Abstract
We tested the hypothesis that cyclic AMP plays a significant role in modulating the growth of embryonic chick cartilage by determining whether cyclic AMP levels change in growing embryonic cartilage and whether cyclic AMP could stimulate embryonic cartilage growth in a long term in vitro organ culture. Cyclic AMP levels were low (0.1 pmol/mg wet wt) in 8-d chick embryo pelvic cartilage, and increased progressively through the 11th d of embryonic development at which time they reached a maximum (1.8 pmol/mg wet weight) and thereafter remained constant. We developed an in vitro organ culture system to determine whether cyclic AMP, a factor known to stimulate radiolabeled precursor incorporation into macromolecules in short-term studies does, in fact, stimulate growth of cartilage. Individual pelvic cartilages were isolated from 9-d chick embryos, placed in serum-free medium (BGJb-FJ modification) and incubated for 3 to 5 d during which time they increased in size (39 and 60% in length, respectively), wet weight (90 and 141%, respectively), and content of total soluble protein (30 and 48%, respectively). N6-monobutyryl cyclic AMP (BtcAMP) added to the medium caused a dose-dependent (0.05 to 1.0 mM) stimulation of growth. After 3 d of incubation, 1.0 mM BtcAMP increased wet weight (125%), [14C]leucine incorporation into protein (75%), and [3H]thymidine incorporation into DNA (48%) compared with control cartilages incubated in medium alone. 1-methyl-3-isobutyl xanthine, a phosphodiesterase inhibitor, also increased cartilage growth above control while sodium butyrate, AMP, and ATP had no effect. Histological examination of cartilage grown in medium was similar to that of cartilage developing in ovo, whereas, cartilage grown in medium containing BtcAMP showed marked hypercellularity with many immature chondrocytes. Our observations are compatible with the hypothesis that cyclic AMP can significantly modulate the growth of embryonic cartilage.
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- Adiga P. R., Murthy P. V., McKenzie J. M. Stimulation by thyrotropin, long-acting thyroid stimulator, and dibutyryl 3',5'-adenosine monophosphate of protein and ribonucleic acid synthesis and ribonucleic acid polymerase activities in porcine thyroid in vitro. Biochemistry. 1971 Feb 16;10(4):702–710. doi: 10.1021/bi00780a023. [DOI] [PubMed] [Google Scholar]
- Audhya T. K., Gibson K. D. Enhancement of somatomedin titers of normal and hypopituitary sera by addition of L-triiodothyronone in vitro at physiological concentrations. Proc Natl Acad Sci U S A. 1975 Feb;72(2):604–608. doi: 10.1073/pnas.72.2.604. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bourret L. A., Goetinck P. F., Hintz R., Rodan G. A. Cyclic 3' ,5'-AMP changes in chondrocytes of the proteoglycan-deficient chick embryonic mutant, nanomelia. FEBS Lett. 1979 Dec 15;108(2):353–355. doi: 10.1016/0014-5793(79)80562-1. [DOI] [PubMed] [Google Scholar]
- Burch W. M., Jr, Lebovitz H. E. Specific nuclear binding of adenosine 3',5'-monophosphate-binding protein complex with subsequent poly(A) RNA synthesis in embryonic chick cartilage. J Clin Invest. 1980 Sep;66(3):532–542. doi: 10.1172/JCI109885. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drezner M. K., Eisenbarth G. S., Neelon F. A., Lebovitz H. E. Stimulation of cartilage amino acid uptake by growth hormone-dependent factors in serum. Mediation by adenosine 3':5'-monophosphate. Biochim Biophys Acta. 1975 Feb 13;381(2):384–396. doi: 10.1016/0304-4165(75)90244-5. [DOI] [PubMed] [Google Scholar]
- Drezner M. K., Lebovitz H. E. Adenosine 3',5'-monophosphate effects on cartilage ribonucleic acid synthesis. Endocrinology. 1979 Feb;104(2):322–328. doi: 10.1210/endo-104-2-322. [DOI] [PubMed] [Google Scholar]
- Drezner M. K., Neelon F. A., Lebovitz H. E. Stimulation of cartilage macromolecule synthesis by adenosine 3',5'-monophosphate. Biochim Biophys Acta. 1976 Apr 2;425(4):521–531. doi: 10.1016/0005-2787(76)90016-2. [DOI] [PubMed] [Google Scholar]
- Eisenbarth G. S., Beuttal S. C., Lebovitz H. E. Inhibition of cartilage macromolecular synthesis by prostaglandin A. J Pharmacol Exp Ther. 1974 Apr;189(1):213–220. [PubMed] [Google Scholar]
- Feinglos M. N., Drezner M. K., Lebovitz H. E. Measurement of plasma adenosine 3',5'-monophosphate. J Clin Endocrinol Metab. 1978 May;46(5):824–829. doi: 10.1210/jcem-46-5-824. [DOI] [PubMed] [Google Scholar]
- Guidotti A., Chuang D. M., Hollenbeck R., Costa E. Nuclear translocation of catalytic subunits of cytosol cAMP-dependent protein kinase in the transsynaptic induction of medullary tyrosine hydroxylase. Adv Cyclic Nucleotide Res. 1978;9:185–197. [PubMed] [Google Scholar]
- Klagsbrun M., Langer R., Levenson R., Smith S., Lillehei C. The stimulation of DNA synthesis and cell division in chondrocytes and 3T3 cells by a growth factor isolated from cartilage. Exp Cell Res. 1977 Mar 1;105(1):99–108. doi: 10.1016/0014-4827(77)90155-0. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lebovitz H. E., Eisenbarth G. S. Hormonal regulation of cartilage growth and metabolism. Vitam Horm. 1975;33:575–648. doi: 10.1016/s0083-6729(08)60973-5. [DOI] [PubMed] [Google Scholar]
- Phillips L. S., Vassilopoulou-Sellin R. Somatomedins (second of two parts). N Engl J Med. 1980 Feb 21;302(8):438–446. doi: 10.1056/NEJM198002213020805. [DOI] [PubMed] [Google Scholar]
- Russell D. H. Type I cyclic AMP-dependent protein kinase as a positive effector of growth. Adv Cyclic Nucleotide Res. 1978;9:493–506. [PubMed] [Google Scholar]
- Ryan W. L., Heidrick M. L. Role of cyclic nucleotides in cancer. Adv Cyclic Nucleotide Res. 1974;4(0):81–116. [PubMed] [Google Scholar]
- Sharma S. K., Talwar G. P. Action of cyclic adenosine 3',5'-monophosphate in vitro on the uptake and incorporation of uridine into ribonucleic acid in ovariectomized rat uterus. J Biol Chem. 1970 Apr 10;245(7):1513–1519. [PubMed] [Google Scholar]
- Solursh M., Reiter R., Ahrens P. B., Pratt R. M. Increase in levels of cyclic AMP during avian limb chondrogenesis in vitro. Differentiation. 1979;15(3):183–186. doi: 10.1111/j.1432-0436.1979.tb01049.x. [DOI] [PubMed] [Google Scholar]
- Tell G. P., Cuatrecasas P., Van Wyk J. J., Hintz R. L. Somatomedin: inhibiton of adenylate cyclase activity in subcellular membranes of various tissues. Science. 1973 Apr 20;180(4083):312–315. doi: 10.1126/science.180.4083.312. [DOI] [PubMed] [Google Scholar]
- Whitfield J. F., Boynton A. L., MacManus J. P., Sikorska M., Tsang B. K. The regulation of cell proliferation by calcium and cyclic AMP. Mol Cell Biochem. 1979 Nov 1;27(3):155–179. doi: 10.1007/BF00215364. [DOI] [PubMed] [Google Scholar]
- Wilson B. D., Wright R. L. Mechanism of TSH action: effects of dibutyryl cyclic AMP on RNA synthesis in isolated thyroid cells. Biochem Biophys Res Commun. 1970 Oct 9;41(1):217–224. doi: 10.1016/0006-291x(70)90491-2. [DOI] [PubMed] [Google Scholar]

