Sequeira M, Pain S J, de Brun V, Meikle A, Kenyon P R, Blair H T
Laboratory of Nuclear Techniques, Veterinary Faculty, University of the Republic of Uruguay, Montevideo, Uruguay.
International Sheep Research Centre, Massey University, Palmerston North, New Zealand.
Theriogenology. 2016 Oct 1;86(6):1557-1565. doi: 10.1016/j.theriogenology.2016.05.015. Epub 2016 May 25.
The objective of this study was to investigate the gene expression of progesterone and estrogen receptor α (PR, ERα), insulin-like growth factor (IGF) 1, IGF-2, their receptor (IGFR1), IGF-binding proteins (BP) 1 to 6, insulin receptor, adiponectin receptors (AdipoR1/2), cyclooxygenase 2 (PTGS2), mucin 1 and to localize PR, ERα, IGF-1, IGFR1, PTGS2, and proliferating cellular nuclear antigen (PCNA) in the endometrium of pregnant (Day 19) Suffolk and Cheviot ewes carrying Suffolk and Cheviot embryos transferred within and reciprocally between breeds. Gene expression was determined by real-time quantitative polymerase chain reaction (RT-qPCR), and antigen determination was measured by immunohistochemistry in the luminal epithelium (LE), superficial and deep glands (SG, DG, respectively) and superficial and deep stroma. Gene expression of PR, IGF-1, IGFBP2, and IGFBP5 was higher in Suffolk than that in Cheviot ewes (P < 0.05). Greater abundance of IGF-2 and IGBP3 expression was found in Cheviot ewes carrying Cheviot embryos than Cheviot ewes carrying Suffolk embryos (P < 0.05). No staining for PR and ERα was observed in the LE, very scarce staining in SG and DG, whereas positive staining was observed in both superficial and deep stroma. No differences were found for PR staining, but Cheviot ewes had higher ERα staining intensity than Suffolk ewes (P < 0.05). Positive staining for IGF-1 was observed in all cell types except DG, and staining of IGFR1 was observed in all cell types. No differences among groups in staining were found for IGF-1 or IGFR1 in any cell type. Positive staining of PTGS2 was observed in LE and SG in all groups. An interaction between ewe and embryo breed affected PTGS2 staining (P < 0.05), whereby Cheviot ewes carrying Suffolk embryos had a lower PTGS2 staining than Suffolk ewes carrying Suffolk embryos. Positive staining of PCNA was found in LE and SG. Suffolk ewes carrying Suffolk embryos showed lower PCNA immunostaining than Cheviot ewes carrying Suffolk embryos (P < 0.05), whereas no differences were observed in ewes carrying Cheviot embryos. This study showed that gestation-related protein expression in the endometrium of Suffolk and Cheviot ewes is affected by both ewe and embryo breed at Day 19 of pregnancy.
本研究的目的是调查孕酮和雌激素受体α(PR、ERα)、胰岛素样生长因子(IGF)1、IGF-2、它们的受体(IGFR1)、IGF结合蛋白(BP)1至6、胰岛素受体、脂联素受体(AdipoR1/2)、环氧化酶2(PTGS2)、黏蛋白1的基因表达,并在妊娠第19天的萨福克母羊和切维厄特母羊的子宫内膜中定位PR、ERα、IGF-1、IGFR1、PTGS2和增殖细胞核抗原(PCNA),这些母羊体内携带了在品种内和品种间相互移植的萨福克和切维厄特胚胎。通过实时定量聚合酶链反应(RT-qPCR)测定基因表达,并通过免疫组织化学法在腔上皮(LE)、浅表和深部腺体(分别为SG、DG)以及浅表和深部基质中测量抗原。PR、IGF-1、IGFBP2和IGFBP5的基因表达在萨福克母羊中高于切维厄特母羊(P < 0.05)。携带切维厄特胚胎的切维厄特母羊中IGF-二和IGBP3的表达丰度高于携带萨福克胚胎的切维厄特母羊(P < 0.05)。在LE中未观察到PR和ERα的染色,在SG和DG中染色非常稀少,而在浅表和深部基质中均观察到阳性染色。PR染色未发现差异,但切维厄特母羊的ERα染色强度高于萨福克母羊(P < 0.05)。除DG外,在所有细胞类型中均观察到IGF-1的阳性染色,在所有细胞类型中均观察到IGFR1的染色。在任何细胞类型中,IGF-1或IGFR1的染色在各实验组之间均未发现差异。在所有组的LE和SG中均观察到PTGS2的阳性染色。母羊和胚胎品种之间的相互作用影响PTGS2染色(P < 0.05),即携带萨福克胚胎的切维厄特母羊的PTGS2染色低于携带萨福克胚胎的萨福克母羊。在LE和SG中发现PCNA的阳性染色。携带萨福克胚胎的萨福克母羊的PCNA免疫染色低于携带萨福克胚胎的切维厄特母羊(P < 0.05),而携带切维厄特胚胎的母羊之间未观察到差异。本研究表明,在妊娠第19天,萨福克和切维厄特母羊子宫内膜中与妊娠相关的蛋白质表达受母羊和胚胎品种的影响。