The role of cyclic AMP (cAMP) as mediator of ACTH action on interrenal steroidogenesis was evaluated in juvenile coho salmon (Oncorhynchus kisutch). Head kidneys (containing the interrenal cells) were incubated in the absence or presence of putative adenylate cyclase activators (forskolin and cholera toxin), ACTH combined with putative adenylate cyclase inhibitors (hydrolysis-resistant ATP analogs), dibutyryl cyclic (dbc) AMP, dbcGMP, or phosphatidylinositol. The cortisol content of the incubation medium was subsequently determined by radioimmunoassay. Forskolin markedly stimulated cortisol secretion by interrenal cells. Adenylate cyclase inhibitors depressed the steroidogenic response to ACTH. Dibutyryl cAMP, but not dbcGMP, enhanced steroid secretion. Thus, cAMP seems to be an important "second messenger" for ACTH action on salmon interrenal cells. In contrast to findings in mammalian adrenocortical cells, exogenous phosphatidylinositol and cholera toxin failed to stimulate corticosteroid secretion in salmon interrenal cells. However, it was unclear whether these negative findings were an artifact resulting from the use of kidney tissue fragments instead of isolated interrenal cells.