In uterobetamethasone affects 3β-hydroxysteroid dehydrogenase and inhibin-α immunoexpression during testis development

Abstract

Prenatal glucocorticoids, commonly used in women at risk of preterm delivery, can predispose the newborn to disease in later life. Since male reproductive function is likely to reflect testis development during fetal life, we studied the effects of prenatal glucocorticoids on two key intra-testicular factors that play roles in cellular proliferation and differentiation, 3β-hydroxysteroid dehydrogenase (3β-HSD) and inhibin-α. Pregnant sheep (n=42) were treated with betamethasone (0.5 mg/kg) or saline (control) at 104, 111 and 118 days of gestation (DG). Testicular tissue was sampled from fetuses at 121 and 132DG, and from lambs at 45 and 90 postnatal days (PD). Within the betamethasone treated group, 3β-HSD immunostaining area was greater at 121DG than at 90PD (P=0.04), but the intensity of immunostaining was higher at 90PD than at 121DG (P=0.04), 132DG (P=0.04) and 45PD (P=0.03). Control animals showed no changes in 3β-HSD area or intensity of immunostaining. No significant differences were observed between treated and control animals in immunostaining area, but immunostaining was more intense in the treated group than in the control group at 90PD (P=0.03). For inhibin-α, the proportion of immunostaining area declined in treated offspring from 121DG to 45PD, in contrast to control values, but recovered fully by 90PD, concomitantly with the onset of spermatogenesis. In conclusion, prenatal betamethasone increased the postnatal testicular expression of inhibin-α but reduced the expression of 3β-HSD. These effects could compromise androgen-mediated testicular development and therefore adult capacity for spermatogenesis.

REPRODUCTIVE TOXICOLOGY: Environmental exposures, fetal testis development and function: phthalates and beyond

“…For example, betamethasone, a glucocorticoid, is used in women at risk of preterm delivery. Recent studies found that in utero betamethasone exposure can disrupt fetal male reproductive development (Borges et al 2016, Pedrana et al 2016, de Barros et al 2018). Similar investigations have been published recently on caffeine, the anti-diabetic drug, metformin, the antiviral drugs, acyclovir and ganciclovir, the anti-epileptic drug, carbamazepine, the antiandrogen and hair loss drug, finasteride, the cholesterol-reducing drug, simvastatin, and others (Bowman et al 2003, Tartarin et al 2012, Andretta et al 2014, Nihi et al 2014, Ogunwole et al 2015, Veroniki et al 2017, Meyer et al 2018, Beverly et al 2019.…”

Maternal undernutrition during pregnancy and lactation affects testicular morphology, the stages of spermatogenic cycle, and the testicular IGF-I system in adult offspring

“…Slides were processed for the detection of immunoreactive IGF-I, inhibin, 3β-HSD, HSF1, and Ki-67 (Table 1) using the streptavidin-biotin-peroxidase protocol, as described previously. 38 In brief, tissue sections were deparaffinized and rehydrated and then underwent heat-induced epitope retrieval; slides were immersed in 0.01-M citrate buffer (pH 6.0) and Tween 20 (5 ml), heated for 5 min (microwave oven), and then washed with distilled water and phosphate-buffered saline (PBS, pH 7.4); endogenous peroxidases were inactivated with 3% hydrogen peroxide (20 min), followed by washing in PBS and incubation with normal rat serum to block non-specific binding proteins; sections were then incubated for 30 min with each primary antibody on different slides (Table 1), for 30 min with biotinylated secondary antibody, and for 30 min with streptavidin-peroxidase enzyme complex (Kit Rabbit Specific HRP/DAB-ABC-Detection IHC, ab 64261; Abcam, Eugene, OR, USA); 3,3 0 diaminobenzidine (DAB) chromogen (30 μl) was added to 1.5-ml DAB substrate, mixed, and incubated for 5 min; testis sections were counterstained with Mayer's hematoxylin, dehydrated, and mounted with Entellan® (Merck, Darmstadt, Germany) and placed under a glass coverslip.…”

Frontiers in sheep reproduction - making use of natural responses to environmental challenges to manage productivity

“…She has shown that the Leydig cells in the ram fetus contain glucocorticoid receptors, and that receptor density is increased by prenatal glucocorticoid treatment (Pedrana et al, 2008). In addition, prenatal glucocorticoid treatment alters dynamics in the expression of 3ß-hydroxysteroid dehydrogenase, an enzyme involved in testosterone synthesis, and decreases the expression of inhibin-α (Pedrana et al, 2016). These effects accompany a reduction in the proliferation of Leydig cells, but not Sertoli cells (Pedrana et al, 2008(Pedrana et al, , 2016, as well as changes the levels of Caspase, Bcl-2, and Bax, shifting the balance from inhibition to promotion of apoptosis (Pedrana et al, 2008).…”

“…In addition, prenatal glucocorticoid treatment alters dynamics in the expression of 3ß-hydroxysteroid dehydrogenase, an enzyme involved in testosterone synthesis, and decreases the expression of inhibin-α (Pedrana et al, 2016). These effects accompany a reduction in the proliferation of Leydig cells, but not Sertoli cells (Pedrana et al, 2008(Pedrana et al, , 2016, as well as changes the levels of Caspase, Bcl-2, and Bax, shifting the balance from inhibition to promotion of apoptosis (Pedrana et al, 2008). Therefore, it seems likely that stress in the pregnant ewe will affect the proliferation and differentiation of cells in the developing testis of ram fetuses.…”