This study examined the DNA methylation profile of endothelial nitric oxide synthase (eNOS) in the placental villous trophoblast in first, second trimester and healthy term placentas. Syncytiotrophoblast DNA revealed a heterogeneous methylation pattern in the first trimester eNOS proximal promoter and transitioned to a densely methylated pattern at term. Healthy, term syncytiotrophoblast and cytotrophoblast obtained through cytotrophoblast isolation technique provided purified cell samples for RNA and DNA extraction. Real-time PCR (rt-PCR) verified the presence and quantity of eNOS mRNA. In summary, the main findings of this thesis are heterogeneous methylation in first trimester compared to hypermethylation at term; greater eNOS expression and variable methylation in term syncytiotrophoblast compared to cytotrophoblast. A heterogeneous methylation pattern in eNOS has been identified in this study and recent stem cell studies in our lab, and we propose it represents plasticity in gene expression, particularly in early development.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/32633 |
| Date | 16 August 2012 |
| Creators | Thompson, Megan Elizabeth |
| Contributors | Marsden, Philip A. |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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