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The intact human fibroblast has been used in clinical and basic research studies of receptor-mediated control of cell function, however there is little information about the relationship between muscarinic receptor density and the regulation of cyclic AMP (cAMP) accumulation. We have compared the muscarinic receptor characteristics of both lung and skin intact fibroblasts at fetal and adult stages of development using carbachol-mediated inhibition of cAMP accumulation and the binding of [3H]quinuclidinyl benzilate (QNB). Prostaglandin E1 (PGE1) stimulated cAMP accumulation in all four cell lines, while carbachol inhibited cAMP accumulation only in the fetal lung, adult lung, and to a lesser extent, the fetal skin. Adult skin fibroblasts did not display significant evidence of inhibitory muscarinic receptor activity. [3H]QNB binding was saturable for the fetal and adult lung, and the fetal skin, yielding Kd values of approximately 0.5 nM for these cell lines. Bmax values were 360 fmol/mg for fetal skin, 600 fmol/mg for adult lung, and 876 fmol/mg for the fetal lung. Specific binding of [3H]QNB to adult skin fibroblasts was found to be low and variable. Comparisons of the Bmax values and maximal inhibitory capacities showed that the receptor density paralleled receptor activity in all cell lines. The lack of muscarinic receptor activity in the adult skin fibroblast was confirmed in several different adult skin cell lines, indicating that the adult skin fibroblast may not be an appropriate model for muscarinic receptor activity in clinical investigations.

Muscarinic receptors on intact human fibroblasts. Absence of receptor activity in adult skin cells.