Corticotropin release-inhibiting factor is preprothyrotropin-releasing hormone-(178-199)

ACTH is the major regulator of the body's adaptive response to stress and the physiological stimulus for glucocorticoid secretion. In addition to the known negative feedback regulation of ACTH by glucocorticoids, a hypothalamic corticotropin release-inhibiting factor (CRIF) that inhibits ACTH synthesis and secretion has been postulated, but not identified. We previously reported that transfection of prepro-TRH complementary DNA into the mouse anterior pituitary tumor cell line AtT-20 results in inhibition of basal and corticotropin-releasing hormone (CRH)-stimulated ACTH synthesis and secretion, suggesting that one or more of the cryptic peptides encoded within the prepro-TRH precursor has CRIF activity. To narrow the choice of peptides responsible for CRIF activity, we first deleted specific sequences within the prepro-TRH complementary DNA and transfected these constructs into AtT-20 cells. Deletion of sequences encoding amino acids 119-229 resulted in the loss of CRIF activity. Of the peptides encoded within this region, prepro-TRH-(178-199), a 22-amino acid peptide, inhibited basal and CRH-stimulated ACTH synthesis and secretion in cultured primary anterior pituitary cells. As this peptide is processed from prepro-TRH in vivo, is found in the external zone of the median eminence, and is secreted from hypothalamic slices in vitro, prepro-TRH-(178-199) fulfills the criteria for a physiological CRIF. The significance of TRH and CRIF sharing a common precursor opens new areas of research in the integrated regulation of pituitary-adrenal and pituitary-thyroid functions.