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Lipopolysaccharide enhances apoptosis of corpus luteum in isolated perfused bovine ovariesin vitro


Lüttgenau, Johannes; Möller, Benjamin; Kradolfer, David; Wellnitz, Olga; Bruckmaier, Rupert M; Miyamoto, Akio; Ulbrich, Susanne E; Bollwein, Heiner (2016). Lipopolysaccharide enhances apoptosis of corpus luteum in isolated perfused bovine ovariesin vitro. Reproduction, 151(1):17-28.

Abstract

Lipopolysaccharide (LPS), the endotoxin of gram-negative bacteria, has detrimental effects on structure and function of the bovine corpus luteum (CL) in vivo. The objective was to investigate whether these effects were mediated directly by LPS or via LPS-induced release of PGF2α. Bovine ovaries with a mid-cycle CL were collected immediately after slaughter and isolated perfused for 240 min. After 60 min of equilibration, LPS (0.5 μg/mL) was added to the medium of five ovaries, whereas an additional six ovaries were not treated with LPS (control). After 210 min of perfusion, all ovaries were treated with 500 iu of hCG. In the effluent perfusate, concentrations of progesterone (P4) and PGF2α were measured every 10 and 30 min, respectively. Punch biopsies of the CL were collected every 60 min and used for RT-qPCR to evaluate mRNA expression of receptors for LPS (TLR2, -4) and LH (LHCGR), the cytokine TNFA, steroidogenic (STAR, HSD3B), angiogenic (VEGFA121, FGF2) and vasoactive (EDN1) factors, and factors of prostaglandin synthesis (PGES, PGFS, PTGFR) and apoptosis (CASP3, -8, -9). Treatment with LPS abolished the hCG-induced increase in P4 (P≤0.05); however, there was only a tendency (P=0.10) for increased release of PGF2α at 70 min after LPS challenge. Furthermore, mRNA abundance of TLR2, TNFA, CASP3, CASP8, PGES, PGFS, and VEGFA121 increased (P≤0.05) after LPS treatment, whereas all other factors remained unchanged (P>0.05). In conclusion, reduced P4 responsiveness to hCG in LPS-treated ovaries in vitro was not due to reduced steroidogenesis, but was attributed to enhanced apoptosis. However, an impact of luteal PGF2α could not be excluded.

Abstract

Lipopolysaccharide (LPS), the endotoxin of gram-negative bacteria, has detrimental effects on structure and function of the bovine corpus luteum (CL) in vivo. The objective was to investigate whether these effects were mediated directly by LPS or via LPS-induced release of PGF2α. Bovine ovaries with a mid-cycle CL were collected immediately after slaughter and isolated perfused for 240 min. After 60 min of equilibration, LPS (0.5 μg/mL) was added to the medium of five ovaries, whereas an additional six ovaries were not treated with LPS (control). After 210 min of perfusion, all ovaries were treated with 500 iu of hCG. In the effluent perfusate, concentrations of progesterone (P4) and PGF2α were measured every 10 and 30 min, respectively. Punch biopsies of the CL were collected every 60 min and used for RT-qPCR to evaluate mRNA expression of receptors for LPS (TLR2, -4) and LH (LHCGR), the cytokine TNFA, steroidogenic (STAR, HSD3B), angiogenic (VEGFA121, FGF2) and vasoactive (EDN1) factors, and factors of prostaglandin synthesis (PGES, PGFS, PTGFR) and apoptosis (CASP3, -8, -9). Treatment with LPS abolished the hCG-induced increase in P4 (P≤0.05); however, there was only a tendency (P=0.10) for increased release of PGF2α at 70 min after LPS challenge. Furthermore, mRNA abundance of TLR2, TNFA, CASP3, CASP8, PGES, PGFS, and VEGFA121 increased (P≤0.05) after LPS treatment, whereas all other factors remained unchanged (P>0.05). In conclusion, reduced P4 responsiveness to hCG in LPS-treated ovaries in vitro was not due to reduced steroidogenesis, but was attributed to enhanced apoptosis. However, an impact of luteal PGF2α could not be excluded.

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Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Veterinary Clinic > Department of Farm Animals
Dewey Decimal Classification:570 Life sciences; biology
630 Agriculture
Language:English
Date:1 June 2016
Deposited On:18 Dec 2015 13:47
Last Modified:25 Feb 2017 08:07
Publisher:BioScientifica Ltd.
ISSN:1470-1626
Additional Information:Disclaimer: this is not the definitive version of record of this article. This manuscript has been accepted for publication in Reproduction, but the version presented here has not yet been copy-edited, formatted or proofed. Consequently, Bioscientifica accepts no responsibility for any errors or omissions it may contain. The definitive version is now freely available at http://doi.org/10.1530/REP-15-0281 [2016].
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1530/REP-15-0281
PubMed ID:26483517

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