PHYSIOLOGICAL EFFECTS OF ISOCALORIC AND MINERAL DEFICIENT MATERNAL MOUSE DIETS ON OFFSPRING GROWTH AND BODY COMPOSITION A THESIS SUBMITTED TO THE FACULTY OF THE UNIVERSITY OF MINNESOTA BY Amanda Palowski IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE CO-ADVISORS Christopher Faulk, Gerald C. Shurson August 2018 Ó Amanda Palowski 2018 Acknowledgements First, I would like to acknowledge and express my thankfulness to my academic advisors, Drs. Gerald Shurson and Christopher Faulk. Both of my advisors have challenged me to think outside of the box, pushed me to keep improving, and provided me with the knowledge and tools to be successful in the research field. I would not be the student and scientist that I am today without their guidance and support. Secondly I would like to thank all of the Integrated Animal Systems Biology professors. It has been a great pleasure working with all of you and I thank you for your guidance and help throughout my graduate career. I would also like to thank Dr. Dan Gallaher for being a part of my committee and providing an outside set of eyes on my project and thesis, I appreciate all of your comments and guidance. I would also like to thank all of the graduate and undergraduate students who helped me with my project; Mathia (Tia) Colwell, Mickie Trudeau, Yuan-tai Hung, Anna Clarke, Melissa Drown, and Chelsea Drown. I also want to thank all of the other graduate students in our department, Marta Ferrandis, Joey Mielke, Leonard Fung, Zhikai Zeng, Jinlong Zhu, Edward Yang, Audrey Walter, and Julia Holen, thank you for showing me friendship and helping me with classes. Lastly I would like to thank my family for the constant support along this journey to further my education, especially my husband Adam. It is never easy buying a house, getting married and starting a family in the middle of grad school, but he showed me constant love and support and I appreciate you pushing me to be a better person. i Dedication This thesis is dedicated to my husband Adam Palowski. Thank you so much for being my best friend and all of the love and support you have shown me through this journey. Thank you for always pushing me and being there to help me pick myself up and dust myself off when I stumble and fall. Thank you for being the best support system ever, I love you so much. ii Abstract Over the last 30 years, researchers have evaluated the various effects of feeding maternal high fat diets on the health of the offspring of various species, but mainly in rodents. Our hypothesis was that dietary source and amount of energy in maternal mouse diets would induce physiological, growth, and body composition changes in offspring independently of maternal body weight. The objective of this study was to evaluate the changes in body weight and composition from feeding maternal isocaloric diets on mouse offspring (1 generation). Parent mice were weaned and fed one of 3 diets consisting of either low fat with controlled calories based on prior ad libitum mouse experiment measurements (LFr), high fat diets restricted in calories to match the low fat diet (HFr), or high fat ad libitum (HFa) diets. Offspring from the first generation were fed either low fat ad libitum (LFa) or high fat ad libitum (HFa) diets and randomly assigned to four different dietary treatment groups: 1) dams fed LFr and offspring fed LFa (LFr:LFa) diets, 2) dams fed LFr and offspring fed HFa (LFr:HFa) diets, 3) dams fed HFr and offspring fed HFa (HFr:HFa) diets, and 4) dams fed HFa and offspring fed HFa (HFa:HFa) diets. Average weekly body weight and composition of offspring was determined for each of the four dietary treatments. Unfortunately, the experimental diets obtained from the manufacturer were severely deficient in all of the essential macro and micro minerals, which resulted in high pup mortality and prevented testing of the original hypothesis. Overall, both dams and offspring fed the LF diet had the greatest body weight gain and average percentage of body fat mass compared with those fed the other dietary treatments. In conclusion, feeding the mineral deficient diets confounded the effects of dietary calorie source and consumption on developmental programming in mice. iii Table of Contents Acknowledgements i Dedication ii Abstract iii List of Figures v List of Tables viii Chapter 1: Literature Review 1 Introduction 1 Developmental Programming 1 Maternal Nutritional Effects of Developmental Programming 4 Maternal Malnutrition and Effects on Offspring 5 Molecular Mechanisms of Maternal Nutritional Effects 7 Effects of Maternal High Fat Diet on Offspring Metabolic Health 9 Maternal High Fat Diet Effects on Milk Production 14 Maternal High Fat Diet Effects on Offspring Hepatic Function and Metabolic Syndrome 15 Mental Health Effects of Maternal Diets 16 Comparison of Dietary Energy Systems 17 Effect of Mineral Deficiencies on Reproduction and Growth of Mice 22 Conclusions 27 Chapter 2: Physiological Effects of Isocaloric and Mineral Deficient Maternal Diets On Offspring Growth and Body Composition 29 Synopsis 29 Introduction 30 Materials and Methods 32 Results and Discussion 45 Conclusion 60 Overall Summary 61 References 63 iv List of Figures Figure 1. Partitioning of dietary energy losses and utilization (National Research Council, 2012)...…………………………………………………………….19 Figure 2. Experimental design of dietary treatments of parent (F0) mice……….33 Figure 3. Daily amount of food provided to female mice fed low fat restricted (LFr) and high fat restricted (HFr) diets compared with the amount of food (g) consumed by the female mice fed the high fat ad libitum (HFa) dietary treatment…...…………………………………………………….………….40 Figure 4. Daily amount of calories offered to female mice fed low fat restricted (LFr) and high fat restricted (HFr) diets compared with the amount of calories consumed by the female mice fed the high fat ad libitum (HFa) dietary treatment.………………..………………….………………………40 Figure 5. Comparison of the weekly calorie consumption by female mice fed the 2018 diet (Atwater calories), the high fat ad libitum diet (HFa; Atwater calories) and HFa (NE calories system calculations)…………………...41 Figure 6. Timeline of significant events that occurred throughout the study…….42 Figure 7. Experimental design of dietary treatments of F1.2 mice offspring (first diet abbreviation represents the dam diet and the second diet abbreviation represents the offspring diet)…………………………………………………43 Figure 8. Average weekly body weight (g) of the dams fed low fat restricted (LFr; n = 8), high fat restricted (HFr; n = 6), of high fat ad libitum (HFa; n = 8) diets………………….….....…………………………………………………...46 Figure 9. Average weekly body weight (g) of the male offspring among the four maternal and offspring dietary treatment combinations with LFr:HFa representing the low fat restricted dam diet and high fat ad libitum offspring diet (n = 9), LFr:LFa representing the low fat restricted dam diet and low fat ad libitum offspring diet (n = 13), HFr:HFa representing the high fat restricted dam diet, high fat ad libitum offspring diet (n = 19), and HFa:HFa representing the high fat ad libitum dam diet and high fat ad libitum offspring diet (n = 12)…………………………………………………47 v Figure 10. Average weekly body weight (g) of the female offspring among the four maternal and offspring dietary treatment combinations with LFr:HFa representing the low fat restricted dam diet and high fat ad libitum offspring diet (n = 7), LFr:LFa representing the low fat restricted dam diet and low fat ad libitum offspring diet (n = 10), HFr:HFa representing the high fat restricted dam diet, high fat ad libitum offspring diet (n = 9), and HFa:HFa representing the high fat ad libitum dam diet and high fat ad libitum offspring diet (n = 16)…………………………………………………48 Figure 11. Average percentage of body fat (A) and fat-free mass (B) of dams at 24 weeks of age among the three maternal dietary treatments with LFr representing low fat restricted diet (n = 8), HFr representing high fat restricted diet (n = 6), HFa representing high fat ad libitum diet (n = 8). Different letters indicate statistically significant differences……………….50 Figure 12. Average percentage of body fat (A) and fat free mass (B) of offspring at 12 weeks of age among the four maternal and offspring dietary treatment combinations with LFr:HFa representing the low fat restricted dam diet and high fat ad libitum offspring diet (n = 16), LFr:LFa representing the low fat restricted dam diet and low fat ad libitum offspring diet (n = 23), HFr:HFa representing the high fat restricted dam diet, high fat ad libitum offspring diet (n = 28), and HFa:HFa representing the high fat ad libitum dam diet and high fat ad libitum offspring diet (n = 28)…….51 Figure 13. Average percentage of fat composition between male and female offspring among the four maternal and offspring dietary treatment groups. LFr:HFa representing the low fat restricted dam diet and high fat ad libitum offspring diet (n = 9 males and 7 females), LFr:LFa representing the low fat restricted dam diet and low fat ad libitum offspring diet (n = 13 males and 10 females), HFr:HFa representing the high fat restricted dam diet, high fat ad libitum offspring diet (n = 19 males and 9 females), HFa:HFa representing the high fat ad libitum dam diet and high fat ad libitum offspring diet (n = 12 males and 16 females)….……………………………52 vi Figure 14. Average percentage of fat free mass composition between male and female offspring among the four maternal and offspring dietary treatment groups. LFr:HFa representing the low fat restricted dam diet and high fat ad libitum offspring diet (n = 9 males and 7 females), LFr:LFa representing the low fat restricted dam diet and low fat ad libitum offspring diet (n = 13 males and 10 females), HFr:HFa representing the high fat restricted dam diet, high fat ad libitum offspring diet (n = 19 males and 9 females), HFa:HFa representing the high fat ad libitum dam diet and high fat ad libitum offspring diet (n = 12 males and 16 females)………………………53 Figure 15.