Estrogen effects on fetal penile and urethral development in organotypic mouse genital tubercle culture.
OBJECTIVE
We developed an organotypic genital tubercle culture system in vitro and used it to investigate the direct effects of the hyperestrogenic state on fetal mouse penile and urethral development.


METHODS
Genital tubercles were dissected from embryonic day 14.5 C57B/L6 male mouse fetuses and cultured using an air-liquid interface on a microporous membrane support soaked in synthetic medium. Cultures were separated into 4 groups. Groups 1 to 3 were supplied with 10 nM dihydrotestosterone, estradiol and 10 nM dihydrotestosterone plus estradiol, respectively. Group 4 was cultured in hormone-free medium. After 36 to 72-hour culture morphological, histological, proliferation, apoptosis, androgen signaling and activating transcription factor 3 analyses were done.


RESULTS
The physiological concentration of 10 nM dihydrotestosterone was essential for genital tubercle growth in vitro. Androgen induced growth and urethral development were significantly suppressed by high dose estrogen. Concurrently we observed increased apoptosis and decreased proliferation in the mesenchyma. Androgen signaling was disrupted and activating transcription factor 3, a factor related to hypospadias genesis, was up-regulated.


CONCLUSIONS
High dose estrogen suppressed male genital tubercle development in vitro. The organotypic genital tubercle culture system in vitro consisting of urethral epithelial and mesenchymal cells can recapitulate the hormonal sensitivity of fetal penile and urethral development. This method is potentially useful for studying the effects of various factors, particularly endocrine disruptors.
Ma
Li-Min
LM
Department of Urology, Ninth People's Hospital Affiliated to Medical College of Shanghai JiaoTong University, Shanghai, China.
Wang
Zhong
Z
Wang
Hao
H
Li
Run-Sheng
RS
Zhou
Juan
J
Liu
Ben-Chun
BC
Baskin
Laurence S
LS
D000818:Animals; D004967:Estrogens / Q000494:pharmacology*; D005837:Genitalia, Male / Q000187:drug effects* / Q000196:embryology*; D008297:Male; D051379:Mice; D009924:Organ Culture Techniques; D010413:Penis / Q000187:drug effects* / Q000196:embryology*; D014521:Urethra / Q000187:drug effects* / Q000196:embryology*
D004967:Estrogens