In utero methoxychlor exposure increases rat fetal Leydig cell number but inhibits its function

The objective of the present study is to determine whether in utero exposure to methoxychlor (MXC) affects rat fetal Leydig cell number, cell size, or functions. Pregnant Sprague Dawley dams were gavaged with corn oil (control, 0 mg/kg/day MXC) or MXC at doses of 10, 50, or 100 mg/kg/day from gestational day (GD) 12 to 21. The results show that MXC increased fetal Leydig cell numbers dose-dependently from 95 ± 8 × 103 cells/testis (control, mean ± SEM) to 101 ± 6, 148 ± 22, and 168 ± 21 × 103 cells/testis, at the doses of 10, 50, and 100 mg/kg, respectively. The increase of Leydig cell number by MXC was contributed by the increase of single cell population of Leydig cells, which increased from 21 ± 2% of the control to 31 ± 4%, 39 ± 3%, or 40 ± 4% at the doses of 10, 50 or 100 mg/kg, respectively. Quantitative PCR results show that MXC increased Lhcgr expression at dose of 10 mg/kg and Scarb1 and Cyp11a1 mRNA levels at doses of 50 and 100 mg/kg. Immunohistochemical staining demonstrated the increase of CYP11A1 protein level from the dose of 10 mg/kg. However, at the highest dose (100 mg/kg) MXC reduced the testicular testosterone level and MXC (1 μM) in vitro treatment also inhibited androgen production from isolated fetal Leydig cells. In conclusion, our findings indicate that at low dose MXC may increase fetal Leydig cell numbers and the expressions of some steroidogenic enzymes, but at high dose it reduces the testicular testosterone level leading to reproductive tract malformations in the male offspring.