Intramammary infusion of lipopolysaccharide promotes inflammation and alters endometrial gene expression in lactating Holstein cows

Abstract

The objective of the current study was to evaluate the effects of 2 intramammary infusions of lipopolysaccharide (LPS) on inflammatory and reproductive parameters and endometrial gene expression of lactating Holstein cows. At 35 ± 7 d in milk, 20 cows were submitted to a Double Ovsynch program and randomly assigned to control (n = 11) and LPS (n = 9) treatments. Cows from the LPS treatment received 2 intramammary infusions of 25 µg of LPS after morning milking on d 5 and 10 post-AI, whereas control cows were infused with only saline. Blood samples were taken and ultrasound scanning of the ovaries was performed during the entire study before and after AI to determine haptoglobin, tumor necrosis factor-α, and progesterone concentrations as well as response to the hormonal protocol and corpus luteum diameter. Milk yield was evaluated and samples were taken for somatic cell count at 0, 10, 24, 34, and 96 h relative to each infusion. Rumen-reticular temperature was recorded using a rumen-reticular bolus logger and summarized hourly. On d 15 post-AI, uterine flushing for conceptus recovery and endometrial biopsies were performed. Samples of endometrium from cows with positive embryo recovery (control = 5; LPS = 6) were submitted to mRNA extraction and quantitative reverse-transcription PCR analysis of 96 target genes. Haptoglobin concentrations in plasma were greater for LPS treatment (control = 0.24 ± 0.07, LPS = 0.89 ± 0.06 optical density), but tumor necrosis factor-α concentrations were similar (control = 0.67 ± 0.11, LPS = 0.46 ± 0.11 ng/mL) between treatments. Lipopolysaccharide reduced milk yield after treatment (control = 34.3 ± 1.5, LPS = 29.4 ± 1.6 kg/d), whereas somatic cell count (log) was greater in LPS-treated cows until 34 h after infusions (control = 2.3 ± 0.1, LPS = 3.3 ± 0.1 cells/mL of milk). Rumen-reticular temperature of LPS cows was elevated between 5 and 10 h after each infusion compared with control cows (control = 39.5 ± 0.1, LPS = 40.1 ± 0.1°C). Progesterone concentration after AI was unaffected by treatment or pregnancy status as well as corpus luteum diameter and conceptus length on d 15. Lipopolysaccharide treatment altered the expression of 13 key genes in the endometrium (mostly upregulated), whereas another 17 tended to be modulated. Modified gene expression included genes related to immune response (PTX3 = 2.34-fold increase; IL6 = 3.42-fold increase; and TCN1 = 2.52-fold increase), adhesion molecules (CADM3 = 1.93-fold increase; MMP19 = 1.49-fold increase; EMMPRIN = 1.20-fold increase; SELL = 1.91-fold increase), Wnt signaling pathway (WNT2, FZD4, and FZD7, all <1.5-fold increase), and interferon-stimulated genes (BMP15 = 0.27-fold decrease; ISG15 = 2.17-fold increase, and MX2 = 2.23-fold increase). In summary, intramammary infusions of LPS were able to trigger an inflammatory response with no effect on corpus luteum diameter and concentration of progesterone in plasma. However, a limited but important set of modulations in the endometrium gene expression at d 15 of gestation was found.

Influence of pathogens causing clinical mastitis on reproductive variables of dairy cows

“…Some authors have been studying this mechanism, trying to understand the underlying pathway. Lipopolysaccharide could delay oocyte development in cows (Soto et al, 2003), decrease the release of estradiol from granulosa cells (Williams et al, 2008), inhibit ovulation (Williams et al, 2008), lead to early apoptosis of oocytes (Zhao et al, 2019), affect oviducts promoting early embryo death (Ibrahim et al, 2015), delay cytoplasm maturation in oocytes causing embryonic developmental changes (Magata and Shimizu, 2017), cause problems in pregnancy maintenance (Bromfield et al, 2015;Campos et al, 2018), and reduce the primordial ovarian follicle pool to cause a long-term effect on fertility (Bromfield and Sheldon, 2013). These findings support the results discussed in this investigation.…”

“…The gram-negative group had greater pregnancy loss compared with the control and gram-positive groups. Apparently, the LPS in the gram-negative bacteria causes a significant disturbance in embryo development, leading to an increase in pregnancy loss in cows with mastitis caused by gram-negative bacteria; this has also been observed by other authors (Bromfield et al, 2015;Ibrahim et al, 2015;Magata and Shimizu, 2017;Campos et al, 2018). No difference in pregnancy loss was observed for cows with subclinical mastitis after embryo transfer (Barbosa et al, 2018).…”

A Clinical Predictive Model of Progesterone Elevation on the Day of HCG Administration Was Establish Based on the Basal P, BMI and Ovarian Induction Regimen

“…More recently microarray-based studies con rmed that the PE induced a change in endometrial gene expression [4,5].…”

Endometrium-on-a-chip reveals the endometrial transcriptome, and protein content of secretome are altered by changes in circulating concentrations of insulin and glucosein vitro

“…Data indicates this metabolic stress alters global gene expression in the embryo/conceptus, the oviduct (9,13), and endometrium (13)(14)(15). Conceptuses derived from early in lactation are less developmentally competent (metabolic stress) compared to late stages of lactation (16). Even if a high-quality embryo is produced exposure to the uterine environment of lactating cows can alter developmental potential (16,17).…”

“…Conceptuses derived from early in lactation are less developmentally competent (metabolic stress) compared to late stages of lactation (16). Even if a high-quality embryo is produced exposure to the uterine environment of lactating cows can alter developmental potential (16, 17).…”