title: Estrogenic compounds inhibit gap junctional intercellular communication in mouse Leydig TM3 cells.
abstract: Some estrogenic compounds are reported to cause testicular disorders in humans and/or experimental animals by direct action on Leydig cells. In carcinogenesis and normal development, gap junctional intercellular communication (GJIC) plays an essential role in maintaining homeostasis. In this study, we examine the effects of diethylstilbestrol (DES, a synthetic estrogen), 17beta-estradiol (E(2), a natural estrogen), and genistein (GEN, a phytoestrogen) on GJIC between mouse Leydig TM3 cells using Lucifer yellow microinjection. The three compounds tested produced GJIC inhibition in the TM3 cells after 24 h. Gradually, 10 microM DES began to inhibit GJIC for 24 h and this effect was observed until 72 h. On the other hand, both 20 microM E(2) and 25 microM GEN rapidly inhibited GJIC in 6 h and 2 h, respectively. The effects continued until 24 h, but weakened by 72 h. Furthermore, a combined effect at microM level between DES and E(2) on GJIC inhibition was observed, but not between GEN and E(2). DES and E(2) showed GJIC inhibition at low dose levels (nearly physiological estrogen levels) after 72 h, but GEN did not. DES-induced GJIC inhibition at 10 pM and 10 microM was completely counteracted by ICI 182,780 (ICl), an estrogen receptor antagonist. On the other hand, the inhibitory effects on GJIC with E(2) (10 pM and 20 microM) and GEN (25 microM) were partially blocked by ICI or calphostin C, a protein kinase C (PKC) inhibitor, and were completely blocked by the combination of ICI and calphostin C. These results demonstrate that DES inhibits GJIC between Leydig cells via the estrogen receptor (ER), and that E(2) and GEN inhibit GJIC via ER and PKC. These estrogenic compounds may have different individual non-genotoxic mechanism including PKC pathway on testicular carcinogenesis or development.
authors.affiliation: Department of Bioenvironmental Medicine, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan. Iwase.Yumiko@mg.m-pharma.co.jp
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Iwase
authors.forename: Hideki
authors.initials: H
authors.lastname: Fukata
authors.forename: Chisato
authors.initials: C
authors.lastname: Mori
mesh_terms: D000818:Animals; D002460:Cell Line; D004054:Diethylstilbestrol / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology / Q000633:toxicity*; D004965:Estrogen Antagonists / Q000494:pharmacology; D004967:Estrogens / Q000633:toxicity*; D005456:Fluorescent Dyes / Q000378:metabolism; D000077267:Fulvestrant; D017629:Gap Junctions / Q000187:drug effects* / Q000378:metabolism; D019833:Genistein / Q000633:toxicity*; D007546:Isoquinolines / Q000378:metabolism; D007985:Leydig Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D009281:Naphthalenes / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011960:Receptors, Estrogen / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D004338:Drug Combinations; D004791:Enzyme Inhibitors; D004965:Estrogen Antagonists; D004967:Estrogens; D005456:Fluorescent Dyes; D007546:Isoquinolines; D009281:Naphthalenes; D011960:Receptors, Estrogen; C061465:calphostin complex; D000077267:Fulvestrant; D004958:Estradiol; D004054:Diethylstilbestrol; C017475:lucifer yellow; D019833:Genistein; D011493:Protein Kinase C