The Effect of Bisphenol A (BPA) on the Expression of Aromatase B
Location
CSU Ballroom
Start Date
21-4-2014 10:00 AM
End Date
21-4-2014 11:30 AM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
Theresa Salerno
Mentor's Email Address
theresa.salerno@mnsu.edu
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
Bisphenol A (BPA) is a common plastic additive that has been shown to have physiological effects that mimic estrogen. Unfortunately, the exact mechanism behind this estrogenic effect is still unclear. The general purpose of this research project was to explore the possibility of post-transcriptional regulation by microRNAs as a possible reason for the estrogenic effect of BPA. The specific objective was to quantify a microRNA that could target and subsequently block expression of the mRNA coding for aromatase, a key enzyme in estrogen synthesis, and to correlate levels of this microRNA with protein expression of aromatase B in a zebra fish system. Computer-based algorithms (BLAST) were used to confirm a very probable match between miR-21 and the 3’-UTR of the aromatase B mRNA. Whole body zebrafish tissues with varying exposure to BPA (0, 0.2, and 20 ppb) were used for the three separate quantification procedures: microRNA, mRNA and protein. Frozen samples were ground and homogenized and microRNAs and mRNAs were isolated simultaneously with the mirVana™ miRNA Isolation Kit (Life Technologies). The short RNAs were converted to cDNA with sequence-specific stem-loop primers from the TaqMan microRNA RT Kit (Life Technologies). The TaqMan microRNA Assay for miR-21 and the endogenous control (U6 snRNA) were used to complete the qPCR quantification. Preliminary research confirmed efficiency of the microRNA qPCR method and showed decreased miR-21 expression with a corresponding increase of BPA exposure. Additional samples will be evaluated with qPCR and Western blotting for levels of miR-21, aromatase B mRNA and protein.
The Effect of Bisphenol A (BPA) on the Expression of Aromatase B
CSU Ballroom
Bisphenol A (BPA) is a common plastic additive that has been shown to have physiological effects that mimic estrogen. Unfortunately, the exact mechanism behind this estrogenic effect is still unclear. The general purpose of this research project was to explore the possibility of post-transcriptional regulation by microRNAs as a possible reason for the estrogenic effect of BPA. The specific objective was to quantify a microRNA that could target and subsequently block expression of the mRNA coding for aromatase, a key enzyme in estrogen synthesis, and to correlate levels of this microRNA with protein expression of aromatase B in a zebra fish system. Computer-based algorithms (BLAST) were used to confirm a very probable match between miR-21 and the 3’-UTR of the aromatase B mRNA. Whole body zebrafish tissues with varying exposure to BPA (0, 0.2, and 20 ppb) were used for the three separate quantification procedures: microRNA, mRNA and protein. Frozen samples were ground and homogenized and microRNAs and mRNAs were isolated simultaneously with the mirVana™ miRNA Isolation Kit (Life Technologies). The short RNAs were converted to cDNA with sequence-specific stem-loop primers from the TaqMan microRNA RT Kit (Life Technologies). The TaqMan microRNA Assay for miR-21 and the endogenous control (U6 snRNA) were used to complete the qPCR quantification. Preliminary research confirmed efficiency of the microRNA qPCR method and showed decreased miR-21 expression with a corresponding increase of BPA exposure. Additional samples will be evaluated with qPCR and Western blotting for levels of miR-21, aromatase B mRNA and protein.
Recommended Citation
Sobota, Sara. "The Effect of Bisphenol A (BPA) on the Expression of Aromatase B." Undergraduate Research Symposium, Mankato, MN, April 21, 2014.
https://cornerstone.lib.mnsu.edu/urs/2014/poster_session_A/14
