Summary
The developing chick limb was studied to determine the ability of parathyroid hormone (PTH) and prostaglandin E2 (PGE2) to increase intracellular cyclic AMP (cAMP) during various stages of development. All developmental stages examined (stages 20–21, 24–25, and 26–28) responded to PGE2 when the cells were assayed immediately following the removal of the limbs from the embryos. In contrast, only stage 26–28 limb cells responded to PTH when assayed in a similar manner. The response to PTH was temporally correlated with the appearance of cartilage matrix in vivo. Undifferentiated limb cells were also cultured and assayed at various times for hormone responsiveness. Stage 24–25 high-density cell cultures responded initially to PGE2 but not to PTH. However, by 36 h and in all subsequent itme intervals tested, the response to PTH was significantly greater than that to PGE2. The PTH receptor, in contrast to that of PGE2, was shown to be sensitive to trypsin treatment, but could be regenerated during subsequent cell culture. The majority of the hormoneresponsive cells were found in cartilaginous regions of the limb, and were shown to respond to both hormones in a dose-dependent manner. The PTH-induced cAMP response was affected by low cell density and mouse serum, both of which significantly inhibit the chondrogenic potential of cultured limb cells. These findings are consistent with a temporal correlation between the development of the PTH response and chondrogenesis in vivo.
Similar content being viewed by others
References
Copp, D. H.: Endocrine regulation of calcium metabolism, Annu. Rev. Physiol.32:61–86, 1970
Reynolds, J. J.: Bone turnover, vitamin D and plasma calcium homeostasis. In R. V. Talmage, M. Owen, J. A. Parsons (eds.): Calcium-Regulating Hormones, pp. 254–261, Excerpta Medica, Amsterdam, 1975
Suzuki, F., Yoneda, T., Shimomura, Y.: Calcitonin and parathyroid hormone stimulation of acid mucopolysaccharide synthesis in cultured chondrocytes isolated from growth cartilage, FEBS Lett.70:155–158, 1976
Smith, D. M., Roth, L. M., Johnston, C. C., Jr.: Hormonal responsiveness of adenylate cyclase activity in cartilage, Endocrinology98:242–246, 1976
Chase, L. R., Aurbach, G. D.: The effect of parathyroid hormone on the concentration of adenosine 3′,5′-monophosphate in skeletal tissue in vitro, J. Biol. Chem.245:1520–1526, 1970
Smith, D. M., Johnston, C. C., Jr.: Hormonal responsiveness of adenylate cyclase activity from separated bone cells, Endocrinology95:130–139, 1974
Zull, J. E., Krug, S., Abel, D., Caplan, A. I.: Development of parathyroid hormone and calcitonin-activated adenylate cyclases in embryonic chick limb and in cultured cells from embryonic chicken limbs, Proc. Natl. Acad. Sci. U.S.A.75:3871–3875, 1978
Zwilling, E.: Morphogenetic phases in development. In M. Locke (ed.): 27th Symposium of the Society for Developmental Biology, pp. 184–207. Academic Press, New York, 1968
Caplan, A. I., Rosenberg, M. J.: Control of chondrogenic and myogenic expression in chick limb mesoderm cells. In H. Slavkin, R. Gruelich (eds.): Extracellular Matrix Influences on Gene Expression, pp. 47–55. Academic Press, New York, 1975
Caplan, A. I., Zwilling, E., Kaplan, N. O.: 3-Acetylpyridine: effect in vitro related to teratogenic activity in chicken embryos, Science160:1009–1010, 1968
Lebovitz, H. E., Eisenbarth, G. S.: Hormonal regulation of cartilage growth and metabolism, Vitam. Horm.33:575–648, 1975
Eisenbarth, G. S., Beuttel, S. C., Lebovitz, H. E.: Inhibition of cartilage macromolecular synthesis by prostaglandin A1, J. Pharmacol. Exp. Ther.189:213–220, 1974
Kuehl, F. A., Jr.: Prostaglandins, cyclic nucleotides and cell function, Prostaglandins5:325–334, 1974
Hamburger, V., Hamilton, H. L.: A series of normal stages in the development of the chick embryo, J. Morphol.88:49–92, 1951
Finch, R. A., Parker, C. L., Walton, S. T.: The lack of an inhibitory effect of hyaluronate on chondrogenesis in chick limb-bud mesoderm cells grown in culture, Cell Differ.7:283–293, 1978
Caplan, A. I.: A simplified procedure for preparing myogenic cells for culture, J. Embryol. Exp. Morphol.36:175–181, 1976
Parker, C. L., Paulsen, D. F., Rosebrock J. A., Hooper, W. C.: Inhibition of chondrogenesis by normal mouse serum in cultured chick limb cells, Exp. Cell Res.130:21–30, 1980
Lowry, O. H., Rosebrough, H. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent, J. Biol. Chem.193:265–275, 1951
Gilman, A. G.: A protein binding assay for adenosine 3′,5′-cyclic phosphate, Proc. Natl. Acad. Sci. U.S.A.67:305–312, 1970
Tovey, K. C., Oldham, K. G., Whelan, J. A. M.: A simple direct assay for cyclic AMP in plasma and other biological samples using an improved competitive binding technique, Clin. Chim. Acta56:221–230, 1974
Mallory, F. B.: Pathological Technique, p. 144. W. B. Saunders Co., Philadelphia, 1942
Meyer, K.: The biological significance of hyaluronic acid and hyaluronidase, Physiol. Rev.27:335–359, 1947
Duncan, D. B.: Multiple range and multiple F-tests, Biometrics11:1–11, 1955
Osdoby, P., Caplan, A. I.: The possible differentiation of osteogenic elements in vitro from chick limb mesodermal cells, Dev. Biol.52:283–299, 1976
Caplan, A. I.: Effects of the nicotinamide-sensitive teratogen 3-acetylpyridine on chick limb cells in culture, Exp. Cell Res.62:341–355, 1970
Chase, L.: Selective proteolysis of the receptor for parathyroid hormone in renal cortex, Endocrinology96:70–76, 1975
Biddulph, D. M., Currie, M. G., Wrenn, R. W.: Effects and interactions of parathyroid hormone and prostaglandins on adenosine 3′, 5′-monophosphate concentrations in isolated renal tubules, Endocrinology104:1164–1171, 1979
Wrenn, R. W., Biddulph, D. M.: Parathyroid hormoneinduced calcium efflux from isolated renal cortical tubules: evidence for cyclic AMP mediation, Mol. Cell. Endocrinol.15:29–40, 1979
Zenser, T. V., Craven, P. A., DeRubertis, F. R., Davis, B. B.: Differential inhibition of cyclic AMP and cyclic GMP hydrolysis in rat renal cortex, Arch. Biochem. Biophys.178:598–606, 1977
Dienstman, S. R., Biehl, J., Holtzer, S., Holtzer, H.: Myogenic and chondrogenic lineages in developing limb buds grown in vitro, Dev. Biol.39:83–95, 1974
Ahrens, P. B., Solursh, M., Reiter, R. S.: Stage-related capacity for limb chondrogenesis in cell culture, Dev. Biol.60:69–82, 1977
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Parker, C.L., Biddulph, D.M. & Ballard, T.A. Development of the cyclic AMP response to parathyroid hormone and prostaglandin E2 in the embryonic chick limb. Calcif Tissue Int 33, 641–648 (1981). https://doi.org/10.1007/BF02409502
Issue Date:
DOI: https://doi.org/10.1007/BF02409502