Background
Fetal Alcohol Spectrum Disorders (FASDs) refer to the range of developmental abnormalities that occur in a fetus following prenatal alcohol exposure (PAE). It is unclear how PAE affects the development of the embryonic heart. Recent data indicates that the Wnt-signaling pathway may be implicated in congenital heart defects caused by PAE. In previous RNA-Sequencing (RNA-Seq) studies, Wnt7a, Wnt7b, and Wnt11 showed significantly changed expression in embryonic mouse hearts after a single maternal binge ethanol dose at embryonic day 9.5 (E9.5).
Hypothesis
We hypothesize that there will be significant change in expression of Wnt7a, Wnt7b, and Wnt11 following maternal ethanol binge at E9.5. We also hypothesize a significant decrease in expression of Wnt7a in C2C12 cells following ethanol exposure.
Experimental Methods
In-vivo, timed pregnant mice were given a single oral gavage of 0.9% saline or 2.5g/kg ethanol at E9.5. RNA from the embryonic heart was quantified and analyzed after 24 hours. Invitro, C2C12 murine myoblasts were cultured and incubated with ethanol or water for 2-24 hours. Cells at 4 different differentiation stages were also exposed to ethanol or water for 24 hours before expression quantification.
Results
Out of our 3 genes, only Wnt7a showed sustained depressed expression after 24 hours. We also concluded there is no significant impact of alcohol on Wnt7a expression in DM6 C2C12 cells exposed to different doses of ethanol from 2 to 24 hours following exposure. There was a significant change between Wnt7a expression in DM0 controls vs. UD, DM3, and DM6 controls.
Conclusion
These results suggest that the stage of differentiation plays a large role in Wnt7a activity and its sensitivity to ethanol. This study creates a greater understanding of the Wnt-signaling pathway's response to alcohol in-vivo and Wnt7a's vulnerability to alcohol at various stages of muscle differentiation.
| Identifer | oai:union.ndltd.org:ucf.edu/oai:stars.library.ucf.edu:honorstheses-2530 |
| Date | 01 January 2023 |
| Creators | Srivatsa, Anagha |
| Publisher | STARS |
| Source Sets | University of Central Florida |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Honors Undergraduate Theses |
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