Do G-protein coupled estrogen receptor and bisphenol A analogs influence on Leydig cell epigenetic regulation in immature boar testis ex vivo?

P. Pawlicki , M. Duliban , Ryszard Tuz , A. Ptak , A. Milon , E. Gorowska-Wojtowicz , W. Tworzydło , B. J. Płachno , B. Bilińska , K. Knapczyk-Stwora , Małgorzata Kotula-Balak


Organotypic culture of testicular fragments from 7-day-old male pigs (Polish White Large) was used. Tissues were treated with an antagonist of G-protein coupled estrogen receptor (GPER) (G-15; 10 nM), and bisphenol A (BPA), and its analogs (TBBPA, TCBPA; 10 nM) alone or in combination and analyzed using electron and light (stainings for collagen fibers, lipid droplet and autophagy markers) microscopes. In addition, mRNA and protein abundances and localization of molecules required for miRNA biogenesis and function (Drosha, Exportin 5; EXPO5, Dicer, and Argonaute 2; AGO2) were assessed together with calcium ion (Ca2+) and estradiol concentrations. Regardless of GPER blockade and/or treatment with BPA, TBBPA and TCBPA, there were no changes in Leydig cell morphology. Also, there were no changes in lipid droplet content and distribution but there were changes in lipid and autophagy protein abundance. In the interstitial tissue, there was an increase of collagen content, especially after treatment with BPA analogs and G-15 + BPA. Independent of the treatment, there was downregulation of EXPO5 and Dicer genes but the Drosha and AGO2 genes were markedly upregulated as a result of treatment with G-15 + BPA and TCBPA, respectively. There was always a lesser abundance of EXPO5 and AGO2 proteins regardless of treatment. There was markedly greater abundances of Drosha after G-15 + BPA treatment, and this also occurred for Dicer after treatment with G-15 + TCBPA. Immunolocalization of miRNA proteins indicated there was a cytoplasmic-nuclear pattern in control and treated cells. There was an increase of Ca2+ concentrations after treatment with G-15 and BPA analogs. Estradiol secretion decreased after antagonist and chemical treatments when these were administered alone, however, there was an increase in estradiol secretion after treatment with combinations of these compounds.
Author P. Pawlicki
P. Pawlicki,,
, M. Duliban
M. Duliban,,
, Ryszard Tuz (FoAS / IoAS)
Ryszard Tuz,,
- Institute of Animal Science
, A. Ptak
A. Ptak,,
, A. Milon
A. Milon,,
, E. Gorowska-Wojtowicz
E. Gorowska-Wojtowicz,,
, W. Tworzydło
W. Tworzydło,,
, B. J. Płachno
B. J. Płachno,,
, B. Bilińska
B. Bilińska,,
, K. Knapczyk-Stwora
K. Knapczyk-Stwora,,
et al.`
Journal seriesAnimal Reproduction Science, ISSN 0378-4320, e-ISSN 1873-2232, (N/A 140 pkt)
Issue year2019
Publication size in sheets0.7
Keywords in EnglishBisphenol A, Boar, G-coupled membrane estrogen receptor, Leydig cell, MicroRNA biogenesis and function controlling, molecules
ASJC Classification2700 General Medicine; 3403 Food Animals; 1103 Animal Science and Zoology; 1310 Endocrinology
Internal identifierWHiBZ/2019/34
Languageen angielski
Score (nominal)140
Score sourcejournalList
Publication indicators WoS Citations = 0; Scopus SNIP (Source Normalised Impact per Paper): 2018 = 1.145; WoS Impact Factor: 2018 = 1.817 (2) - 2018=1.998 (5)
Citation count*
Additional fields
FinansowanieThis research was supported by a grant SONATA BIS5 2015/18/E/NZ4/00519 (M.K-B) from National Science Centre, Poland.
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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