Our previous research has identified granulin (grn) and p130 genes as sex steroidinducible genes in the rat hypothalamus, which might be involved in sexual differentiation of the brain. Phthalate esters that are used as plasticizers and also found at low levels in foods such as dairy products are often mentioned as suspected endocrine disrupters. The purpose of the present study is to elucidate whether perinatal exposure to dinbutyl phthalate (DBP), diisononyl phthalate (DINP) and di2ethylhexyl adipate (DEHA) affects hypothalamic sex steroidinducible genes. The present study assessed the effects of perinatal exposure to DBP, DINP and DEHA on sex steroid hormones levels and hypothalamic grn and p130 mRNA expressions at postnatal day (PND) 3 and 7. Pregnant rats were fed a soyfree diet containing 20, 200, 2,000 and 10,000 ppm of DBP, 40, 400, 4,000 and 20,000 ppm of DINP, or 480, 2,400 and 12,000 ppm of DEHA from gestational day (GD) 15 to GD 3 or 7. At PND 3 and 7, perinatal exposure to these chemicals did not substantially affect serum concentrations of testosterone and estradiol. At PND 3, the expression of grn mRNA levels in males was decreased by DEHA, and that of p130 was decreased by DBP, DINP and DEHA, though the effects were not dosedependent. At PND 7, the expression of grn gene in female pups was increased by higher doses of DBP and all the doses, except for 4,000 ppm, of DINP, while that in male pups decreased by 480 and 12,000 ppm of DEHA. Hypothalamic expression of p130 mRNA in males was increased by lower doses of DBP and all the doses of DINP, whereas that of females was decreased by 480 and 2,400 ppm of DEHA. These results suggest that these chemicals may affect the expression of grn and p130 genes by directly acting on the hypothalamus, thus leading to inappropriate expression of these genes.

Di-n-butyl phthalate (DBP), diisononyl phthalate (DINP) and di-(2-ethylhexyl) adipate (DEHA) are ubiquitously distributed chemicals that are widely used as plasticizers and also found at low levels in foods. The aims of this study were to determine whether perinatal exposure to DBP, DINP and DEHA could alter normal patterns of neonatal development. Dams were provided with pulverized soy-free diet containing 20, 200, 2,000 and 10,000 ppm of DBP, 40, 400, 4,000 and 20,000 ppm of DINP, or 480, 2,400 and 12,000 ppm of DEHA from gestational day 15 to postnatal day 21. Exposure to the high doses of DBP, DINP and DEHA during gestational period significantly decreased food consumption and body weight gain of dams. These chemicals reduced neonatal body weight as well as that of the after maturation. Also, exposure to DINP of all the doses used and the higher doses (2,400 and 12,000 ppm) of DEHA decreased AGD at PND 1 in male neonates, though that to DBP did not affect AGD in males. In female neonates, an increase in AGD was observed in DBP- and DINP-exposed animals at the highest doses. Moreover, these chemicals affected survival rate of pups at PND 5, and delayed onset of eye opening in all chemical-exposed groups at PND 17. These results suggest that perinatal exposure to these chemicals may affect the normal development and / or growth of offspring.

As an endocrine disruptor, bisphenol-A (BPA) causes several functional and behavioral abnormalities related to reproduction. The current study was design to evaluate the effect of perinatal exposure of female mice to BPA on sperm function of adult F(1) offspring. Pregnant female mice F(0) were gavaged with three different concentration of BPA, such as 50 μg/kg/day (tolerable daily intake value by the European Food Safety Authority), 5 mg/kg/day (no-observed-adverse-effect level; NOAEL), and 50 mg/kg/day (lowest-observed-adverse-effect level; LOAEL) and corn oil (7 mg/kg/day; vehicle control). The functional parameters of F(1) spermatozoa were studied both before and after capacitation, whereas the fertility assessment was evaluated by in vitro and in vivo assay using unexposed females. Our results showed that spermatozoa hyperactivated motility, capacitation, intracellular ATP, Ca2+, and ROS levels after capacitation were significantly affected using NOAEL and LOAEL concentration of BPA. However, the sperm motility was only affected by LOAEL dose after capacitation. All of the tested parameters were potentially unaffected by BPA before capacitation, except intracellular ATP that decreased by all concentrations. Although both NOAEL and LOAEL concentration were effectively reduced the rate of fertilization and embryonic development in vitro, however the average litter size was only affected by LOAEL dose. Our finding suggested that perinatal exposure of 50 μg/kg/day did not produce significant effects; however both NOAEL and LOAEL affects overall sperm function after capacitation, leading to impairments in the fertility of F(1) male offspring.

Although fenarimol is a widely used chlorinated fungicide applied to fruits and vegetables and considered as a suspected endocrine disrupter (ED), transgeneration studies of fenarimol at its low doses are not available. Objectives: The aims of this study are to address the effect of perinatal exposure to low doses of fenarimol on reproductive performance and to investigate molecular and cellular mechanisms which are associated with. Methods: The body and organ weights and anogenital distance (AGD) of mice offspring (F1) maternally exposed to fenarimol were determined, and their reproductive performances were assessed by mating and ovarian follicular and sperm analyses. In addition, differentially expressed genes (DEGs) in F1 ovaries were identified by DNA microarray. Up-regulated genes were confirmed by quantitative real-time PCR (qRT-PCR) and immunohistochemical analysis. Results: Fenarimol-exposed F1 mice showed the shortened AGD, increased body weight with altered organ weights, increased number of pub, abundant follicles, and enhanced sperm count and quality. Microarray data showed 82 up-regulated and 742 down-regulated genes on the ovaries of fenarimol-exposed mice, in which Cyp17a1 and Cyp19a1 were up-regulated. Conclusions: Low doses of perinatal fenarimol exposure caused reproductive dysfunction in mice and thus can possibly impose risks on reproductive activities of human and wild-life.