title: Calcitonin responsiveness and receptor expression in porcine and murine osteoclasts: a comparative study.
abstract: The presence of the calcitonin (CT) receptor is a distinguishing characteristic of osteoclasts; however, species variability exists with respect to functional responsiveness to CT. In the present study, CT responsiveness and temporal expression of the CT receptor in differentiating cultures of porcine osteoclasts was examined and compared to murine osteoclasts. In vitro porcine osteoclast differentiation was evaluated using bone marrow cultures from neonatal pigs. Murine osteoclast differentiation was studied using cocultures of murine bone marrow and BALC cells, a calvarial-derived cell line. In the presence of 1,25 (OH)2D3, a time-dependent increase in osteoclast differentiation was observed in porcine and murine cultures. Salmon CT (sCT) and porcine CT (pCT) inhibited 1,25 (OH)2D3-stimulated porcine osteoclast differentiation at 10(-8) and 10(-7) mol/L (60% with 10(-7) mol/L sCT and 85% inhibition with 10(-7) mol/L pCT). Treatment of murine cocultures with sCT (10(-17)-10(-7) mol/L) resulted in a concentration-dependent decrease in osteoclast differentiation with a maximal inhibition of 70%. Osteoclast differentiation was inhibited in a concentration-dependent manner by recombinant human transforming growth factor-beta1 (rhTGF-beta1) in both species. The effects of CT on resorption lacunae formation were determined by culturing in vitro generated porcine or murine osteoclasts on bovine cortical bone slices for 18 h in the presence or absence of CT. With both porcine and murine osteoclasts, a concentration-dependent decrease in resorption lacunae formation was observed between 10(-13) and 10(-7) mol/L sCT with the highest concentrations completely abolishing resorption. However, pCT only inhibited porcine osteoclastic resorption at 10(-7) mol/L. CT receptor messenger ribonucleic acid (mRNA) expression was determined at different time points during in vitro osteoclast differentiation. In porcine cultures, expression of CT receptor mRNA correlated with the presence of osteoclasts. In murine cocultures, mRNA for the CT receptor was observed at each time point examined and was independent of the presence of multinucleated osteoclasts. Thus, porcine and murine differentiating osteoclast cultures express CT receptor mRNA; however, receptor expression correlates with osteoclast formation only in the porcine cultures. In summary, porcine and murine osteoclasts express CT receptor mRNA and functional responsiveness to CT. These findings suggest that the effects of sCT on osteoclast resorption are similar in murine and porcine cells, but that sCT is a less potent inhibitor of porcine than murine osteoclast differentiation.
authors.affiliation: Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA. galvin_rachelle_j_s@lilly.com
authors.forename: R J
authors.initials: RJ
authors.lastname: Galvin
authors.forename: P
authors.initials: P
authors.lastname: Bryan
authors.forename: M
authors.initials: M
authors.lastname: Venugopalan
authors.forename: D P
authors.initials: DP
authors.lastname: Smith
authors.forename: J E
authors.initials: JE
authors.lastname: Thomas
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D001862:Bone Resorption / Q000517:prevention & control; D002116:Calcitonin / Q000494:pharmacology*; D002117:Calcitriol / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D006801:Humans; D051379:Mice; D010010:Osteoclasts / Q000187:drug effects* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D018003:Receptors, Calcitonin / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D012886:Skull / Q000378:metabolism; D013045:Species Specificity; D013552:Swine; D016212:Transforming Growth Factor beta / Q000494:pharmacology
chemical_list: D012333:RNA, Messenger; D018003:Receptors, Calcitonin; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C028815:salmon calcitonin; D002116:Calcitonin; D002117:Calcitriol