Which bovine endometrial cells are the source of and target for lysophosphatidic acid?

doi:10.1016/j.repbio.2013.01.166

Reproductive Biology

Volume 13, Issue 1, March 2013, Pages 100-103

https://doi.org/10.1016/j.repbio.2013.01.166 Get rights and content

Abstract

The objective of the study was to examine which cultured endometrial cells are the source and which are the target for lysophosphatidic acid (LPA) in the bovine uterus. LPA concentration as well as mRNA and protein expressions of the enzymes responsible for LPA synthesis (phospholipase A2: PLA₂, autotaxin: AX) were greater in epithelial than in stromal cells (P < 0.05). In turn, mRNA and protein expression of LPA receptor (LPAR1) was lower in epithelial than in stromal cells (P < 0.05). We suggest that LPA in bovine endometrium is produced mainly by epithelial cells and affects mostly stromal cells acting via LPAR1.

Introduction

Recently, we have found that LPA is locally produced and released from the bovine endometrium, both during the estrous cycle and early pregnancy. In addition, a specific receptor type 1 (LPAR1) was demonstrated in the endometrium [1]. LPA preferentially stimulated in vivo PGE₂ output from bovine endometrium during the luteal phase of the estrous cycle [1], [2]. Moreover, LPA exerted a stimulatory effect on in vitro PGE₂ production by stromal cells via the induction of prostaglandin 2 synthase (PTGS2) and prostaglandin E₂ synthase (PGES) expression [3]. These findings indicate that LPA of a uterine origin may, through the modulation of PG synthesis, play an autocrine and/or paracine role in the bovine uterus. Since it is not known which bovine endometrial cells are source and target for LPA, the aim of the present study was to determine: (1) localization of phospholipase A2 (PLA₂) and autotaxin (AX), the enzymes responsible for LPA synthesis as well as LPAR1 in the endometrial tissue during different phases of the estrous cycle; (2) LPA concentration in the epithelial and stromal cells of bovine endometrium; and (3) mRNA and protein expression of PLA₂, AX and LPAR1 in epithelial and stromal cells of bovine endometrium.

Section snippets

Materials and methods

All experimental procedures were approved by the Local Animal Care and Use Committee in Olsztyn, Poland (Agreement No. 34/2005/N). For all experiments, normally cycling Holstein/Polish Black and White (75/25%; respectively) cows (n = 22) were used (2007–2009). The animals were culled because of their low milk production. The estrus of the animals was synchronized, determined and confirmed as described previously [1]. The day of estrus was designated Day 0 of the cycle. At sequential times after

Results and discussion

In the present study we demonstrated that enzymes involved in LPA synthesis (AX and PLA₂) as well as active LPAR1 are localized in bovine endometrium. AX and PLA₂ were immunoexpressed both in epithelial and stromal cells (Fig. 1B and D). On the other hand, LPAR1 protein was mainly present in stromal cells (Fig. 1F). The staining intensity of the examined proteins did not differ among the phases of the estrous cycle. We found that LPA concentration was significantly higher in epithelial than in

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TXA2 mediates LPA1-stimulated uterine contraction in late pregnant mouse
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Lysophosphatidic acid (LPA) is known to increase uterine contraction in the estrus cycle and early pregnancy, however, the effect of LPA in late pregnant uterus and its mechanisms are not clear. In the present study, we show the LPA receptor subtypes expressed and the mechanism of LPA-induced contractions in late pregnant mouse uterus. We determined the relative mRNA expression of LPA receptor genes by quantitative PCR and elicited log concentration-response curves to oleoyl-L-α-LPA by performing tension experiments in the presence and absence of nonselective and selective receptor antagonists and inhibitors of the TXA2 pathway. LPA1 was the most highly expressed receptor subtype in the late pregnant mouse uterus and LPA1/2/3 agonist (Oleoyl-L-α LPA) elicited increased contractions in this tissue that had lesser efficacy compared to oxytocin. LPA1/3 antagonist, Ki-16425, and a potent LPA1 antagonist (AM-095) significantly inhibited the LPA-induced contractions. Further, the nonselective COX inhibitor, indomethacin, and potent thromboxane A2 synthase inhibitor, furegrelate significantly impaired LPA-induced contractions. Moreover, selective thromboxane receptor (TP) antagonist, SQ-29548, and Rho kinase inhibitor, Y-27632 almost eliminated LPA-induced uterine contractions. LPA1 stimulation elicits contractions in the late pregnant mouse uterus using the contractile prostanoid, TXA2 and may be targeted to induce labor in uterine dysfunctions/ dystocia.
Study of lysophosphatidic acid receptors (LPARs) in buffalo uterus demonstrated upregulation of LPAR1 and LPAR6 in early pregnancy
2017, Theriogenology
Citation Excerpt :
Stromal cells exhibited scarce patches of immunopositivity. Boruszewska et al. [36] demonstrated LPA1 receptor expression in stromal endometrial cells of the bovine species by mRNA and protein expression studies. In ovine endometrium, immunohistochemical localization exhibited the presence of LPA1 in the endometrial stroma as well as in glandular epithelial cells.
Lysophosphatidic acid (LPA) is an important factor involved in embryo implantation and pregnancy establishment in humans and domestic livestock. LPA exerts its action through six G-protein-coupled receptors (LPA1-LPA6). We investigated the types of LPA receptors expressed in buffalo uterus and also their differential expression in the nonpregnant, and early-pregnant endometrium. The nonpregnant, and early-pregnant (<42 days) uteri were collected from the local slaughterhouse. RT-PCR experiments detected mRNAs of all the six LPA receptors (LPAR1-LPAR6) in both nonpregnant, and early-pregnant endometrial tissues. Their comparative profiling by real-time PCR revealed that the early pregnant endometrium expressed more mRNAs of LPAR1 and LPAR6. All the mRNA fragments were sequenced and submitted to Genbank, NCBI. Western blot studies also showed a similar expression pattern of these two receptor proteins, including higher expression of both LPA1 and LPA6 proteins during early pregnancy. And between these two receptors, LPA6 upregulation was more pronounced than LPA1. In immunohistochemistry, these receptors were found to be localized in the endometrial glandular epithelial cells of both types of uterus. Level of LPA was also higher in early pregnant endometrial tissues. In summary, our study demonstrated expression of all the six LPAR mRNAs in buffalo uterus, wherein the early-pregnant uterus did express comparatively higher mRNA as well as protein of LPA1 and LPA6, indicating their role in pregnancy. The more pronounced expression of LPA6 possibly indicates its greater contribution to mediating LPA signaling in early pregnancy (29–42 days) of buffalo.
Studies on lysophosphatidic acid action during in vitro preimplantation embryo development
2016, Domestic Animal Endocrinology
Citation Excerpt :
In turn, in bovine ovary, the expression of 4 LPARs was detected in the corpus luteum (CL) [29] and in granulosa cells of the follicle [55]. In bovine endometrial tissue, the expression of only LPAR1 was found [18], with higher expression in stromal cells than in epithelial cells [56]. The different expression pattern of specific LPARs during the estrous cycle and early pregnancy accounts for the involvement of LPA in diverse events during the estrous cycle and early pregnancy [18,29].
Assisted reproductive technologies, including in vitro embryo production (IVP), have been successfully used in animal reproduction to optimize breeding strategies for improved production and health in animal husbandry. Despite the progress in IVP techniques over the years, further improvements in in vitro embryo culture systems are required for the enhancement of oocyte and embryo developmental competence. One of the most important issues associated with IVP procedures is the optimization of the in vitro culture of oocytes and embryos. Studies in different species of animals and in humans have identified important roles for receptor-mediated lysophosphatidic acid (LPA) signaling in multiple aspects of human and animal reproductive tract function. The data on LPA signaling in the ovary and uterus suggest that LPA can directly contribute to embryo–maternal interactions via its influence on early embryo development beginning from the influence of the ovarian environment on the oocyte to the influence of the uterine environment on the preimplantation embryo. This review discusses the current status of LPA as a potential supplement in oocyte maturation, fertilization, and embryo culture media and current views on the potential involvement of the LPA signaling pathway in early embryo development.
Influence of lysophosphatidic acid on estradiol production and follicle stimulating hormone action in bovine granulosa cells
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Citation Excerpt :
We also previously indicated the possibility of LPA synthesis in bovine endometrium [9] and CL [10] during the estrous cycle and early pregnancy. Moreover, the presence of ATX and PLA2 in stromal and epithelial cells of the bovine endometrium was also demonstrated [14]. Thus, expression of ATX and PLA2 genes in granulosa cells suggests that LPA can be produced in bovine follicles, with ATX playing the major role in this process.
The objective of the study was to examine the effect of lysophosphatidic acid (LPA) on 17β-estradiol (E₂) synthesis and follicle stimulating hormone (FSH) action in bovine granulosa cells. We found that granulosa cells in the bovine antral follicle, in addition to the uterus and the CL, are also the site of LPA synthesis and the target for LPA action in the bovine reproductive tract. Our findings suggest that LPA stimulates E₂ synthesis, probably via increased expression of FSHR and 17β-HSD genes.
Analysis of the transcriptome of bovine endometrial cells isolated by laser micro-dissection (1): specific signatures of stromal, glandular and luminal epithelial cells
2021, BMC Genomics
Lysophosphatidic acid and sphingosine 1-phosphate metabolic pathways and their receptors are differentially regulated during decidualization of human endometrial stromal cells
2014, Molecular Human Reproduction

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^☆: This research was supported by Grants-in-Aid for Scientific Research from the Polish Ministry of Sciences and Higher Education DPN/DWM/MZ/5751/08/09.

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Copyright © 2013 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

Short CommunicationWhich bovine endometrial cells are the source of and target for lysophosphatidic acid?☆

Abstract

Introduction

Section snippets

Materials and methods

Results and discussion

Journal of Lipid Research

Prostaglandins and Other Lipid Mediators

The Journal of Biological Chemistry

The Journal of Biological Chemistry

Lysophosphatidic acid modulates prostaglandin secretion in the bovine uterus

Reproduction

Lysophosphatidic acid action during early pregnancy in the cow: in vivo and in vitro studies

The Journal of Reproduction and Development

Short Communication
Which bovine endometrial cells are the source of and target for lysophosphatidic acid?☆