Abstract
In healthy term human newborns a unique hemostatic balance exists with reduced plasma concentrations of several coagulant and anticoagulant proteins, including antithrombin III (AT III). In preterm newborns even lower AT III concentrations are observed, with an associated thromboembolic risk. As part of our study program on the gene regulation of AT III, we investigated whether the increase in plasma AT III activity during fetal and neonatal development is particularly controlled at the transcriptional level. Plasma AT III activity and liver AT III mRNA content between the 8th wk of gestation and the 4th wk after birth were determined in sheep. AT III activity gradually increased from 34% of the mean adult level at 8-10 wk of gestation to 86%(2.5-fold) at term (21 wk), and remained in the adult range after birth. The mean body weight, and thus plasma volume, increased 57-fold. Therefore, the total plasma AT III activity increased 140-fold. The total liver AT III mRNA content increased only 14-fold between these fetal stages, mainly due to increased liver weight. Therefore, the total plasma AT III activity increased 10-fold more than the liver AT III mRNA content. In the neonatal period between d 1-3 and 28, the total plasma AT III activity increased only 2-fold more than the liver AT III mRNA content. We conclude that the increase in plasma AT III activity during the fetal period, and similarly the neonatal period, is not regulated at the transcriptional level. Furthermore, a unique fetal isoform of AT III was detected in sheep. This isoform had a 2500-D higher molecular mass compared with the other fetal, neonatal, and adult AT III isoform, and disappeared from the circulation between d 2 and 7 after birth. These AT III isoforms differ in their carbohydrate moiety.
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Abbreviations
- AT III:
-
antithrombin III
- PNGase F:
-
peptide:N-glycosidase F
References
Rosenberg RD, Damus PS 1973 The purification and mechanism of action of human antithrombin-heparin cofactor. J Biol Chem 248: 6490–6505.
Kurachi K, Fujikawa K, Schmer G, Davie EW 1976 Inhibition of bovine factor IXa and factor Xa by antithrombin III. Biochemistry 15: 373–377.
Koide T 1979 Isolation and characterization of antithrombin III from human, porcine, and rabbit plasma, and rat serum. J Biochem 86: 1841–1850.
Jordan RE 1983 Antithrombin in vertebrate species: conservation of the heparin-dependent anticoagulant mechanism. Arch Biochem Biophys 227: 587–595.
Damus PS, Hicks M, Rosenberg RD 1973 Anticoagulant action of heparin. Nature 246: 355–357.
Egeberg O 1965 Inherited antithrombin deficiency causing thrombophilia. Thromb Diath Haemorrh 13: 516–530.
Hathaway WE, Bonnar J 1978 Physiology of coagulation in the fetus and newborn infant. In: Oliver T (ed) Perinatal Coagulation-Monographs in Neonatology. Grune & Stratton, New York, 53–80.
Peters M, Jansen E, ten Cate JW, Kahle LH, Ockelford P, Breederveld C 1984 Neonatal antithrombin III. Br J Haematol 58: 579–587.
Andrew M, Paes B, Milner R, Johnston M, Mitchell L, Tollefsen DM, Castle V, Powers P 1988 Development of the human coagulation system in the healthy premature infant. Blood 72: 1651–1657.
Peters M, ten Cate JW, Breederveld C, de Leeuw R, Emeis JJ, Koppe JG 1984 Low antithrombin III levels in neonates with idiopathic respiratory distress syndrome: poor prognosis. Pediatr Res 18: 273–276.
Kisker CT, Perlman S, Bohlken D, Wicklund B 1988 Measurement of prothrombin mRNA during gestation and early neonatal development. J Lab Clin Med 112: 407–412.
Manco-Johnson MJ, Spedale S, Peters M, Townsend SF, Jacobson LJ, Christian J, Krugman SD, Hay WW, Sparks JW 1995 Identification of a unique form of protein C in the ovine fetus: developmentally linked transition to the adult form. Pediatr Res 37: 365–372.
Lamontagne L, Gauldie J 1980 Ontogeny of mouse and rat antithrombin III. Thromb Res 20: 417–424.
D'Souza SE, Mercer JFB 1987 Antithrombin III mRNA in adult rat liver and kidney and in rat liver during development. Biochem Biophys Res Commun 142: 417–421.
Amrani DL, Mosesson MW, Koide T 1985 Evidence that chicken antithrombin III is a developmentally regulated glycoprotein synthesized by hepatocytes. Biochim Biophys Acta 847: 324–334.
Amrani DL, Rosenberg J, Samad F, Bergtrom G, Banfield DK 1993 Developmental expression of chicken antithrombin III is regulated by increased RNA abundance and intracellular processing. Biochim Biophys Acta 1171: 239–246.
Andrew M, O'Brodovich H, Mitchell L 1988 Fetal lamb coagulation system during normal birth. Am J Hematol 28: 116–118.
Moalic P, Gruel Y, Foloppe P, Delahousse B, Leclerc M-H, Leroy J 1989 Hemostasis development in the lamb fetus and neonate. Am J Vet Res 50: 59–63.
Hedner U, Nilsson IM 1973 Antithrombin III in a clinical material. Thromb Res 3: 631–641.
Petersen TE, Dudek-Wojciechowska G, Sottrup-Jensen L, Magnusson S 1979 Primary structure of antithrombin III (heparin cofactor). Partial homology between α1-antitrypsin and antithrombin III. In: Collen D, Wiman B, Verstraete M (eds) The Physiological Inhibitors of Blood Coagulation and Fibrinolysis. Elsevier Science Publishers, Amsterdam, 43–54.
Franzen L-E, Svensson S, Larm O 1980 Structural studies on the carbohydrate portion of human antithrombin III. J Biol Chem 255: 5090–5093.
Carlson TH, Atencio AC 1982 Isolation and partial characterization of two distinct types of antithrombin III from rabbit. Thromb Res 27: 23–34.
Peterson CB, Blackburn MN 1985 Isolation and characterization of an antithrombin III variant with reduced carbohydrate content and enhanced heparin binding. J Biol Chem 260: 610–615.
Brennan SO, George PM, Jordan RE 1987 Physiological variant of antithrombin-III lacks carbohydrate side chain at Asn 135. FEBS Lett 219: 431–436.
Naaktgeboren C, Stegeman JHJ 1966 Untersuchungenuber den Einflu des Uterus und der Placenta auf das fetale Wachstum und das Geburtsgwicht, mit besonderer Berucksichtigung des Schafes. Z Tierz Zuechtungsbiol 85: 245–290.
Chomczynski P, Sacchi N 1987 Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159.
Niessen RWLM, Sturk A, Hordijk PL, Michiels F, Peters M 1992 Sequence characterization of a sheep cDNA for antithrombin III. Biochim Biophys Acta 1171: 207–210.
Laudien Gonzalez I, Gorski JL, Campen TJ, Dorney DJ, Erickson JM, Sylvester JE, Schmickel RD 1985 Variation among human 28S ribosomal RNA genes. Proc Natl Acad Sci USA 82: 7666–7670.
Munro HN, Fleck A 1966 The determination of nucleic acids. Methods Biochem Anal 14: 113–176.
Feinberg AP, Vogelstein B 1984 A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–267.
Mc Kay EJ, Laurell C-B 1980 The interaction of heparin with plasma proteins. Demonstration of different binding sites for antithrombin III complexes and antithrombin III. J Lab Clin Med 95: 69–80.
Laemmli UK 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Johnstone A, Thorpe R 1987 Purification of immunoglobulins, constituent chains and fragments. In: Johnstone A, Thorpe R.(eds) Immunochemistry in Practice. Blackwell Scientific Publications, Oxford 48–85.
Towbin H, Staehelin T, Gordon J 1979 Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354.
Peters M, Breederveld C, Kahle LH, ten Cate JW 1982 Rapid microanalysis of coagulation parameters by automated chromogenic substrated methods-application in neonatal patients. Thromb Res 28: 773–781.
Odegard OR, Lie M, Abildgaard U 1976 Antifactor Xa activity measured with amidolytic methods. Haemostasis 5: 265–275.
de Groot CJ, Zonneveld D, de Laaf RTM, Dingemanse MA, Mooren PG, Moorman AFM, Lamers WH, Charles R 1986 Developmental and hormonal regulation of carbamoylphosphate synthetase gene expression in rat liver: evidence for control mechanisms at different levels in the perinatal period. Biochim Biophys Acta 866: 61–67.
Prothero JW 1980 Scaling of blood parameters in mammals. Comp Biochem Physiol 67A: 649–657.
Andrew M, Mitchell L, Berry LR, Schmidt B, Hatton MWC 1988 Fibrinogen has a rapid turnover in the healthy newborn lamb. Pediatr Res 23: 249–252.
Witmer MR, Hatton MWC 1991 Antithrombin III- associates more readily than antithrombin III- with uninjured and de-endothelialized aortic wall in vitro and in vivo. Arterioscler Thromb 11: 530–539.
Bierhuizen MF, Mattei MG, Fukuda M 1993 Expression of the developmental I antigen by a cloned human cDNA encoding a member of a-1,6-N-acetylglucosaminyltransferase gene family. Genes Dev 7: 468–478.
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Niessen, R., Lamping, R., Peters, M. et al. Fetal and Neonatal Development of Antithrombin III Plasma Activity and Liver Messenger RNA Levels in Sheep. Pediatr Res 39, 685–691 (1996). https://doi.org/10.1203/00006450-199604000-00021
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DOI: https://doi.org/10.1203/00006450-199604000-00021
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