Neonatal exposure to agonists and antagonists of sex steroid receptors induces changes in the expression of oocyte-derived growth factors and their receptors in ovarian follicles in gilts

Neonatal Exposure to Agonists and Antagonists of Sex Steroid Receptors Affects AMH and FSH Plasma Level and Their Receptors Expression in the Adult Pig Ovary

“…150/2013, 122/2014, 123/2014, 187/2014 and 188/2014). The experimental design was as previously described [11]: twenty-five newborn female pigs (Large White × Polish Landrace) were included in the study. Piglets were separated randomly into five groups and injected subcutaneously with: (1) testosterone propionate (20 mg/kg body weight [bw]; N = 5; Sigma-Aldrich, St. Louis, MO, USA), (2) flutamide (50 mg/kg bw; N = 5; Sigma-Aldrich), (3) 4-tert-octylphenol (100 mg/kg bw; N = 5; Sigma-Aldrich), (4) ICI 182,780 (400 µg/kg bw; N = 5; Sigma-Aldrich), or (5) vehicle only (corn oil), which served as a control group (CTR, N = 5).…”

“…To purchase preantral follicles (primordial, primary, and early secondary), we used a protocol involving enzymatic digestion [11]. Briefly, ovarian cortical tissue was rinsed in Dulbecco's PBS medium (PAA The Cell Culture Company) and put into a tissue sectioner (Tissue Slicer Coronal, World Precision Instruments, Sarasota, FL, USA) to obtain uniform-size pieces of 1 × 1 × 1 mm.…”

“…IHC was conducted as before [11]. Goat serum (5%, for BMPR1A, AMHR2, and FSHR) or horse serum (5%, for ACVR1) was used to block unspecific binding sites followed by an incubation with the primary antibodies listed in Table 1, overnight at 4 • C. Next respective biotinylated secondary antibodies (anti-rabbit or anti-goat IgGs; 1:300; 1.5 h; room temperature; Vector Laboratories, Burlingame, CA, USA) and avidin-biotinylated horseradish peroxidase complex (1:100; 40 min; room temperature; Dako, Glostrup, Denmark) were applied in succession.…”

“…Our previous results demonstrated that neonatal exposure to androgen and estrogen agonists or antagonists affected the number of primordial and primary follicles in piglets [10]. Recently, we have revealed that the exposure to the same compounds, i.e., testosterone propionate (TP; a synthetic androgen), flutamide (FLU; a nonsteroidal antiandrogen), 4-tert-octylphenol (OP; compound with estrogenic activity), and ICI 182,780 (ICI; a pure antiestrogen), during neonatal window affected GDF9 and BMP15 signalling in ovaries of adult pigs suggesting the acceleration of initial follicle recruitment and disruption of small antral follicles development [11]. Since AMH shares with BMP15 its type I receptor, BMPR1B, whose expression was altered by neonatal EACs treatment [11], it seems that disruption of steroid signalling may also affect AMH signalling in the ovary of adult pigs.…”

“…Recently, we have revealed that the exposure to the same compounds, i.e., testosterone propionate (TP; a synthetic androgen), flutamide (FLU; a nonsteroidal antiandrogen), 4-tert-octylphenol (OP; compound with estrogenic activity), and ICI 182,780 (ICI; a pure antiestrogen), during neonatal window affected GDF9 and BMP15 signalling in ovaries of adult pigs suggesting the acceleration of initial follicle recruitment and disruption of small antral follicles development [11]. Since AMH shares with BMP15 its type I receptor, BMPR1B, whose expression was altered by neonatal EACs treatment [11], it seems that disruption of steroid signalling may also affect AMH signalling in the ovary of adult pigs. Interestingly, we have also demonstrated the altered expression of FSH receptor (FSHR) in the neonatal porcine ovary following prenatal exposure to FLU pointing out the function of FSH in the promotion of early follicular development in pigs [12].…”