Endogenous estrogen receptor beta is transcriptionally active in primary ovarian cells from estrogen receptor knockout mice.
The estrogen receptor (ER) alpha is a hormone-inducible transcription factor that has a pivotal physiological role. Intriguingly, a clear and undisputed physiological function of the recently described ERbeta remains elusive, with the exception of the ovary where a cooperative role of ERalpha and ERbeta has been demonstrated. We have, therefore, investigated whether endogenous ERs, in particular ERbeta, act as ligand-inducible transcription factors in primary ovarian cells derived from wild-type, ERalpha or ERbeta knockout mice. Granulosa-enriched cell fractions naturally expressing ERbeta and thecal cell fractions that express ERalpha were analyzed in transactivation assays using the vitellogenin A2 consensus estrogen response element and potent ER agonists diethylstilbestrol and S-indenestrol A. We studied also the potency-selective ERbeta agonist R-indenestrol A, the pure ERalpha agonist and ERbeta antagonist R,R-diethyl-tetrahydrochrysene and the pure ERalpha agonist propylpyrazole-triol. Using ER subtype-specific physiological cell models and these ER subtype-specific structural probes, we analyzed trans-activation of ERalpha and ERbeta. This analysis revealed that endogenously expressed ERbeta is indeed functional as a transcription factor, that it responds to estrogens appropriately, and that the ligands used are true ER subtype-specific probes in primary ovarian cells. In conclusion, this study demonstrates that endogenously expressed ERbeta is capable of regulating gene transcription independent of ERalpha.
Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA.
Stefan O
SO
Mueller
John A
JA
Katzenellenbogen
Kenneth S
KS
Korach
D000818:Animals; D002478:Cells, Cultured; D002911:Chrysenes / Q000494:pharmacology; D004054:Diethylstilbestrol / Q000494:pharmacology; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000819:agonists / Q000235:genetics; D047629:Estrogen Receptor beta / Q000819:agonists / Q000235:genetics / Q000502:physiology*; D004967:Estrogens / Q000494:pharmacology; D005260:Female; D000077267:Fulvestrant; D005786:Gene Expression Regulation / Q000187:drug effects; D006107:Granulosa Cells / Q000378:metabolism*; D007192:Indenes / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010053:Ovary / Q000166:cytology; D010636:Phenols; D011720:Pyrazoles / Q000494:pharmacology; D011960:Receptors, Estrogen / Q000819:agonists / Q000235:genetics / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D013799:Theca Cells / Q000378:metabolism*; D015533:Transcriptional Activation; D014162:Transfection; D014819:Vitellogenins / Q000235:genetics
C120076:5,11-diethyl-5,6,11,12-tetrahydrochrysene-2,8-diol; D002911:Chrysenes; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D004967:Estrogens; D007192:Indenes; D010636:Phenols; D011720:Pyrazoles; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D014819:Vitellogenins; C486184:4,4',4''-(4-propyl-((1)H)-pyrazole-1,3,5-triyl) tris-phenol; D000077267:Fulvestrant; C047437:indenestrol; D004958:Estradiol; D004054:Diethylstilbestrol; D008156:Luciferases