The secretion of peptide products derived from pro-ACTH/endorphin was examined with several radioimmunoassays and with polyacrylamide gel analyses of immunoprecipitates of radioactively labeled peptides . In studies using a mouse pituitary tumor cell line the accumulation of each of the four molecular forms of adrenocorticotropic hormone (ACTH) in tissue culture medium was shown to be a linear function of time . No evidence for selfinhibition of secretion by accumulated, secreted peptides (i .e ., ultra-short feedback) was found . Furthermore, synthetic human ACTH and synthetic camel /3-endorphin did not alter secretion of peptides when added to the culture medium at levels up to 10,000 times physiological . Stimulation of the release of ACTH-, endorphin-, lipotropin-, and 16k fragment immunoreactive material by norepinephrine was fully blocked by cobalt ; by this criterion, stimulated release was calcium dependent. All the smaller molecules derived from the pro-ACTH/endorphin common precursor were secreted in equimolar amounts under all circumstances tested, within the precision of these studies (±11%) . Norepinephrine and cobalt did not significantly alter the secretion of pro-ACTH/endorphin and ACTH biosynthetic intermediate . The stimulation of secretion by norephinephrine and inhibition of secretion by cobalt was restricted to the lower molecular weight products derived from pro-ACTH/endorphin : glycosylated and nonglycosylated ACTH(1-39) ; a-lipotropin,,-endorphin, and y-lipotropin ; and 16k fragment .The AtT-20/D-16v mouse pituitary tumor cell line, which was derived from anterior pituitary tissue, provided the first system in which the common precursor to adrenocorticotropic hormone (ACTH)' and /3-lipotropin (#LPH)/,B-endorphin was demonstrated (38) . There is now considerable support for the existence of a similar common precursor to ACTH-related and endorphin-related peptides (pro-ACTH/endorphin) in many species and in several different tissues (reviewed in references 5, 16, and 22). The ACTH/endorphin biosynthetic pathway that operates in AtT20/D-16v cells is summarized in Fig. 1 . 'Abbreviations used in thispaper: ACTH, adrenocorticotropic hormone or corticotropin; 8LPH, /3-lipotropin; yLPH, y-lipotropin or roughly the NH 2-terminal two-thirds of /8LPH; aMSH ; a-melanotropin or Nacetyl-ACTH(1-13)NH2 ; CLIP, corticotropin-like intermediate lobe peptide or ACTH(18-39) ; 16k fragment, the NH2-terminal non-ACTH, non-/3LPH region of pro-ACTH/endorphin, containing a melanotropin-like region called yMSH; 31k, 16k, etc., molecules with apparent molecular weights of 31,000, 16,000, etc., when determined by SDS polyacrylamide gel electrophoresis using a borate/acetate buffer system.