A precisely regulated sequence of molecular and cellular changes occurs in the female reproductive tract in early pregnancy to facilitate development of the embryo and its successful implantation. Cytokine–leukocyte networks are integral in the tissue remodelling and immuno-regulatory processes underpinning successful implantation. Seminal factors are implicated in activating expression of cytokine genes and inflammatory leukocyte recruitment in the uterus, but whether semen-induced effects extend to the oviduct to influence blastocyst development has not been examined. The aim of this study was to quantitate the expression of mRNA encoding epidermal growth factor (EGF), granulocyte-macrophage colony-stimulating factor (GM-CSF), heparin-binding epidermal growth factor-like growth factor (HB-EGF), insulin-like growth factor (IGFII), leukaemia inhibitory factor (LIF), tumour necrosis factor (TNFα), transforming growth factor (TGFα) and TGFβ from oviducts collected from mice at oestrus and on day 1 of pregnancy, after mating with intact, seminal plasma deficient (svx) and vasectomised (vas) mice. Total RNA was extracted, DNAse treated, reverse transcribed into cDNA, and quantified by real-time PCR using SYBR Green chemistry. All cytokine-specific primers were designed using GenBank sequences and data were normalised to β-actin mRNA expression. Expression of LIF mRNA was induced following mating with intact or vas males, but not svx males, showing that LIF mRNA is induced by factors present in seminal plasma. mRNAs encoding EGF, GM-CSF, HB EGF, IGFII, TNFα, TGFα and TGFβ were all detected in oviduct cDNA collected from oestrus and day 1 mice. These data support the proposal that cytokine-leukocyte networks shown previously to be operative in the uterus extend to the oviduct, and are influenced by exposure to seminal plasma at mating. The cytokines expressed in the oviduct during early pregnancy are likely to be key regulators of embryo growth and development.
Effect of seminal plasma infusion into the vagina on the normalization of endometrial epidermal growth factor concentrations and fertility in repeat breeder dairy cows
