Strength of nonhuman primate studies of developmental programming: review of sample sizes, challenges, and steps for future work.

Nonhuman primate (NHP) studies are crucial to biomedical research. NHPs are the species most similar to humans in lifespan, body size, and hormonal profiles. Planning research requires statistical power evaluation, which is difficult to perform when lacking directly relevant preliminary data. This is especially true for NHP developmental programming studies, which are scarce. We review the sample sizes reported, challenges, areas needing further work, and goals of NHP maternal nutritional programming studies. The literature search included 27 keywords, for example, maternal obesity, intrauterine growth restriction, maternal high-fat diet, and maternal nutrient reduction. Only fetal and postnatal offspring studies involving tissue collection or imaging were included. Twenty-eight studies investigated maternal over-nutrition and 33 under-nutrition; 23 involved macaques and 38 baboons. Analysis by sex was performed in 19; minimum group size ranged from 1 to 8 (mean 4.7 ± 0.52, median 4, mode 3) and maximum group size from 3 to 16 (8.3 ± 0.93, 8, 8). Sexes were pooled in 42 studies; minimum group size ranged from 2 to 16 (mean 5.3 ± 0.35, median 6, mode 6) and maximum group size from 4 to 26 (10.2 ± 0.92, 8, 8). A typical study with sex-based analyses had group size minimum 4 and maximum 8 per sex. Among studies with sexes pooled, minimum group size averaged 6 and maximum 8. All studies reported some significant differences between groups. Therefore, studies with group sizes 3-8 can detect significance between groups. To address deficiencies in the literature, goals include increasing age range, more frequently considering sex as a biological variable, expanding topics, replicating studies, exploring intergenerational effects, and examining interventions.

Antenatal Synthetic Glucocorticoid Exposure at Human Therapeutic Equivalent Doses Predisposes Middle-Age Male Offspring Baboons to an Obese Phenotype That Emerges With Aging.

INTRODUCTION: Women threatening premature delivery receive synthetic glucocorticoids (sGC) to accelerate fetal lung maturation, reducing neonatal mortality and morbidity. Few investigations have explored potential long-term offspring side effects. We previously reported increased pericardial fat and liver lipids in 10-year-old (human equivalent 40 years) male baboons exposed to 3 antenatal sGC courses. We hypothesized middle-aged sGC male offspring show obesity-related morphometric changes. METHODS: Pregnant baboons received courses of 2 betamethasone injections (175 µg·kg-1·d-1 intramuscular) at 0.6, 0.64, and 0.68 gestation. At 10 to 12.5 years, we measured morphometrics and serum lipids in 5 sGC-exposed males and 10 age-matched controls. We determined whether morphometric parameters predicted amount of pericardial fat or lipids. Life-course serum lipids were measured in 25 males (7-23 years) providing normal regression formulas to compare sGC baboons' lipid biological and chronological age. RESULTS: Birth weights were similar. When studied, sGC-exposed males showed a steeper weight increase from 8 to 12 years and had increased waist and hip circumferences, neck and triceps skinfolds, and total and low-density lipoprotein cholesterol. Triceps skinfold correlated with apical and midventricular pericardial fat thickness, hip and waist circumferences with insulin. CONCLUSIONS: Triceps skinfold and waist and hip circumferences are useful biomarkers for identifying individuals at risk for obesity and metabolic dysregulation following fetal sGC exposure. Prenatal sGC exposure predisposes male offspring to internal adiposity, greater body size, and increased serum lipids. Results provide further evidence for developmental programming by fetal sGC exposure and call attention to potential emergence of adverse life-course effects.

Effect of 30% nutrient restriction in the first half of gestation on maternal and fetal baboon serum amino acid concentrations.

Mechanisms linking maternal nutrient restriction (MNR) to intra-uterine growth restriction (IUGR) and programming of adult disease remain to be established. The impact of controlled MNR on maternal and fetal amino acid metabolism has not been studied in non-human primates. We hypothesised that MNR in pregnant baboons decreases fetal amino acid availability by mid-gestation. We determined maternal and fetal circulating amino acid concentrations at 90 d gestation (90dG, term 184dG) in control baboons fed ad libitum (C, n 8) or 70% of C (MNR, n 6). Before pregnancy, C and MNR body weights and circulating amino acids were similar. At 90dG, MNR mothers had lower body weight than C mothers (P< 0·05). Fetal and placental weights were similar between the groups. MNR reduced maternal blood urea N (BUN), fetal BUN and fetal BUN:creatinine. Except for histidine and lysine in the C and MNR groups and glutamine in the MNR group, circulating concentrations of all amino acids were lower at 90dG compared with pre-pregnancy. Maternal circulating amino acids at 90dG were similar in the MNR and C groups. In contrast, MNR fetal ß-alanine, glycine and taurine all increased. In conclusion, maternal circulating amino acids were maintained at normal levels and fetal amino acid availability was not impaired in response to 30% global MNR in pregnant baboons. However, MNR weight gain was reduced, suggesting adaptation in maternal-fetal resource allocation in an attempt to maintain normal fetal growth. We speculate that these adaptive mechanisms may fail later in gestation when fetal nutrient demands increase rapidly, resulting in IUGR.

Metabolic adjustments to moderate maternal nutrient restriction.

Reduced food availability in pregnancy influences fetal growth, obstetric outcomes and offspring health in both developing and developed countries. The objective of the present study was to determine responses to moderate global maternal nutrient restriction (MNR) during pregnancy in baboons (Papio hamadryas) - an established non-human primate model for pregnancy-related research. Starting at 30 d gestation (dG), twelve pregnant baboons received 70 % of food (MNR group) consumed by twenty ad libitum-fed pregnant controls. Maternal body weight, BMI, food intake and physical activity were measured before pregnancy, at 90 dG and at 165 dG (full-term 180 dG). Fetal and placental weights were recorded at the time of Caesarean section (90 and 165 dG). Activity patterns were also evaluated in fourteen non-pregnant female baboons. Behavioural observations were made in five non-pregnant, six control and four MNR animals. Pregnant baboons decreased overall physical activity and energy-expensive behaviours compared with non-pregnant baboons. In the MNR group, maternal weight, weight gain and maternal physical activity were reduced compared with the control animals. MNR decreased placental weight and volume compared with control, while fetal weight and length were unaffected. We conclude that decreased physical activity and increased usage of maternal available body stores play an important role in the maternal response to pregnancy. Also, adaptations in maternal behaviour and energy utilisation protect fetal growth during moderate MNR.

Three weekly courses of betamethasone administered to pregnant baboons at 0.6, 0.65, and 0.7 of gestation alter fetal and maternal lymphocyte populations at 0.95 of gestation.

The hypothalamic-pituitary-adrenal (HPA) axis plays a major role in the communication between the immune and neuroendocrine systems. Glucocorticoids are potent immunomodulatory hormones. In the present study, we evaluated the effect of three weekly courses of betamethasone, administered to pregnant baboons at 0.6, 0.65, and 0.7 of gestation, on maternal hematological parameters during treatment, maternal and fetal hematological parameters and lymphocyte populations at 0.95 of gestation, and fetal lymphoid organs and placental structure. Each weekly betamethasone course resulted in decreased granulocytes and increased lymphocytes and monocytes in maternal circulation (by percentage, p < 0.05). The percentage and absolute number of CD8+ T-cells in the maternal circulation were lower and CD4+ T-cells higher (p < 0.05) in treated pregnant animals at 0.95 gestation. The percentage of proliferating CD3- CD8+ cells was lower in blood obtained from the fetal heart of betamethasone-treated animals. In the betamethasone group, the number of CD8+ T-cells and NK cells were elevated and the number of T and CD4+ T-cells were reduced in fetal heart blood compared with the umbilical vein blood. The number of placental macrophages (CD68+ cells) per visual field in betamethasone-treated and control animals were not different. Taken together, our data show that betamethasone treatment of pregnant females with no indication of preterm labor affects some components of the fetal and maternal immune system, altering the maternal CD4+/CD8+ ratio and absolute number of fetal NK cell and maternal CD8+ T-cell.

Ontogeny of hematological cell and biochemical profiles in maternal and fetal baboons (Papio species).

The normal ranges of hematological cell profiles and biochemistry are documented in adult non-pregnant, pregnant, juvenile, and neonatal baboons. Despite the extensive use of the baboon as a model for the study of various aspects of pregnancy, there is no data from paired mothers and their fetuses at different stages of gestation. Hematologic and biochemical profile data were obtained from eight non-pregnant female baboons, 37 mothers and 38 fetal baboons at 30 +/- 2, 90 +/- 2, 125 +/- 2, and 175 +/- 2 days of gestation (mean +/- range; dGA; term, 180 dGA). Changes observed in fetal and maternal blood during normal baboon pregnancy were similar to those reported in human pregnancy. The level of alkaline phosphatase was two times higher in fetal blood circulation than that reported in human pregnancy.

Development of a system for individual feeding of baboons maintained in an outdoor group social environment.

We developed a system that allows individual feeding of adult baboons, 8-15 years of age, maintained in an outdoor group social environment. The purpose of the system is to allow careful monitoring and control of individual diet. Baboons were housed in two group cages, 16 females and a single male in one and 12 females and a single male in the other. Baboons exited the group cage once daily and passed along a chute and over a scale into individual cages where they received their individual diets. Food intake was monitored during their 2-hour stay in the individual cages. Baboons rapidly learned to use this system. Food intake and weight were stable within 20 days. Food consumed decreased during the period of sexual receptivity. The maintenance of the group social environment allowed observations on the group's dominance structure and the relationship of dominance to food consumption. Speed of food access in the group cage was related to dominance. Dominance was not related to food consumed in individual cages. The system permits study of many variables related to behavior and food intake while still retaining critical social interactions.

The effects of photoperiod on the switching of myometrial contractility patterns of pregnant baboons: relationship to surgery and parturition.

OBJECTIVES: Pregnant baboons were studied to determine the precise time of the switch from myometrial contractures to contractions in relation to photoperiod after laparotomy and at parturition. We compared the patterns recorded in baboons to those we have previously reported in pregnant rhesus monkeys to determine fundamental primate characteristics. METHODS: Seven pregnant baboons (126-160 days' gestation) were instrumented with femoral arterial and venous catheters and electrodes for myometrial electromyogram. All animals were subjected to a 14-hour light:10-hour dark photoperiod. Myometrial activity was monitored using a computer-based data acquisition system. Onset time for all switches was noted and standardized against time of lights off. Animals were studied at three stages of pregnancy (stage 1, first 10 days after laparotomy; stage 2, more than 10 days after laparotomy and more than 10 days before cesarean; and stage 3, 10 days before cesarean section or vaginal delivery). RESULTS: All baboons demonstrated myometrial switches for a variable number of days preceding parturition. Onset of darkness was 0 hours. Average time of stage 1 switch onset was 2.17 +/- 0.60 hours and was not different from stage 3 switch onset, which was -1.00 +/- 0.27 hours. CONCLUSIONS: Myometrial contractile patterns showed clear photoperiodicity in the switch from contractures to contractions in late pregnancy in the baboon. The relationship of the switch from contractures to contractions was not altered by surgical laparotomy. There was a significant difference in the time of switch in relation to photoperiod between pregnant rhesus monkeys and baboons. However, the fact that a significant photoperiod exists in both species indicates a fundamental similarity in the switch from contractures to contractions in primate pregnancy.