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Gene transcription analysis of Daphnia magna multigenerations exposed to carbendazim

ABSTRACT: Pesticides are continuously released into the environment, with possible long-term consequences on aquatic organisms. One of the pesticides still applied in several crops in some countries is the fungicide carbendazim, ending up in surface waters with concentrations reaching 5 µg/L. Daphnia magna (clone k6) was used in this study as a model organism and it was exposed to a sub-lethal concentration of carbendazim (5 µg/L) for twelve generations. Gene expression alterations induced by this compound were assessed in the F0 and F12 generations using D. magna custom microarrays. Results revealed that carbendazim caused changes at the gene expression level in both generations. Genes involved in response to stress, DNA replication/repair, neurotransmission, protein biosynthesis, ATP production, lipids and carbohydrates metabolism were the most affected in both F0 and F12, although a lower number of differentially expressed genes were observed in the F12 generation exposed to carbendazim. The exposure of daphnids to carbendazim did not cause a stable change in gene expression from F0 to F12 generations. Effects at the gene expression level were early detected at the F0 generation after a short-time exposure (10 days), highlighting the advantages of using high throughput tools as early warning analysis but also providing information on chemical mode of action, which can add value in risk assessment procedures.

ORGANISM(S): Daphnia magna

PROVIDER: GSE78120 | GEO | 2017/05/01

SECONDARY ACCESSION(S): PRJNA312654

REPOSITORIES: GEO

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Project description:A reduced level of DNA methylation has recently been described in both Zn-exposed and non-exposed offspring of Daphnia magna exposed to Zn. The hypothesis examined in this study is that DNA hypomethylation has an effect on gene transcription. A second hypothesis is that accumulative epigenetic effects can affect gene transcription in non-exposed offspring from parents with an exposure history of more than one generation. Transcriptional gene regulation was studied with a cDNA microarray. In the exposed and non-exposed hypomethylated daphnids, a large proportion of common genes were similarly up- or downregulated, indicating a possible effect of the DNA hypomethylation. Two of these genes can be mechanistically involved in DNA methylation reduction. The similar transcriptional regulation of two and three genes in the F0 and F1 exposed daphnids on one hand and their non-exposed offspring on the other hand, could be the result of a one-generation temporary transgenerational epigenetic effect, which was not accumulative. The experimental design used in the current study is as follows. A set of neonates (0-24h) taken from the laboratory culture was divided into two batches. One batch was transferred to modified standard M4 medium (Elendt and Bias, 1990) and cultured for three generations (CtrlF0–CtrlF2). A second batch of neonates was transferred into the same medium, with the Zn concentration adjusted to 388 µg/L. A group of F1 neonates born from this ZnF0 generation was then cultured in this Zn-contaminated medium [Zn(+)F1]. Another group of these F1 neonates was transferred back to the control medium [Zn(-)F1]. In this way, Zn(-)F1 daphnids were only briefly exposed to Zn during the first hours of their life-cycle. Offspring of the F1Zn(-) organisms were also cultured in the control medium [Zn(-/-)F2, which were named F2Zn(-) in Vandegehuchte et al. (2009))]. Offspring of the Zn(+)F1 daphnids were in turn divided in a similar manner into two batches, one of which was cultured further in Zn-contaminated medium [Zn(+)F2] while the other one was transferred to the control medium [Zn(+/-)F2]. Each combination of generation and exposure history is termed a ‘treatment’. Three replicates of ten adult daphnids each per treatment were sampled for RNA extraction on the day the next generation was started. In general, a universal reference design was used in which the reference sample was a pool composed of differently aged daphnids cultured in aerated carbon-filtered tap water, enriched with selenium (1 µg/L) and vitamins (7.5 mg/L thiamine, 100 µg/L cyanocobalamin and 75 µg/L biotin). Dye-bias effects were accounted for by regularly swapping Cy3 and Cy5 for labelling treatments/reference pool samples.

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Gene transcription analysis of Daphnia magna multigenerations exposed to carbendazim

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