Proteomics complementation of the rat uterotrophic assay for estrogenic endocrine disruptors: A roadmap of advancing high resolution mass spectrometry-based shotgun survey to targeted biomarker quantifications

Laszlo Prokai, Fatima Rahlouni, Khadiza Zaman, Vien Nguyen, Katalin Prokai-Tatrai

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Article number1686
Pages (from-to)1-13
Number of pages13
JournalInternational journal of molecular sciences
Volume22
Issue number4
DOIs
StatePublished - 2 Feb 2021

Keywords

  • 17β-estradiol
  • Bisphenol A
  • Endocrine disruption
  • Estrogen-regulated proteins
  • High resolution mass spectrometry
  • Label-free proteomics
  • Liquid chromatography–mass spectrometry
  • Protein networks
  • Rat uterus
  • Targeted proteomics

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