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Matt Dean's presentation 111
- 1. Matthew Dean
- 2. Early Embryonic Loss (55%) (Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988) Fertilization Failure (5%) Live Birth (33%) Late Embryonic/ Fetal Loss (7%)
- 3. Early Embryonic Loss (55%) Fertilization Failure (5%) Live Birth (33%) Late Embryonic/ Fetal Loss (7%) (Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)
- 4. Early Embryonic Loss (55%) Fertilization Failure (5%) Live Birth (33%) Late Embryonic/ Fetal Loss (7%) (Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)
- 5. Early Embryonic Loss (55%) Fertilization Failure (5%) Live Birth (33%) Late Embryonic/ Fetal Loss (7%) (Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)
- 6. Early Embryonic Loss (55%) Fertilization Failure (5%) Live Birth (33%) Late Embryonic/ Fetal Loss (7%) (Zinaman et al. 1996; Diskin et al. 2006; Wilcox et al. 1988)
- 7. Zygote Uterus Cervix Vagina Oviduct (Hansson 1947; Senger 2005) Ovary
- 12. EndometriumUterine Lumen (Bazer et al. 2012; Yánez et al. 2003) Glucose Glycogen Pyruvate X
- 13. Glycogen
- 14. Glycogen glucose
- 19. -6P -UDP -1P Hexokinase Glycogen Synthase Glycogen Phosphorylase Glycogen glucose Cellular Metabolism
- 20. -6P -UDP -1P Hexokinase Glycogen Synthase Glycogen Phosphorylase -6P Glycogen glucose Cellular Metabolism
- 21. -6P -UDP -1P Hexokinase Glycogen Synthase Glycogen Phosphorylase -6P Glucose-6- Phosphatase Glycogen glucose Cellular Metabolism
- 22. -6P -UDP -1P Hexokinase Glycogen Synthase Glycogen Phosphorylase -6P Glucose-6- Phosphatase Glycogen glucose Cellular Metabolism
- 23. Estrus/ Mated GlycogenConcentration (mg/100gwettissue) (modified from Demers et al. 1972: Gupta et al. 2013) Day Post Mating Implantation Estrus
- 24. 0 1 2 3 4 Estrus 1 2 3 4 5 1 2 3 4 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 … 50 Implantation Estrus Implantation Day Day
- 25. 1) Characterize changes in uterine glycogen metabolism during estrus, embryonic diapause, and pregnancy in mink. 2) Determine if the glycogenic effect of estradiol on uterine epithelia is direct.
- 26. Feb AprilMarch May 15 20 25 5 10 15 20 25 30 5 10 15 20 25 30 5 10 15 Estrus Implantation Parturition Estrus Diapause Pregnancy 3 Collections Objective 1. Characterize changes in uterine glycogen metabolism during estrus, embryonic diapause, and pregnancy in mink.
- 27. 0 5 10 15 20 25 30 (Nested ANOVA A-CP<0.05) Estrus IIS IS A B C GlycogenConcentration (mg/mgtissue) Diapause Pregnancy
- 28. Myometrium Endometrium Periodic Acid-Schiff- stain for carbohydrates
- 29. 0 10 20 30 40 A B C (ANOVA and Tukey’s, P<0.05) PercentArea Estrus Diapause Pregnancy Estrus Diapause Pregnancy
- 31. 0 10 20 30 40 50 Glandular Epithelia Lumenal Epithelia Stroma A B B [ [ * (Nested ANOVA A-B,* P<0.05) GlycogenContent (PercentArea) Estrus Diapause Pregnancy Estrus Diapause Pregnancy *
- 32. Estrus Diapause Pregnancy Enzyme= Blue Counterstain=red/pink -6P
- 33. Estrus Diapause Pregnancy Glycogen Synthase Glycogen Phosphorylase Enzyme= Blue Counterstain=red/pink
- 35. Estradiol??? Cont Estradiol PercentArea 0.0 0.1 0.2 Glandular Epithelial Glycogen * Estrus
- 36. Estradiol Estrus
- 38. 1) Characterize changes in uterine glycogen metabolism during estrus, embryonic diapause, and pregnancy in mink. 2) Determine if the glycogenic effect of estradiol on uterine epithelia is direct.
- 39. Mink uterine epithelia Immortalized with SV40 large T antigen Positive for cytokeratin 18 (Moreau et al. 1995; http://micro.magnet.fsu.edu)
- 41. 0 0.1 1 10 100 0.0 0.1 0.2 0.3 Glycogen(g/106cells) Estradiol (nM)
- 42. Insulin (g ml-1) 0 1.8 5.5 16.6 50 Glycogen(g/106cells) 0.0 0.2 0.4 0.6 * * (*P<0.05 relative to 0 μg ml-1)
- 43. PPPT- Igf1r antagonist (*P<0.05 relative to 0 μg ml-1) Glycogen(g/106 cells) 0.0 0.1 0.2 0.3 Cont PPPT Insulin Insulin +PPPT *
- 44. Insulin Estradiol 0 0 50 0 50 0.1 50 1.0 50 10 50 100 Glycogen(g/106 cells) 0.0 0.2 0.4 * * (*P<0.05 relative to 0 μg ml-1 insulin & 0 nM estradiol)
- 45. ICI= Estrogen Receptor antagonist (*P<0.05 relative to cont Glycogen C Glycogen(g/106 cells) 0.0 0.2 0.4 0.6 * EstradiolICICont Estradiol + ICI
- 46. 1)Endometrial glycogen contributes to histotroph. localized to epithelial at estrus mobilized before implantation high glucose-6-phosphatase during diapause 2) Estradiol stimulates glycogenesis through both paracrine and direct mechanisms. activation of IGF-1 receptor increases glycogen estradiol increases glycogen (in presence of insulin)
- 51. Committee Dr. Rose Dr. Bearden Dr. Cretekos Dr. Pfau Lab Mates Jason Hunt Kole Bowman Lisa McDougall Josh Astle Ayokunle Hodonu Evan Stegelmeier Other Grad Students Other Professors Dr. Rodnick Dr. Aho Dr. Thomas Funding Sources Moyle Mink and Tannery Fur Commission USA Idaho INBRE ISU Research Council MCRF Seed Grant Sigma Xi
- 52. Thank you
Notas do Editor
- CDC estimates: 6.7 M women -> reduced fertility 1.5 M women -> clinically infertile human fertility- maximal at 30% per menstrual cycle
- 40% humans pregnancies lost after hCG is detectable
- Pre-Implantation Period mice and rats: 4-5 days humans: 10 days mink: variable, up to 50 days
- Embryo require glucose4 Glucose uptake increases exponentially Too much glucose is toxic
- uterus cannot make glucose
- Glycogen Stores up to 55,000 glucose residues 25% of infertile women had diminished endometrial glycogen reserves
- Mink Mating Feb-March Implantation April Birth- May
- Swigart et al. (1962)- neither insulin or glucose increased uterine glycogen in rats Hay et al. (1984)- glucose clamp did not increase glucose uptake by uterus in sheep Casalino-Matsuda et al. (2000)- not consistent results OVX and diet restricted = insulin ↑ glycolysis Intact and diet restricted = insulin ↓ glycolysis
- Swigart et al. (1962)- neither insulin or glucose increased uterine glycogen in rats Hay et al. (1984)- glucose clamp did not increase glucose uptake by uterus in sheep Casalino-Matsuda et al. (2000)- not consistent results OVX and diet restricted = insulin ↑ glycolysis Intact and diet restricted = insulin ↓ glycolysis
- Swigart et al. (1962)- neither insulin or glucose increased uterine glycogen in rats Hay et al. (1984)- glucose clamp did not increase glucose uptake by uterus in sheep Casalino-Matsuda et al. (2000)- not consistent results OVX and diet restricted = insulin ↑ glycolysis Intact and diet restricted = insulin ↓ glycolysis