristics that make them advantageous for compound screening are their small size, transparency and ability to absorb compounds through the water. Furthermore, gene function analysis with antisense technology is now routine procedure. Thus, it is relatively simple to assess whether compounds or gene knockdowns cause toxic effects in zebrafish. Assays are being developed to exploit the unique characteristics of zebrafish for pharmacological toxicology. This review discusses assays that may be used to assess invivo toxicity and provides examples of compounds known to be toxic to humans that have been demonstrated to function similarly in zebrafish.

[1]  D A Kane,et al.  Genes controlling and mediating locomotion behavior of the zebrafish embryo and larva. , 1996, Development.

[2]  M W Hornung,et al.  Early life stage toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in zebrafish (Danio rerio). , 1997, Toxicology and applied pharmacology.

[3]  S. Ekker,et al.  Effective targeted gene ‘knockdown’ in zebrafish , 2000, Nature Genetics.

[4]  M. Pack,et al.  Unique and conserved aspects of gut development in zebrafish. , 2003, Developmental biology.

[5]  M. Carvan,et al.  Ethanol effects on the developing zebrafish: neurobehavior and skeletal morphogenesis. , 2004, Neurotoxicology and teratology.

[6]  U. Strähle,et al.  The use of zebrafish mutants to identify secondary target effects of acetylcholine esterase inhibitors. , 2004, Toxicological sciences : an official journal of the Society of Toxicology.

[7]  D. Stainier,et al.  A window to the heart: can zebrafish mutants help us understand heart disease in humans? , 2002, Trends in genetics : TIG.

[8]  V. Cunliffe.  Histone deacetylase 1 is required to repress Notch target gene expression during zebrafish neurogenesis and to maintain the production of motoneurones in response to hedgehog signalling , 2004, Development.

[9]  A. Ghysen,et al.  Development of the zebrafish lateral line , 2004, Current Opinion in Neurobiology.

[10]  A. Rubinstein,et al.  Neuroprotection of MPTP-induced toxicity in zebrafish dopaminergic neurons. , 2005, Brain research. Molecular brain research.

[11]  M. Halpern,et al.  The parapineal mediates left-right asymmetry in the zebrafish diencephalon , 2003, Development.

[12]  V. Prince,et al.  Constructing the hindbrain: Insights from the zebrafish , 2002, Developmental dynamics : an official publication of the American Association of Anatomists.

[13]  V. Castellucci,et al.  A novel actin isoform is expressed in the ovotestis of Aplysia californica. , 2005, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[14]  Gabriele Vacun,et al.  Zebrafish embryos express an orthologue of HERG and are sensitive toward a range of QT-prolonging drugs inducing severe arrhythmia. , 2003, Toxicology and applied pharmacology.

[15]  Giuseppe Curigliano,et al.  Drug-induced prolongation of the QT interval. , 2004, The New England journal of medicine.

[16]  M. Farrell,et al.  Promoter analysis in living zebrafish embryos identifies a cis-acting motif required for neuronal expression of GATA-2. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[17]  P. Konstantinopoulos,et al.  The Cardiovascular Toxicity of Selective and Nonselective Cyclooxygenase Inhibitors: Comparisons, Contrasts, and Aspirin Confounding , 2005, Journal of clinical pharmacology.

[18]  H. Takeda,et al.  Transgenic zebrafish for detecting mutations caused by compounds in aquatic environments , 2000, Nature Biotechnology.

[19]  Jay S. Fine,et al.  Chronic Treatment with the γ-Secretase Inhibitor LY-411,575 Inhibits β-Amyloid Peptide Production and Alters Lymphopoiesis and Intestinal Cell Differentiation* , 2004, Journal of Biological Chemistry.

[20]  Francois Pognan,et al.  Modulation of notch processing by gamma-secretase inhibitors causes intestinal goblet cell metaplasia and induction of genes known to specify gut secretory lineage differentiation. , 2004, Toxicological sciences : an official journal of the Society of Toxicology.

[21]  Wolfgang Rottbauer,et al.  High-throughput assay for small molecules that modulate zebrafish embryonic heart rate , 2005, Nature chemical biology.

[22]  M. Halpern,et al.  Genetic Analysis of Digestive Physiology Using Fluorescent Phospholipid Reporters , 2001, Science.

[23]  Su Guo,et al.  Sensitivity of zebrafish to environmental toxins implicated in Parkinson's disease. , 2004, Neurotoxicology and teratology.

[24]  R. Peterson,et al.  Induction of cytochrome P450 1A is required for circulation failure and edema by 2,3,7,8-tetrachlorodibenzo-p-dioxin in zebrafish. , 2003, Biochemical and biophysical research communications.

[25]  T. Braunbeck,et al.  Effects of triphenyltin acetate on survival, hatching success, and liver ultrastructure of early life stages of zebrafish (Danio rerio). , 1999, Ecotoxicology and environmental safety.

[26]  J. Eisen,et al.  Patterning the zebrafish central nervous system. , 2002, Results and problems in cell differentiation.

[27]  R. Pillai,et al.  Histone deacetylase 1 (HDAC‐1) required for the normal formation of craniofacial cartilage and pectoral fins of the zebrafish , 2004, Developmental dynamics : an official publication of the American Association of Anatomists.

[28]  Michael R. Taylor,et al.  Pentylenetetrazole induced changes in zebrafish behavior, neural activity and c-fos expression , 2005, Neuroscience.

[29]  V. Korzh,et al.  Zebrafish embryos are susceptible to the dopaminergic neurotoxin MPTP , 2005, The European journal of neuroscience.

[30]  G. Breithardt,et al.  Experimental models of torsade de pointes. , 1998, Cardiovascular research.

[31]  C. Kimmel,et al.  Inhibition of zebrafish fgf8 pre‐mRNA splicing with morpholino oligos: A quantifiable method for gene knockdown , 2001, Genesis.

[32]  W. Driever,et al.  Mutations in the zebrafish unmask shared regulatory pathways controlling the development of catecholaminergic neurons. , 1999, Developmental biology.

[33]  D A Kane,et al.  The identification of genes with unique and essential functions in the development of the zebrafish, Danio rerio. , 1996, Development.

[34]  P. Drapeau,et al.  Time course of the development of motor behaviors in the zebrafish embryo. , 1998, Journal of neurobiology.

[35]  E. Linney,et al.  Expression of cyp26b1 during zebrafish early development. , 2005, Gene expression patterns : GEP.

[36]  W. Talbot,et al.  Targeted gene knockdown in zebrafish using negatively charged peptide nucleic acid mimics , 2003, Developmental dynamics : an official publication of the American Association of Anatomists.

[37]  J. Ramsdell,et al.  Developmental toxicity of domoic acid in zebrafish (Danio rerio). , 2005, Neurotoxicology and teratology.

[38]  D. Buhler,et al.  Constitutive and xenobiotics-induced expression of a novel CYP3A gene from zebrafish larva. , 2005, Toxicology and applied pharmacology.

[39]  U. Strähle,et al.  Ethanol impairs migration of the prechordal plate in the zebrafish embryo. , 1998, Developmental biology.

[40]  Ben S. Wittner,et al.  Association of valproate‐induced teratogenesis with histone deacetylase inhibition in vivo , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[41]  F. Vaglini,et al.  Species differences in the role of excitatory amino acids in experimental parkinsonism , 1997, Neuroscience & Biobehavioral Reviews.

[42]  G. Callard,et al.  Differential tissue distribution, developmental programming, estrogen regulation and promoter characteristics of cyp19 genes in teleost fish , 2001, The Journal of Steroid Biochemistry and Molecular Biology.

[43]  J. Hendricks,et al.  Neoplasia in Zebrafish (Danio rerio) Treated with N-methyl-N'nitro-N-nitrosoguanidine by Three Exposure Routes at ifferent Developmental Stages , 2000, Toxicologic pathology.

[44]  M. Fishman,et al.  Convergence of distinct pathways to heart patterning revealed by the small molecule concentramide and the mutation heart-and-soul , 2001, Current Biology.

[45]  S. Laurenson,et al.  Chemical discovery and global gene expression analysis in zebrafish , 2003, Nature Biotechnology.

[46]  S. Saszik,et al.  Effects of embryonic exposure to ethanol on zebrafish visual function. , 2002, Neurotoxicology and teratology.

[47]  C. Haass,et al.  A γ‐secretase inhibitor blocks Notch signaling in vivo and causes a severe neurogenic phenotype in zebrafish , 2002, EMBO reports.

[48]  W. Heideman,et al.  Heart malformation is an early response to TCDD in embryonic zebrafish. , 2005, Toxicological sciences : an official journal of the Society of Toxicology.

[49]  B. Naudts,et al.  Development and application of a brain-specific cDNA microarray for effect evaluation of neuro-active pharmaceuticals in zebrafish (Danio rerio). , 2005, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[50]  W. H. Jordan,et al.  Adipsin, a Biomarker of Gastrointestinal Toxicity Mediated by a Functional γ-Secretase Inhibitor* , 2003, Journal of Biological Chemistry.

[51]  M. Westerfield,et al.  CYP2K6 from zebrafish (Danio rerio): cloning, mapping, developmental/tissue expression, and aflatoxin B1 activation by baculovirus expressed enzyme. , 2005, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[52]  Y. Aoki,et al.  MNNG-induced mutations in the adult gill and hepatopancreas and in embryos of rpsL transgenic zebrafish. , 2004, Mutation research.

[53]  Mark C. Fishman,et al.  Cardiomyopathy in zebrafish due to mutation in an alternatively spliced exon of titin , 2002, Nature Genetics.

[54]  L. Zon,et al.  Mutation in the transcriptional coactivator EYA4 causes dilated cardiomyopathy and sensorineural hearing loss , 2005, Nature Genetics.

[55]  M. Kent,et al.  The State of the Art of the Zebrafish Model for Toxicology and Toxicologic Pathology Research—Advantages and Current Limitations , 2003, Toxicologic pathology.

[56]  D. Stainier,et al.  Formation of the digestive system in zebrafish. II. Pancreas morphogenesis. , 2003, Developmental biology.

[57]  Jules C Hancox,et al.  Troubleshooting problems with in vitro screening of drugs for QT interval prolongation using HERG K+ channels expressed in mammalian cell lines and Xenopus oocytes. , 2002, Journal of pharmacological and toxicological methods.

[58]  A. Schier,et al.  A genetic screen for mutations affecting embryogenesis in zebrafish. , 1996, Development.

[59]  C. Ton,et al.  The use of zebrafish for assessing ototoxic and otoprotective agents , 2005, Hearing Research.

[60]  Taise Bresolin,et al.  Expression of PXR, CYP3A and MDR1 genes in liver of zebrafish. , 2005, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[61]  F. Hsieh,et al.  Germ‐line transmission of a myocardium‐specific GFP transgene reveals critical regulatory elements in the cardiac myosin light chain 2 promoter of zebrafish , 2003, Developmental dynamics : an official publication of the American Association of Anatomists.

[62]  Wolfgang Driever,et al.  Dopamine transporter expression distinguishes dopaminergic neurons from other catecholaminergic neurons in the developing zebrafish embryo , 2001, Mechanisms of Development.

[63]  Tobias Roeser,et al.  Formation of the digestive system in zebrafish. I. Liver morphogenesis. , 2003, Developmental biology.

[64]  Shuo Lin,et al.  Pdx‐1 knockdown reduces insulin promoter activity in zebrafish , 2001, Genesis.

[65]  Didier Y. R. Stainier,et al.  Cardiac troponin T is essential in sarcomere assembly and cardiac contractility , 2002, Nature Genetics.

[66]  K. Cheng,et al.  Ethylnitrosourea Induces Neoplasia in Zebrafish (Danio rerio) , 2000, Laboratory Investigation.

[67]  K. Lewis,et al.  From cells to circuits: development of the zebrafish spinal cord , 2003, Progress in Neurobiology.

[68]  S. P. Chambers,et al.  Short interfering RNA‐mediated gene targeting in the zebrafish , 2004, FEBS letters.

[69]  C. V. Van Leeuwen,et al.  Fish embryos as teratogenicity screens: a comparison of embryotoxicity between fish and birds. , 1990, Ecotoxicology and environmental safety.

[70]  A. Setini,et al.  Molecular characterization of monoamine oxidase in zebrafish (Danio rerio). , 2005, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[71]  F. Checler,et al.  JLK isocoumarin inhibitors: Selective γ‐secretase inhibitors that do not interfere with notch pathway in vitro or in vivo , 2003, Journal of neuroscience research.

[72]  Zuo-yan Zhu,et al.  Efficient RNA interference in zebrafish embryos using siRNA synthesized with SP6 RNA polymerase , 2005, Development, growth & differentiation.

[73]  D. Nebert,et al.  Transgenic Zebrafish as Sentinels for Aquatic Pollution , 2000, Annals of the New York Academy of Sciences.

[74]  J. Dowling,et al.  Small molecule developmental screens reveal the logic and timing of vertebrate development. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[75]  J. Langston,et al.  Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis. , 1983, Science.

[76]  M. Fishman,et al.  Defective "pacemaker" current (Ih) in a zebrafish mutant with a slow heart rate. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[77]  H Okamoto,et al.  Visualization of Cranial Motor Neurons in Live Transgenic Zebrafish Expressing Green Fluorescent Protein Under the Control of the Islet-1 Promoter/Enhancer , 2000, The Journal of Neuroscience.

[78]  E. Linney,et al.  Zebrafish as a neurotoxicological model. , 2004, Neurotoxicology and teratology.

[79]  Sung-Kook Hong,et al.  Analysis of upstream elements in the HuC promoter leads to the establishment of transgenic zebrafish with fluorescent neurons. , 2000, Developmental biology.

[80]  Calum A. MacRae,et al.  Drugs That Induce Repolarization Abnormalities Cause Bradycardia in Zebrafish , 2003, Circulation.