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  • Title: Effects of a putative prostaglandin E2 antagonist, AH6809, on chondrogenesis in serum-free cultures of chick limb mesenchyme.
    Author: Capehart AA, Biddulph DM.
    Journal: J Cell Physiol; 1991 Jun; 147(3):403-11. PubMed ID: 1648563.
    Abstract:
    In the present study, we have examined the effects of a putative antagonist of prostaglandin E2 (PGE2), AH6809, on chondrogenesis in serum-free cultures of mesenchyme from distal tips of stage 25 chick limb buds in order to test the hypothesis that endogenous PGE2, through receptor-linked adenylate cyclase (AC), initiates differentiation of cartilage in limb mesenchyme. Daily addition of 10(-4) M concentrations of AH6809 produced marked inhibition of chondrogenesis over a 5-day period of cell culture as evaluated by Alcian green binding to cartilage matrix components. Inhibition of chondrogenesis by this compound was further shown to be reversible and treatment of cells with the antagonist limited to periods when chondrocytes had differentiated and were actively secreting cartilage-specific matrix components had little effect. Preincubation of control cells in 10(-4) M concentrations of AH6809 inhibited PGE2-induced activation of AC by greater than 80% without significant (P greater than .05) inhibition of basal activity by the antagonist. Responses to parathyroid hormone, which increased AC activity by 7-fold, and forskolin which increased AC activity by 23-fold in control cells, were also uninhibited by preincubation in AH6809. The results demonstrate that blockade of PGE2-AC linked receptors in prechondrogenic limb mesenchyme inhibits chondrogenesis supporting the hypothesis that endogenous PGE2 concentrations in undifferentiated limb mesenchyme play an initiating role in the differentiation of cartilage.
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