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Abstract
Interferon tau (IFNT) is the pregnancy recognition signal in ruminants and is secreted by trophoblast cells. Paracrine action in the endometrium is well established by inhibiting luteolytic pulses of prostaglandin F2 alpha. Recently, endocrine action was documented in the corpus luteum, blood cell and liver. It was hypothesized that conditioned medium (CM) obtained from days 7, 9 and 12 parthenogenetic embryos alters luteal cell gene expression. The aim was to establish a bovine mixed luteal cell culture to evaluate cellular response associated to interferon stimulated genes, steroidogenesis and apoptosis. Conditioned medium was obtained from Days 7, 9 and 12 parthenogenetic (PA) embryos culture. Moreover, antiviral assay was performed on CM from Days 7, 9 and 12 to verify Type I interferon activity. Luteal cell culture was validated by steroidogenic and apoptotic genes (CYP11A1 , HSD3B1, BAX, BCL2, AKT and XIAP mRNA expression), and concentration of progesterone as endpoint. Luteal cell culture was treated with interferon alpha (IFNA) and CM from parthenogenetic embryos. Antiviral assay revealed Type I interferon activity on CM from embryos increasing on Days 9 and 12. ISG15 mRNA was greater in the mixed luteal cells culture treated with 1, 10 and 100ng/ml of interferon alpha (IFNA) and also on Days 7, 9 and 12 CM treatments. Concentration of progesterone was not altered in luteal cell culture regardless of treatments. Steroidogenic and apoptotic genes were similar among groups in luteal cell culture treated with different doses of IFNA or CM from PA embryos. In conclusion, parthenogenetic embryo-derived CM has antiviral activity, luteal cell culture respond to Type I interferon by expressing IGS15. These data indicate this model can be used for IFNT endocrine signaling studies.
Highlights
Embryonic losses in early pregnancy result in premature return to estrus on dairy and beef cattle
Interferon-stimulated gene 15 (ISG15) is a protein expressed in the uterine endometrium on early pregnancy in response to the paracrine action of the embryo-derived interferon tau (IFNT) in ruminants (Austin et al, 2004)
Luteal cells ISG15 mRNA expression increased in response to recombinant ovine (Antoniazzi et al, 2013) or bovine (Shirasuna et al, 2015) IFNT in a dose-dependent manner, up to 1ng/ml
Summary
Embryonic losses in early pregnancy result in premature return to estrus on dairy and beef cattle. Interferon-stimulated gene 15 (ISG15) is stimulated by Type 1 interferon (IFN), called ubiquitin cross-reactive protein (Haas et al, 1987), and it is upregulated in endometrial cells of the mouse (Austin et al, 2003), human (Bebington et al, 1999) and ruminants (Austin et al, 1996) during early pregnancy. For this reason, ISG15 can be used as an indirect evidence of IFNT-dependent cellular response. IFNT binds to Type 1 interferon receptor (IFNAR1 and IFNAR2) to induce a response, and conjugates to intracellular proteins (Loeb and Haas, 1992) in a mechanism similar to ubiquitin (Narasimhan et al, 1996)
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