Effect of phorbol and bryostatin I on chondrogenic expression of chick limb bud, in vitro.
The ability of phorbol esters to promote tumor formation and alter cell differentiation has been attributed to its action on a number of critical cellular functions, in particular, on protein phosphorylation, through the activation of protein C kinase. The present paper describes the effects of PMA (phorbol 12-myristate 13 acetate) on in vitro chondrogenesis in non-passaged, embryonic limb bud cells, relative to the effects of Bryostatin I. This compound also activates C kinase and binds competitively to the phorbol ester receptor, yet does not affect cell differentiation. Levels of PMA as low as 10(-7) M markedly reduced cartilage formation in 4-day cultures, as indicated by nodule count and Alcian blue staining for chondroitin sulfate. Coadministration of Bryostatin I at equimolar concentration prevented the PMA inhibitory effect on chondrocytic expression. This confirms other findings that phorbol activation of C kinase cannot exclusively account for the activity of phorbol on cell expression, i.e., that other pathway(s) must also be involved. Altering the time of PMA exposure demonstrated that PMA inhibited chondrocyte phenotypic expression, rather than cell commitment: early (0-48 h) exposure to PMA (during chondrocytic commitment in vitro) had little inhibitory effect on the staining index, whereas, exposure from 49-96 h (presumably post-commitment) and 0-96 h had moderate and strong inhibitory effects, respectively, on cartilage synthesis. Further research on the phorbol/Bryostatin I interaction should add to our knowledge of the control processes involved in tumor promotion and cell differentiation.
D000818:Animals; D054713:Bryostatins; D002356:Cartilage / Q000166:cytology / Q000187:drug effects / Q000196:embryology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D002809:Chondroitin Sulfates / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D005121:Extremities / Q000196:embryology; D007783:Lactones / Q000494:pharmacology*; D018942:Macrolides; D011493:Protein Kinase C / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013997:Time Factors
D054713:Bryostatins; D007783:Lactones; D018942:Macrolides; C046785:bryostatin 1; D002809:Chondroitin Sulfates; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate