Francis Bloomfield

Fetal exposure to excess glucocorticoid is unlikely to explain the effects of periconceptional undernutrition in sheep

Periconceptional undernutrition alters fetal growth, metabolism and endocrinology in late gestation. The underlying mechanisms remain uncertain, but fetal exposure to excess maternal glucocorticoids has been hypothesized. We investigated the effects of periconceptional undernutrition on maternal hypothalamic–pituitary–adrenal axis function and placental 11β‐hydroxysteroid dehydrogenase type 2 (11βHSD2) activity. Ewes received maintenance feed (N, n= 20) or decreased feed from −60 to +30 days from mating to achieve 15% weight loss after an initial 2‐day fast (UN, n= 21). Baseline plasma samples and arginine vasopressin (AVP)–corticotrophin‐releasing hormone (CRH) challenges were performed on days −61, −57, −29, −1, +29, 33, and 49 from mating (day 0). Maternal adrenal and placental tissue was collected at 50 days. Baseline plasma levels of adrenocorticotrophic hormone (ACTH) and cortisol decreased in the UN group (P < 0.0001). ACTH response to AVP–CRH was greater in UN ewes during undernutrition (P= 0.03) returning to normal levels after refeeding. Cortisol response to AVP–CRH was greater in UN ewes after the initial 2‐day fast, but thereafter decreased and was lower in UN ewes from mating until the end of the experiment (P= 0.007). ACTH receptor, StAR and p450c17 mRNA levels were down‐regulated in adrenal tissue from UN ewes. Placental 11βHSD2 activity was lower in UN than N ewes at 50 days (P= 0.014). Moderate periconceptional undernutrition results in decreased maternal plasma cortisol concentrations during undernutrition and after refeeding, and adrenal resistance to ACTH for at least 20 days after refeeding. Fetal exposure to excess maternal cortisol is unlikely during the period of undernutrition, but could occur later in gestation if maternal plasma cortisol levels return to normal while placental 11βHSD2 activity remains low.

Effects of twin pregnancy and periconceptional undernutrition on maternal metabolism, fetal growth and glucose–insulin axis function in ovine pregnancy

Although twins have lower birthweights than singletons, they may not experience the increased disease risk in adulthood reportedly associated with low birthweight. In contrast, another periconceptional event, maternal undernutrition, does not reduce birthweight but does affect fetal and postnatal physiology in sheep. We therefore studied maternal and fetal metabolism, growth and glucose–insulin axis function in late gestation in twin and singleton sheep pregnancies, either undernourished from 60 days before until 30 days after conception or fed ad libitum. We found that twin‐bearing ewes had decreased maternal food intake in late gestation and lower maternal and fetal plasma glucose and insulin levels. Twin fetuses had fewer everted placentomes, grew slower in late gestation, and had a greater insulin response to a glucose challenge, but lesser response to arginine. In contrast, periconceptional undernutrition led to increased maternal food intake and a more rapid fall in maternal glucose levels in response to fasting. Periconceptional undernutrition increased the number of everted placentomes, and abolished the difference in insulin responses to glucose between twins and singletons. Thus, the physiology of twin pregnancy is quite different from that of singleton pregnancy, and is probably determined by a combination of factors acting in both early and late gestation. The inconsistency of the relationships between low birthweight and postnatal disease risk of twins may lie in their very different fetal development. These data suggest that twin pregnancy may be another paradigm of developmental programming, and indicate that twins and singletons must be examined separately in any study of fetal or postnatal physiology.

Effects of sex, litter size and periconceptional ewe nutrition on offspring behavioural and physiological response to isolation

Maternal periconceptional undernutrition alters fetal hypothalamic-pituitary-adrenal (HPA) axis development. However, the effects of this early nutritional insult on postnatal HPA axis function and stress-related behaviours are unknown. We investigated in sheep the effects of different periods of undernutrition, and of sex and litter size, on offspring behavioural and cortisol responses to isolation stress. We studied four nutritional groups: controls well nourished throughout pregnancy (n=39), or ewes undernourished (UN, 10-15% body weight reduction) before mating (-60 to 0d, n=26), after mating (-2 to +30d, n=20) or both (-60 to +30d, n=36). At 4 and 18months of age, offspring were isolated for 5min, their behaviour video recorded, and plasma cortisol concentrations measured. Offspring of all undernourished groups demonstrated 50% fewer escape attempts than controls at 4 months of age, and offspring of UN-60+30 ewes had 20% lower plasma cortisol area under the curve in response to isolation at 18months. Females had higher cortisol concentrations and vocalised more than males at 4 and 18months, and were more active at 18months. After isolation, UN-2+30 males had higher cortisol concentrations than UN-2+30 females whereas in all other groups males had lower concentrations than females. Singleton males made more escape attempts than females, whereas in twins females made more escape attempts than males. These findings suggest that maternal periconceptional undernutrition in sheep can suppress behavioural reactions and cortisol secretion in response to isolation stress in the offspring into adulthood, and that these effects differ between males and females.

The late effects of fetal growth patterns

The immediate prenatal and postnatal consequences of reduced fetal growth have long been known. The longer term associations between reduced birth weight and adult disease risk are also now well established. Reduced fetal growth is usually detected late in gestation, and the assumption has been that this is the time when factors regulating fetal growth have their greatest effect. However, recent evidence suggests that both the growth trajectory of the fetus and its adaptive responses to the prenatal and postnatal environment may be determined in the period around the time of conception.

The effects of maternal nutrition around the time of conception on the health of the offspring.

The incidence of prematurity, diabetes and cardiovascular disease have been increasing in both the developed and developing world. Increasing numbers of human studies suggest that these serious health outcomes may have developmental origins originating from nutritional deficits in the periconceptional period, with maternal nutrition around the time of conception now shown to have important effects on the length of gestation, trajectory of fetal growth and on postnatal growth and health. Biomedical research using the pregnant sheep has been widely employed to gain a deeper understanding of the underlying mechanisms involved. There is growing awareness that this field of research has major implications for the livestock production industry. From our own studies on sheep we have evidence that maternal undernutrition during the periconceptional period results in altered fetal hypothalamic-pituitary-adrenal axis (HPAA) development, an increased rate of premature birth, altered fetal pancreatic function, insulin signalling and amino acid metabolism, and also alterations in maternal adaptation to pregnancy. We are currently studying the postnatal consequences of these changes. Other research groups have shown that restricted nutrition of sheep in the early part of pregnancy alters postnatal muscle development, fat deposition, cardiovascular regulation and HPAA function. One aim of this review is to illustrate how biomedical research using animals such as the sheep has been used to gain a better understanding of the consequences of reduced maternal nutrition during the periconceptional period. We suggest that there are equally important consequences of this research for the livestock production industries.

Experimental aspects of nutrition and fetal growth

Intrauterine growth restriction (IUGR) remains a major cause of perinatal morbidity and mortality and, as yet, there is no effective treatment. Most fetuses with ultrasound evidence of moderate to severe IUGR do not grow better out of the womb than in, despite early enteral feeds and subsequent calorie supplementation. Research into possible therapies for growth restricted babies has thus also been directed towards the fetus. Major advances have been made in recent years in the understanding of the physiology of fetal growth, and it has become clear that fetal nutrition is the determining factor.

Size at birth and adult fat mass in twin sheep are determined in early gestation

•  Reduced size at birth and shorter gestation length are both associated with increased risks of non‐communicable diseases (NCD) in later adult life. •  Twins are born both smaller and earlier than singletons and adult twins also are reported to be at increased risk of common NCDs such as diabetes. •  The smaller size and shorter gestation length of twins has been presumed to be due to a lack of intrauterine space and/or limitations of placental nutrient supply in late gestation, but there are few data to support this. •  We show that size at birth and adult fat mass in twin sheep are determined largely in early gestation. •  Knowledge of the mechanisms underlying early pregnancy determination of fetal growth and gestation length in twins are likely to increase understanding of how early pregnancy factors influence lifelong health for offspring from all pregnancies.

Fetal growth factors and fetal nutrition

Optimal fetal growth is important for a healthy pregnancy outcome and also for lifelong health. Fetal growth is largely regulated by fetal nutrition, and mediated via the maternal and fetal glucose/insulin/insulin-like growth factor axes. Fetal nutrition may reflect maternal nutrition, but abnormalities of placental function can also affect fetal growth, as the placenta plays a key intermediary role in nutritional signalling between mother and fetus. Fetal nutrition also impacts on the development of key fetal endocrine systems such as the glucose-insulin and insulin-like growth factor axes. This is likely to contribute to the link between both fetal growth restriction and fetal overgrowth, and increased risks of obesity and impaired glucose tolerance in later life. This review focuses on the associations between maternal and fetal nutrition, fetal growth and later disease risk, with particular emphasis on the role of insulin-like growth factors and the importance of the periconceptional period.

A chronic low dose infusion of insulin-like growth factor I alters placental function but does not affect fetal growth.

Knowledge of the anabolic effects of insulin-like growth factor I (IGF-I) on fetal growth and feto-placental metabolism are derived from studies using large doses of IGF-I. Low doses of enteral IGF-I have trophic effects on the fetal gut, but there are no data on the effects of systemic low doses of IGF-I on fetal growth and feto-placental metabolism. We therefore compared the effects of a chronic infusion of low dose IGF-I (50 microg day(-1), n = 7) with vehicle-infused controls (n = 7) on fetal growth, metabolism and placental transfer capacity in the chronically instrumented late gestation ovine fetus (121-132 days of gestation; term = 145 days). Insulin-like growth factor I infusion did not affect fetal growth or the size of individual organs, including liver, spleen and bone. Placental morphology was altered, and placental clearances of 3-O-[methyl-3H]D-glucose (a non-metabolizable glucose analogue) and [methyl(14C)]aminoisobutyric acid (a non-metabolizable analogue of amino acids utilizing the system A transporter), were reduced in IGF-I-treated fetuses (P < 0.05 v. control). However, fetal and placental metabolite uptake was not significantly different between groups. We conclude that, despite altering placental transfer capacity and morphology, a chronic low dose infusion of IGF-I does not alter fetal growth or metabolism.

Periconceptional undernutrition in sheep leads to decreased locomotor activity in a natural environment

Maternal undernutrition during pregnancy increases offspring obesity and metabolic disease risk. We hypothesized that periconceptional undernutrition in sheep from 60 days before conception through to day 30 of gestation (UN) would decrease voluntary locomotor activity in adult offspring. Distance travelled was measured at 18 months of age for ∼48 h in the paddock. Data were analysed using multiple regression analysis, with explanatory variables including sex, nutrition group, birth weight, average time between GPS measurements and percentage of time during the measurement period spent in daylight. Mean (±s.e.) distance walked (m/h) was greater for control (CON) than UN animals, and greater for females than males [110.2 (6.5), CON females; 110.7 (6.3), CON males; 105.1 (5.3), UN females and 95.5 (5.8), UN males; P = 0.02 for nutrition group effect and for sex effect]. Periconceptional undernutrition may lead to a significant decrease in voluntary physical activity in adult offspring.