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Mapping of estradiol binding sites through receptor micro-autoradiography in the endometrial stroma of early pregnant mice

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Abstract

Estradiol triggers key biological responses in the endometrium, which rely on the presence and levels of its cognate receptors on target cells. Employing the receptor micro-autoradiography (RMAR) technique, we aimed to provide a temporal and spatial map of the functional binding sites for estradiol in the mouse endometrial stroma during early pregnancy. Uterine samples from days 1.5 to 7.5 of pregnancy were collected 1 h after tritiated- (3H-) estradiol administration and prepared for RMAR analysis. Autoradiographic incorporation of 3H-thymidine (after 1-h pulse) was evaluated over the same gestational interval. Combined RMAR with either histochemistry with Dolichus biflorus (DBA) lectin or immunohistochemistry for detection of the desmin further characterized 3H-estradiol binding pattern in uterine Natural Killer (uNK) and decidual cells, respectively. 3H-estradiol binding levels oscillated in the pregnant endometrial stroma between the mesometrial and antimesometrial regions as well as the superficial and deep domains. Although most of the endometrial stromal cells retained the hormone, a sub-population of them, as well as endothelial and uNK cells, were unable to do so. Rises in the levels of 3H-estradiol binding preceded endometrial stromal cell proliferation. 3H-estradiol binding and 3H-thymidine incorporation progressively decreased along the development of the antimesometrial decidua. Endothelial proliferation occurred regardless of 3H-estradiol binding, whereas pericytes proliferation was associated with high levels of hormone binding. Endometrial cell populations autonomously control their levels of 3H-estradiol binding and retention, a process associated with their proliferative competence. Collectively, our results illustrate the intricate regulatory dynamic of nuclear estrogen receptors in the pregnant mouse endometrium.

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Acknowledgements

The authors would like to thank Mrs. Cleusa Pellegrini for her enormous and precious support in the preparation of the samples for autoradiographic analysis. Grants (99/00098-1 and 99/00097-5; TMTZ) were provided by São Paulo Research Foundation-FAPESP. Fellowships were provided by FAPESP (11/22429-3; RRF) and by Coordination for the Improvement of Higher Education Personnel—CAPES (20131525; RRF).

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  1. Mauricio Soto-Suazo

    Present address: Medical School, Finis Terrae University, Santiago, Chile

  2. Telma M. T. Zorn and Rodolfo R. Favaro contributed equally.

Authors and Affiliations

  1. Laboratory of Reproductive and Extracellular Matrix Biology, Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, Av. Prof. Lineu Prestes 1524; Room 429, São Paulo, SP, 05508-000, Brazil

    Telma M. T. Zorn, Rodolfo R. Favaro, Mauricio Soto-Suazo & Walter E. Stumpf

  2. University of North Carolina at Chapel Hill, Chapel Hill, USA

    Walter E. Stumpf

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  1. Telma M. T. Zorn
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Correspondence to Telma M. T. Zorn.

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This article is dedicated to Professor Walter E. Stumpf in memoriam for his valuable contributions to the localization and mechanisms of action of hormones and drugs on tissues, particularly through the development of autoradioautographic approaches, and also for his generosity to share his knowledge with the scientific community.

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Zorn, T.M.T., Favaro, R.R., Soto-Suazo, M. et al. Mapping of estradiol binding sites through receptor micro-autoradiography in the endometrial stroma of early pregnant mice. Histochem Cell Biol 148, 257–272 (2017). https://doi.org/10.1007/s00418-017-1568-2

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