Impact of Maternal Undernutrition on the Hypothalamic–Pituitary– Adrenal Axis Responsiveness in Sheep at Different Ages Postnatal
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495 Impact of maternal undernutrition on the hypothalamic–pituitary– adrenal axis responsiveness in sheep at different ages postnatal S E Chadio1, B Kotsampasi1, G Papadomichelakis2, S Deligeorgis3, D Kalogiannis1, I Menegatos1 and G Zervas2 1Laboratory of Anatomy and Physiology of Domestic Animals, 2Laboratory of Animal Nutrition, 3Laboratory of Animal Breeding, Department of Animal Science, Agricultural University of Athens, 75, Iera odos, 11855 Athens, Greece (Requests for offprints should be addressed to S E Chadio; Email: [email protected]) Abstract Epidemiological and experimental data support the lower (P!0.05) in group R2 compared with control. Birth hypothesis of ‘fetal programming’, which proposes that weight of lambs was not affected by the maternal nutritional alterations in fetal nutrition and endocrine status lead to manipulation. The area under the curve for ACTH and permanent adaptations in fetal homeostatic mechanisms, cortisol response to CRH challenge was greater (P!0.05) in producing long-term changes in physiology and determine lambs of group R1 at two months of age, whereas no susceptibility to later disease. Altered hypothalamic–pituitary– difference was detected at the ages of 5.5 and 10 months. adrenal (HPA) axis function has been proposed to play an However, significantly higher (P!0.01) basal cortisol levels important role in programming of disease risk. The aim of the were observed in lambs of R1 group at 5.5 months of age. present study was to examine the effects of maternal nutrient There was no interaction between treatment and sex for both restriction imposed during different periods of gestation on pituitary and adrenal responses to the challenge. A significant the HPA axis function in sheep, at different ages postnatal. sex effect was evident with females responding with higher Pregnant ewes were fed a 50% nutrient-restricted diet from ACTH and cortisol levels at the age of 5.5 months (P!0.01, days 0–30 (group R1, nZ7), or from days 31–100 of gestation ! . Z P 0 001 respectively) and with higher cortisol levels (group R2, n 7) or a control 100% diet throughout (P!0.01) at 10 months of age than males. It is concluded pregnancy, (Control, nZ8). Blood samples were collected that the HPA axis is programmable by altered nutrition in at 10-day intervals from day 40 of gestation to term. Lambs utero. The sensitivity of the axis to exogenous stimulation is were born naturally and fed to appetite throughout the study enhanced during early postnatal life and attenuated with age, period. At 2, 5.5, and 10 months of age lambs were given an suggesting a role for the postnatal influences in resetting of the i.v. injection of corticotrophin-releasing hormone (CRH) and blood samples were collected at K15, 0, 15, 30, 60, 120, HPA axis and emphasizing the importance of identifying and 180 min postinjection. Maternal cortisol levels were the impact of maternal undernutrition at several time points significantly higher (P!0.05) in group R1 compared with after birth. the other two groups, whereas maternal insulin levels were Journal of Endocrinology (2007) 192, 495–503 Introduction programming sequelae of maternal nutrient restriction (Langley-Evans et al. 1996). In sheep, maternal cortisol A series of experimental and epidemiological studies had led changes are related to the timing when feed restriction is to the fetal programming hypothesis, which implies that imposed. Late-gestation undernutrition resulted in elevated adverse environmental factors, acting in utero program the maternal cortisol concentrations (Edwards & McMillen development of fetal tissues, producing dysfunctions and 2001), while early- to mid-gestation or periconceptional diseases in adults (Barker et al. 1993). Such programming undernutrition resulted in reduced or unchanged cortisol reflects the action of a factor during a sensitive period or levels respectively (Bispham et al. 2003, Gardner et al. 2006). window of development to exert organizational effects that It has been shown that maternal nutrient restriction, which persist throughout life (Seckl 2001). The triggers operative has no effect on birth weight, can alter the function of the during fetal life that have been studied most extensively are hypothalamic–pituitary–adrenal (HPA) axis (Hawkins et al. undernutrition and glucocorticoid exposure (Seckl 2001, 1999, 2000), which has been suggested to play a role in Symonds et al. 2001). The glucocorticoids are proposed to act programming of the later disease (Seckl 1997). Studies as intermediary factors that transcribe the development examining the impact of maternal nutrient restriction on Journal of Endocrinology (2007) 192, 495–503 DOI: 10.1677/JOE-06-0172 0022–0795/07/0192–495 q 2007 Society for Endocrinology Printed in Great Britain Online version via http://www.endocrinology-journals.org Downloaded from Bioscientifica.com at 10/01/2021 12:05:37AM via free access 496 S E CHADIO and others . Maternal undernutrition in sheep offspring HPA axis have been mostly limited to the late-gestation fetus concentrate mixture (in pellets) containing barley, corn, or young lambs (Hawkins et al. 2000, Edwards et al. 2001, wheat bran, cottonseed meal, limestone, bicalcium phos- Edwards & McMillen 2002, Bloomfield et al. 2004) and only a phate, sodium chloride, and mineral–vitamin premix. The few have been undertaken to establish the long-term effects of lucerne chaff, wheat straw and pelleted concentrate provided maternal undernutrition on HPA function in the adult sheep. 7.2, 5.5, 10 MJ metabolizable energy/kg and contained 135, Brief undernutrition for 10 but not 20 days during late 32 and 115 g crude protein/kg respectively. gestation altered the HPA function in adult offspring, further Pregnancy and fetal number were confirmed by ultrasound emphasizing the fact that the timing, type, and duration of at day 70 of gestation. Only twin-bearing ewes were used, fetal nutrient restriction are each important in determining while two singleton-bearing ewes from group R2 were the nature of the fetal adaptive responses and their excluded from the study. All nutritional regimens were pathophysiological sequelae in later life (Bloomfield et al. adjusted for gestational age and fetal number, while 2003). More recently periconceptional undernutrition was maintaining the treatment difference and taking into account shown to exert a minor influence on HPA axis function in the increasing fetal burden. young adult sheep (Gardner et al. 2006). Ewes were weighed weekly before morning feeding It has been suggested that HPA axis function may undergo throughout gestation. Blood samples were collected at developmental changes, since prenatal glucocorticoid 10-day intervals before feeding for the measurement of exposure in sheep resulted in different HPA axis responsive- cortisol and insulin concentrations, from day 40 until ness between 6 months and 1 year postnatal age, which parturition. All blood samples were centrifuged at highlights the importance of studying outcome at several 2800 r.p.m. for 15 min and plasma was separated into aliquots different times postnatal (Sloboda et al. 2002). and stored at K20 8C for subsequent analysis. To our knowledge, no study has investigated the age- Lambs were delivered naturally. After delivery they were related changes on stimulated HPA axis function in sheep dried, ear tagged, and weighed and the date of birth and sex offspring as a result of maternal undernutrition at different were recorded. Two days after birth, newborns were discrete times during pregnancy. separated from their mothers and reared on artificial milk Therefore, the present study was undertaken in order to containing coconut oil, whey powder, wheat starch, soya characterize the effects of 50% maternal nutrient restriction protein, and vitamin–antioxidants premix (Capragno, Serval during two (0–30 and 31–100 days) periods of gestation on S.A., France). Milk was offered ad libitum, in order to ensure the HPA axis responsiveness to exogenously administered that lambs met their daily requirements, until the age of corticotropin-releasing hormone at three different ages in 45 days. After this age, lambs’ diet consisted of lucerne hay postnatal life in sheep. and a commercial pellet mixture containing barley, corn, soya meal, sunflower meal, wheat bran, powder milk, limestone, bicalcium phosphate, and mineral–vitamin pre- Materials and Methods mix. Lambs were weighed biweekly from birth to the end of experiment. Three lambs from the control and one from the Animal management and nutritional treatments R2 group were weighed until weaning and therefore Z Z Twenty-two ewes (Ovies aries) of the Chios breed, of similar treatment groups were: C (male (m) 7/female (f) 6), R1 Z Z Z Z age (2.0G0.3 years) and body weight (63.0G0.6 kg), were (m 7/f 7), R2: (m 5/f 4). used. Housing and care of animals conformed to Faculty of Animal Science guidelines. Ewes were housed indoors and Corticotropin releasing hormone (CRH) challenges were mated following estrus synchronization with intrava- At three different ages (2, 5.5, and 10 months) lambs were ginal progestagen sponges (Chronogest, Intervet, Holland). At the time of mating, they were randomly allocated to one of injected with 0 5 mg ovine CRH/kg body weight (Sigma the following treatments. Control group (C, nZ8) fed 100% Chemical). Blood samples were collected at 15 and 0 min of the recommended metabolizable energy (ME) and crude before and at 15, 30, 60, and 180 min after the CRH protein (CP) requirements (9 MJ ME/day and 104 g CP/day challenge, in heparinized tubes kept on ice, centrifuged at for the maintenance of a 60 kg non pregnant ewe, National 2800 r.p.m. for 15 min and stored at K20 8C until hormone Research Council (NRC 1985) for the whole of gestational analysis. All challenges were administered between 0800 and period, nutrient restricted group 1 (R1, nZ7) offered 50% of 0900 h in order to minimize the impact of circadian the control nutrient allowance during the first 30 days of variability on measurements of plasma adrenocorticotropic pregnancy, and nutrient restricted group 2 (R2, nZ7) fed to hormone (ACTH) and cortisol.