Effects of maternal stress and obesity on human feto-placental glucocorticoid exposure

1! Abstracts! from! this! thesis! 4! s! from! this! thesis! 4! Abbreviations! 5! List! of! Figures! 7! List! of! Tables! 10! Chapter! 1:! Introduction! 12! 1.1! Fetal! Programming! 13! 1.1.1! Programming! and! Glucocorticoids! 16! 1.2! Pregnancy! 18! 1.3! The! HypothalamicHPituitaryHAdrenal! Axis! 18! 1.3.1! Overview! 18! 1.3.2! Cortisol! 22! 1.3.3! Intracellular! Action! of! Glucocorticoids! 23! 1.3. 4! The! HPACAxis! During! Pregnancy! 27! 1.3. 5! The! Placental! Glucocorticoid! Barrier:! 11βHSD2! 31! 1.4! Stress! and! Obesity! as! Programming! Factors! 34! 1.5! Obesity! 36! 1.6! Obesity! during! Pregnancy! 39! 1.7! Obesity! and! the! HPA! Axis! 40! 1.7. 1! Maternal! Obesity! and! Programming! 43! 1.7. 2! Programming! versus! Postnatal! Obesogenic! Lifestyle! 44! 1.7. 3! Stress! and! the! HPA! Axis! 45! 1.7. 4! Acute! and! Chronic! Stress! 46! 1.7. 5! Maternal! Stress! and! Programming! 47! Effects of maternal stress and obesity on human feto-placental glucocorticoid exposure v 1.8! Programming! and! the! Placenta! 50! 1.8. 1! Placental! Structure! and! Development! 50! 1.8. 2! Placental! function! 52! 1.8. 3! The! Programming! Effects! of! Placental! 11βHSD2! 53! 1.9! Epigenetics! 56! 1.9. 1! IGF2! 58! 1.9. 2! Methylation! and! the! regulation! of! glucocorticoid! exposure:! GR! and! 11βHSD2! 61! Aims! and! Hypothesis! 63! Chapter! 2:! Materials! and! Methods! 64! 2.1! Clinical! Methods! 65! 2.1.1! Longitudinal! study! of! very! severely! obese! pregnancy! 65! 2.1.2! First! Trimester! Placenta! Study! 66! 2.1.3! Second! Trimester! Placental! and! Liver! Study! 67! 2.1.4! Term! placental! samples! 68! 2.1.5! PREDO! Stress! In! Pregnancy! Study! 68! 2.2! Materials! 71! 2.2.1! General! Chemicals! 71! 2.2.2! Equipment! 72! 2.2.3! Software! 73! 2.2.4! Solutions! and! Buffers! 73! 2.3! Laboratory! Materials! and! Methods! 75! 2.3.1! Quantitative! Real! Time! PCR! 75! 2.3.2! DNA! Methylation! Analysis! 82! 2.3.3! Protein! Analysis! 92! 2.3. 4! Immunohistochemistry! 94! 2.3. 5! Radioimmunoassays! 95! Effects of maternal stress and obesity on human feto-placental glucocorticoid exposure vi 2.3. 6! CRH! 100! 2.4! Statistical! Analysis! 101! Chapter! 3:! The! Maternal! HPA! Axis! in! Obese! Pregnancy! and! Associations! with! Gestational! Weight! Gain,! Birth! Weight! and! Gestation! at! Delivery! 102! 3.1! Introduction! 103! 3.1.1! Hypotheses! 104! 3.1.2! Aims! 104! 3.2! Methods! 105! 3.2.1! Study! Design! 105! 3.2.2! Subject! Recruitment! 105! 3.2.3! Laboratory! methods! 106! 3.2.4! Statistics! 106! 3.3! Results! 108! 3.3.1! Demographics! 108! 3.3.2! Total! Serum! Cortisol! Profiles! Differ! Between! Lean! and! Obese! Pregnant! Women! 108! 3.3.3! Salivary! diurnal! patterns! 109! 3.3. 4! Serum! CBG! Is! Lower! in! Obese! Women! than! Lean! 113! 3.3. 5! Calculated! Free! Cortisol! Differed! Only! at! 36! Weeks! 113! 3.3. 6! CRH! 117! 3.3. 7! Gestational! Weight! Gain! 117! 3.3. 8! Gestational! Weight! Gain! and! Birth! Weight! 119! 3.3. 9! Associations! Between! Maternal! Cortisol! Concentrations! and! Gestational! Weight! Gain! 119! 3.3.10! Birth! weight! and! Total! Maternal! Cortisol! 1