Alternative RNA processing of the calcitonin gene primary transcript results in production of two peptides, calcitonin and calcitonin gene-related peptide (CGRP). We have used the TT cell line, which produces both peptides, to ascertain whether secretion of peptides produced by alternative RNA processing is under identical regulatory control. Short-term treatment of TT cells with phorbol esters and cAMP analogs caused a rapid and parallel release of both calcitonin and CGRP. The measurement of calcitonin and CGRP mRNA levels during treatment revealed that new RNA synthesis was not required for secretion. Four potential regulators of phorbol ester-mediated and five of cAMP-mediated secretion were identified by incorporation of radioactive phosphate into protein as analysed by two-dimensional polyacrylamide gel electrophoresis and autoradiography. From these results we conclude that calcitonin and CGRP secretion in this human C cell model is not differentially affected by alternative RNA processing for the phorbol ester-, and cAMP-dependent secretory pathways.