Bovine oocyte exposure to perfluorohexane sulfonate (PFHxS) induces phenotypic, transcriptomic, and DNA methylation changes in resulting embryos in vitro

Hallberg, Ida; Persson, S. G. B.; Olovsson, Matts; Moberg, Mikaela; Laskowski, Denise; Damdimopoulou, Pauliina; Sirard, Marc‐André; Rüegg, Joëlle

doi:10.1016/j.reprotox.2022.02.004

Cited by 6 publications

(15 citation statements)

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“…For exposure, PFOS (potassium salt >98%, CAS 1763-23-1) and PFHxS (potassium salt >98%, CAS 355-46-4) dissolved in molecular grade water was added to the maturation media of the respective exposed groups to reach the concentration of 0.1 μg/mL (PFOS) or 40 μg/mL (PFHxS). These concentrations were chosen based on results from previous experiments [ 29 , 34 ]. In the control, vehicle (molecular grade water) was added in the same volume as the stock solutions containing PFOS/PFHxS.…”

Section: Methodsmentioning

confidence: 99%

“…Due to persistence of the compounds used, significant degradation was not expected during storage. The stock solution concentrations of PFOS and PFHxS were validated using mass-spectrometry which is described elsewhere [ 29 , 34 ].…”

Section: Methodsmentioning

confidence: 99%

“…An apoptotic nucleus was defined as: a nucleus within the blastocyst area, stained with both DAPI and TUNEL stains. TUNEL-positive cells were segmented and identified using a macro developed for ImageJ [ 38 ] as previously described [ 29 ]. In brief, the basis for the apoptosis segmentation was an iterative 3D version of the Per Object Ellipsefit (POE) algorithm.…”

Section: Methodsmentioning

confidence: 99%

“…In animal models, oocyte toxicity has been observed in the mouse [ 27 ] and pig [ 28 ]. It has also previously been shown that development of blastocysts is affected upon exposure during oocyte maturation in the bovine using an in vitro model [ 29 ].…”

Section: Introductionmentioning

confidence: 99%

“…Apoptosis is commonly described as programmed cell death and can, for instance, occur as a response to toxicity [ 33 ]. In the bovine blastocyst, PFASs appear to alter genes associated with apoptotic pathways [ 29 ]. Therefore, we also wished to assess the effects of PFASs on cell count and proportion of apoptotic cells in resulting blastocysts, by using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining assay and objective image analysis.…”

Section: Introductionmentioning

confidence: 99%

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Occurrence of late-apoptotic symptoms in porcine preimplantation embryos upon exposure of oocytes to perfluoroalkyl substances (PFASs) under in vitro meiotic maturation

Leclercq

Hallberg

2022

PLoS ONE

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The objectives of this study were to evaluate the effect of perfluoroalkyl substances on early embryonic development and apoptosis in blastocysts using a porcine in vitro model. Porcine oocytes (N = 855) collected from abattoir ovaries were subjected to perfluorooctane sulfonic acid (PFOS) (0.1 μg/ml) and perfluorohexane sulfonic acid (PFHxS) (40 μg/ml) during in vitro maturation (IVM) for 45 h. The gametes were then fertilized and cultured in vitro, and developmental parameters were recorded. After 6 days of culture, resulting blastocysts (N = 146) were stained using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and imaged as stacks using confocal laser scanning microscopy. Proportion of apoptotic cells as well as total numbers of nuclei in each blastocyst were analyzed using objective image analysis. The experiment was run in 9 replicates, always with a control present. Effects on developmental parameters were analyzed using logistic regression, and effects on apoptosis and total numbers of nuclei were analyzed using linear regression. Higher cell count was associated with lower proportion of apoptotic cells, i.e., larger blastocysts contained less apoptotic cells. Upon PFAS exposure during IVM, PFHxS tended to result in higher blastocyst rates on day 5 post fertilization (p = 0.07) and on day 6 post fertilization (p = 0.05) as well as in higher apoptosis rates in blastocysts (p = 0.06). PFHxS resulted in higher total cell counts in blastocysts (p = 0.002). No effects attributable to the concentration of PFOS used here was seen. These findings add to the evidence that some perfluoroalkyl substances may affect female reproduction. More studies are needed to better understand potential implications for continued development as well as for human health.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%