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Increased apoptosis, reduced Wnt/β-catenin signaling, and altered tail development in zebrafish embryos exposed to a human-relevant chemical mixture
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organismal Biology, Environmental toxicology. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Centre for Reproductive Biology in Uppsala (CRU).ORCID iD: 0000-0001-5512-2214
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organismal Biology, Environmental toxicology. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Centre for Reproductive Biology in Uppsala (CRU).ORCID iD: 0000-0002-7389-8849
Uppsala University, Science for Life Laboratory, SciLifeLab. Uppsala University, Disciplinary Domain of Science and Technology, Mathematics and Computer Science, Department of Information Technology, Division of Visual Information and Interaction. Uppsala University, Disciplinary Domain of Science and Technology, Mathematics and Computer Science, Department of Information Technology, Computerized Image Analysis and Human-Computer Interaction.ORCID iD: 0000-0002-6699-4015
Public Health Sciences, Karlstad University; Icahn School of Medicine at Mount Sinai.
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2021 (English)In: Chemosphere, ISSN 0045-6535, E-ISSN 1879-1298, Vol. 264, no 1, article id 128467Article in journal (Refereed) Published
Abstract [en]

A wide variety of anthropogenic chemicals is detected in humans and wildlife and the health effects of various chemical exposures are not well understood. Early life stages are generally the most susceptible to chemical disruption and developmental exposure can cause disease in adulthood, but the mechanistic understanding of such effects is poor. Within the EU project EDC-MixRisk, a chemical mixture (Mixture G) was identified in the Swedish pregnancy cohort SELMA by the inverse association between levels in women at around gestational week ten with birth weight of their children. This mixture was composed of mono-ethyl phthalate, mono-butyl phthalate, mono-benzyl phthalate, mono-ethylhexyl phthalate, mono-isononyl phthalate, triclosan, perfluorohexane sulfonate, perfluorooctanoic acid, and perfluorooctane sulfonate. In a series of experimental studies, we characterized effects of Mixture G on early development in zebrafish models. Here, we studied apoptosis and Wnt/β-catenin signaling which are two evolutionarily conserved signaling pathways of crucial importance during development. We determined effects on apoptosis by measuring TUNEL staining, caspase-3 activity, and acridine orange staining in wildtype zebrafish embryos, while Wnt/β-catenin signaling was assayed using a transgenic line expressing an EGFP reporter at β-catenin-regulated promoters. We found that Mixture G increased apoptosis, suppressed Wnt/β-catenin signaling in the caudal fin, and altered the shape of the caudal fin at water concentrations only 20–100 times higher than the geometric mean serum concentration in the human cohort. These findings call for awareness that pollutant mixtures like mixture G may interfere with a variety of developmental processes, possibly resulting in adverse health effects.

Place, publisher, year, edition, pages
Elsevier, 2021. Vol. 264, no 1, article id 128467
Keywords [en]
Mixtures, Zebrafish, Apoptosis, Wnt/beta-catenin, PFOS
National Category
Environmental Sciences Developmental Biology
Identifiers
URN: urn:nbn:se:uu:diva-409016DOI: 10.1016/j.chemosphere.2020.128467ISI: 000599817400057PubMedID: 33032226OAI: oai:DiVA.org:uu-409016DiVA, id: diva2:1424543
Funder
EU, Horizon 2020, 634880Swedish Institute
Note

Title in thesis list of papers: Increased apoptosis, reduced Wnt/β-catenin signaling, and altered tail development in zebrafish embryos exposed to a chemical mixture that has been inversely associated with birth weight in humans

Available from: 2020-04-17 Created: 2020-04-17 Last updated: 2024-01-15Bibliographically approved
In thesis
1. Developmental exposure to mixtures of environmental pollutants: Studies on metabolism, developmental processes, and reproductive organs in zebrafish and chicken embryos
Open this publication in new window or tab >>Developmental exposure to mixtures of environmental pollutants: Studies on metabolism, developmental processes, and reproductive organs in zebrafish and chicken embryos
2020 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Humans and wildlife are continuously exposed to mixtures of environmental pollutants. Mixture toxicity can be challenging to predict due to interactions between chemicals and thus whole-mixture approaches are crucial in toxicology. Developing organisms are generally more sensitive to chemical insult than adults and early exposure has been linked to metabolic and reproductive disorders later in life. It is thus imperative to clarify how mixtures of environmental pollutants affect early development.

Within this thesis, consequences of early exposure to human-relevant chemical mixtures have been demonstrated using zebrafish and chicken embryos. The mixtures were designed previously based on negative associations with birth weight (mixture G) or anogenital distance (mixture S) in Swedish children. Mixture G consist of phthalate monoesters, perfluoroalkyl acids, and triclosan (TCS). It was assessed for effects on developmental processes (apoptosis and wnt/β-catenin signaling) and lipid metabolism in zebrafish. Two components of mixture G were assessed as single compounds: perfluorooctane sulfonate (PFOS) and TCS. Exposure to mixture G induced apoptosis, reduced wnt/β-catenin signalling, increased visceral adiposity, and reduced blood- and whole body-lipid levels in developing zebrafish. PFOS induced apoptosis but not Wnt/β-catenin signaling and TCS had similar effects on lipid levels as the mixture, although the effect of TCS on adipogenesis was not as pronounced. Mixture S, which consists of four phthalate monoesters, and a suggested bisphenol A metabolite (4-Methyl-2,4-bis(4-hydroxyphenyl)pent-1-ene; MBP) were assessed for disruption of reproductive organ development in chicken embryos. No effects were observed by mixture S. MBP caused feminization in male embryos (left ovotestis, feminized gonadal mRNA expression pattern, and Müllerian duct retention). MBP-exposed females displayed smaller left ovaries, malformed left Müllerian ducts, and right Müllerian duct retention.

In conclusion, a mixture that has been implicated in altered intrauterine metabolism and growth in Swedish children caused developmental and metabolism disrupting effects in larval zebrafish. PFOS and TCS most likely contribute to the effects by the mixture. Furthermore, the suggested bisphenol A metabolite MBP, but not a mixture of phthalate monoesters, altered both male and female reproductive organ development in chicken embryos. The results were generated using models of both environmental and human relevance. The results in this thesis demonstrate the value of combining epidemiological and experimental studies to assess mixture toxicity. 

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2020. p. 57
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1937
Keywords
Mixtures, Developmental toxicology, Endocrine disrupting chemicals, Metabolism, Feminization, Zebrafish, Chicken embryo
National Category
Pharmacology and Toxicology Environmental Sciences Developmental Biology
Research subject
Biology with specialization in Environmental Toxicology; Biology with specialization in Environmental Toxicology; Biology with specialization in Environmental Toxicology; Biology with specialization in Environmental Toxicology
Identifiers
urn:nbn:se:uu:diva-409018 (URN)978-91-513-0954-5 (ISBN)
Public defence
2020-06-10, Ekmansalen, Evolutionsbiologiskt centrum, Norbyvägen 14, Uppsala, 13:00 (English)
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Supervisors
Available from: 2020-05-19 Created: 2020-04-17 Last updated: 2020-06-16

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Smirnova, AnnaMentor, AnnaRanefall, PetterBrunström, BjörnMattsson, AnnaJönsson, Maria

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