Anatomy and Embryology

, Volume 184, Issue 2, pp 141–152 | Cite as

Uteroglobin in the developing rabbit conceptus in vivo and in vitro

  • D. R. Dannhorn
  • S. Gierhake
  • C. Kirchner
Article

Summary

Uteroglobin (UGL) was measured in day- 4 to day-10 rabbit conceptuses by a competitive ELISA. Levels in blastocyst fluid, tissues, coverings and in the early fetus were determined separately. The total amount of UGL increased from 18.4 ng to 6.8 μg per conceptus. The UGL content of individual day-6 blastocysts was studied in vitro. Culturing was carried out up to 60 h in Ham's F10 medium with polyvinylpyrrolidone as macromolecular component, with and without progesterone, and with progesterone plus estradiol. UGL was determined in the blastocyst fluids, tissues with coverings and in the culture media. After labelling with [35S]-methionine, protein patterns of total blastocysts and of culture media were analysed by two-dimensional gel electrophoresis and fluorography. The morphology of cultured blastocysts was examined by electron microscopy. During 60 h of culture, the blastocysts expanded in diameter by 84%, and released 19% of their initial UGL content into the medium, independent of the hormonal substitution. Neither de novo synthesis, nor degradation of UGL was found: the protein remained unlabelled in fluorography, and its total quantity was not significantly different from that of non-cultured controls. Trophoblast, endoderm and embryoblast cells showed well preserved cell organelles and intercellular junctions, while the morphological differentiation of the germ layer was inhibited.

Key words

Blastocyst Uteroglobin In vitro culture Progesterone ELISA Fluorography Rabbit 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Angle MJ, Mead RA (1979) The source of progesterone in preimplantation rabbit blastocysts. Steroids 33:625–637Google Scholar
  2. Anthony RV, Helmer SD, Sharif SF, Roberts RM, Hansen PJ, Thatcher WW, Bazer FW (1988) Synthesis and processing of ovine trophoblast protein-1 and bovine trophoblast protein-1 conceptus secretory proteins involved in the maternal recognition of pregnancy. Endocrinology 123:1274–1280Google Scholar
  3. Arthur AJ, Daniel JC (1972) Progesterone regulation of blastokinin production and maintenance of rabbit blastocysts transferred into uteri of castrate recipients. Fertil Steril 23:115–122Google Scholar
  4. Atger M, Milgrom E (1977) Progesterone-induced messengerRNA: translation, purification, and preliminary characterization of uteroglobin mRNA. J Biol Chem 252:5412–5418Google Scholar
  5. Beato M (1977) Physico-chemical characterization of uteroglobin and its interaction with progesterone. In: Johnson MH (ed) Development in mammals. vol 2 Elsevier/North Holland, Amsterdam, pp 173–198Google Scholar
  6. Beato M, Arnemann J (1975) Hormone-dependent synthesis and secretion of uteroglobin in isolated rabbit uterus. FEBS Lett 58:126–129Google Scholar
  7. Beato M, Baier R (1975) Binding of progesterone to the proteins of the uterine luminal fluid. Identification of uteroglobin as the binding protein. Biochim Biophys Acta 392:346–356Google Scholar
  8. Beier HM (1968) Biochemisch-entwicklungsphysiologische Untersuchungen am Proteinmilieu für die Blastocystenentwicklung des Kaninchens (Oryctolagus cuniculus). Zool Jb Anat 85:72–190Google Scholar
  9. Beier HM (1976) Uteroglobin and related biochemical changes in the reproductive tract during early pregnancy in the rabbit. J Reprod Fertil [Suppl 25]:53–69Google Scholar
  10. Beier HM (1978) Physiology of uteroglobin. Reprod Physiol 8:219–248Google Scholar
  11. Beier HM, Maurer RR (1975) Uteroglobin and other proteins in rabbit blastocyst fluid after development in vivo and in vitro. Cell Tissue Res 159:1–10Google Scholar
  12. Blum H, Beier H, Gross HJ (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8:93–99Google Scholar
  13. Bochskanl R, Kirchner C (1981) Uteroglobin and the accumulation of progesterone in the uterine lumen of the rabbit. Wilhelm Roux' Arch Entwicklungsmech Org 190:127–131Google Scholar
  14. Bochskanl R, Thie M, Kirchner C (1984) Progesterone-dependent uptake of uteroglobin by rabbit endometrium. Histochemistry 80:581–589Google Scholar
  15. Bochskanl R, Thie M, Wirth B, Kirchner C (1988) Uteroglobin as progesterone-binding protein in the preimplantation uterine epithelium of the rabbit: histochemical studies. Cell Tissue Res 252:625–630Google Scholar
  16. Borland RM, Erickson GF, Ducibella T (1977) Accumulation of steroids in rabbit preimplantation blastocysts. J Reprod Fertil 49:219–224Google Scholar
  17. Bullock DW (1980) Uterine proteins as markers of progesterone action. In: Beato M (ed) Steroid induced proteins. Developments in endocrinology. vol 8, Elsevier/North Holland, Amsterdam, pp 315–318Google Scholar
  18. Bullock DW, Woo SLC, O'Malley BW (1976) Uteroglobin messenger RNA: translation in vitro. Biol Reprod 15:435–443Google Scholar
  19. Cowan BD, Manes C, Hagerman DD (1976) Progesterone concentrations in rabbit uterine flushings before implantation. J Reprod Fertil 47:459–461Google Scholar
  20. Dannhorn DR, Kirchner C (1990a) Uptake of tritiated uteroglobin by the endometrium of the rabbit during peri-implantation. Cell Tissue Res 259:519–528Google Scholar
  21. Dannhorn DR, Kirchner C (1990b) Uptake and accumulation of tritiated uteroglobin by day-6 rabbit blastocysts. Cell Tissue Res 262:569–577Google Scholar
  22. Dannhorn DR, Henkel R, Kirchner C (1988) Synthese von Uteroglobinin der Blastozyste des Kaninchens? Autoradiographische und immuncytochemische Untersuchungen. Fertilität 4:223–226Google Scholar
  23. Darmhorn DR, Wirth B, Kirchner C (1989) Purification of uteroglobin using monospecific antibodies coupled to divinylsulphone-activated agarose. J Immunol Methods 119:223–230Google Scholar
  24. El-Shershaby AM, Hinchliffe HJ (1974) Cell redundancy in the zona-intact preimplantation mouse blastocyst: a light and electron microscope study of dead cells and their fate. J Embryol Exp Morphol 31:643–654Google Scholar
  25. Enders AC (1971) The fine structure of the blastocyst. In: Blandau RJ (ed) The biology of the blastocyst. University of Chicago Press, Chicago London, pp 71–94Google Scholar
  26. Fowler RE, Johnson MH, Walters DE, Pratt HPM (1976) The progesterone and protein composition of rabbit uterine flushings. J Reprod Fertil 46:427–430Google Scholar
  27. Fowler RE, Johnson MH, Walters DE, Eager DD (1977) The progesterone content of rabbit uterine flushings. J Reprod Fertil 50:301–308Google Scholar
  28. Fridlansky F, Milgrom E (1976) Interaction of uteroglobin with progesterone, 5 α-pregnane-3,20-dione and estrogens. Endocrinology 99:1244–1251Google Scholar
  29. Fuchs AR, Beling C (1974) Evidence of early ovarian recognition of blastocysts in rabbits. Endocrinology 95:1054–1058Google Scholar
  30. Fujimoto S, Sundaram K (1978) The source of progesterone in rabbit blastocysts. J Reprod Fertil 52:231–233Google Scholar
  31. Godkin JD, Bazer FW, Sessions F, Roberts RM (1982) Purification and properties of a major, low molecular weight protein released by the trophoblast of sheep blastocysts at day 13–21. J Reprod Fertil 65:141–150Google Scholar
  32. Hamana K, Hafez ESE (1970) Disc electrophoretic patterns of uteroglobin and serum proteins in rabbit blastocoelic fluid. J Reprod Fertil 21:555–558Google Scholar
  33. Hegele-Hartung C, Fischer B, Beier HM (1988a) Development of preimplantation rabbit embryos after in vitro culture and embryo transfer: an electron microscopic study. Anat Rec 220:31–42Google Scholar
  34. Hegele-Hartung C, Schumacher A, Fischer B (1988b) Ultrastructure of preimplantation rabbit embryos exposed to visible light and room temperature. Anat Embryol 178:229–241Google Scholar
  35. Hegele-Hartung C, Dreiner U, Beier HM (1991) Effect of in vitro culture on the dynamics of uteroglobin distribution in rabbit blastocysts. Anat Embryol 183:119–128Google Scholar
  36. Helmer SD, Hansen PJ, Anthony RV, Thatcher WW, Bazer FW, Roberts RM (1987) Identification of bovine trophoblast protein-1, a secretory protein immunologically related to ovine trophoblast protein-1. J Reprod Fertil 79:83–91Google Scholar
  37. Henkel R, Dannhorn DR, Petzoldt U, Kirchner C (1990) Ultrastructure, protein synthesis and secretion of day-6 rabbit blastocysts cultured in a chemically defined, protein-free medium. Anal Embyrol 182:465–472Google Scholar
  38. Kirchner C (1969) Untersuchungen an uterusspezifischen Glykoproteinen während der frühen Gravidität des KaninchensOryctolagus cuniculus. Wilhelm Roux' Arch Entwicklungsmech Org 164:97–133Google Scholar
  39. Kirchner C (1972) Immune histologic studies on the synthesis of a uterine-specific protein in the rabbit and its passage through the blastocyst coverings. Fertil Steril 23:131–136Google Scholar
  40. Kirchner C (1976) Uteroglobin in the rabbit. 1. Intracellular localization in the oviduct, uterus and preimplantation blastocyst. Cell Tissue Res 170:415–471Google Scholar
  41. Kirchner C, Seitz KA (1972) Elektronenmikroskopische Untersuchungen über die Blastozyste des Kaninchens vor der Implantation in bezug auf ihre Wechselbeziehung zur uterinen Umgebung. Wilhelm Roux' Arch Entwicklungsmech Org 170:221–233Google Scholar
  42. Laskey RA, Mills AD (1975) Quantitative film detection of3H and14C in polyacrylamide gels by fluorography. Eur J Biochem 56:335–341Google Scholar
  43. Malsky ML, Bullock DW, Willard JJ, Darrell NW (1979) Progesterone-induced secretory protein: NH2-terminal sequence of pre-uteroglobin. J Biol Chem 254:1580–1585Google Scholar
  44. Märki F, Pfeilschifter J, Rink H, Wiesenberg I (1990) “Antiflammins”: two nonapeptide fragments of uteroglobin and lipocortin I have no phospholipase AZ inhibitory and anti-inflammatory activity. FEBS Lett 264:171–175Google Scholar
  45. Mayol RF, Longenecker DE (1974) Development of a radioimmunoassay for blastokinin. Endocrinology 95:1534–1542Google Scholar
  46. Miele L, Cordella-Miele E, Mukherjee AB (1987) Uteroglobin: structure, molecular biology, and new perspectives on its function as a phospholipase AZ inhibitor. Endocr Rev 8:474–490Google Scholar
  47. Nieder GL (1990) Protein secretion by the mouse trophoblast during attachment and outgrowth in vitro. Biol Reprod 43:251–259Google Scholar
  48. Nieder GL, Jennes L (1990) Production of mouse placental lactogen-I by trophoblast giant cells in utero and in vitro. Endocrinology 126:2809–2814Google Scholar
  49. O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021Google Scholar
  50. Pierce GB, Lewellyn AL, Parchment RE (1989) Mechanism of programmed cell death in the blastocyst. Proc Natl Acad Sci USA 86:3654–3658Google Scholar
  51. Rajkumar K, Bigsby R, Lieberman R, Gershenson LE (1983) Effect of progesterone and 17β-estradiol on the production of uteroglobin by cultured rabbit uterine epithelial cells. Endocrinology 112:1499–1505Google Scholar
  52. Robinson DH, Kirk KL, Benos DJ (1989) Macromolecular transport in rabbit blastocysts: evidence for a specific uteroglobin transport system. Mol Cell Endocrinol 63:227–237Google Scholar
  53. Schumacher A, Fischer B (1988) The influence of visible light and room temperature on cell proliferation in preimplantation rabbit embryos. J Reprod Fertil 84:197–204Google Scholar
  54. Seamark RF, Lutwak-Mann C (1972) Progestins in rabbit blastocysts. J Reprod Fertil 29:147–148Google Scholar
  55. Sharif SF, Francis H, Keisler DH, Roberts RM (1989) Correlation between the release of ovine trophoblast protein-1 by the conceptus and the production of polypeptides by the maternal endometrium of ewes. J Reprod Fertil 85:471–476Google Scholar
  56. Singh MM, Booth WD (1979) Origin of oestrogen in preimplantation rabbit blastocysts. J Steroid Biochem 11:723–728Google Scholar
  57. Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43Google Scholar
  58. Torkkeli T, Krusius T, Jänne O (1978) Uterine and lung uteroglobins in the rabbit. Two similar proteins with differential hormonal regulation. Biochim Biophys Acta 544:578–592Google Scholar
  59. Van Blerkom J, Manes C, Daniel JC (1973) Development of preimplantation rabbit embryos in vivo and in vitro. 1. An ultrastructural comparison. Dev Biol 35:262–282Google Scholar
  60. Warembourg M, Tranchant O, Atger M, Milgrom E (1986) Uteroglobin messenger ribonucleic acid: localization in rabbit uterus and lung by in situ hybridization. Endocrinology 119:1632–1640Google Scholar
  61. Williams BS, Biggers JD (1990) Polar trophoblast (Rauber's layer) of the rabbit blastocyst. Anat Rec 211:222Google Scholar
  62. Wu JT (1987) Metabolism of progesterone by preimplantation mouse blastocysts in culture. Biol Reprod 36:549–556Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • D. R. Dannhorn
    • 1
  • S. Gierhake
    • 1
  • C. Kirchner
    • 1
  1. 1.Department of BiologyPhilipps UniversityMarburgGermany

Personalised recommendations