Gestation-related gene expression and protein localization in endometrial tissue of Suffolk and Cheviot ewes at gestation Day 19, after transfer of Suffolk or Cheviot embryos.
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.
Laboratory of Nuclear Techniques, Veterinary Faculty, University of the Republic of Uruguay, Montevideo, Uruguay.
M
M
Sequeira
International Sheep Research Centre, Massey University, Palmerston North, New Zealand.
S J
SJ
Pain
Laboratory of Nuclear Techniques, Veterinary Faculty, University of the Republic of Uruguay, Montevideo, Uruguay.
V
V
de Brun
Laboratory of Nuclear Techniques, Veterinary Faculty, University of the Republic of Uruguay, Montevideo, Uruguay. Electronic address: meikleana@gmail.com.
A
A
Meikle
International Sheep Research Centre, Massey University, Palmerston North, New Zealand.
P R
PR
Kenyon
International Sheep Research Centre, Massey University, Palmerston North, New Zealand.
H T
HT
Blair
D000818:Animals; D051546:Cyclooxygenase 2 / Q000032:analysis / Q000235:genetics; D004624:Embryo Transfer / Q000662:veterinary*; D047108:Embryonic Development / Q000235:genetics; D004717:Endometrium / Q000737:chemistry* / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000032:analysis / Q000235:genetics; D005260:Female; D015870:Gene Expression; D005865:Gestational Age; D007150:Immunohistochemistry; D018969:Insulin-Like Growth Factor Binding Proteins / Q000235:genetics; D007334:Insulin-Like Growth Factor I / Q000032:analysis; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D018396:Mucin-1 / Q000235:genetics; D011247:Pregnancy; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D017526:Receptor, IGF Type 1 / Q000032:analysis / Q000235:genetics; D011972:Receptor, Insulin / Q000235:genetics; D054419:Receptors, Adiponectin / Q000235:genetics; D011980:Receptors, Progesterone / Q000032:analysis / Q000235:genetics; D012756:Sheep / Q000235:genetics*; D013045:Species Specificity
D047628:Estrogen Receptor alpha; D018969:Insulin-Like Growth Factor Binding Proteins; D018396:Mucin-1; D018809:Proliferating Cell Nuclear Antigen; D054419:Receptors, Adiponectin; D011980:Receptors, Progesterone; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D051546:Cyclooxygenase 2; D017526:Receptor, IGF Type 1; D011972:Receptor, Insulin
Embryo growth; Endometrium; Gene expression; Growth factors; Sheep