Skip to main content
Log in

Oxytocinase-immunohistochemical demonstration in the immature and term human placenta

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

Oxytocinase (cystine aminopeptidase) was purified from human retroplacental serum by a combination of fractional precipitation, hydroxylapatite chromatography and gel exlusion chromatography on Sephadex G-200. The purified enzyme possessed a specific activity of 980 mIU/mg using L-cystine-di-p-nitroanilide as substrate. This represented a 3200 fold concentration from the starting material in an overall yield of 12%. Antibodies against oxytocinase were raised in rabbits and the γ-globulin fraction labelled with fluorescein isothiocyanate prior to its use in the immunofluorescence histochemical localization of the enzyme in human placental tissue. Oxytocinase was confined to the syncytiotrophoblastic cells of normal term, and immature placentas as well as in placentas from patients suffering from severe toxaemia. Specific immunofluorescence was also present in the outer margins of the chorion and to a lesser extent in the amnion.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Earth, T., Pliška, V., Rychlík, I., Šorm, F.: Enzymatic inactivation of oxytocin V. Purification and some properties of enzymes from human retroplacental serum. Coll. Czech, chem. Comm. 32, 2327–2336 (1967)

    Google Scholar 

  • Barth, T., Rychlík, I., Mannsfeldt, H. G.: Human pregnancy oxytocinase isolation and sub-strate characteristics. Coll. Czech, chem. Comm. 36, 2540–2546 (1971)

    Google Scholar 

  • Beck, J. S., Gordon, R. L., Donald, D., Melvin, J. M. O.: Characterisation of antisera to a growth-hormone-like placental antigen (human placental lactogen): Immunofluorescence studies with these sera on normal and pathological syncytiotrophoblast. J. Path. 97, 545–555 (1969)

    Google Scholar 

  • Fekete, K.: Beiträge zur Physiologie der Gravidität. Endokrinologie 7, 364–369 (1930)

    Google Scholar 

  • Fekete, K.: Gibt es während der Schwangerschaft ein aktives Hypophysen-Hinterlappenhormon im Blute? Endokrinologie 10, 16–23 (1932)

    Google Scholar 

  • Hashimoto, T.: Studies on 1-cystine-aminopeptidase. Part II. Clinical observations. J. Jap. obstet. gynaec. Soc. d8, 96–104 (1961)

  • Hawker, R. W.: Inactivation of antidiuretic hormone and oxytocin during pregnancy. Quart. J. exp. Physiol. 41, 301–308 (1956)

    Google Scholar 

  • Hempel, Von E., Geyer, G.: Submikroskopische Verteilung der alkalischen Phosphatase in der menschlichen Placenta. Acta histochem. (Jena) 34, 138–147 (1969)

    Google Scholar 

  • James, N. T.: Histochemical demonstration of oxytocinase in the human placenta. Nature (Lond.) 210, 1276–1277 (1966)

    Google Scholar 

  • Kulling, B., Yman, L.: Purification and some properties of leucine aminopeptidases from retroplacental serum. Acta pharma. suecica 7, 65–74 (1970)

    Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin Phenol reagent. J. biol. Chem. 193, 265–275 (1951)

    CAS  PubMed  Google Scholar 

  • Mathur, V. S., Walker, J. M.: The origin of human placental oxytocinase. J. Physiol. (Lond.) 208, 291–298 (1970)

    Google Scholar 

  • Müller-Hartburg, W. H., Nesvabda, H., Tuppy, H.: Die Anwendung einer chemischen Methode zur Bestimmung des Oxytocinasespiegels im Schwangerenserum. Arch. Gynäk. 191, 442–456 (1959)

    Google Scholar 

  • Nairn, R. C.: Fluorescent protein tracing, 3rd edn. London: Livingstone 1969

    Google Scholar 

  • Page, E. W.: The value of plasma pitocinase determinations in obstetrics. Amer. J. Obstet. Gynec. 52, 1014–1022 (1946)

    Google Scholar 

  • Riad, A. M.: Studies on pregnancy serum cystine aminopeptidase activity “oxytocinase”. J. Obstet. Gynaec. Brit. Comm. 69, 409–416 (1962)

    Google Scholar 

  • Rydén, G.: Cystine aminopeptidase and oxytocinase activity in pregnancy. Acta. obstet. gynaec. scand. Vol. 45, Suppl. No. 3 (1966)

  • Seelig, H.-P., Roemheld, R.: Untersuchungen zur histochemischen Lokalisation der Leucin- und Cystinaminopeptidase (Oxytocinase) in der Placenta. Histochemie 18, 30–39 (1969)

    Google Scholar 

  • Semm, K., Waidl, U. E.: Histochemische Untersuchungen über die Serumoxytocinase-Bildung im menschlichen Trophoblasten. Z. Geburtsh. Gynäk. 158, 165–171 (1962)

    Google Scholar 

  • Sjöholm, L, Yman, L.: Preparation of highly purified oxytocinase (cystine aminopeptidase) from retroplacental serum. Acta pharma. suecica 3, 377–388 (1966)

    Google Scholar 

  • Small, C. W., Watkins, W. B.: Immunochemical localisation of some placental enzymes. N. Z. Med. J. 74, 338 (1971a)

    Google Scholar 

  • Small, C. W., Watkins, W. B.: An improved method for the determination of human pregnancy serum oxytocinase activity. Enzymologia 41, 121–128 (1971b)

    Google Scholar 

  • Small, C. W., Watkins, W. B.: S-benzyl-L-cysteine-p-nitroanilide. A new substrate for the determination of oxytocinase with improved specificity. Biochem. Med. 9, 103–112 (1974)

    Google Scholar 

  • Tovey, J. E.: Serum oxytocinase. Clin. Biochem. 2, 289–310 (1969)

    Google Scholar 

  • Tuppy, H.: The influence of enzymes on neurohypophysial hormones and similar peptides. In: Handbook of experimental pharmacology, ed. Berde, B., p. 67–129. Berlin-Heidelberg-New York: Springer 1968

    Google Scholar 

  • Tuppy, H., Nesvadba, U. H.: deÜber die Aminopeptidaseaktivität des Schwangerenserums und ihre Beziehung zu dessen Vermögen, Oxytocin zu inaktivieren. Mh. Chem. 88, 977–988 (1957)

    Google Scholar 

  • Tuppy, H., Wintersberger, E.: Reinigung und Eigenschaften der Serumoxytocinase. Mh. Chem. 91, 1001–1010 (1960)

    Google Scholar 

  • Watkins, W. B., Anderson, S. R. A.: The origin of heat-stable placental alkaline phosphatase. J. Obstet. Gynaec. Brit. Comm. 78, 691–693 (1971)

    Google Scholar 

  • Watkins, W. B., Small, C. W.: Immunologic inactivation of human pregnancy serum oxytocinase activity. Amer. J. Obstet. Gynec. 113, 973–978 (1972)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This work was financed by a grant from The Medical Research Council of New Zealand.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Small, C.W., Watkins, W.B. Oxytocinase-immunohistochemical demonstration in the immature and term human placenta. Cell Tissue Res. 162, 531–539 (1975). https://doi.org/10.1007/BF00209352

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00209352

Key words

Navigation