Molecular Markers for the Murine Uterus

O'Hara, Laura (2007). Molecular Markers for the Murine Uterus. MPhil thesis The Open University.



The uterus of a mature female mammal undergoes hormonally initiated morphological and transcriptional changes during early pregnancy, preparing it for the implantation and subsequent nurturing of the developing embryo. Implantation involves a global change in gene expression that has not been fully elucidated. The uterus consists of layers of different types of tissue: the luminal epithelium confronts the uterine lumen and acts as a transducer between the blastocyst and the underlying uterine stroma. The stroma contains areas of glandular epithelium and is surrounded by two myometriai layers: one circular and one longitudinal. Each layer of the uterus is undergoing different transcriptional changes and these changes often involve communicating with the other uterine layers and the embryo.

The aim of this work was to identify and characterise molecular markers for the luminal epithelial, glandular epithelial and stromal layers of the uterus. Samples of uterine luminal epithelium and stroma/glands were obtained from mice on day 4 of pseudopregnancy using an enzyme treatment and mechanical extraction technique. Maternally controlled molecular markers of peri-implantation receptivity in the luminal epithelium (EP2), glandular epithelium (LIF) and stroma (EP3 and Tn-c) were characterised using real-time RT-PCR with Taqman® fluorogenic probes. Pure samples of each tissue were then obtained using laser microdissection, and were used first to verify the specificity of the markers, then to run a microarray study to identify more putative markers. Finally, the enzyme treatment and mechanical extraction technique with subsequent real-time RT-PCR using Taqman® was used to investigate the expression of canonical circadian genes (Bmal, Clock, Perl, Per2 and Cryl) in the luminal epithelium and stroma of the uterus. All genes were found to be expressed in the luminal epithelium and the stroma, but did not appear to have a circadian rhythm.

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