RNA-seq based gene expression analysis of ovarian granulosa cells exposed to zearalenone in vitro: significance to steroidogenesis

Abstract

Zearalenone (ZEA) is a natural contaminant of various food and feed products representing a significant problem worldwide. Since the occurrence of ZEA in grains and feeds is frequent, the present study was carried out to evaluate the possible effects of ZEA on steroid production and gene expression of porcine granulosa cells, using RNA-seq analysis. Porcine granulosa cells were administered 10 μM and 30 μM ZEA during 72 h of culture in vitro. Following ZEA treatment the gene expression profile of control and exposed granulosa cells was compared using RNA-seq analysis. The results showed that in the exposed granulosa cells ZEA significantly altered the transcript levels, particularly steroidogenesis associated genes. Compared with the control group, 10 μM and 30 μM ZEA treatment significantly increased the mRNA expression of EDN1, IER3, TGFβ and BDNF genes and significantly reduced the mRNA expression of IGF-1 and SFRP2 genes. In particular, ZEA significantly decreased the expression of genes essential for estrogen synthesis including FSHR, CYP19A1 and HSD17β in granulosa cells. Furthermore, Q-PCR and Western-blot analysis also confirmed reduced expression of these genes in ZEA exposed granulosa cells. These effects were associated with a significant reduction of 17β-estradiol concentrations in the culture medium of granulosa cells. Collectively, these results demonstrated a concretely deleterious effect of ZEA exposure on the mRNA expression of steroidogenesis related genes and the production of steroid hormones in porcine ovarian granulosa cells in vitro.

Zearalenone Exposure Enhanced the Expression of Tumorigenesis Genes in Donkey Granulosa Cells via the PTEN/PI3K/AKT Signaling Pathway

“…However, the authors stated that mares fed with the naturally contaminated oats had a high incidence of follicular hematomas, which did not occur in the control ( Cortinovis et al, 2013 ). Similar to the findings in estrous pig ( Zhang et al, 2017b ), ZEA and its derivatives induced in vitro apoptosis of the granulosa cells (GCs), which were collected from estrous mares’ ovaries ( Minervini et al, 2006 ). Hence, the authors suggested that ZEA could induce follicular atresia in domestic animals.…”

“…Numerous studies have proven ZEA’s cytotoxic effect on the reproductive ( Kiang et al, 1978 ; Mehmood et al, 2000 ; Nikov et al, 2000 ), immune ( Abbes et al, 2006a , b ; Luongo et al, 2006 ), and endocrine ( Mueller et al, 2004 ) systems, as well as on heredity ( Kouadio et al, 2005 ). ZEA indirectly affects mammalian fertility by impairing the formation of primordial follicle ( Zhang et al, 2017a ), thereby causing changes in gene expression; it also induces DNA damage in ovarian GCs ( Zhang et al, 2017b ). Not much information is available on ZEA-induced alteration in dGC proliferation and development and impairment of mammalian fertility.…”

“…During follicle development, GCs replicate, secrete hormones, and support the growth of the oocyte ( Hamel et al, 2008 ). Previous investigations demonstrated that ZEA may alter GC’s function in swine ( Ranzenigo et al, 2008 ; Zhang et al, 2017b ). This study aims to evaluate the in vitro toxicity of 10 and 30 μM ZEA in donkey ovarian GCs through transcriptome analysis.…”

Transcriptional analysis of deoxynivalenol‐induced apoptosis of sow ovarian granulosa cell

“…The apoptosis of ovarian granulosa cells directly or indirectly results in the reduction of germ cells, which plays a crucial role in the determination of the number of follicles destined to ovulate in each reproductive cycle (Gebauer, Peter, Onesime, & Dhanasekaran, ). However, there are many toxins of fodder that were found to have a negative effect on ovarian granulosa cells such as 2,5‐hexanedione (2,5‐HD), T‐2 toxin and zearalenone (ZEN) (Wu, Tu, Yuan, Yuan, & Wen, ; Zhang et al, ; Zhang et al, ).…”

Zearalenone: A Mycotoxin With Different Toxic Effect in Domestic and Laboratory Animals’ Granulosa Cells

“…E. asinus , that were fed naturally ZEA-contaminated oats, had a high incidence of follicular hematomas, which did not occur during control cycles (Cortinovis et al, 2013). ZEA and its derivatives induced the apoptosis of GCs collected from the ovaries of cycling mares in vitro (Minervini et al, 2006) and from cycling pigs (Zhang et al, 2017b). Considering the contemporaneous presence of both processes, authors suggested that ZEA could induce follicular atresia in domestic animals.…”