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Thesis Title Goes Here High Vitamin Intakes during Pregnancy and Characteristics of Metabolic Syndrome in Wistar Rat Dams and their Offspring by Sandra Alicia Reza López A thesis submitted in conformity with the requirements for the degree of Doctor of Philosophy Department of Nutritional Sciences University of Toronto © Copyright by Sandra Alicia Reza López, 2011 High Vitamin Intakes during Pregnancy and Characteristics of Metabolic Syndrome in Wistar Rat Dams and their Offspring Sandra Alicia Reza López Doctor of Philosophy Department of Nutritional Sciences University of Toronto 2011 Abstract High vitamin (HV), AIN-93G diet with ten-fold the regular amount of vitamins (RV), consumed by pregnant Wistar rats increases characteristics of metabolic syndrome (MetS) in their first litters. Therefore, the effects of the maternal HV-diet on tissue mechanisms regulating insulin resistance in offspring (Part 1) and on characteristics of MetS in the dams and their second litter (L2) offspring (Part 2) were examined. Part 1 (studies 1 and 2) hypothesis was that the maternal HV-diet alters tissue fatty acid (FA) concentrations, expression of peroxisome-proliferator-activated receptors (PPARs) genes, and their regulation of metabolism in the offspring, favoring insulin resistance. Part 2 (studies 3 and 4) hypothesis was that high-vitamin intakes during the first pregnancy increase weight gain, food intake and markers of MetS in both the dams and their litters. In all experiments, dams were fed the RV or HV-diet. In study 4, a high-folic-acid-diet (HFol, RV+10-fold folic acid) was added. In studies 1 and 2, the offspring were weaned to an obesogenic diet. The HV-diet affected tissue FA concentrations (study 1), increased muscle PPAR-ɑ mRNA levels and ii uncoupled relationships between hepatic PPAR-γ mRNA levels and insulin resistance (study 2) in male offspring. In study 3, dams fed the HV-diet during the first pregnancy were maintained on the RV-diet and then mated again after 12wk. Their litters were fed the RV- diet. The HV-diet increased weight gain and food intake of both dams and L2, and insulin resistance in their offspring. In study 4, both HV and HFol-diets increased post-weaning weight gain, but differed in their effects on biomarkers of food intake regulation. In conclusion, feeding the HV-diet during the first pregnancy increases post-weaning body weight and food intake in Wistar rat dams, uncouples tissue regulation of glucose metabolism and promotes characteristics of MetS in their litters. Folic acid is not the only vitamin involved. iii Acknowledgments All the way along the PhD program, I have received advice and encouragement from many people, faculty, staff, and students in the Department of Nutritional Sciences. Thank you all. I am sincerely and heartily grateful to Dr. G. Harvey Anderson, for his thoughtful guidance and support in developing this thesis and the work behind it. My sincere thanks also go to Dr. David Ma for his advice and encouragement during all these years. I would also like to thank Dr. Richard Bazinet for his valuable advice and suggestions for improving this thesis and the studies it contains. Thanks to all the members of Anderson and Ma Laboratories. My special gratitude to Dr. Bohdan Luhovyy for his help and encouragement, to Pedro Huot for sharing some of his knowledge, from analytical techniques to electronic devices, and to Ignatius Szeto for providing some data and samples from previous experiments. Special thanks to the research assistants and volunteers for their help in the laboratory work, in particular to Abraham Poon and Daniel Cho, for their assistance in gene expression analysis. My gratitude extends to all the people from other laboratories and the departamental staff that enriched my experience along these years in the Department. Thanks to Dr. Thompson, Dr. Ward, Dr. Archer, Dr. El-Sohemy and Dr. Bazinet’s groups for their feedback and comments during lab meetings that helped me to improve seminar and other presentations. Also thanks to Dr. Elena Comelli for her advice on RT-PCR. My appreciation also extends to Louisa Kung and Vijay Chetty, and members of the staff, for their guidance and help in administrative matters and their kindness all along the program. My sincere appreciation also to Nita Prayitno, Tina Akhavan, Rebecca Mollard, Maria Fernanda Nunez and Diana Sanchez, for their kindness, friendship and for sharing too many moments along these years. I truly thank my husband Santiago for his unconditional help and support. iv My gratitude extends to the Consejo Nacional de Ciencia y Tecnologia (CONACyT) in Mexico, for the scholarship provided during the first 4 years of the PhD program. Also special thanks to the Delta-Kappa-Gamma Society International for the World Fellowship, along with my profound respect and admiration for their contribution in improving opportunities for women in education, and to the University of Toronto for the Doctoral Thesis Completion Grant awarded during my last year of the program. This research was supported by the Canadian Institute of Health Research, Institute of Nutrition, Metabolism and Diabetes Strategic Initiative (CIHR-INMD): “Excellence, Innovation and Advancement in the Study of Obesity and Healthy Body Weight” – Grant Number OOP-77980, and Institute of Nutrition, reference MOP-93624; and the Bristol- Myers Squibb / Mead Johnson Freedom to Discover Grant. v Table of Contents Abstract ............................................................................................................................................ ii Acknowledgments .......................................................................................................................... iv List of Tables ................................................................................................................................. xii List of Figures ................................................................................................................................ xv List of Abbreviations ................................................................................................................... xvii List of Appendices ........................................................................................................................ xxi List of Publications Arising from this Thesis .............................................................................. xxii Chapter 1. Introduction .................................................................................................................... 1 Chapter 2. Literature Review........................................................................................................... 4 2.1. Introduction .......................................................................................................................... 4 2.2. Maternal Nutrition and Offspring Phenotype ...................................................................... 5 2.2.1. The developmental origins of health and disease ...................................................... 5 2.2.1.1. Maternal nutrition and overweight and adiposity in offspring ................... 7 2.2.1.2 Maternal nutrition and lipid metabolism in offspring ............................... 10 2.2.1.3 Maternal nutrition and glucose metabolism in offspring........................... 12 2.2.2. Maternal characteristics and developmental origins of disease............................... 15 2.2.3. Diet during pregnancy and effects on the mother .................................................... 17 vi 2.2.3.1. Maternal physiology during pregnancy .................................................... 17 2.2.3.2. Diet components and maternal metabolism.............................................. 19 2.2.4. Animal models ......................................................................................................... 20 2.2.5. Potential mechanisms of developmental origins of disease .................................... 20 2.2.5.1. Alterations in organ/tissue structure ......................................................... 21 2.2.5.2. Hormone concentrations and early development ..................................... 22 2.2.5.3. Epigenetic regulation of gene expression ................................................. 25 2.3. Maternal Vitamin Intake and Offspring Phenotype ........................................................... 30 2.3.1. General vitamin functions ....................................................................................... 30 2.3.2. Consequences of vitamin deficiencies during pregnancy ........................................ 31 2.3.2.1. Studies in humans ..................................................................................... 32 2.3.2.2. Studies in animals ..................................................................................... 34 2.3.3. Vitamin supplementation during pregnancy............................................................ 36 2.3.3.1. Birth weight .............................................................................................. 36 2.3.3.2. Postnatal growth, lipid and glucose metabolism in offspring .................. 39 2.3.4. Consequences of excessive vitamin intakes ............................................................ 40 2.3.4.1. Studies in humans ..................................................................................... 41 2.3.4.2. Studies in animals ..................................................................................... 42 2.3.5.
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