Skip to main content

Advertisement

SpringerLink
Log in

Localization of prostaglandin synthetase in chicken epiphyseal cartilage

  • Published:
Calcified Tissue Research Aims and scope Submit manuscript

Summary

Prostaglandin synthetase activity in highspeed particulate fractions of chick epiphyseal cartilage has been characterized with respect to cofactor requirements, pH optimum, buffer-ion effects, types of prostaglandins formed, and the distribution of prostaglandin synthetase activity in zones of the epiphyseal plate.

Direct homogenization of cartilage was found to be more efficacious than releasing chondrocytes by enzymatic digestion for preparation of prostaglandin synthetase, a homogenization time of 4 min yielding maximal activity. The optimal incubation medium contained 50 mM Tris buffer (pH 7.5), 2.5 mM epinephrine, 1 μM hemoglobin, 3.25 mM glutathione, 200 μg/ml enzyme protein, and 5 μM substrate. Glutathione was effective only if present during homogenization. Rates of PGE2 biosynthesis were linear up to 15 min and then rapidly declined, indicative of self-deactivation. The low levels of PGF2α formed, and their decrease after 20 min incubation, suggests the possible presence of degradative enzymes.

Prostaglandin synthetase was inhibited by aspirin, indomethacin, and vitamin E, but not vitamin K1. Cation concentrations in the physiological range had only modest effects on prostaglandin biosynthesis, and then only if present during tissue homogenization. In the presence of phosphate buffer, Ca2+ was somewhat inhibitory. Since in the absence of phosphate Ca2+ had no deleterious effect, it is probably that the inhibitory effect was caused by precipitation of calcium phosphate.

Hypertrophic and calcified cartilage exhibited significantly higher prostaglandin synthetase activity than the proliferating and maturing zones. The increased synthesis of prostaglandins in the low layers of the growth plate may indicate a role of these factors in chondrocyte differentiation and/or calcification.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Samuelsson, B., Granström, E., Green, K., Hamberg, M., Hammarström, S.: Prostaglandins, Annu. Rev. Biochem.44:669–695, 1975

    PubMed  Google Scholar 

  2. Wong, P.Y.-K., Wuthier, R.E.: Isolation and identification of prostaglandin PGB1 in growth cartilage, Prostaglandins8:125–132, 1974

    PubMed  Google Scholar 

  3. Wong, P.Y.-K., Majeska, R.J., Wuthier, R.E.: Biosynthesis and metabolism of prostaglandins in chick epiphyseal cartilage, Prostaglandins14:839–851, 1977

    PubMed  Google Scholar 

  4. Wuthier, R.E.: The role of phospholipids in biological calcification. Distribution of phospholipase activity in calcifying epiphyseal cartilage. Clin. Orthop.90:191–200, 1973

    PubMed  Google Scholar 

  5. Wuthier, R.E.: Lipids of matrix vesicles, Fed. Proc.35:117–121, 1976

    PubMed  Google Scholar 

  6. Anderson, H.C.: Electron microscopic studies of induced cartilage development and calcification. J. Cell Biol.35:81–101, 1967

    PubMed  Google Scholar 

  7. Anderson, H.C.: Vesicles associated with calcification in the matrix of epiphyseal cartilage, J. Cell Biol.41:59–72, 1969

    PubMed  Google Scholar 

  8. Bonucci, E.: Fine structure of early cartilage calcification, J. Ultrastruct. Res.20:33–50, 1969

    Google Scholar 

  9. Wuthier, R.E.: Electrolytes of isolated epiphyseal chondrocytes, matrix vesicles and extracellular fluid, Calcif. Tissue Res.23:125–133, 1977

    PubMed  Google Scholar 

  10. Ramwell, P.W., Shaw, J.E.: Biological significance of the prostaglandins, Recent Prog. Horm. Res.26:139–173, 1970

    PubMed  Google Scholar 

  11. Horrobin, D.F.: Interactions between prostaglandins and calcium: The importance of bell-shaped dose-response curves, Prostaglandins14:667–677, 1977

    PubMed  Google Scholar 

  12. Klein, D.C., Raisz, L.G.: Stimulation of bone resorption in tissue culture, Endocrinology86:1436–1440, 1970

    PubMed  Google Scholar 

  13. Tashjian, A.H., Jr., Voelkel, E.F., Levine L., Goldhaber, P.: Evidence that bone resorption-stimulating factor produced by mouse fibrosarcoma cells is prostaglandin E2. A new model for the hypercalcemia of cancer. J. Exp. Med.136:1329–1343, 1972

    PubMed  Google Scholar 

  14. Voelkel, E.F., Tashjian, A.H., Jr., Franklin, R., Wasserman, E., Levine, L.: Hypercalcemia and tumor prostaglandins. The VX2 carcinoma model in the rabbit. Metab. Clin. Exp.24:973–986, 1975

    PubMed  Google Scholar 

  15. Robinson, D.R., Tashjian, A.H., Jr., Levine, L.: Prostaglandin-stimulated bone resorption by rheumatoid synovia. A possible mechanism for bone destruction in rheumatoid arthritis, J. Clin. Invest.56:1181–1188, 1975

    PubMed  Google Scholar 

  16. Gomes, B.C., Hausmann, E., Weinfeld, N., DeLuca, C.: Prostaglandins: Bone resorption stimulating factors released from monkey gingiva, Calcif. Tissue Res.19:285–293, 1976

    PubMed  Google Scholar 

  17. Raisz, L.G., Sandberg, A.L., Goodson, J.M., Simmons, H.A., Mergenhagen, S.E.: Complement-dependent stimulation of prostaglandin synthesis and bone resorption, Science185:789–791, 1974

    PubMed  Google Scholar 

  18. Tashjian, A.H., Jr., Voelkel, E.F., Goldhaber, P., Levine, L.: Prostaglandins, calcium metabolism and cancer, Fed. Proc.33:81–86, 1974

    PubMed  Google Scholar 

  19. Robertson, R.P., Baylink, D.J.: Hypercalcemia induced by prostaglandin E2 in thyroparathyroidectomized, but not intact rats, Prostaglandins,13:1141–1145, 1977

    PubMed  Google Scholar 

  20. Roos, B.A., Bundy, L.L., Bailey, R., Deftos, L.J.: Calcitonin secretion in vitro. I. Preparation of monolayer C-cell cultures. Endocrinology95:1142–1149, 1974

    PubMed  Google Scholar 

  21. Roos, B.A., Deftos, L.J.: Radioimmunoassay of calcitonin in plasma normal thyroid, and medullary thyroid carcinoma of the rat, J. Lab. Clin. Med.88:173–182, 1976

    PubMed  Google Scholar 

  22. Malmström, K., Carafoli, E.: Effects of prostaglandins on the interaction of Ca2+ with mitochondria, Arch. Biochem. Biophys.171:418–423, 1975

    PubMed  Google Scholar 

  23. Carsten, M.E., Miller, J.D.: Effects of prostaglandins and oxytocin on calcium release from a uterine microsomal fraction. Hypothesis for ionophoretic action of prostaglandins, J. Biol. Chem.252:1576–1581, 1977

    PubMed  Google Scholar 

  24. Majeska, R.J., Wuthier, R.E.: Studies on matrix vescles isolated from chick epiphyseal cartilage, Biochim, Biophys. Acta391:51–60, 1975

    Google Scholar 

  25. Newcombe, D.S., Ishikawa, Y.: The effect of anti-inflammatory agents on human synovial fibroblast prostaglandin synthetase, Prostaglandins12:849–869, 1976

    PubMed  Google Scholar 

  26. 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

    PubMed  Google Scholar 

  27. Wuthier, R.E.: A zonal analysis of inorganic and organic constitutents of the epiphysis during endochondral calcification, Calcif. Tissue Res.4:20–38, 1969

    PubMed  Google Scholar 

  28. Sigma Technical Bulletin No. 104 (10–74), Sigma Chemical Co., St. Louis, Mo.

  29. Hamberg, M., Samuelsson, B.: Prostaglandins in human seminal plasma. J. Biol. Chem.241:257–263, 1966

    PubMed  Google Scholar 

  30. Pace-Asciak, C.R.: Catecholamine-induced increase in prostaglandin E biosynthesis in homogenates of the rat stomach fundus, Adv. Biosci.9:23–34, 1973

    Google Scholar 

  31. Flower, R.J.: Drugs which inhibit prostaglandin biosynthesis, Pharmacol. Rev.26:33–67, 1974

    PubMed  Google Scholar 

  32. Lands, W.E.M., LeTellier, P.R., Rome, L.H., Vanderhoek, J.Y.: Inhibition of prostaglandin biosynthesis, Biosci.9:15–28, 1973

    Google Scholar 

  33. Panganamala, R.V., Miller, J.S., Gwebu, E.T., Sharma, H.M., Cornwell, D.G.: Differential inhibitory effects of vitamin E and other antioxidants on prostaglandin synthetase, platelet aggregation and lipoxidase, Prostaglandins14:261–271, 1977

    PubMed  Google Scholar 

  34. Christ, E.J., Van Dorp, D.A.: Comparative aspects of prostaglandin biosynthesis in animal tissues, Biochim. Biophys. Acta270:537–545, 1972

    Google Scholar 

  35. Egan, R.W., Paxton, J., Kuehl, F.A., Jr.: Mechanism for irreversible self-deactivation of prostaglandin synthetase, J. Biol. Chem.251:7329–7335, 1976

    PubMed  Google Scholar 

  36. Yoshimoto, A., Ito, H., Tomita, K.: Cofactor requirements of the enzyme synthesizing prostaglandins in bovine seminal vesicles, J. Biochem.68:487–499, 1970

    PubMed  Google Scholar 

  37. Thomas, D.R., Philpott, G.W., Jaffe, B.M.: The relationship between concentration of prostaglandin E and rates of cell replication. Exp. Cell Res.84:40–46, 1974

    PubMed  Google Scholar 

  38. Brighton, C.T., Hunt, R.M.: Histochemical localization of calcium in growth plate mitochondria and matrix vesicles, Fed. Proc.35:143–147, 1976

    PubMed  Google Scholar 

  39. Eisenbarth, G., Beuttel, S., Lebovitz, H.E.: Inhibition of cartilage macromolecular synthesis by prostaglandin A1, J. Pharmacol. Exp. Ther.189:213–220, 1974

    PubMed  Google Scholar 

  40. Raisz, L.G., Koolesman-Beynen, A.: Inhibition of bone collagen synthesis by PGE2 in organ culture, Prostaglandins8:377–385, 1974

    PubMed  Google Scholar 

  41. Malemüd, C.J., Sokoloff, L.: The effect of prostaglandins on cultured lapine articular chondrocytes, Prostaglandins13:845–860, 1977

    PubMed  Google Scholar 

  42. Chang, W.C., Abe, M., Murota, S.: Stimulation by PGF2α of acidic glycosaminoglycans produced in cultured fibroblasts, Prostaglandins13:55–60, 1977

    PubMed  Google Scholar 

  43. Cottee, F., Flower, R.J., Moncada, S., Salmon, J.A., Vane, J.R.: Synthesis of 6-keto-PGF1α by ram seminal vesicle microsomes, Prostaglandins14:413–423, 1977

    PubMed  Google Scholar 

  44. Needleman, P., Minkes, M., Raz, A.: Thromboxanes: Selective biosynthesis and distinct biological properties, Science193:163–165, 1976

    PubMed  Google Scholar 

  45. Moncada, S., Bunting, S., Mullane, K., Thorogood, P., Vane, J.R., Raz, A., Needleman, P.: Imidazole: A selective inhibitor of thromboxane synthetase, Prostaglandins13:611–618, 1977

    PubMed  Google Scholar 

Download references

Author information

Author notes

  1. F. K. Northington

    Present address: Department of Biochemistry, Medical University of South Carolina, 29401, Charleston, South Carolina

Authors and Affiliations

  1. Department of Chemistry, University of South Carolina, 29208, Columbia, South Carolina

    F. K. Northington, T. D. Oglesby, Y. Ishikawa & R. E. Wuthier

Authors
  1. F. K. Northington
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. D. Oglesby
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y. Ishikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. E. Wuthier
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Northington, F.K., Oglesby, T.D., Ishikawa, Y. et al. Localization of prostaglandin synthetase in chicken epiphyseal cartilage. Calc. Tis Res. 26, 227–236 (1978). https://doi.org/10.1007/BF02013263

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02013263

Key words

Search

Navigation