Juanita K. Jellyman

Fetal cardiovascular, metabolic and endocrine responses to acute hypoxaemia during and following maternal treatment with dexamethasone in sheep

In sheep, direct fetal treatment with dexamethasone alters basal cardiovascular function and the cardiovascular response to acute hypoxaemia. However, in human clinical practice, dexamethasone is administered to the mother, not to the fetus. Hence, this study investigated physiological responses to acute hypoxaemia in fetal sheep during and following maternal treatment with dexamethasone in doses and at dose intervals used in human clinical practice. Under anaesthesia, 18 fetal sheep were instrumented with vascular and amniotic catheters, a carotid flow probe and a femoral flow probe at 118 days gestation (term ca 145 days). Following 6 days recovery at 124 days gestation, 10 ewes received dexamethasone (2 × 12 mg daily i.m. injections in saline). The remaining animals were saline‐injected as age‐matched controls. Two episodes of hypoxaemia (H) were induced in all animals by reducing the maternal FIO2for 1 h (H1, 8 h after the second injection; H2, 3 days after the second injection). In fetuses whose mothers received saline, hypoxaemia induced significant increases in fetal arterial blood pressure, carotid blood flow and carotid vascular conductance and femoral vascular resistance, significant falls in femoral blood flow and femoral vascular conductance and transient bradycardia. These cardiovascular responses were accompanied by a fall in arterial pH, increases in blood glucose and blood lactate concentrations and increased plasma concentrations of catecholamines. In fetuses whose mothers were treated with dexamethasone, bradycardia persisted throughout hypoxaemia, the magnitude of the femoral vasoconstriction, the glycaemic, lactacidaemic and acidaemic responses and the plasma concentration of neuropeptide Y (NPY) were all enhanced during H1. However, during H2, all of these physiological responses were similar to saline controls. In dexamethasone fetuses, the increase in plasma adrenaline was attenuated during H1 and the increase in carotid vascular conductance during hypoxaemia failed to reach statistical significance both during H1 and during H2. These data show that maternal treatment with dexamethasone in doses and intervals used in human obstetric practice modified the fetal cardiovascular, metabolic and endocrine defence responses to acute hypoxaemia. Furthermore, dexamethasone‐induced alterations to these defences depended on whether the hypoxaemic challenge occurred during or following maternal dexamethasone treatment.

Effect of dexamethasone on pulmonary and renal angiotensin-converting enzyme concentration in fetal sheep during late gestation ☆

OBJECTIVES The effect of dexamethasone on tissue angiotensin-converting enzyme (ACE) was investigated in fetal sheep. STUDY DESIGN Pulmonary and renal ACE concentrations were measured in 16 sheep fetuses at between 127 and 131 days of gestation (term 145+/-2 days): 6 were untreated, whereas 10 were chronically catheterized and infused intravenously with either saline solution (0.9%, n=4) or dexamethasone (45-60 microg. kg(-1). d(-1), n=6) for the previous 2 days. The dexamethasone dose increased plasma dexamethasone to around one fifth of that measured in newborn human infants delivered after maternal dexamethasone treatment. RESULTS Over the period of infusion, arterial blood pressure increased significantly in the dexamethasone (+6.8+/-1.5 mm Hg, P<.05) but not saline-treated fetuses (+1.6+/-0.6 mm Hg). At delivery, pulmonary ACE in the dexamethasone-infused fetuses (1.24+/-0.26 nmoles hippurate. min(-1). mg protein(-1)) was significantly greater than in the control fetuses (0.50+/-0.07 nmoles. min(-1). mg protein(-1), P<.005); renal ACE was unchanged by dexamethasone treatment. Overall, pulmonary ACE and blood pressure were correlated on the last day of infusion (r=0.70, P<.05). CONCLUSION The rise in pulmonary ACE seen in dexamethasone-treated sheep fetuses may contribute, in part, to the glucocorticoid-induced increase in blood pressure.

Effects of Cortisol and Dexamethasone on Insulin Signalling Pathways in Skeletal Muscle of the Ovine Fetus during Late Gestation

Before birth, glucocorticoids retard growth, although the extent to which this is mediated by changes in insulin signalling pathways in the skeletal muscle of the fetus is unknown. The current study determined the effects of endogenous and synthetic glucocorticoid exposure on insulin signalling proteins in skeletal muscle of fetal sheep during late gestation. Experimental manipulation of fetal plasma glucocorticoid concentration was achieved by fetal cortisol infusion and maternal dexamethasone treatment. Cortisol infusion significantly increased muscle protein levels of Akt2 and phosphorylated Akt at Ser473, and decreased protein levels of phosphorylated forms of mTOR at Ser2448 and S6K at Thr389. Muscle GLUT4 protein expression was significantly higher in fetuses whose mothers were treated with dexamethasone compared to those treated with saline. There were no significant effects of glucocorticoid exposure on muscle protein abundance of IR-β, IGF-1R, PKCζ, Akt1, calpastatin or muscle glycogen content. The present study demonstrated that components of the insulin signalling pathway in skeletal muscle of the ovine fetus are influenced differentially by naturally occurring and synthetic glucocorticoids. These findings may provide a mechanism by which elevated concentrations of endogenous glucocorticoids retard fetal growth.

Glucocorticoid programming of intrauterine development

Glucocorticoids (GCs) are important environmental and maturational signals during intrauterine development. Toward term, the maturational rise in fetal glucocorticoid receptor concentrations decreases fetal growth and induces differentiation of key tissues essential for neonatal survival. When cortisol levels rise earlier in gestation as a result of suboptimal conditions for fetal growth, the switch from tissue accretion to differentiation is initiated prematurely, which alters the phenotype that develops from the genotype inherited at conception. Although this improves the chances of survival should delivery occur, it also has functional consequences for the offspring long after birth. Glucocorticoids are, therefore, also programming signals that permanently alter tissue structure and function during intrauterine development to optimize offspring fitness. However, if the postnatal environmental conditions differ from those signaled in utero, the phenotypical outcome of early-life glucocorticoid receptor overexposure may become maladaptive and lead to physiological dysfunction in the adult. This review focuses on the role of GCs in developmental programming, primarily in farm species. It examines the factors influencing GC bioavailability in utero and the effects that GCs have on the development of fetal tissues and organ systems, both at term and earlier in gestation. It also discusses the windows of susceptibility to GC overexposure in early life together with the molecular mechanisms and long-term consequences of GC programming with particular emphasis on the cardiovascular, metabolic, and endocrine phenotype of the offspring.

Pituitary-adrenal responses to acute hypoxemia during and after maternal dexamethasone treatment in sheep.

The effects of maternal dexamethasone treatment on hypothalamic-pituitary-adrenal axis function were determined during basal and hypoxemic conditions in maternal and fetal sheep. Under halothane, ewes and their fetuses were catheterized at 117 d gestation (term = 145 d). Starting at 124 d, the ewes received i.m. injections of two doses of either dexamethasone (12 mg) or saline at 24-h intervals. All animals experienced one episode of hypoxemia when the dexamethasone was present in the maternal and fetal circulations [125 ± 1 d (H1)] and a second episode of hypoxemia when the steroid was no longer detectable in either the maternal or fetal circulations [128 ± 1 d (H2)]. The fall in partial pressure of oxygen in arterial blood in response to hypoxia was similar in the two episodes in both the fetal and the maternal blood. Maternal dexamethasone treatment diminished maternal and fetal basal plasma cortisol but not ACTH during the normoxic period of H1 but not H2. In control animals, hypoxemia induced increases in fetal but not maternal ACTH and cortisol concentrations. In dexamethasone-treated animals, maternal ACTH and cortisol concentrations also remained unchanged from baseline in both H1 and H2. In contrast, fetal plasma ACTH and cortisol responses to hypoxemia were significantly suppressed during H1 but not H2. Correlation of fetal plasma ACTH and cortisol concentrations suggested diminished cortisol output without a change in adrenocortical responsiveness in dexamethasone-treated fetuses during H1 but not H2. Maternal treatment with dexamethasone transiently suppressed maternal and fetal basal hypothalamic-pituitary-adrenal axis function and the fetal plasma ACTH and cortisol responses to acute hypoxemia in sheep.

Antenatal glucocorticoid therapy increases glucose delivery to cerebral circulations during acute hypoxemia in fetal sheep during late gestation

OBJECTIVE To determine the effects of 2 maternal injections with dexamethasone on the calculated oxygen and glucose deliveries to fetal cerebral and peripheral circulations during acute hypoxemia in sheep. STUDY DESIGN Beginning at 124 days, ewes received 2 intramuscular injections of either dexamethasone (2 x 12 mg, n = 10) or saline solution (2 x 2 mL, n = 12) 24 hours apart. Hypoxemia (1 hour) was induced 32 hours after the first injection (H1) and 3 days after the second (H2). RESULTS In saline solution-treated fetuses, glucose delivery was unchanged or increased in femoral and carotid circulations, respectively, during H1 and H2. In dexamethasone-treated fetuses, the increase in glucose delivery to the head tended to be greater during H1 and was significantly enhanced in dexamethasone- vs saline solution-treated fetuses during H2. CONCLUSION Two maternal injections with dexamethasone significantly enhanced glucose delivery to the head during acute hypoxemia in the ovine fetus.

Sex‐associated differences in pancreatic β cell function in healthy preweaning pony foals

REASONS FOR PERFORMING STUDY Pancreatic β cells are responsive to a range of stimuli during early post natal life in healthy pony foals. However, little is known about whether these responses are sex-linked. OBJECTIVES To determine pancreatic β cell responses to the insulin secretagogues, glucose, arginine and tolbutamide, in fillies and colts during the first 3 months after birth. STUDY DESIGN In vivo experiment examining sex differences in pancreatic β cell function in foals. METHODS Female (n = 8) and male (n = 5) pony foals were infused i.v. with glucose (0.5 g/kg bwt 40% dextrose), arginine (100 mg/kg bwt) or tolbutamide (10 or 20 mg/kg bwt) over 5 min, at 48 h intervals, to assess pancreatic β cell function at ages 2 and 12 weeks. Blood samples (4 ml) were taken through a jugular catheter at -30, -15 and 0 min (immediately before) and 5, 15, 30, 45, 60, 90 and 120 min after glucose, arginine and tolbutamide administration for measurements of plasma glucose, α-amino-nitrogen and insulin concentrations. RESULTS The maximum increment in plasma insulin concentration in response to glucose was significantly higher in female (395 ± 58 ng/l) than male (172 ± 37 ng/l, P<0.05) pony foals 2 weeks after birth and the area under the insulin curve was significantly greater in females at this age. At 12 weeks, the insulin increment in response to glucose was significantly greater in fillies 45 min post infusion. The β cell responses to arginine and tolbutamide were not sex-linked at either age. CONCLUSIONS These data show that in ponies, fillies have a greater β cell response to glucose than colts in early post natal life. Since glucose clearance was unaffected by sex, the results suggest that fillies may be less insulin sensitive than colts shortly after birth. Innate sex differences in the secretion and action of insulin in early post natal life may influence tissue development and growth with potentially more long-term metabolic consequences.

Renal growth retardation following angiotensin II type 1 (AT1) receptor antagonism is associated with increased AT2 receptor protein in fetal sheep

The actions of angiotensin II on type 1 (AT₁) and type 2 (AT₂) receptor subtypes are important for normal kidney development before birth. This study investigated the effect of AT₁ receptor antagonism on renal growth and growth regulators in fetal sheep during late gestation. From 125 days of gestation (term 145±2 days), chronically catheterised sheep fetuses were infused intravenously for 5 days with either an AT₁-specific receptor antagonist (GR138950, 2-4 mg/kg per day, n=5) or saline (0.9% NaCl, n=5). Blockade of the AT₁ receptor decreased arterial blood oxygenation and pH and increased blood pCO₂, haemoglobin and lactate, and plasma cortisol and IGF-II. Blood glucose and plasma thyroid hormones and IGF-I were unchanged between the treatment groups. On the 5th day of infusion, the kidneys of the GR-treated fetuses were lighter than those of the control fetuses, both in absolute and relative terms, and were smaller in transverse cross-sectional width and cortical thickness. In the GR-infused fetuses, renal AT₂ receptor protein concentration and glomerular density were significantly greater than in the saline-infused fetuses. Blockade of the AT₁ receptor had no effect on relative cortical thickness, fractional or mean glomerular volumes, or renal protein levels of the AT₁ receptor, IGF type 1 receptor, insulin receptor or protein kinase C ζ. Therefore, in the ovine fetus, AT₁ receptor antagonism causes increased renal protein expression of the AT₂ receptor subtype, which, combined with inhibition of AT₁ receptor activity, may be partly responsible for growth retardation of the developing kidney.

Effects of maternal dexamethasone treatment on pancreatic β cell function in the pregnant mare and postnatal foal

Reasons for performing study: Synthetic glucocorticoids are used to treat inflammatory conditions in horses. In other pregnant animals, glucocorticoids are given to stimulate fetal maturation with long‐term metabolic consequences for the offspring if given preterm. However, their metabolic effects during equine pregnancy remain unknown. Objective: Thus, this study investigated the metabolic effects of dexamethasone administration on pregnant pony mares and their foals after birth. Study design: Experimental study. Methods: A total of 3 doses of dexamethasone (200 &mgr;g/kg bwt i.m.) were given to 6 pony mares at 48 h intervals beginning at ≈270 days of pregnancy. Control saline injections were given to 5 mares using the same protocol. After fasting overnight, pancreatic &bgr; cell responses to exogenous glucose were measured in the mares before, during and after treatment. After birth, pancreatic &bgr; cell responses to exogenous glucose and arginine were measured in the foals at 2 and 12 weeks. Results: In mares during treatment, dexamethasone but not saline increased basal insulin concentrations and prolonged the insulin response to exogenous glucose. Basal insulin and glucose concentrations still differed significantly between the 2 groups 72 h post treatment. Dexamethasone treatment significantly reduced placental area but had little effect on foal biometry at birth or subsequently. Foal &bgr; cell function at 2 weeks was unaffected by maternal treatment. However, by 12 weeks, pancreatic &bgr; cell sensitivity to arginine, but not glucose, was less in foals delivered by dexamethasone‐ than saline‐treated mares. Conclusions: Dexamethasone administration induced changes in maternal insulin‐glucose dynamics, indicative of insulin resistance and had subtle longer term effects on post natal &bgr; cell function of the foals. The programming effects of dexamethasone in horses may be mediated partially by altered maternal metabolism and placental growth.

Maternal Dexamethasone Treatment Alters Tissue and Circulating Components of the Renin-Angiotensin System in the Pregnant Ewe and Fetus.

Antenatal synthetic glucocorticoids promote fetal maturation in pregnant women at risk of preterm delivery and their mechanism of action may involve other endocrine systems. This study investigated the effect of maternal dexamethasone treatment, at clinically relevant doses, on components of the renin-angiotensin system (RAS) in the pregnant ewe and fetus. From 125 days of gestation (term, 145 ± 2 d), 10 ewes carrying single fetuses of mixed sex (3 female, 7 male) were injected twice im, at 10–11 pm, with dexamethasone (2 × 12 mg, n = 5) or saline (n = 5) at 24-hour intervals. At 10 hours after the second injection, maternal dexamethasone treatment increased angiotensin-converting enzyme (ACE) mRNA levels in the fetal lungs, kidneys, and heart and ACE concentration in the circulation and lungs, but not kidneys, of the fetuses. Fetal cardiac mRNA abundance of angiotensin II (AII) type 2 receptor decreased after maternal dexamethasone treatment. Between the two groups of fetuses, there were no significant differences in plasma angiotensinogen or renin concentrations; in transcript levels of renal renin, or AII type 1 or 2 receptors in the lungs and kidneys; or in pulmonary, renal or cardiac protein content of the AII receptors. In the pregnant ewes, dexamethasone administration increased pulmonary ACE and plasma angiotensinogen, and decreased plasma renin, concentrations. Some of the effects of dexamethasone treatment on the maternal and fetal RAS were associated with altered insulin and thyroid hormone activity. Changes in the local and circulating RAS induced by dexamethasone exposure in utero may contribute to the maturational and tissue-specific actions of antenatal glucocorticoid treatment.