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Published on: August 3, 2012
Determining the LIF-sensitive period for implantation using a LIF-receptor antagonist
L Mohamet1, J K Heath, S J Kimber
1Faculty of Life Sciences, The University of Manchester, Core Technology Facility, Manchester, UK.
Abstract:
Uteri of Lif null mice do not support embryo implantation. Since deletion of some genes often prevents the survival of null mice to adulthood, we have used a proven inhibitor of leukaemia inhibitory factor (LIF) signalling to identify the precise window of time during which LIF is required in vivo, and assessed the cellular expression of several LIF-associated targets. On day 4 of pregnancy, mice were injected with hLIF-05 (inhibitor) into the uterine lumen, with corresponding volumes of PBS (vehicle) injected into the contralateral horn. On days 5 and 6, the number of implantation sites was recorded and the uteri processed for immunohistochemistry. Blockade of LIF on day 4 reduced embryo implantation by 50% (P
Insights
Leukaemia inhibitory factor (LIF) is crucial for embryo implantation, with its signalling required during a specific window on day 4 of pregnancy. Inhibiting LIF replicates many effects seen in LIF-deficient mice.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Endocrinology
Background:
- Leukaemia inhibitory factor (LIF) is essential for successful embryo implantation in mice.
- LIF-deficient (null) mice exhibit uterine failure to support implantation.
- Investigating the precise temporal requirements of LIF signaling in vivo is crucial.
Purpose of the Study:
- To determine the critical time window for leukaemia inhibitory factor (LIF) signaling in uterine embryo implantation.
- To assess the impact of LIF signaling blockade on implantation rates and associated molecular markers.
- To compare the effects of LIF inhibition with the phenotype of LIF null mice.
Main Methods:
- Administration of a LIF signaling inhibitor (hLIF-05) or vehicle (PBS) into the uterine lumen of pregnant mice on day 4.
- Quantification of embryo implantation sites on days 5 and 6.
- Immunohistochemical analysis of uterine tissues to evaluate the expression of LIF-associated targets, including phosphorylated STAT3, amphiregulin, H-type-1 antigen, interleukin-1alpha, oncostatin M, and PTGS2.
Main Results:
- LIF blockade on day 4 significantly reduced embryo implantation by 50%, with maximal effect observed between 09:30 and 12:30 h.
- Inhibition of LIF signaling led to a lack of phosphorylated STAT3 in the luminal epithelium and altered expression of amphiregulin, H-type-1 antigen, interleukin-1alpha, and oncostatin M.
- PTGS2 expression in the stroma remained unaffected by LIF inhibition, contrasting with findings in LIF null mice.
Conclusions:
- LIF signaling is critical for uterine receptivity during a discrete temporal window on day 4 of pregnancy.
- Pharmacological antagonism of LIF signaling effectively recapitulates key aspects of the uterine environment observed in LIF null mice.
- The use of cytokine antagonists is validated as a method to study tissue-specific and temporal signaling in reproductive processes.
