Parallel increase in secretory activity between N-glycosylated and nonglycosylated α-amylase without protein synthesis after short-term β-adrenergic receptor activation in isolated rat parotid acinar cells.

Abstract

Objective: To investigate comparisons of the secretory activity between N-glycosylated and nonglycosylated α-amylase, and α-amylase synthetic activity, after β-adrenergic receptor activation in rat parotid acinar cells in vitro.

Study design: Rat parotid acinar cells were incubated in the presence or absence of (-)-isoproterenol. For β-adrenergic blocking experiments, acinar cells were pretreated with (±)-propranolol prior to adding agonist. After the time indicated, the "released amylase" and "total amylase" were obtained. Western blotting was applied to identify and quantify the N-glycosylated and nonglycosylated α-amylase. Amylase activity was also measured.

Results: The potent β-adrenergic agonist (-)-isoproterenol induced a dramatic increase (2-3-fold) of α-amylase secretion for 30 minutes (p < 0.05 vs. control), while the effect was completely abolished when cells were pretreated with (±)-propranolol for 15 minutes. Moreover, the N-glycosylated level of released and total amylase among groups was measured accordingly. Our data showed the N-glycosylated level ratios (released amylase/total amylase) did not differ among groups, which indicated that the N-glycosylated form of α-amylase was not secreted more easily than the nonglycosylated one after stimulation. Interestingly, the total amylase concentration remained unchanged after stimulation within 30 minutes, which might indicate no α-amylase synthesized within the time indicated.

Conclusion: Our findings suggest a parallel increase in secretory activity between N-glycosylated and nonglycosylated α-amylase after β-adrenergic receptor activation. There seems to be a dissociation of α-amylase synthesis from secretion within 30 minutes.