Epiphyseal cartilage cAMP changes produced by electrical and mechanical perturbations.

Epiphyseal cartilage from chick embryo was subjected in vitro for one to 15 minutes to static compressive forces or oscillating electric fields. The same stimuli were applied to suspensions of cells isolated from the epiphyses. At the end of the perturbation cAMP was measured in the tissue or cell extracts by radioimmunoassay.

Evidence for preferential effects of parathyroid hormone, calcitonin and adenosine on bone and periosteum.

In order to explore the distribution of hormone-responsive cells in skeletal tissues, we have examined the effects of synthetic bovine parathyroid hormone N-terminal peptide (bPTH 1-34) and salmon calcitonin (sCT) on cyclic AMP levels in periosteum-free rat calvaria, segments of periosteum, and in isolated cells dispersed from each tissue by collagenase digestion. Synthetic bovine PTH increased cyclic AMP levels to a greater degree in calvaria and in isolated bone cells than in the periosteal segments and cells, whereas sCT was more effective in the periosteal than in the bone systems. Primary cultures prepared from bone and periosteal cell populations exhibited progressive increases in their responsiveness to bPTH (1-34) and progressive decreases in responsiveness to sCT.

Membrane changes during cartilage maturation. Increase in 5'-nucleotidase and decrease in adenosine inhibition of adenylate cyclase.

To examine the potential participation of the plasma membrane in differentiation, we studied the enzymatic activities of 5'-nucleotidase and adenylate cyclase as a function of chondrocyte maturation. 16-day-old chick embryo tibiae epiphyses were dissected into proliferative, growing and hypertrophying zones. Partially purified membrane fractions prepared by differential centrifugation from the respective tissue segments were assayed for enzymatic activity.

The role of calcium in the inhibition of cAMP accumulation in epiphyseal cartilage cells exposed to physiological pressure.

A hydrostatic pressure of 60g/cm sq (0.85 psi) inhibits the accumulation of cAMP in cells isolated from the proliferative zone of chick-tibia epiphyseal cartilage. The following findings indicate that this effect is mediated by a translocation of calcium: (i) the pressure enhances the cellular uptake of radiocalcium; (ii) the pressure effect on cAMP can be simulated by the calcium-ionophore A23187; (iii) the effects of pressure and A23187 are non-additive; (iv) the pressure effect is not produced in the presence of ethylenebis-(oxyethylenenitrilo)-pressure effect is not produced in the presence of ethylenebis-(oxyethylenenitrilo)-tetraacetic-acid (EGTA); (v) the particulate adenyl cyclase activity of the proliferative zone is susceptible to non-competitive calcium inhibition.

Interrelationships between Ca2+ and adenylate and guanylate cyclases in the control of platelet secretion and aggregation.

Ca2+ is a powerful inhibitor (Ki is congruent to 16 muM) of basal and prostaglandin E1 (PGE1)-stimulated adenylate cyclase [ATP pyrophosphate-lyase (cyclizing); EC 4.6.1.

Demonstration of adenylate cyclase activity in human red blood cell ghosts.

The presence of adenylate cyclase (ATP pyrophosphate-lyase (cyclizing) EC 4.6.1.

Cyclic AMP and cyclic GMP: mediators of the mechanical effects on bone remodeling.

Compressive forces of physiological magnitude (60 grams per square centimeter) reduce the adenosine 3',5'-monophosphate and guanosine 3',5'-monophosphate content of the epiphyses of tibiae from 16-day-old chick embryos. An equivalent hydrostatic pressure applied directly to cells isolated from this tissue also affects cyclic nucleotide accumulation. The tissue response is uniform throughout the epiphysis, whereas the cell response varies according to the area of origin.

A quantitative method for the application of compressive forces to bone in tissue culture.

A quantitative method for the application of compressive forces to long bones in culture is described. Using this method it was found that a physiological pressure of 80g/cm sq. applied to tibiae of 16-day-old chick embryos reduced glucose consumption to 50% of controls.