Altering n-3 to n-6 polyunsaturated fatty acid ratios affects prostaglandin production by ovine uterine endometrium

Cheng, Zhangrui; Abayasekara, D R E; Ward, Freya; Preece, Daniel M.W.; Raheem, Kabir Ayobami; Wathes, D C

doi:10.1016/j.anireprosci.2013.10.015

Cited by 15 publications

(13 citation statements)

References 57 publications

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“…Uterine endometrial cells (a mixture of primary epithelial and stromal cells) were isolated and cultured following the methods described previously (Cheng et al 2013, Oguejiofor et al 2015b. Briefly, under sterile conditions, strips of intercaruncular endometrium were separated and put into serum-free Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-12 Ham (DMEM/F12 medium) (Sigma) and chopped into 1 mm 3 cubes.…”

Section: Animal Cell Isolation and Culturementioning

confidence: 99%

“…The concentrations of all the ten target genes (PTGS1, PTGS2, mPGES1, AKR1B1, PTGER2, PTGER3, PTGFR, OXTR, PGR and ESR1) and four reference genes (GAPDH, RPL19, ACTB and 18SrRNA) were quantified using qPCR via an absolute quantification approach following the method described previously (Cheng et al 2013). The DNA amplified from cDNA used for standards in the qPCR assay was purified using a QIAquick PCR purification kit (Qiagen), and their quality and concentrations were determined with the NanoDrop ND-1000 spectrophotometer.…”

Section: Qpcr Analysis For Gene Expressionmentioning

confidence: 99%

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BVDV alters uterine prostaglandin production during pregnancy recognition in cows

Cheng¹,

Abudureyimu²,

Oguejiofor³

et al. 2016

Reproduction

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Embryonic mortality in cows is at least in part caused by failure of pregnancy recognition (PR). Evidence has shown that bovine viral diarrhoea virus (BVDV) infection can disrupt pregnancy. Prostaglandins (PG) play important roles in many reproductive processes, such as implantation. The aim of this study was to investigate the effect of BVDV infection on uterine PG production and PR using an in vitro PR model. Bovine uterine endometrial cells isolated from ten BVDV-free cows were cultured and treated with 0 or 100 ng/mL interferon-τ (IFNT) in the absence or presence of non-cytopathic BVDV (ncpBVDV). PGF 2α and PGE 2 concentrations in the spent medium were measured using radioimmunoassays, and in the treated cells expression of the genes associated with PG production and signalling was quantified using qPCR. The results showed that the IFNT challenge significantly stimulated PTGS1 and PTGER3 mRNA expression and PGE 2 production; however, these stimulatory effects were neutralised in the presence of ncpBVDV infection. ncpBVDV infection significantly increased PTGS1 and mPGES1 mRNA expression and decreased AKR1B1 expression, leading to increased PGE 2 and decreased PGF 2α concentrations and an increased PGE 2 :PGF 2α ratio. The other tested genes, including PGR, ESR1, OXTR, PTGS2, PTGER2 and PTGFR, were not significantly altered by IFNT, ncpBVDV or their combination. Our study suggests that BVDV infection may impair PR by (1) inhibiting the effect of IFNT on uterine PG production and (2) inducing an endocrine switch of PG production from PGF 2α to PGE 2 to decrease uterine immunity, thereby predisposing the animals to uterine disease.Reproduction (2016) 151 605-614

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Section: Animal Cell Isolation and Culturementioning

confidence: 99%

Section: Qpcr Analysis For Gene Expressionmentioning

confidence: 99%

BVDV alters uterine prostaglandin production during pregnancy recognition in cows

Cheng¹,

Abudureyimu²,

Oguejiofor³

et al. 2016

Reproduction

Self Cite

View full text Add to dashboard Cite

show abstract

“…The qPCR amplification was performed following the method described previously (Cheng et al, 2013) on a CFX 96 real-time PCR detection system (BioRad) using a Sygreen mix kit (PCR Biosystems Ltd.) following the protocol supplied. A melting curve analysis was included to confirm the specificity of amplification and avoid primer dimers.…”

Section: Quantitative Real-time Reverse Transcription-polymerase Chaimentioning

confidence: 99%

“…A melting curve analysis was included to confirm the specificity of amplification and avoid primer dimers. GAPDH was initially quantified using qPCR via an absolute quantification approach following the method described previously (Cheng et al, 2013). As GAPDH expression was not affected by LPS treatments (P>0.05) (data not shown) it was used as an appropriate reference gene…”

Section: Quantitative Real-time Reverse Transcription-polymerase Chaimentioning

confidence: 99%

Bovine P-selectin mediates leukocyte adhesion and is highly polymorphic in dairy breeds

Chen

Cheng

Werling

et al. 2016

Research in Veterinary Science

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“…The procedures have been described in an earlier study [30]. The concentration 152 and purity of the isolated RNA samples was determined with a NanoDrop ND-1000 153 spectrophotometer (NanoDrop Technologies Inc., Wilmington, DE).…”

Section: Aspx) Sequence Information Accession Numbers and 148mentioning

confidence: 99%

In vivo and in vitro studies of MUC1 regulation in sheep endometrium

et al. 2016

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Altering n-3 to n-6 polyunsaturated fatty acid ratios affects prostaglandin production by ovine uterine endometrium

Cited by 15 publications

References 57 publications

BVDV alters uterine prostaglandin production during pregnancy recognition in cows

BVDV alters uterine prostaglandin production during pregnancy recognition in cows

Bovine P-selectin mediates leukocyte adhesion and is highly polymorphic in dairy breeds

In vivo and in vitro studies of MUC1 regulation in sheep endometrium

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