Temporal and spatial expression of adrenomedullin and its receptors in the porcine uterus and peri-implantation conceptuses

Abstract Adrenomedullin (ADM) is an evolutionarily conserved multifunctional peptide hormone that regulates implantation, embryo spacing, and placentation in humans and rodents. However, the potential roles of ADM in implantation and placentation in pigs, as a litter-bearing species, are not known. This study determined abundances of ADM in uterine luminal fluid, and the patterns of expression of ADM and its receptor components (CALCRL, RAMP2, RAMP3, and ACKR3) in uteri from cyclic and pregnant gilts, as well as conceptuses (embryonic/fetus and its extra-embryonic membranes) during the peri-implantation period of pregnancy. Total recoverable ADM was greater in the uterine fluid of pregnant compared with cyclic gilts between Days 10 and 16 post-estrus and was from uterine luminal epithelial (LE) and conceptus trophectoderm (Tr) cells. Uterine expression of CALCRL, RAMP2, and ACKR3 were affected by day (P < 0.05), pregnant status (P < 0.01) and/or day x status (P < 0.05). Within porcine conceptuses, the expression of CALCRL, RAMP2, and ACKR3 increased between Days 10 and 16 of pregnancy. Using an established porcine trophectoderm (pTr1) cell line, it was determined that 10–7 M ADM stimulated proliferation of pTr1 cells (P < 0.05) at 48 h, and increased phosphorylated mechanistic target of rapamycin (p-MTOR) and 4E binding protein 1 (p-4EBP1) by 6.1- and 4.9-fold (P < 0.0001), respectively. These novel results indicate a significant role for ADM in uterine receptivity for implantation and conceptus growth and development in pigs. They also provide a framework for future studies of ADM signaling to affect proliferation and migration of Tr cells, spacing of blastocysts, implantation, and placentation in pigs. Summary sentence Our study suggests that adrenomedullin plays a significant role in uterine receptivity and conceptus growth and development, via the stimulation of cell proliferation and activation of MTORC1 signaling pathway in the porcine conceptus trophectoderm. Graphical Abstract

[1]  G. Tomes,et al.  Factors Affecting Reproductive Efficiency of the Breeding Herd , 2020, Bioscientifica Proceedings.

[2]  G. Ladds,et al.  RAMP3 determines rapid recycling of atypical chemokine receptor-3 for guided angiogenesis , 2019, Proceedings of the National Academy of Sciences.

[3]  S. Young,et al.  SOX17 regulates uterine epithelial–stromal cross-talk acting via a distal enhancer upstream of Ihh , 2018, Nature Communications.

[4]  Guoyao Wu,et al.  Innate differences and colostrum-induced alterations of jejunal mucosal proteins in piglets with intra-uterine growth restriction. , 2018, The British journal of nutrition.

[5]  S. Young,et al.  Adrenomedullin improves fertility and promotes pinopodes and cell junctions in the peri-implantation endometrium† , 2017, Biology of Reproduction.

[6]  Guoyao Wu,et al.  Physiological alterations associated with intrauterine growth restriction in fetal pigs: Causes and insights for nutritional optimization , 2017, Molecular reproduction and development.

[7]  Guoyao Wu,et al.  Uterine Histotroph and Conceptus Development. II. Arginine and Secreted Phosphoprotein 1 Cooperatively Stimulate Migration and Adhesion of Ovine Trophectoderm Cells via Focal Adhesion-MTORC2 Mediated Cytoskeleton Reorganization1 , 2016, Biology of reproduction.

[8]  Guoyao Wu,et al.  mTOR: The Master Regulator of Conceptus Development in Response to Uterine Histotroph During Pregnancy in Ungulates , 2016 .

[9]  Guoyao Wu,et al.  Select nutrients and their effects on conceptus development in mammals , 2015, Animal nutrition.

[10]  Guoyao Wu,et al.  Functional Roles of Arginine During the Peri-Implantation Period of Pregnancy. III. Arginine Stimulates Proliferation and Interferon Tau Production by Ovine Trophectoderm Cells via Nitric Oxide and Polyamine-TSC2-MTOR Signaling Pathways1 , 2015, Biology of reproduction.

[11]  Guoyao Wu,et al.  Uterine Histotroph and Conceptus Development. I. Cooperative Effects of Arginine and Secreted Phosphoprotein 1 on Proliferation of Ovine Trophectoderm Cells via Activation of the PDK1-Akt/PKB-TSC2-MTORC1 Signaling Cascade1 , 2015, Biology of reproduction.

[12]  J. Miles,et al.  Placental accommodations for transport and metabolism during intra-uterine crowding in pigs , 2014, Journal of Animal Science and Biotechnology.

[13]  J. Ross,et al.  Cytokines from the pig conceptus: roles in conceptus development in pigs , 2014, Journal of Animal Science and Biotechnology.

[14]  Guoyao Wu,et al.  Putrescine Stimulates the mTOR Signaling Pathway and Protein Synthesis in Porcine Trophectoderm Cells1 , 2014, Biology of reproduction.

[15]  Samantha L. Hoopes,et al.  Decoy receptor CXCR7 modulates adrenomedullin-mediated cardiac and lymphatic vascular development. , 2014, Developmental cell.

[16]  U. Yallampalli,et al.  Adrenomedullin Promotes Rat Trophoblast Stem Cell Differentiation1 , 2014, Biology of reproduction.

[17]  Guoyao Wu,et al.  Functional Role of Arginine During the Peri-implantation Period of Pregnancy. II. Consequences of Loss of Function of Nitric Oxide Synthase NOS3 mRNA in Ovine Conceptus Trophectoderm1 , 2014, Biology of reproduction.

[18]  Guoyao Wu,et al.  Temporal proteomic analysis reveals defects in small-intestinal development of porcine fetuses with intrauterine growth restriction. , 2014, The Journal of nutritional biochemistry.

[19]  Guoyao Wu,et al.  Functional role of arginine during the peri‐implantation period of pregnancy. I. Consequences of loss of function of arginine transporter SLC7A1 mRNA in ovine conceptus trophectoderm , 2014, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[20]  Guoyao Wu,et al.  Arginine Decarboxylase and Agmatinase: An Alternative Pathway for De Novo Biosynthesis of Polyamines for Development of Mammalian Conceptuses1 , 2014, Biology of reproduction.

[21]  F. Bazer,et al.  Pig blastocyst-uterine interactions. , 2014, Differentiation; research in biological diversity.

[22]  Guoyao Wu,et al.  Impacts of arginine nutrition on embryonic and fetal development in mammals , 2013, Amino Acids.

[23]  Guoyao Wu,et al.  Intrauterine growth restriction alters the hepatic proteome in fetal pigs. , 2013, The Journal of nutritional biochemistry.

[24]  Defa Li,et al.  IUGR alters muscle fiber development and proteome in fetal pigs. , 2013, Frontiers in bioscience.

[25]  I. Brown,et al.  18S rRNA is a reliable normalisation gene for real time PCR based on influenza virus infected cells , 2012, Virology Journal.

[26]  K. Caron,et al.  Adrenomedullin and pregnancy: perspectives from animal models to humans , 2012, Trends in Endocrinology & Metabolism.

[27]  Guoyao Wu,et al.  Select nutrients in the uterine lumen of sheep and pigs affect conceptus development. , 2012, The Journal of reproduction and development.

[28]  Samantha L. Hoopes,et al.  Adrenomedullin Function in Vascular Endothelial Cells: Insights from Genetic Mouse Models , 2011, Current hypertension reviews.

[29]  Y. Takei,et al.  Shared and separate functions of the RAMP-based adrenomedullin receptors , 2011, Peptides.

[30]  J. Miles,et al.  Effect of empty uterine space on birth intervals and fetal and placental development in pigs. , 2011, Animal reproduction science.

[31]  Rong Li,et al.  Mitotic progression becomes irreversible in prometaphase and collapses when Wee1 and Cdc25 are inhibited , 2011, Molecular biology of the cell.

[32]  D. Hay,et al.  Adrenomedullin and calcitonin gene-related peptide receptors in endocrine-related cancers: opportunities and challenges. , 2010, Endocrine-related cancer.

[33]  Guoyao Wu,et al.  Secreted phosphoprotein 1 binds integrins to initiate multiple cell signaling pathways, including FRAP1/mTOR, to support attachment and force-generated migration of trophectoderm cells. , 2010, Matrix biology : journal of the International Society for Matrix Biology.

[34]  O. Gavet,et al.  Progressive activation of CyclinB1-Cdk1 coordinates entry to mitosis. , 2010, Developmental cell.

[35]  Guoyao Wu,et al.  Temporal proteomic analysis reveals continuous impairment of intestinal development in neonatal piglets with intrauterine growth restriction. , 2010, Journal of proteome research.

[36]  Guoyao Wu,et al.  Proteomic analysis reveals altered expression of proteins related to glutathione metabolism and apoptosis in the small intestine of zinc oxide-supplemented piglets , 2009, Amino Acids.

[37]  Guoyao Wu,et al.  Select Nutrients in the Ovine Uterine Lumen. I. Amino Acids, Glucose, and Ions in Uterine Lumenal Flushings of Cyclic and Pregnant Ewes1 , 2009, Biology of reproduction.

[38]  Guoyao Wu,et al.  Select Nutrients in the Ovine Uterine Lumen. II. Glucose Transporters in the Uterus and Peri-Implantation Conceptuses1 , 2009, Biology of reproduction.

[39]  K. Caron,et al.  Haploinsufficiency for Adrenomedullin Reduces Pinopodes and Diminishes Uterine Receptivity in Mice1 , 2008, Biology of reproduction.

[40]  G. Wu,et al.  Proline metabolism in the conceptus: implications for fetal growth and development , 2008, Amino Acids.

[41]  Guoyao Wu,et al.  Intrauterine growth restriction affects the proteomes of the small intestine, liver, and skeletal muscle in newborn pigs. , 2008, The Journal of nutrition.

[42]  K. Caron,et al.  Receptor activity-modifying proteins: RAMPing up adrenomedullin signaling. , 2007, Molecular endocrinology.

[43]  D. Erbaş,et al.  Maternal and umbilical venous adrenomedullin and nitric oxide levels in intrauterine growth restriction , 2007, The journal of maternal-fetal & neonatal medicine : the official journal of the European Association of Perinatal Medicine, the Federation of Asia and Oceania Perinatal Societies, the International Society of Perinatal Obstetricians.

[44]  D. Hay,et al.  The pharmacology of adrenomedullin receptors and their relationship to CGRP receptors , 2004, Journal of Molecular Neuroscience.

[45]  T. Magnuson,et al.  Reduced maternal expression of adrenomedullin disrupts fertility, placentation, and fetal growth in mice. , 2006, The Journal of clinical investigation.

[46]  Guoyao Wu,et al.  Board-invited review: intrauterine growth retardation: implications for the animal sciences. , 2006, Journal of animal science.

[47]  W. Lamberson,et al.  Genetic and phenotypic relationships of farrowing and weaning survival to birth and placental weights in pigs. , 2006, Journal of animal science.

[48]  F. Bazer,et al.  Functional effects of transforming growth factor beta on adhesive properties of porcine trophectoderm. , 2005, Endocrinology.

[49]  G. Foxcroft,et al.  Embryonic and fetal development in a commercial dam-line genotype. , 2005, Animal reproduction science.

[50]  J. Wallace,et al.  Effect of nutrient intake during pregnancy on fetal and placental growth and vascular development. , 2004, Domestic animal endocrinology.

[51]  T. Spencer,et al.  Progesterone and Placental Hormone Actions on the Uterus: Insights from Domestic Animals1 , 2004, Biology of reproduction.

[52]  N. Minamino,et al.  The role of adrenomedullin and receptors in glomerular hyperfiltration in streptozotocin-induced diabetic rats. , 2004, Kidney international.

[53]  T. Spencer,et al.  Uterine and placental factors regulating conceptus growth in domestic animals. , 2004, Journal of animal science.

[54]  V. Han,et al.  Adrenomedullin messenger ribonucleic acid expression in the placentae of normal and preeclamptic pregnancies. , 2003, The Journal of clinical endocrinology and metabolism.

[55]  K. Kanenishi,et al.  Adrenomedullin Concentrations in Early 2nd-Trimester Amniotic Fluid: Relation to Preterm Delivery and Fetal Growth at Birth , 2003, Gynecologic and Obstetric Investigation.

[56]  W. Rascher,et al.  Adrenomedullin gene expression in human placental tIssue and leukocytes: a potential marker of severe tIssue hypoxia in neonates with birth asphyxia. , 2002, European journal of endocrinology.

[57]  R. Geisert,et al.  Early embryonic survival in the pig: Can it be improved? , 2002 .

[58]  T. Spencer,et al.  Keratinocyte growth factor is up-regulated by estrogen in the porcine uterine endometrium and functions in trophectoderm cell proliferation and differentiation. , 2001, Endocrinology.

[59]  E. Knol,et al.  Progress of farrowing and early postnatal pig behavior in relation to genetic merit for pig survival. , 2001, Journal of animal science.

[60]  K. Hayashi,et al.  Plasma adrenomedullin levels in pregnancies with appropriate for gestational age and small for gestational age infants , 2001, Journal of perinatal medicine.

[61]  O. Smithies,et al.  Extreme hydrops fetalis and cardiovascular abnormalities in mice lacking a functional Adrenomedullin gene. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[62]  A Tuchscherer,et al.  Early identification of neonates at risk: traits of newborn piglets with respect to survival. , 2000, Theriogenology.

[63]  L. Zhang,et al.  Expression of the hypoxically regulated angiogenic factor adrenomedullin correlates with uterine leiomyoma vascular density. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[64]  J. Vallet.  Fetal Erythropoiesis and Other Factors which Influence Uterine Capacity in Swine , 2000 .

[65]  D. Gazzolo,et al.  Adrenomedullin is increased in the fetoplacental circulation in intrauterine growth restriction with abnormal umbilical artery waveforms. , 2000, American journal of obstetrics and gynecology.

[66]  H. Leis,et al.  Colorimetric determination of cell numbers by Janus green staining. , 1999, Analytical biochemistry.

[67]  R. Johnson,et al.  Responses in ovulation rate, embryonal survival, and litter traits in swine to 14 generations of selection to increase litter size. , 1999, Journal of animal science.

[68]  H. Nishimatsu,et al.  Role of nitric oxide-cGMP pathway in adrenomedullin-induced vasodilation in the rat. , 1999, Hypertension.

[69]  M. Ko,et al.  Expression of adrenomedullin, a hypotensive peptide, in the trophoblast giant cells at the embryo implantation site in mouse. , 1998, Developmental biology.

[70]  C. Letizia,et al.  Immunoreactive adrenomedullin in human fetoplacental tissues. , 1998, American journal of obstetrics and gynecology.

[71]  U. Zwiener,et al.  Body weight distribution and organ size in newborn swine (sus scrofa domestica) -- a study describing an animal model for asymmetrical intrauterine growth retardation. , 1998, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[72]  Lyna Zhang,et al.  PCR display identifies tamoxifen induction of the novel angiogenic factor adrenomedullin by a non estrogenic mechanism in the human endometrium , 1998, Oncogene.

[73]  L. Montuenga,et al.  Expression of Adrenomedullin and Its Receptor during Embryogenesis Suggests Autocrine or Paracrine Modes of Action. , 1997, Endocrinology.

[74]  M. Rothschild.  Genetics and reproduction in the pig , 1996 .

[75]  R. Geisert,et al.  Embryonic Mortality in Domestic Species , 1994 .

[76]  K. Kangawa,et al.  Adrenomedullin: a novel hypotensive peptide isolated from human pheochromocytoma. , 1993, Biochemical and biophysical research communications.

[77]  R. Roberts,et al.  Establishment of pregnancy in the pig: II. Cellular remodeling of the porcine blastocyst during elongation on day 12 of pregnancy. , 1982, Biology of reproduction.

[78]  R. Roberts,et al.  Establishment of pregnancy in the pig: I. Interrelationships between preimplantation development of the pig blastocyst and uterine endometrial secretions. , 1982, Biology of reproduction.

[79]  G. Tomes.  25 – FACTORS AFFECTING REPRODUCTIVE EFFICIENCY OF THE BREEDING HERD , 1982 .

[80]  F. Bazer,et al.  Conceptus development in intact and unilaterally hysterectomized-ovariectomized gilts: interrelations among hormonal status, placental development, fetal fluids and fetal growth. , 1977, Journal of animal science.

[81]  L. C. Ulberg,et al.  Effect of quantity of uterus on uterine capacity in gilts. , 1970, Journal of animal science.

[82]  L. C. Ulberg,et al.  Uterine capacity at two stages of gestation in gilts following embryo superinduction. , 1969, Journal of animal science.

[83]  L. C. Ulberg,et al.  Uterine capacity in gilts. , 1969, Journal of reproduction and fertility.

[84]  P. Dziuk.  Effect of number of embryos and uterine space on embryo survival in the pig. , 1968, Journal of animal science.