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Development and evaluation of gene expressions in Bufo bufo- tadpoles as biomarkers for exposure of chemical pollutants
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences, Toxicology.
2019 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Introduction: Occurrence and emission of chemical pollutants such as organic compounds, heavy metals and pharmaceuticals are high in the aquatic environment and a concern worldwide due to the risk of affecting organisms negatively. Biomarkers can indicate the magnitude of organism response to environmental pollutants and provide the causal link between the presence of a chemical and ecological effect. These pollution indicators can be used in effect-based environmental monitoring. Cytochrome p450 1A (CYP1A), metallothionein (MT) and cytochrome p450 3A (CYP3A) are examples which can be used as biomarkers for identification of exposure of pollutants.

The aims of this project were to find functioning primers for MT, CYP1A and CYP3A, and examine whether mRNA levels of these genes are relevant to use as biomarkers to demonstrate exposure of metals, dioxin-like chemicals and ethinyl estradiol (EE2).

Methods: The tadpoles were exposed to Cadmium chloride (CdCl2), β-naphthoflavone (BNF) and EE2. RNA of the liver tissue samples was isolated, and concentration and integrity were measured. The primers were designed based on different strategies, A, existing gene mRNA sequences, B, Short Read Archive (SRA) database of NBCI and C, extended RNA-sequencing. qRTPCR was used for RNA-primer-testing and mRNA expression level measurement.

The results show significantly differences between sites and individuals, expression level of CYP1A was significantly induced but no induction of CYP3A and MT was achieved.

Conclusions: Applying CYP1A gene as biomarker on wild tadpoles looks promising. For the use of CYP3A and MT as biomarkers, further efforts are needed.

Place, publisher, year, edition, pages
2019. , p. 26
Keywords [en]
Bufo bufo, CYP1A, CYP3A, Metallothionein, Biomarker
National Category
Pharmacology and Toxicology
Identifiers
URN: urn:nbn:se:uu:diva-391084OAI: oai:DiVA.org:uu-391084DiVA, id: diva2:1349224
External cooperation
Sveriges Lantbruksuniversitet
Subject / course
Toxicology
Educational program
Master of Science Programme in Pharmacy
Presentation
(English)
Supervisors
Examiners
Available from: 2019-09-10 Created: 2019-09-06 Last updated: 2019-09-10Bibliographically approved

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