Temporal changes in the transcriptomic profile of granulosa cells of pigs treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin

The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the transcriptome of aryl hydrocarbon receptor (AhR) knock-down porcine granulosa cells

“…The AVG-16 cell line was derived from granulosa cells of medium porcine follicles (Horisberger, 2006), and the cells were reported to exhibit AhR expression, both at mRNA and protein level (Sadowska et al, 2015). Previously, we employed the intact (untransfected) AVG-16 cells to study the impact of TCDD on the gene (Sadowska et al, 2017;Nynca et al, 2019) and lncRNA expression profiles (Ruszkowska et al, 2018) as well as on the cell proteome (Orlowska et al, 2018). The AhR gene and protein abundance in the cultured AhR knock-down AVG-16 cells was significantly reduced in comparison to control untransfected cells.…”

“…The changes in the gene expression profile of AhR knock-down granulosa cells were examined after 3, 12 and 24 h of TCDD treatment. Due to the fact that the expression of particular genes reach a peak or a nadir at different times after a treatment, a one-timepoint-study does not allow for ascertaining some important changes in gene expression as well as does not allow for proper investigation of changes overtime in the transcriptomic profile (Nynca et al, 2019). In consequence, a total of 360 DEGs was identified in AhR knock-down cells treated with TCDD for 3, 12 and 24 h. Similarly to previously reported TCDD-induced changes in gene expression level of intact porcine granulosa cells (Sadowska et al, 2017), most of the presently identified DEGs (354 out of 360) were detected after three hours of TCDD treatment.…”

“…In the present study, we were focused on identifying all possible pathways (molecules) involved in the mechanism of TCDD action in porcine granulosa cells and, therefore, the selected dose of TCDD (100 nM) moderately exceeded its environmentally relevant concentrations. The concentration of 100 nM of TCDD was previously reported to affect porcine granulosa cells steroidogenesis (Gregoraszczuk et al, 2000;Gregoraszczuk, 2002;Jablonska et al, 2014), gene expression (Piasecka-Srader et al, 2016;Sadowska et al, 2017;Nynca et al, 2019), lncRNA expression (Ruszkowska et al, 2018) and proteome (Orlowska et al, 2018) without affecting cell viability and morphology. Moreover, 100 nM was one of the two most frequently used TCDD concentrations in in vitro experiments performed on porcine ovarian cells.…”

“…Since TCDD influences the production of estrogens and progesterone by porcine granulosa cells (Gregoraszczuk, 2002;Jablonska et al, 2011;Jablonska et al, 2014), it is of importance to identify its molecular targets in follicular cells. The results of our previous studies, performed on porcine granulosa cells, demonstrated, among others, that TCDD affected the expression of transcripts involved in the follicular atresia as well as cell proliferation and cell cycle regulation (Sadowska et al, 2017;Ruszkowska et al, 2018;Nynca et al, 2019). The granulosa cells of pigs were found to express the Ah receptor (Sadowska et al, 2015).…”

“…In the current study, we compared the transcriptomes of the TCDD-treated AhR knockdown porcine granulosa cells and the untreated AhR knock-down cells. To better recognize the TCDD mode of action, the comparison was made at three time points: 3, 12 and 24 h of cell incubations with TCDD (Sadowska et al, 2017;Nynca et al, 2019). Due to the fact that two examined cell types (TCDD-treated and untreated cells) were both AhR knock-down, the effect of AhR knock-down was excluded.…”