Time- and dose-related effects of di-(2-ethylhexyl) phthalate and its main metabolites on the function of the rat fetal testis in vitro.
BACKGROUND
Endocrine-disrupting effects of phthalates are understood primarily from in utero exposures within the fetal rat testis. Nevertheless, their path of action, dose-response character, and cellular target(s) within the fetal testis are not known.


OBJECTIVE
In this study we investigated the effects of di-(2-ethylhexyl) phthalate (DEHP), mono-(2-ethylhexyl) phthalate (MEHP), and several of their metabolites on the development of organo-cultured testes from rat fetus.


METHODS
We removed testes from 14.5-day-old rat fetuses and cultured them for 1-3 days with or without DEHP, MEHP, and the metabolites.


RESULTS
DEHP (10(-5) M) produced a proandrogenic effect after 3 days of culture, whereas MEHP disrupted testis morphology and function. Leydig cells were the first affected by MEHP, with a number of them being inappropriately located within some seminiferous tubules. Additionally, we found a time- and dose-dependent reduction of testosterone. By 48 hr, gonocyte proliferation had decreased, whereas apoptosis increased. Sertoli cell number was unaffected, although some cells appeared vacuolated, and production of anti-Müllerian hormone decreased in a time- and dose-dependent manner. The derived metabolite mono-(2-ethyl-5-hydroxyhexyl) phthalate was the only one to cause deleterious effects to the rat fetal testis in vitro.


CONCLUSIONS
We hope that this in vitro method will facilitate the study of different phthalate esters and other endocrine disruptors for direct testicular effects.
Chauvigné
François
F
INSERM (Institut National de la Santé et de la Recherche Médicale), U625, GERHM, Université Rennes I, Campus de Beaulieu, Rennes, Bretagne, France.
Menuet
Arnaud
A
Lesné
Laurianne
L
Chagnon
Marie-Christine
MC
Chevrier
Cécile
C
Regnier
Jean-François
JF
Angerer
Jürgen
J
Jégou
Bernard
B
D000818:Animals; D017209:Apoptosis; D004051:Diethylhexyl Phthalate / Q000031:analogs & derivatives* / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005333:Fetus; D066298:In Vitro Techniques; D008297:Male; D008940:Mitotic Index; D010968:Plasticizers / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012671:Seminiferous Tubules / Q000187:drug effects; D012708:Sertoli Cells / Q000187:drug effects / Q000502:physiology; D013737:Testis / Q000166:cytology / Q000187:drug effects* / Q000196:embryology; D013739:Testosterone / Q000378:metabolism; D013997:Time Factors
D010968:Plasticizers; D013739:Testosterone; D004051:Diethylhexyl Phthalate; C016599:mono-(2-ethylhexyl)phthalate