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Preliminary study on adverse effects of phenanthrene and its methyl and phenyl derivatives in larval zebrafish, Danio rerio

Wolińska L., Brzuzan P., Woźny M., Góra M., Łuczyński M. K., Podlasz P., Kolwicz S., Piasecka A.

Environmental Biotechnology

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2011

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Vol. 7, No. 1

26-33

EN

Toxic effects of polycyclic aromatic hydrocarbons (PAHs) have been extensively studied in fish, although knowledge concerning biological activities of phenanthrene and its derivatives remains still incomplete. The aim of this work was to evaluate lethal and sublethal effects of benzo(a)pyrene, phenanthrene and phenanthrene derivatives (1-methylphenanthrene, 4-methylphenanthrene, 1-phenylphenanthrene and 4-phenylphenanthrene) on zebrafish (Danio rerio) larvae. We conducted acute toxicity test, using 96h static renewal exposure to a series of the PAH concentrations (0.05, 0.50, 5.00, 50.00µmol*l-1), to determine the No Effect Concentration (NEC) value for each compound examined. The mean NEC estimates obtained in the study were 5.16۪.45µmol*l-1 (B[a]P), 4.88۪.13µmol*l-1 (Ph), 40.24䔰.93µmol*l-1 (1P-Ph), 47.92ۭ.61µmol*l-1 (1M-Ph), 24.31۱.33µmol*l-1 (4P-Ph) and 3.11۫.01µmol*l-1 (4M-Ph) and suggested the following order of PAH toxicities on Danio rerio larvae: 4M-Ph>Ph˜B[a]P>4PPhᲙP-Ph>1M-Ph. To gain insight into possible molecular mechanisms of apparent toxicity of phenanthrene derivatives on zebrafish larvae, we examined mRNA expression of cyp1a, cyp1b1, and vtg genes in the larvae exposed for 48h to a PAH concentration of 0.50µmol*l-1. Whereas the larvae exposed to each tested PAH displayed many developmental abnormalities (i.e. pericardial and yolk sac edema, dorsal curvature, or tail malformations), no significant upregulation of cyp1a and cyp1b1 mRNA was observed, except for zebrafish exposed to B[a]P. However, significant reduction of vtg mRNA was observed in the larvae exposed to phenanthrene and its 4P- derivative. The results may contribute to the development of a new knowledge about effects of structurally diverse phenanthrene derivatives on vertebrate organisms.

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Preliminary evaluation of ER- and AhR-mediated gene expression patterns in rainbow trout (Oncorhynchus mykiss) liver after short-term exposure to zearalenone in binary mixtures

Woźny M., Brzuzan P., Wolińska L., Góra M., Łuczyński M. K.

Environmental Biotechnology

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2010

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Vol. 6, No. 1

16-23

EN

While estrogenic properties of mycotoxin zearalenone (ZEA) has been an extensively studied issue, little is known about molecular background of its biological responses that cannot be simply explained by the estrogenic potential. The present study describes effects of ZEA (10mg*kg-1 body weight) in binary mixtures either with benzo[a]pyrene (B[a]P), or with 17β-estradiol (E2) on ER- and AhR-dependent gene expression in juvenile rainbow trout liver, evaluated using Real-Time qPCR. The study revealed dual nature of ZEA, as the treatment with this compound alone increased mRNA levels of both ER- and AhR-mediated gene expression. However, our results did not show any synergistic or additive effect of ZEA in binary mixures with E2 or B[a]P on studied gene expression levels. Whether the intriguing potential of ZEA to elicit distinct signals was a result of binding affinity to AhR or/and ER and AhR mutual receptor interactions, should be investigated in further experiments.

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MicroRNA expression in liver of whitefish (Coregonus lavaretus) exposed to microcystin-LR

Brzuzan P., Woźny M., Wolińska L., Piasecka A., Łuczyński M. K.

Environmental Biotechnology

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2010

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Vol. 6, No. 2

53-60

EN

MicroRNAs (miRNAs) are small, highly conserved, non-coding RNAs that regulate gene expression of target mRNAs through cleavage or translational inhibition. In the field of toxicology, the relationship between toxicity and microRNA expression is poorly understood. In the present study we analyzed the abundance of 9 selected miRNAs (omy-miR-21, omy-miR-21t, omy-miR-122, omy-miR-125a, omy-miR-125b, omy-miR-125t, omy-miR-199-5a, omy-miR-295, omy-let-7a) and mRNA of 3 genes (histone H2A, ribosome protein rpl19, and Dicer which is a miRNA processing enzyme) in liver samples of whitefish exposed to Microcystin-LR (MC-LR) at a dose of 100µg*kg-1body weight for 24 or 48h. In the examined liver tissue, omymiR-122 showed the highest relative constitutive level, what is consistent with data obtained from fish and mammals. Unexpectedly, the reference H2A mRNA level was consistently up-regulated (over 20-fold; P<0.05) in fish liver after both 24 and 48h of exposure to MC-LR. The result may suggest that MC-LR acts as an initiator of specific cell-physiologic signals triggering DNA replication in fish liver cells. MC-LR treatment had no effect on the examined miRNAs levels, except for omy-miR-125a and omy-let-7a. Whereas omy-miR-125a was up-regulated (ER=2.68; S.E. 1.61-6.78; P<0.05), omy-let-7a was down-regulated (ER=0.55; S.E. 0.32-0.79; P<0.05) in whitefish liver after 48h of the treatment with MC-LR, when compared to controls. More work with the fish is essential for understanding the crosstalk of the regulatory network controlled by the two miRNAs in the context of MC-LR toxicity.