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Gene expression profiling of three different stressors in the water flea Daphnia magna

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Abstract

Microarrays are an ideal tool to screen for differences in gene expression of thousands of genes simultaneously. However, often commercial arrays are not available. In this study, we performed microarray analyses to evaluate patterns of gene transcription following exposure to two natural and one anthropogenic stressor. cDNA microarrays compiled of three life stage specific and three stressor-specific EST libraries, yielding 1734 different EST sequences, were used. We exposed juveniles of the water flea Daphnia magna for 48, 96 and 144 h to three stressors known to exert strong selection in natural populations of this species i.e. a sublethal concentration of the pesticide carbaryl, infective spores of the endoparasite Pasteuria ramosa, and fish predation risk mimicked by exposure to fish kairomones. A total of 148 gene fragments were differentially expressed compared to the control. Based on a PCA, the exposure treatments were separated into two main groups based on the extent of the transcriptional response: a low and a high (144 h of fish or carbaryl exposure and 96 h of parasite exposure) stress group. Firstly, we observed a general stress-related transcriptional expression profile independent of the treatment characterized by repression of transcripts involved in transcription, translation, signal transduction and energy metabolism. Secondly, we observed treatment-specific responses including signs of migration to deeper water layers in response to fish predation, structural challenge of the cuticle in response to carbaryl exposure, and disturbance of the ATP production in parasite exposure. A third important conclusion is that transcription expression patterns exhibit stress-specific changes over time. Parasite exposure shows the most differentially expressed gene fragments after 96 h. The peak of differentially expressed transcripts came only after 144 h of fish exposure, while carbaryl exposure induced a more stable number of differently expressed gene fragments over time.

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Acknowledgments

We thank Claudia Buser for help during the exposures. A special thank also to John Colbourne for very useful suggestions to ameliorate the first drafts of the manuscript. Carbaryl analyses were done in the laboratorium of Prof. Dr. W.P. de Voogt (University of Amsterdam, Faculteit der Natuurwetenschappen, Wiskunde en Informatica—Instituut voor Biodiversiteit en Ecosystem Dynamica (IBED)). M.J. enjoyed a PhD fellowship of the Agency for Innovation by Science and Technology in Flanders (IWT, Flanders). This research was financially supported by projects GOA/08/06 and PF/2010/07 of the KULeuven Research Fund and project G.229.09 of the fund for scientific research FWO, Flanders. Our work benefits from and contributes to the “Daphnia Genomics Consortium”.

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The authors declare that they have no conflict of interest.

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  1. Tine Vandenbrouck

    Present address: Apeiron-Team nv, Pluyseghemstraat 69, 2550, Kontich (Antwerpen), Belgium

Authors and Affiliations

  1. Laboratory of Aquatic Ecology, Evolution and Conservation, Catholic University of Leuven, Charles Deberiotstraat 32, 3000, Leuven, Belgium

    Mieke Jansen, Katina I. Spanier, Anke Cielen & Luc De Meester

  2. Systemic Physiological and Ecotoxicological Research (SPHERE), Department of Biology, University of Antwerp, Groenenborgerlaan 171, 2020, Antwerpen, Belgium

    Lucia Vergauwen, Tine Vandenbrouck, Dries Knapen & Nathalie Dom

  3. Department of Veterinary Sciences, Gamete Research Center (GRC), Veterinary Physiology and Biochemistry, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk, Belgium

    Lucia Vergauwen & Dries Knapen

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Table S1: General overview of the differentially expressed genes. Supplementary material 1 (DOCX 54 kb)

10646_2013_1072_MOESM2_ESM.docx

Figure S2: Heatmaps of identified GO classes in the fisher exact test of treatment-specific differentially expressed transcripts compared to the total set of transcript differentially expressed in any of the treatments. Supplementary material 2 (DOCX 14677 kb)

Figure S3: The experimental design. Supplementary material 3 (DOCX 81 kb)

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Jansen, M., Vergauwen, L., Vandenbrouck, T. et al. Gene expression profiling of three different stressors in the water flea Daphnia magna . Ecotoxicology 22, 900–914 (2013). https://doi.org/10.1007/s10646-013-1072-y

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