Vitamins are readily available in the modern diet due to liberalized fortification and supplementation policies. This research tested the hypothesis that high multi-vitamin intake by Wistar rats during pregnancy leads to the development of obesity and characteristics of the metabolic syndrome in the offspring. Pregnant Wistar rats were fed the AIN-93G diet containing either the recommended (RV) or 10-fold increase (HV) in vitamin content. Pups were weaned to the RV diet (Study 1), an obesogenic liquid diet (Ob, Study 2), low vitamin diets (1/3RV or 1/6RV, Study 3), or a nutrient selection paradigm (NSP) with 10% and 60% casein diets (Study 4). Body weight (BW), food intake (FI), glucose and insulin responses, appetite hormones, abdominal fat pad mass (FPM) and systolic blood pressure (SBP) was measured. Expressions of mRNA for hypothalamic serotonin (5-HT) receptors and proopiomelanocortin (POMC) were measured in Study 4. Males, but not females, born to HV dams had higher post-weaning BW and FI when weaned to the RV or 1/3RV diet, and exhibited components of metabolic syndrome, including higher FPM, hyperglycemia, insulin resistance and elevated SBP compared with those born to RV dams. The Ob diet led to exaggerated weight gain and expressions of components of metabolic syndrome in both sexes born to dams fed the HV diet. Female pups on the 1/6RV diet from HV dams had two-fold higher glucose response and lower insulin response, but no difference in post-weaning BW and daily FI compared to those from RV dams. In contrast to the pups born to HV dams and fed a single diet, those from the HV dams and on the NSP gained less weight and ate less, and had lower hypothalamic mRNA expressions of 5-HT receptors and POMC. In conclusion, high multi-vitamin intake during pregnancy may lead to obesity, and result in a higher risk of developing characteristics of metabolic syndrome in the offspring. However, sex, weaning diet composition, and the presence of diet choice alter the outcomes.

The hypothesis that source of protein in a nutritionally adequate diet during gestation, lactation and weaning alters food intake and characteristics of metabolic syndrome in the offspring was investigated. Pregnant Wistar rats were randomized to either the AIN 93-G casein (C) or soy protein (S) diets (n=12/group) during gestation only or during gestation and lactation. Male offspring in each dams’ diet group were weaned to either C or S diets (n=12/group). Food intake, body weight (BW), fat pad mass, systolic (SBP) and diastolic (DBP) blood pressure, and plasma homocysteine (p<0.05) were higher in offspring born to dams fed the S diet. Fasting blood glucose (BG), BG in response to a glucose gavage and Homeostatic Model of Assessment of Insulin Resistance (HOMA-IR) index were higher only in male offspring born to dams fed the S diet. Moreover, gene expression of Agouti Related Protein (AgRP) was higher in offspring born to dams fed the S diet at weaning. Extending the dams’ diet during gestation and lactation magnified the effect of the gestational S diet on BW and composition and glucose metabolism in male offspring. Although composition of the weaning diets interacted with that of the dams’ diets, the latter was the dominant factor in determining metabolic outcomes in the offspring. In conclusion, the soy protein diet, compared to the casein diet when consumed during gestation or throughout gestation and lactation increased food intake and the presence of characteristics of metabolic syndrome in the offspring.

Vitamins are readily available in the modern diet due to liberalized fortification and supplementation policies. This research tested the hypothesis that high multi-vitamin intake by Wistar rats during pregnancy leads to the development of obesity and characteristics of the metabolic syndrome in the offspring. Pregnant Wistar rats were fed the AIN-93G diet containing either the recommended (RV) or 10-fold increase (HV) in vitamin content. Pups were weaned to the RV diet (Study 1), an obesogenic liquid diet (Ob, Study 2), low vitamin diets (1/3RV or 1/6RV, Study 3), or a nutrient selection paradigm (NSP) with 10% and 60% casein diets (Study 4). Body weight (BW), food intake (FI), glucose and insulin responses, appetite hormones, abdominal fat pad mass (FPM) and systolic blood pressure (SBP) was measured. Expressions of mRNA for hypothalamic serotonin (5-HT) receptors and proopiomelanocortin (POMC) were measured in Study 4. Males, but not females, born to HV dams had higher post-weaning BW and FI when weaned to the RV or 1/3RV diet, and exhibited components of metabolic syndrome, including higher FPM, hyperglycemia, insulin resistance and elevated SBP compared with those born to RV dams. The Ob diet led to exaggerated weight gain and expressions of components of metabolic syndrome in both sexes born to dams fed the HV diet. Female pups on the 1/6RV diet from HV dams had two-fold higher glucose response and lower insulin response, but no difference in post-weaning BW and daily FI compared to those from RV dams. In contrast to the pups born to HV dams and fed a single diet, those from the HV dams and on the NSP gained less weight and ate less, and had lower hypothalamic mRNA expressions of 5-HT receptors and POMC. In conclusion, high multi-vitamin intake during pregnancy may lead to obesity, and result in a higher risk of developing characteristics of metabolic syndrome in the offspring. However, sex, weaning diet composition, and the presence of diet choice alter the outcomes.

The hypothesis that source of protein in a nutritionally adequate diet during gestation, lactation and weaning alters food intake and characteristics of metabolic syndrome in the offspring was investigated. Pregnant Wistar rats were randomized to either the AIN 93-G casein (C) or soy protein (S) diets (n=12/group) during gestation only or during gestation and lactation. Male offspring in each dams’ diet group were weaned to either C or S diets (n=12/group). Food intake, body weight (BW), fat pad mass, systolic (SBP) and diastolic (DBP) blood pressure, and plasma homocysteine (p<0.05) were higher in offspring born to dams fed the S diet. Fasting blood glucose (BG), BG in response to a glucose gavage and Homeostatic Model of Assessment of Insulin Resistance (HOMA-IR) index were higher only in male offspring born to dams fed the S diet. Moreover, gene expression of Agouti Related Protein (AgRP) was higher in offspring born to dams fed the S diet at weaning. Extending the dams’ diet during gestation and lactation magnified the effect of the gestational S diet on BW and composition and glucose metabolism in male offspring. Although composition of the weaning diets interacted with that of the dams’ diets, the latter was the dominant factor in determining metabolic outcomes in the offspring. In conclusion, the soy protein diet, compared to the casein diet when consumed during gestation or throughout gestation and lactation increased food intake and the presence of characteristics of metabolic syndrome in the offspring.

Environmental stress during embryonic development could lead to growth restriction of the embryo, and act as a risk factor for the development of cardiovascular disease in adult life. A common environmental stressor that causes growth restriction is prenatal hypoxia, which has been shown to adversely affect adult health in mammalian models. Prenatal hypoxia causes an increase in catecholamines which results in over stimulation of the cardiac β-adrenergic receptors. Previous work on chickens has shown that prenatal hypoxia causes an increase in the sensitivity of β-adrenergic receptors to epinephrine in the embryonic heart. The sensitivity of these receptors was found to be decreased in prenatal hypoxic juvenile. Prenatal hypoxia has no significant effect on the density of these receptors in neither the embryo nor the juvenile. The lack of change in receptor density implies that the effects of hypoxia are further down stream in the signalling cascade. The β2 adrenergic receptor can couple to both the stimulatory Gα subunit (Gsα) and the inhibitory Gα subunit (Giα). We hypothesized that prenatal hypoxia would cause an increase in the Gsα in the sensitized embryos, while increasing Giα in the desensitized juveniles. This study evaluated the relative levels of Gsα and Giα in the hypoxic chicken embryo, and in the prenatally hypoxic juvenile, Using western blotting. Hypoxia considerably increased Giα in the chicken embryo while having no effect on Gsα. In the prenatally hypoxic juvenile Gsα was significantly increased while no changes were found in Giα. This dissociation between the levels of Gα subunit and receptor sensitivity implies that that hypoxia affects the signaling cascade downstream of the Gα subunit.

The developmental origins of health and disease hypothesis suggests that exposure to adverse prenatal environmental influences can determine an individual’s susceptibility to obesity in adult life. However, the specific causal mechanisms which underlie this hypothesis have yet to be identified. Focusing on the potential mechanistic role of the leptin endocrine axis, the main objective of this thesis was to investigate the long term effects of prenatal undernutrition and different levels of postnatal nutrition on leptin sensitivity and the development of diet-induced obesity (DIO) in the Wistar rat. A well established animal model of maternal undernutrition during pregnancy was used to induce prenatal undernutrition in experimental offspring. To investigate the interaction between prenatal nutrition and postnatal diet, and its effects on obesity development, female offspring were placed on three different diets: standard chow, a high fat diet or a calorie restricted diet. The effects of prenatal undernutrition and postnatal diet on leptin sensitivity were investigated, in adult offspring, by measuring the response to 14 days of peripheral leptin treatment. Changes in gene expression in the liver, retroperitoneal adipose tissue and soleus muscle were then characterised by custom microarray and quantitative real-time RT-PCR (QPCR) analysis. Adult female offspring exposed to prenatal undernutrition (UN offspring) were found to exhibit leptin resistance in adulthood, independent of postnatal DIO. This result demonstrates for the first time that exposure to prenatal undernutrition has a long term effect on adult leptin sensitivity. In UN offspring fed on a high-fat diet, leptin resistance significantly accelerated the development of DIO while in contrast, offspring maintained on calorie restriction remained lean. These findings suggest that prenatal nutrition can shape future susceptibility to DIO by altering postnatal leptin sensitivity. An analysis of gene expression suggests that prenatal undernutrition causes the development of peripheral tissue-specific leptin resistance, and may also further enhance an offspring’s susceptibility to DIO by altering the regulation of peripheral tissue lipogenesis, mitochondrial function, glucocorticoid metabolism and insulin sensitivity. In conclusion, these studies identify peripheral leptin resistance as a key mechanism that can influence postnatal susceptibility to DIO in female offspring exposed to prenatal undernutrition. Furthermore, the identification of specific changes in peripheral gene expression highlights four additional metabolic mechanisms which may also facilitate the development of DIO in leptin resistant UN offspring.

The developmental origins of health and disease hypothesis suggests that exposure to adverse prenatal environmental influences can determine an individual’s susceptibility to obesity in adult life. However, the specific causal mechanisms which underlie this hypothesis have yet to be identified. Focusing on the potential mechanistic role of the leptin endocrine axis, the main objective of this thesis was to investigate the long term effects of prenatal undernutrition and different levels of postnatal nutrition on leptin sensitivity and the development of diet-induced obesity (DIO) in the Wistar rat. A well established animal model of maternal undernutrition during pregnancy was used to induce prenatal undernutrition in experimental offspring. To investigate the interaction between prenatal nutrition and postnatal diet, and its effects on obesity development, female offspring were placed on three different diets: standard chow, a high fat diet or a calorie restricted diet. The effects of prenatal undernutrition and postnatal diet on leptin sensitivity were investigated, in adult offspring, by measuring the response to 14 days of peripheral leptin treatment. Changes in gene expression in the liver, retroperitoneal adipose tissue and soleus muscle were then characterised by custom microarray and quantitative real-time RT-PCR (QPCR) analysis. Adult female offspring exposed to prenatal undernutrition (UN offspring) were found to exhibit leptin resistance in adulthood, independent of postnatal DIO. This result demonstrates for the first time that exposure to prenatal undernutrition has a long term effect on adult leptin sensitivity. In UN offspring fed on a high-fat diet, leptin resistance significantly accelerated the development of DIO while in contrast, offspring maintained on calorie restriction remained lean. These findings suggest that prenatal nutrition can shape future susceptibility to DIO by altering postnatal leptin sensitivity. An analysis of gene expression suggests that prenatal undernutrition causes the development of peripheral tissue-specific leptin resistance, and may also further enhance an offspring’s susceptibility to DIO by altering the regulation of peripheral tissue lipogenesis, mitochondrial function, glucocorticoid metabolism and insulin sensitivity. In conclusion, these studies identify peripheral leptin resistance as a key mechanism that can influence postnatal susceptibility to DIO in female offspring exposed to prenatal undernutrition. Furthermore, the identification of specific changes in peripheral gene expression highlights four additional metabolic mechanisms which may also facilitate the development of DIO in leptin resistant UN offspring.

The developmental origins of health and disease hypothesis suggests that exposure to adverse prenatal environmental influences can determine an individual’s susceptibility to obesity in adult life. However, the specific causal mechanisms which underlie this hypothesis have yet to be identified. Focusing on the potential mechanistic role of the leptin endocrine axis, the main objective of this thesis was to investigate the long term effects of prenatal undernutrition and different levels of postnatal nutrition on leptin sensitivity and the development of diet-induced obesity (DIO) in the Wistar rat. A well established animal model of maternal undernutrition during pregnancy was used to induce prenatal undernutrition in experimental offspring. To investigate the interaction between prenatal nutrition and postnatal diet, and its effects on obesity development, female offspring were placed on three different diets: standard chow, a high fat diet or a calorie restricted diet. The effects of prenatal undernutrition and postnatal diet on leptin sensitivity were investigated, in adult offspring, by measuring the response to 14 days of peripheral leptin treatment. Changes in gene expression in the liver, retroperitoneal adipose tissue and soleus muscle were then characterised by custom microarray and quantitative real-time RT-PCR (QPCR) analysis. Adult female offspring exposed to prenatal undernutrition (UN offspring) were found to exhibit leptin resistance in adulthood, independent of postnatal DIO. This result demonstrates for the first time that exposure to prenatal undernutrition has a long term effect on adult leptin sensitivity. In UN offspring fed on a high-fat diet, leptin resistance significantly accelerated the development of DIO while in contrast, offspring maintained on calorie restriction remained lean. These findings suggest that prenatal nutrition can shape future susceptibility to DIO by altering postnatal leptin sensitivity. An analysis of gene expression suggests that prenatal undernutrition causes the development of peripheral tissue-specific leptin resistance, and may also further enhance an offspring’s susceptibility to DIO by altering the regulation of peripheral tissue lipogenesis, mitochondrial function, glucocorticoid metabolism and insulin sensitivity. In conclusion, these studies identify peripheral leptin resistance as a key mechanism that can influence postnatal susceptibility to DIO in female offspring exposed to prenatal undernutrition. Furthermore, the identification of specific changes in peripheral gene expression highlights four additional metabolic mechanisms which may also facilitate the development of DIO in leptin resistant UN offspring.

[Truncated abstract] Saltbush is one of the few types of forage that will grow on salt affected land but, sheep struggle to maintain weight when grazing saltbush mainly because of its high salt content. Therefore, a strategy to improve salt tolerance of sheep would be beneficial to the profitable use of revegetated saline land. This could be done by manipulating the dietary salt load of pregnant or lactating ewes which could 'program', or permanently alter the physiology of their offspring to allow them to cope better with a high-salt diet as adults. When rat dams consume a high amount of salt during pregnancy, the salt balance mechanisms of their offspring are 'programmed' due to suppression of the offspring's renin-angiotensin system in early development. If this occurs in offspring from ewes grazing saltbush, beneficial adaptations may be programmed in these offspring which could allow them to better cope with the high-salt content of saltbush. I tested the general hypothesis that offspring born to ewes that consumed a high-salt or saltbush diet from mid-pregnancy to early lactation would have an increased capacity to cope with salt that would allow them gain weight when grazing saltbush in later life. To test this hypothesis, I pair-fed ewes either a high-salt diet (14% NaCl) or control diet (2% NaCl) in an animal house from day 60 of gestation until day 21 of lactation. During the same period, I also conducted a field experiment where ewes grazed on saltbush (supplemented with barley) or on pasture (supplemented with lupins). ... This led to the high-salt offspring retaining more salt than control animals. In contrast, the renin activity of saltbush was consistently lower than pasture offspring which allowed them to excrete salt more rapidly. In experiment three, the saltbush offspring gained tissue weight after grazing saltbush for 8 weeks, whereas the offspring in the other three treatments lost weight. High-salt and saltbush offspring also had higher greasy fleece weights at 22 months of age than their respective control groups. Feeding saltbush to ewes from mid-pregnancy to early lactation induces physiological adaptations in their offspring that allow them to cope better with salt and gain weight when grazing saltbush as adults, supporting my hypothesis. However, contrary to expectations, the high-salt offspring did not gain weight when grazing saltbush because their physiological adaptations, such as salt retention, did not allow them to cope better with a salt load. The reason that saltbush offspring showed different adaptations to highsalt offspring is likely to be because saltbush contains not only NaCl but also high amounts of other minerals such as potassium, and other plant compounds, which may influence the adaptive responses of the offspring. This research has direct implications for farmers because it shows they could utilize otherwise unproductive saltland by grazing pregnant ewes on saltbush to 'program' their offspring to gain weight when they graze saltbush later in life.

The developmental origins of health and disease hypothesis suggests that exposure to adverse prenatal environmental influences can determine an individual’s susceptibility to obesity in adult life. However, the specific causal mechanisms which underlie this hypothesis have yet to be identified. Focusing on the potential mechanistic role of the leptin endocrine axis, the main objective of this thesis was to investigate the long term effects of prenatal undernutrition and different levels of postnatal nutrition on leptin sensitivity and the development of diet-induced obesity (DIO) in the Wistar rat. A well established animal model of maternal undernutrition during pregnancy was used to induce prenatal undernutrition in experimental offspring. To investigate the interaction between prenatal nutrition and postnatal diet, and its effects on obesity development, female offspring were placed on three different diets: standard chow, a high fat diet or a calorie restricted diet. The effects of prenatal undernutrition and postnatal diet on leptin sensitivity were investigated, in adult offspring, by measuring the response to 14 days of peripheral leptin treatment. Changes in gene expression in the liver, retroperitoneal adipose tissue and soleus muscle were then characterised by custom microarray and quantitative real-time RT-PCR (QPCR) analysis. Adult female offspring exposed to prenatal undernutrition (UN offspring) were found to exhibit leptin resistance in adulthood, independent of postnatal DIO. This result demonstrates for the first time that exposure to prenatal undernutrition has a long term effect on adult leptin sensitivity. In UN offspring fed on a high-fat diet, leptin resistance significantly accelerated the development of DIO while in contrast, offspring maintained on calorie restriction remained lean. These findings suggest that prenatal nutrition can shape future susceptibility to DIO by altering postnatal leptin sensitivity. An analysis of gene expression suggests that prenatal undernutrition causes the development of peripheral tissue-specific leptin resistance, and may also further enhance an offspring’s susceptibility to DIO by altering the regulation of peripheral tissue lipogenesis, mitochondrial function, glucocorticoid metabolism and insulin sensitivity. In conclusion, these studies identify peripheral leptin resistance as a key mechanism that can influence postnatal susceptibility to DIO in female offspring exposed to prenatal undernutrition. Furthermore, the identification of specific changes in peripheral gene expression highlights four additional metabolic mechanisms which may also facilitate the development of DIO in leptin resistant UN offspring.