Differential gene expression in bovine endometrial epithelial cells after challenge with LPS; specific implications for genes involved in embryo maternal interactions

Abstract

Lipopolysaccharide (LPS) expressed on the surface of Gram-negative bacteria activates pro-inflammatory pathways, dys-regulates the function of endometrial cells and is a key player in the mechanisms involved in endometritis. This study aimed to investigate the effects of LPS on bovine endometrial epithelial cells (bEEC) from whole transcriptome with a special focus on genes involved in embryo-maternal interactions. Following in vitro culture, bEEC from three cows were exposed to 0, 2, and 8 μg/mL LPS for 24h. RNA samples extracted at 0 and 24 hours were analyzed by RNA sequencing (RNA-seq). At 24h, 2035 differentially expressed genes (DEGs) were identified between controls and samples treated with 2 μg/mL LPS. Gene ontology analysis showed that over-expressed DEGs were associated to immune response, response to stress and external stimuli, catalytic activity, and cell cycle. Genes associated with cell membrane and cell adhesion pathways were under-expressed. LPS induced changes in expression of specific genes related to embryo-maternal interactions including under-expression of eight members of the cadherin superfamily, over-expression of six members of the mucin family, and differential expression of a large set of genes binding the above molecules and of more than 20 transcripts coding for cytokines and their receptors. Type I interferon-τ dependent genes were also over-expressed. From a sub-set of 19 genes, (biological replicates of bEEC from cows taken at time 6 (n = 3), 24 (n = 6) and 48 hours (n = 3), and 2 technical replicates per sample) differential gene expression was confirmed by RT 2 -qPCR (r 2 between fold changes at 24 hours by RT 2 -qPCR and RNA-seq = 0.97). These results indicate that LPS affects the function of bEEC in many ways by differential transcription, glycolytic metabolism and oxidative stress. Many transcriptomic signatures related to implantation and embryo maternal interactions were strongly affected by LPS. These results pave the way for further studies to investigate the duration of these changes and their possible impact on endometrial function and fertility.

Construction and Analysis for Dysregulated lncRNAs and mRNAs in Lipopolysaccharide-Induced Porcine Peripheral Blood Mononuclear Cells

“…It was well recognized that a series of genes involved in LPS-induced in ammation [18][19][20]. In this study, the DE mRNAs functional enrichment results showed that these genes were related to some biological processes, including in ammatory response, immune response, cytokine activity, chemokine activity, cytokine-cytokine receptor interaction, TNF signal pathway, NF-κB signal pathway, JAK-STAT signal pathway, NOD-like receptor signal pathway and TLR signal pathway, which were closely associated with LPS-induced in ammation [21][22][23][24][25].…”

Analysis of transcriptomic changes in bovine endometrial stromal cells treated with lipopolysaccharide

“…We chose P6 stromal cells. Previous study using P5 bovine endometrial epithelial cells to investigate effects of LPS on whole transcriptome with a special focus on genes involved in embryo-maternal interactions [23]. And transcriptome profiling analysis of on P6 bovine vaginal epithelial cell response to an isolated lactobacillus strain [24].…”

“…Other studies have been performed on RNA-seq analysis of effects of LPS on whole transcriptome of bovine endometrial epithelial cells with focus on genes involved in embryo-maternal interactions. They identified 2,035 and 2,073 DEGs in control cells and cells treated with LPS [23]. However, comprehensive transcriptomic analysis of response of bovine endometrial stromal cells to LPS by RNA-seq remains to be elucidated.…”

Functional profiling of the endometrium transcriptome during preimplantation development in Finnsheep, Texel and their F1 crosses

“…Transcriptome analyses of pretransfer embryos and the endometrium identified several genes with potential indicator to pregnancy success in cattle (Salilew-Wondim et al 2010). Another study reported over expression of several immune-related genes including cytokines in the bovine endometrium as a result of lipopolysaccharide (LPS) treatment (Guo et al 2019). Brooks et al (2016) evaluated gene expression profiles of the endometrium during early pregnancy in sheep.…”