Molecular impact of propiconazole on Daphnia magna using a reproduction-related cDNA array.

We have developed a first version cDNA microarray of the cladoceran Daphnia magna. Through Suppression Subtractive Hybridisation PCR (SSH-PCR) 855 life stage-specific cDNAs were collected and used to document the toxicological mode of action of the pesticide propiconazole. DNA sequencing analysis revealed gene fragments related to important functional classes such as embryo development, energy metabolism, molting and cell cycle. Major changes in transcription were observed in organisms exposed for 4 and 8 days to 1 microg/mL. After 4 days a 3-fold down-regulation of the gene encoding the yolk protein, vitellogenin, was observed indicating impaired oocyte maturation. Moreover, genes such as a larval-specific gene and chaperonin were repressed, whereas the heat shock 90 protein and ATP synthase were induced. Organismal effects clearly confirmed the major molecular findings: at the highest concentration (1 microg/mL) adult growth was significantly (p < 0.05) impaired and increased developmental effects in the offspring could be noted. We have demonstrated the potential of microarray analysis in toxicity screening with D. magna. The use of vitellogenin mRNA as a rapid biomarker of reproductive effects in chronic toxicity studies with cladocerans is suggested.

[1]  S. Dodson,et al.  Commentary on effects of anthropogenic and natural organic chemicals on development, swimming behavior, and reproduction of Daphnia, a key member of aquatic ecosystems. , 1995, Environmental health perspectives.

[2]  T. Snell,et al.  Gene Expression Profiling in Ecotoxicology , 2003, Ecotoxicology.

[3]  D. Yamamoto,et al.  Filamin Is Required for Ring Canal Assembly and Actin Organization during Drosophila Oogenesis , 1999, The Journal of cell biology.

[4]  T. Hunt,et al.  New B-type cyclin synthesis is required between meiosis I and II during Xenopus oocyte maturation. , 2001, Development.

[5]  N. Denslow,et al.  Differential gene expression analysis in fish exposed to endocrine disrupting compounds. , 2003, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[6]  K. Yamaguchi,et al.  Chicken chromobox proteins: cDNA cloning of CHCB1, -2, -3 and their relation to W-heterochromatin. , 1998, Experimental cell research.

[7]  E. Werle,et al.  Convenient single-step, one tube purification of PCR products for direct sequencing. , 1994, Nucleic acids research.

[8]  Yasuhiko Kato,et al.  A vitellogenin chain containing a superoxide dismutase-like domain is the major component of yolk proteins in cladoceran crustacean Daphnia magna. , 2004, Gene.

[9]  A. Cossins,et al.  Application of microarray technology in environmental and comparative physiology. , 2003, Annual review of physiology.

[10]  C. Rider,et al.  Embryotoxicity of the alkylphenol degradation product 4-nonylphenol to the crustacean Daphnia magna. , 2000, Environmental health perspectives.

[11]  Ronald W. Davis,et al.  Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA Microarray , 1995, Science.

[12]  S. Dudoit,et al.  Normalization for cDNA microarray data: a robust composite method addressing single and multiple slide systematic variation. , 2002, Nucleic acids research.

[13]  E. Linney,et al.  Microarray gene expression profiling during the segmentation phase of zebrafish development. , 2004, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[14]  J. Schlatter,et al.  Azole fungicides affect mammalian steroidogenesis by inhibiting sterol 14 alpha-demethylase and aromatase. , 2002, Environmental health perspectives.

[15]  Colin R. Janssen,et al.  The use of biomarkers in Daphnia magna toxicity testing V. In vivo alterations in the carbohydrate metabolism of Daphnia magna exposed to sublethal concentrations of mercury and lindane. , 2001 .

[16]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[17]  C. Mothersill,et al.  In vitro methods in aquatic toxicology , 2003 .

[18]  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.

[19]  C. Rider,et al.  The fungicide propiconazole interferes with embryonic development of the crustacean Daphnia magna , 2001, Environmental toxicology and chemistry.

[20]  T. Yamori,et al.  Development of cDNA microarray for expression profiling of estrogen-responsive genes. , 2002, Journal of molecular endocrinology.

[21]  S. Dodson,et al.  Sublethal toxic effects of cyanobacteria and nonylphenol on environmental sex determination and development in Daphnia , 1997 .

[22]  S. Minchin,et al.  A DNA expression array to detect toxic stress response in European flounder (Platichthys flesus). , 2003, Aquatic toxicology.

[23]  Michael A. Thomas,et al.  Genomics for the ecological toolbox. , 2004, Trends in ecology & evolution.

[24]  Thomas H Hutchinson,et al.  Ecotoxicogenomics: the challenge of integrating genomics into aquatic and terrestrial ecotoxicology. , 2004, Aquatic toxicology.

[25]  M. Depledge,et al.  Stress proteins (HSP's): Methods of Detection and Their Use as an Environmental Biomarker , 1999 .

[26]  J. Changeux,et al.  Control of neurulation by the nucleosome assembly protein-1–like 2 , 2000, Nature Genetics.

[27]  C. Janssen,et al.  The missing biomarker link: Relationships between effects on the cellular energy allocation biomarker of toxicant‐stressed Daphnia magna and corresponding population characteristics , 2003, Environmental toxicology and chemistry.

[28]  A. Oikari,et al.  Response of rainbow trout transcriptome to model chemical contaminants. , 2004, Biochemical and biophysical research communications.

[29]  R. Livesey.  Have microarrays failed to deliver for developmental biology? , 2002, Genome Biology.

[30]  Samuel Granjeaud,et al.  Expression scanning of an array of growth control genes in human tumor cell lines , 1999, Oncogene.

[31]  J. Kelso,et al.  Gene expression analysis of largemouth bass exposed to estradiol, nonylphenol, and p,p'-DDE. , 2002, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[32]  Karl Mechtler,et al.  Methylation of histone H3 lysine 9 creates a binding site for HP1 proteins , 2001, Nature.

[33]  M. Feder,et al.  Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. , 1999, Annual review of physiology.

[34]  G. Horgan,et al.  Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR , 2002 .

[35]  M. Nemer.  Histone deacetylase mRNA temporally and spatially regulated in its expression in sea urchin embryos , 1998, Development, growth & differentiation.

[36]  P. Grandi,et al.  Dimerization of the largest subunit of chromatin assembly factor 1: importance in vitro and during Xenopus early development , 2001, The EMBO journal.

[37]  R. Wooster,et al.  Gene expression microarray analysis in cancer biology, pharmacology, and drug development: progress and potential. , 2001, Biochemical pharmacology.

[38]  Gerald T Ankley,et al.  Evaluation of the aromatase inhibitor fadrozole in a short-term reproduction assay with the fathead minnow (Pimephales promelas). , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[39]  R. Gibble,et al.  The effects of 4-nonylphenol and ethanol on acute toxicity, embryo development, and reproduction in Daphnia magna. , 2003, Ecotoxicology and environmental safety.

[40]  A. García‐Gasca,et al.  Partial cloning and expression of the cyclin B gene in the ovary of the bullseye puffer (Sphoeroides annulatus). , 2003, Journal of experimental zoology. Part A, Comparative experimental biology.

[41]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.