Activating transcription factor 3 is estrogen-responsive in utero and upregulated during sexual differentiation.
OBJECTIVE
Synthetic estrogens induce hypospadias, an anomaly of genital tubercle/urethral development. Activating transcription factor 3 (ATF3), which is estrogen-responsive in vitro, is upregulated in hypospadiac human tissue. We used a mouse model of steroid-dependent genital tubercle development to elucidate the ontogeny of ATF3 expression and the developmental response of ATF3 in vivo to estrogen exposure.


METHODS
We used quantitative RT-PCR to assess ontogenic expression of ATF3 and its response to estrogen treatment in utero. Immunohistochemistry was used to localize the protein.


RESULTS
Quantitative RT-PCR showed that ATF3 mRNA is upregulated in all estrogen-exposed fetal genital tubercles compared to controls (p = 0.024), including specifically in males exposed in utero (p = 0.049). Additionally, its expression increases significantly during the period of sexual differentiation in both sexes and significantly correlates with female development (p = 0.004), a phenomenon that appears to be attributable to higher levels at birth in females. The protein localizes in the nucleus, as expected.


CONCLUSIONS
ATF3 is estrogen-responsive in vivo. The response of ATF3 to estrogenic stimulation in utero at an earlier stage may contribute to urethral abnormalities observed in estrogen-exposed male fetuses, although it is likely not the only gene involved, which supports the general understanding that hypospadias is subject to multifactorial influences. ATF3 may therefore be an appropriate gene for further investigations in an endocrine context.
Liu
Benchun
B
Center for the Study and Treatment of Hypospadias, Department of Urology, University of California, San Francisco, Calif. 94143, USA.
Agras
Koray
K
Willingham
Emily
E
Vilela
Marcelo L B
ML
Baskin
Laurence S
LS
D051700:Activating Transcription Factor 3 / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D004967:Estrogens / Q000494:pharmacology*; D004997:Ethinyl Estradiol / Q000494:pharmacology*; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D007021:Hypospadias / Q000139:chemically induced / Q000235:genetics* / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012733:Sex Differentiation; D015854:Up-Regulation; D014521:Urethra / Q000187:drug effects / Q000196:embryology
D051700:Activating Transcription Factor 3; C103002:Atf3 protein, mouse; D004967:Estrogens; D012333:RNA, Messenger; D004997:Ethinyl Estradiol