The rate of synthesis of secretory proteins increases significantly in rat parotid glands after stimulated discharge of stored proteins. How any difference in the amount of secretory protein discharge affects the rate of subsequent protein synthesis, and whether this post-secretory synthesis is regulated at the level of messenger RNA, was now examined. One group of rats was stimulated to secrete 97% of stored secretory proteins by an intraperitoneal injection of isoproterenol. The other group received a much smaller dose to induce the discharge of about 40% of the proteins. Despite this difference in secretion, the subsequent rates of total protein synthesis, as well as of amylase, were increased to about the same extent. The amylase messenger RNA (mRNA) was identified and quantified by hybridization with a 32P-labelled amylase complementary DNA (cDNA) probe. The amylase mRNA in stimulated and unstimulated rats was of the same molecular size (Northern blot analyses). The amount of amylase mRNA, determined by dot blot analyses, were also increased in stimulated rats, although this increase was not as great as that in the rate of amylase protein synthesis. The implications of this discrepancy concern the possibility that the mechanism of regulation of secretory protein synthesis in parotid glands is at the translational level.