Protein Biomakers of Estrogenic Impact on Mouse and Rat Uterus




Thai, Stephen Duy-An


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Understanding the structural and morphological impact that occur in the uterus upon exposure to estrogens can reveal how these hormones elicit changes in the organ. This study seeks to revalidate the data obtained in studies of ovariectomized mice and rats treated with 17β-estradiol, followed by comparative analysis of the protein expression between the two animal models (the mice model studied by Prokai et al. [1] and the rat model presented in a dissertation by Rahlouni [2]). During these studies, female Swiss-Webster mice and Sprague-Dawley rats were ovariectomized (OVX) and divided respectively into two groups: 1) The control was treated with a corn oil vehicle while 2) The treatment group received 17β-estradiol injection for 5 days. Raw data gathered from the earlier proteomics studies were reanalyzed using Maxquant version 1.6.17, which utilized extracted ion chromatography technique (XIC). LFQ analyst and Ingenuity Pathway Analysis (IPA) version 5.0 were used to identify proteins that were differentially regulated by 17β-estradiol and perform comparative analyses of the protein expression between OVX mice and rats. Reanalysis of OVX mice identified 59 proteins of interest at 95% confidence with 29 upregulated and 30 downregulated significantly. Reanalysis of the OVX rats identified 126 differentially expressed proteins at 95% confidence with 98 upregulated and 27 downregulated significantly. Comparative analysis using IPA® found 27 proteins unique to mice and 85 proteins unique to rats. Conversely, the OVX mice and rats shared 19 proteins regulated by 17β-estradiol in the uterus. Ingenuity Pathway Analysis® also created networks in OVX mouse and rat models showing a relationship with estrogen receptors in the nucleus, which can bind 17β-estradiol and then initiate gene transcription. Although there is overlap between OVX mice and rat protein expression, the proteins that were found to be unique to each animal demonstrate that a complementary model using both animals provides a much broader view of uterine protein expression in OVX animals treated with 17?-estradiol. Additionally, this study illustrates the merit of reanalyzing older data with improved computational and bioinformatic tools to pinpoint proteins of interest for future analysis as potential markers of estrogenic effects in the uterus.