Impact of periconceptional undernutrition on adrenal growth and adrenal insulin-like growth factor and steroidogenic enzyme expression in the sheep fetus during early pregnancy.

Periconceptional undernutrition (PCUN) results in an earlier prepartum activation of the pituitary-adrenal axis in twin compared with singleton fetuses. We have tested the hypotheses that the functional development of the fetal sheep adrenal is delayed in twins compared with singletons in early gestation and that PCUN accelerates adrenal growth and increases the expression of intraadrenal IGF-I and -II and cytochrome P450 17-hydroxylase (CYP17) as early as 55 d gestation. We have investigated the effect of PCUN in the ewe (restricted at 70% of control allowance, n=21; control, n=24) from at least 45 d before mating until d 7 after mating on maternal cortisol and progesterone concentrations, fetal adrenal weight, adrenal IGF-I, IGF-I receptor (IGF-IR), IGF-II, IGF-IIR, and CYP17 mRNA expression and placental 11beta-hydroxysteroid dehydrogenase-1 and -2 mRNA and protein expression at d 53-56 pregnancy. The relative weight of the fetal adrenal and adrenal IGF-I, IGF-IR, IGF-II, IGF-IIR, and CYP17 mRNA expression were lower in twin compared with singleton fetuses. In singleton fetuses of PCUN ewes, there was a loss of the relationship between adrenal IGF-II/IGF-IIR expression and either adrenal weight or CYP17 mRNA, which was present in controls. Similarly in twin fetuses, PCUN resulted in the loss of the relationships between adrenal weight and IGF-I expression and between adrenal CYP17 and IGF-II expression, which were present in controls. Our findings suggest that differences in the timing of the prepartum activation of the fetal adrenal in twins and singletons have their origins in early gestation and highlight the importance of the interaction between the periconceptional environment and embryo number in setting the growth trajectory of the fetal adrenal.

Fetal adrenal development: insight gained from adrenal tumors.

Precise and coordinated mechanisms control the growth and functional differentiation of fetal organ systems. Conversely, tumor progression and the development of cancer probably occur through a process of dysregulation and dedifferentiation. Similarities exist between normal human fetal adrenal cortex and adrenal cancers, such as high expression of growth factors, including insulin-like growth factor II. Therefore, we might gain insight into factors involved in adrenocortical development through better understanding the development and progression of adrenocortical tumors. This review is prompted by recent gene profiling studies that have identified genes differentially expressed between normal and abnormal adrenal glands. Several of these genes are specific growth factors or key cell cycle regulators, in addition to genes not previously associated with adrenal growth or function.

Adrenal TGFbeta1 mRNA levels fall during late gestation and are not regulated by cortisol in the sheep fetus.

During mammalian development there are periods when the fetal adrenal is either relatively refractory or increasingly sensitive to trophic stimulation. This pattern of regulation of adrenal growth and function ensures that the fetal lungs, liver, brain and kidney are exposed in a programmed temporal sequence to the genomic actions of circulating glucocorticoids. The factors which act to maintain periods of adrenal quiescence are not known. In the present study we have measured the level of messenger RNA (mRNA) expression of a putative inhibitor of adrenal steroidogenesis, transforming growth factor beta 1 (TGFbeta1), and a key steroidogenic enzyme, cytochrome P450 17alpha hydroxylase (CYP17), during periods of adrenal quiescence and activation in the sheep fetus. We have also investigated the relative roles of the fetal hypothalamic-pituitary axis and cortisol in the regulation of expression of adrenal TGFbeta1 and CYP17 mRNA during late gestation. Adrenal expression of TGFbeta1 was greatest at around 100 days gestation, at a time when the fetal sheep adrenal is relatively refractory to trophic stimulation and there was an inverse relationship between the expression of TGFbeta1 and CYP17 mRNA in the adrenal gland during the peripartum period. Whilst disconnection of the fetal hypothalamic-pituitary disconnection (HPD) axis resulted in a decrease in adrenal CYP 17 mRNA expression, there was no effect of fetal HPD, with or without cortisol replacement, on adrenal TGFbeta1 mRNA expression in late gestation. Thus TGFbeta1 may play a role in inhibiting adrenal steroidogenesis and ensuring that the adrenal remains relatively refractory to trophic stimulation during mid gestation. The maintenance of low adrenal TGFbeta1 expression during late gestation is not dependent, however, on stimulation by the fetal hypothalamic-pituitary axis.

Functional biology of the primate fetal adrenal gland: advances in technology provide new insight.

1. The main aim of the present review is to summarize recent experimental data from human and non-human primate models that have identified factors essential for adrenal development and other factors that may determine the regulation of the specific structural organization and function of the adrenal gland. 2. The fetal adrenal cortex has two morphologically distinct zones, with the outer definitive zone being comprised of tightly packed small cells, which appear to be steroidogenically quiescent until late gestation, and the inner fetal zone, which appears to be steroidogenically active throughout gestation. 3. In the primate fetus, growth of the adrenal gland involves hyperplasia, hypertrophy, migration and senescence. Cells appear to proliferate in the external portion of the definitive zone and then move centripetally and become non-proliferative in the fetal zone, where they acquire their steroidogenic capacities. 4. A variety of new technologies has been used to identify zonal-specific markers of the cortical zones within the developing human fetal adrenal gland. On microarray, 67 transcripts showed a minimum of a 2.5-fold difference between the fetal and adult adrenal gland. The vast majority of these genes had not been studied in relation to adrenal gland development or function. In combination with techniques such as laser capture microdissection, which has allowed the isolation of fairly pure zone-specific cell populations from the human fetal adrenal cortex, we can begin to unravel the complex interactions regulating adrenal growth and functional differentiation.

Metyrapone infusion stimulates adrenal growth without activating the cell cycle or the IGF system in the late gestation fetal sheep.

Recently, we have demonstrated that administration of metyrapone, to suppress cortisol synthesis and decrease negative feedback at the pituitary, results in an increase in circulating ACTH and adrenal growth in the late gestation sheep fetus. In these studies, we demonstrated a 2-fold increase in adrenocortical growth using morphometric techniques. To elucidate the potential molecular mechanisms leading to the increase in adrenal growth, we examined adrenal expression of the cell cycle regulatory proteins (cyclin D1) and cyclin-kinase inhibitory proteins (p16ink, p21Cip), and insulin-like growth factor-II (IGF-II), IGF-binding protein 2 (IGFBP-2) and IGF-I type 1 Receptor (IGF1R) from fetuses infused with metyrapone or vehicle for 15 days. There was a significant decrease in adrenal expression of cyclin D1 in metyrapone-(472.0 +/- 29.7) compared with vehicle-infused (662.7 +/- 29.2) fetuses. There was no significant difference, however, in the adrenal expression of the cyclin-kinase inhibitory proteins (p16ink or p21Cip) or in the IGF system (IGF-II, IGFBP-2 or IGF1R) mRNA between metyrapone- and vehicle-infused. In summary, in this model of metyrapone-activated adrenal cortical hypertrophy, growth occurs with a suppression of the rate-limiting cell cycle protein and without activation of the IGF system.

Steroidogenic acute regulatory protein expression is decreased in the adrenal gland of the growth-restricted sheep fetus during late gestation.

Functional development of the adrenal cortex is critical for fetal maturation and postnatal survival. In the present study, we have determined the developmental profile of expression of the mRNA and protein of an essential cholesterol-transporting protein, steroidogenic acute regulatory protein (StAR), in the adrenal of the sheep fetus. We have also investigated the effect of placental restriction (PR) on the expression of StAR mRNA and protein in the growth-restricted fetus. Adrenal glands were collected from fetal sheep at 82-91 days (n = 10), 125-133 days (n = 10), and 140-144 days (n = 9) and from PR fetuses at 141-145 days gestation (n = 9) (term = 147 +/- 3 days gestation). The adrenal StAR mRNA:18S rRNA increased (P < 0.05) between 125 days (7.44 +/- 1.61) and 141-144 days gestation (13.76 +/- 1.88). There was also a 13-fold increase (P < 0.05) in the amount of adrenal StAR protein between 133 and 144 days gestation in these fetuses. However, the amount of StAR protein (6.9 +/- 1.7 arbitrary densitometric units [AU]/microg adrenal protein) in the adrenal of the growth-restricted fetal sheep was significantly reduced, when compared with the expression of StAR protein (17.1 +/- 1.9 AU/microg adrenal protein) in adrenals from the age-matched control group. In summary, there is a developmental increase in the expression of StAR mRNA and protein in the fetal sheep adrenal during the prepartum period when adrenal growth and steroidogenesis is dependent on ACTH stimulation. We have found that, while the level of expression of StAR protein is decreased in the adrenal gland of the growth-restricted fetus during late gestation, this does not impair adrenal steroidogenesis. Our data also suggest that the stimulation of adrenal growth and steroidogenesis in the growth-restricted fetus may not be ACTH dependent.

Cortisol infusion decreases renin, but not PGHS-2, EP2, or EP4 mRNA expression in the kidney of the fetal sheep at days 109-116.

Renal prostaglandins (PG), renin, and cortisol are necessary for normal kidney development and function during fetal life. We examined the effects of cortisol infusion before completion of nephrogenesis (d 109-116 gestation; 2.0-3.0 mg hydrocortisone succinate/24 h) on the renal mRNA expression of PGHS-2, the PGE(2) receptors, EP(2) and EP(4), and renin in fetal sheep. Cortisol infusion raised plasma cortisol levels to 42.8 +/- 6.0 nmol/L compared with saline infusion levels of 1.5 +/- 0.5 nmol/L (p < 0.001), but had no effect on fetal body weight, proportional kidney mass, or blood gases. Cortisol decreased significantly the relative expression of renin mRNA (saline: 0.93 +/- 0.06 units; cortisol: 0.32 +/- 0.03 units, p < 0.05), however it had no effect upon the expression of PGHS-2, EP(2), or EP(4) mRNA in fetal sheep kidney. Although there is substantial evidence that PGE(2) acting through either the EP(2) or EP(4) receptor stimulates renin synthesis in the adult kidney, our results have demonstrated that before the completion of nephrogenesis, cortisol down-regulation of renin mRNA expression is independent of any change in the expression of PGHS-2, EP(2), or EP(4) mRNA expression. During nephrogenesis, the insensitivity of PGHS-2, EP(2), and EP(4) expression to down-regulation by cortisol may permit continued PG regulation of renal development and urine formation.