Changes in beta-adrenoceptor binding properties and receptor-cyclase coupling during in vitro maturation of rat...

The loss of beta-adrenergic responsiveness during reticulocyte maturation was studied under tissue culture conditions in a defined cell population from rats. Initial beta-receptor density and receptor-mediated cAMP formation in culture medium (RPMI 1640) exceeded the values obtained in isotonic KC1-buffer by 56 and 120% respectively. During cell cultivation receptor density and hormonal responsiveness decreased rapidly by 40-50% of their initial values within the first 20 h. In the following 3 days the rate of loss varied between 10 and 15%/d. The same time course was observed for the maturation-dependent decrease in forskolin-stimulated cAMP formation. ATP depletion of cultivated cells caused a complete and irreversible loss of cAMP response within 90 min. Our results indicate that cell metabolism regulates the strength of the hormonal response. A defect in adenylate cyclase or in cyclase N-protein interaction seems to be rate-limiting for the functional inactivation of the beta-adrenergic system during reticulocyte maturation.