The effect of lysophosphatidic acid together with interferon tau on the global transcriptomic profile in bovine endometrial cells
Introduction
Lysophosphatidic acid (LPA), which exerts a variety of physiological and pathological actions in various animal species [1], [2], [3], [4], [5], [6], has been documented to regulate reproductive performance in cows [7], [8]. We have recently found that LPA can be locally produced and released from the bovine endometrium [9]. We have also documented that LPA concentrations and gene expression for its active receptor LPAR1 in the bovine endometrium were significantly higher during early pregnancy than during the estrous cycle [9]. Moreover, LPA stimulated progesterone (P4) and prostaglandin (PG) E2 secretion in vivo, and mRNA expression of LPAR1 was positively correlated with the expression of the enzyme responsible for luteotropic PGE2 production (PGES) in the bovine endometrium during the estrous cycle and early pregnancy [9], [10]. Bovine embryos at different stages of development are also able to synthesize and secrete LPA; additionally, LPA-mediated cell signaling during early embryonic development may be relevant in early embryo-maternal interactions that lead to the embryonic survival [11]. These data indicate that LPA may play autocrine and/or paracrine roles in the uterus and serve as an important factor in the maintenance of early pregnancy not only in mouse [3], [12], pig [5] and sheep [6] but also in cow [10].
There is much evidence showing that after fertilization, between days 10 and 21 to 25 in ruminants, the conceptus starts to produce interferon tau (IFNτ), which is the pregnancy recognition signal that prevents development of the endometrial luteolytic mechanism [13], [14]. The maximal production of IFNτ falls on days 14–18 [15]. However, Kimura et al. [16] reported that IFNτ production during culture of in vivo-derived embryos increased dramatically (150,000-fold) from day 9–14. The signals from the conceptus occurring after fertilization are not only local, but spread throughout the entire female as measured in the systemic blood a few days before implantation [17], [18]. This peri-implantation period during early pregnancy represents the time when the blastocyst is hatching; from this time, it becomes a free-floating embryo within the lumen of the uterus that is totally dependent on the uterine environment for survival [15]. At this time, early embryo survival mostly depends on the appropriate function of corpus luteum (CL)-adequate P4 synthesis, as well as on IFNτ-dependent inhibition of the development of the endometrial luteolytic mechanism [13], [14]. In contrast, LPA also serves as a luteotropic factor during early pregnancy by stimulating P4 and PGE2 secretion, thus protecting the bovine CL and early embryo development [9].
Studies to date on the endometrial response to various early pregnancy modulating factors, including LPA, have been based on a candidate gene approach. Therefore, a better understanding is needed of all the internal signaling pathways involved in the potential modulation of LPA action by IFNτ. This study will provide a deeper understanding of uterine functions at the time of early pregnancy recognition in cow and can directly influence further investigations.
Section snippets
Bovine endometrial cell culture
All of the animal procedures were approved by the Local Animal Care and Use Committee in Olsztyn, Poland (Agreement No. 79/2008/N). For all experiments, normally cycling Holstein/Polish Black and White (75/25%, respectively) cows (n = 9) were chosen. Bovine uteri were obtained at a local slaughterhouse within 20 min of exsanguinations and were transported on ice to the laboratory within 40 min. The animals were slaughtered on days 2–5 of the estrous cycle to isolate the endometrial cells as
Microarray quality control
The large-scale analysis of the endometrial transcriptome was conducted using a microarray, which is comprised of over 23 000 bovine transcripts. Hierarchical clustering and principal component analysis revealed sufficient differences in the gene expression profiles from endometrial cells stimulated with PBS, LPA and LPA + IFNτ that allowed the genes to cluster into three separate groups based on the treatment conditions (Fig. 1). The total principal component (PC) accounts for 86.8% of the
Discussion
In this study, we used microarray technology to investigate the effects of LPA on the gene expression profile in bovine endometrial cells. Previous reports showed that LPA stimulated PGE2 synthesis via the enhanced mRNA expression of PGE2 synthase in stromal cells on days 8–10 and 16 to 18 of the estrous cycle and pregnancy in the bovine endometrium [10], [24]. Moreover, LPA inhibited PGF2α synthesis by reducing PGF2α synthase mRNA expression in epithelial cells on days 8–10 and 16 to 18 of
Conclusions
In summary, the transcriptional profiling studies identified a number of genes that are regulated by LPA and LPA together with IFNτ in endometrial cells from the bovine uterus. The results revealed that 10 of the 16 analyzed genes showed the same expression patterns between the microarray and real-time PCR analysis, indicating considerable consistency between both techniques. Lysophosphatidic acid differentially regulates the expression of genes that are probably important for uterine
Funding
This research was supported by Grants-in-Aid for Scientific Research from the Polish Ministry of Sciences and Higher Education (MNiSW-DPN/DWM/MZ/5751/08/09).
Acknowledgements
Not applicable.
References (59)
- et al.
Lysophosphatidic acid influences the morphology and motility of young, postmitotic cortical neurons
Mol Cell Neurosci
(2002) - et al.
Lysophosphatidic acid stimulates nuclear and cytoplasmic maturation of golden hamster immature oocytes in vitro via cumulus cells
Life Sci.
(2002) - et al.
Studies on lysophosphatidic acid action during in vitro preimplantation embryo development
Domest Anim Endocrinol
(2016) - et al.
Prostaglandins and reproduction in female farm animals
Vet J
(2006) - et al.
Cumulus and granulosa cell markers of oocyte and embryo quality
Fertil Steril
(2013) - et al.
Expression of betacellulin and epiregulin genes in the mouse uterus temporally by the blastocyst solely at the site of its apposition is coincident with the “window” of implantation
Dev Biol.
(1997) - et al.
Molecular cloning and chromosomal mapping of a bone marrow stromal cell surface gene, BST2, that may be involved in pre-B-cell growth
Genomics
(1995) - et al.
LPA3-mediated lysophosphatidic acid signalling in embryo implantation and spacing
Nature
(2005) - et al.
Hypoxia enhances lysophosphatidic acid responsiveness in ovarian cancer cells and lysophosphatidic acid induces ovarian tumor metastasis in vivo
Cancer Res.
(2006) - et al.
Analysis of lysophosphatidic acid (LPA) receptor and LPA-induced endometrial prostaglandinendoperoxide synthase 2 expression in the porcine uterus
Endocrinology
(2008)
Lysophosphatidic acid signaling during embryo development in sheep: involvement in prostaglandin synthesis
Endocrinology
Lysophosphatidic acid (LPA) signaling in human and ruminant reproductive tract
Mediat Inflamm
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
J Reprod Dev.
Lysophosphatidic acid signaling in late cleavage and blastocyst stage bovine embryos
Mediat Inflamm
Embryo spacing and implantation timing are differentially regulated by LPA3-mediated lysophosphatidic acid signaling in mice
Biol Reprod
Burghardt RC Fetal–maternal interactions during the establishment of pregnancy in ruminants
Soc Reprod Fertil Suppl.
Bayless K Novel pathways for implantation and establishment and maintenance of pregnancy in mammals
Mol Hum Reprod
In vivo expression of interferon tau mRNA by the embryonic trophoblast and uterine concentrations of interferon tau protein during early pregnancy in the cow
Mol Reprod Dev.
Sexual dimorphism in interferon-production by in vivo-derived bovine embryos
Mol Reprod Dev.
Uterine vein infusion of interferon tau (IFNT) extends luteal life span in ewes
Biol Reprod
Expression of interferon (IFN)-stimulated genes in extrauterine tissues during early pregnancy in sheep is the consequence of endocrine IFN-tau release from the uterine vein
Endocrinology
Specific properties of epithelial and stromal cells from the endometrium of cows
J Reprod Fertil
Lysophosphatidic acid stimulates prostaglandin E2 production in cultured stromal endometrial cells through LPAR1 receptor
Exp Biol Med
Is tumor necrosis factor-a a trigger for the initiation of prostaglandin F2a release at luteolysis in cattle?
Biol Reprod
Normalization of real-time quantitative RT-PCR data: a model based variance estimation approach to identify genes suited for normalization- applied to bladder- and colon-cancer data-sets
Cancer Res.
Comprehensive algorithm for quantitative real-time polymerase chain reaction
J Comput Biol.
Lysophosphatic acid modulates prostaglandin secretion in the bovine endometrial cells differently on days 8-10 of the estrous cycle and early pregnancy
J Reprod Dev.
Relationship between maternal endocrine environment, early embryo development and inhibition of the luteolytic mechanism in cows
Reproduction
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