TY - JOUR
T1 - Proteomics complementation of the rat uterotrophic assay for estrogenic endocrine disruptors
T2 - A roadmap of advancing high resolution mass spectrometry-based shotgun survey to targeted biomarker quantifications
AU - Prokai, Laszlo
AU - Rahlouni, Fatima
AU - Zaman, Khadiza
AU - Nguyen, Vien
AU - Prokai-Tatrai, Katalin
N1 - Funding Information:
Funding: This research was funded by The Welch Foundation (endowment BK-0031 to L.P.), the National Institutes of Health (grants AG031535 to L.P., and EY027005 to K.P.-T.) and a predoctoral bridge funding by the Graduate School of Biomedical Sciences at the University of North Texas Health Science Center (to F.R.).
Funding Information:
This research was funded by The Welch Foundation (endowment BK-0031 to L.P.), the National Institutes of Health (grants AG031535 to L.P., and EY027005 to K.P.-T.) and a predoctoral bridge funding by the Graduate School of Biomedical Sciences at the University of North Texas Health Science Center (to F.R.).
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/2/2
Y1 - 2021/2/2
N2 - The widely used rat uterotrophic assay to assess known and potential estrogenic compounds only considers uterine weight gain as endpoint measurement. To complement this method with an advanced technology that reveals molecular targets, we analyzed changes in protein expression using label-free quantitative proteomics by nanoflow liquid chromatography coupled with high-resolution mass spectrometry and tandem mass spectrometry from uterine protein extracts of ovariectomized rats after daily 17β-estradiol exposure for five days in comparison with those of vehicle-treated control animals. Our discovery-driven study revealed 165 uterine proteins significantly regulated by estrogen treatment and mapped by pathway analyses. Estrogen-regulated proteins represented cell death, survival and development, cellular growth and proliferation, and protein synthesis as top molecular and cellular functions, and a network found with the presence of nuclear estrogen receptor(s) as a prominent molecular node confirmed the relevance of our findings to hormone-associated events. An exploratory application of targeted proteomics to bisphenol A as a well-known example of an estrogenic endocrine disruptor is also presented. Overall, the results of this study have demonstrated the power of combining untargeted and targeted quantitative proteomic strategies to identify and verify candidate molecular markers for the evaluation of endocrine-disrupting chemicals to complement a conventional bioassay.
AB - The widely used rat uterotrophic assay to assess known and potential estrogenic compounds only considers uterine weight gain as endpoint measurement. To complement this method with an advanced technology that reveals molecular targets, we analyzed changes in protein expression using label-free quantitative proteomics by nanoflow liquid chromatography coupled with high-resolution mass spectrometry and tandem mass spectrometry from uterine protein extracts of ovariectomized rats after daily 17β-estradiol exposure for five days in comparison with those of vehicle-treated control animals. Our discovery-driven study revealed 165 uterine proteins significantly regulated by estrogen treatment and mapped by pathway analyses. Estrogen-regulated proteins represented cell death, survival and development, cellular growth and proliferation, and protein synthesis as top molecular and cellular functions, and a network found with the presence of nuclear estrogen receptor(s) as a prominent molecular node confirmed the relevance of our findings to hormone-associated events. An exploratory application of targeted proteomics to bisphenol A as a well-known example of an estrogenic endocrine disruptor is also presented. Overall, the results of this study have demonstrated the power of combining untargeted and targeted quantitative proteomic strategies to identify and verify candidate molecular markers for the evaluation of endocrine-disrupting chemicals to complement a conventional bioassay.
KW - 17β-estradiol
KW - Bisphenol A
KW - Endocrine disruption
KW - Estrogen-regulated proteins
KW - High resolution mass spectrometry
KW - Label-free proteomics
KW - Liquid chromatography–mass spectrometry
KW - Protein networks
KW - Rat uterus
KW - Targeted proteomics
UR - http://www.scopus.com/inward/record.url?scp=85100505683&partnerID=8YFLogxK
U2 - 10.3390/ijms22041686
DO - 10.3390/ijms22041686
M3 - Article
C2 - 33567512
AN - SCOPUS:85100505683
SN - 1661-6596
VL - 22
SP - 1
EP - 13
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 4
M1 - 1686
ER -