Recent studies have pointed to membrane-bound guanylyl cyclases (GCs) type A and type B in the rat pineal gland, which augment levels of cyclic GMP (cGMP) in response to atrial natriuretic peptide (ANP), brain-type natriuretic peptide (BNP), and C-type natriuretic peptide (CNP). The present report demonstrates for the first time the expression of CNP in the bovine pineal gland. The CNP prohormone transcript (unlike pre-pro-ANP) was found by reverse transcriptase (RT)-PCR in bovine pineal extracts. CNP immunoreactivity (ir) was revealed in a subpopulation of pinealocytes in situ and in nearly all pinealocytes in culture. Electron microscopic immunohistochemical investigations showed the presence of CNP-ir in cytoplasmic vesicles, providing evidence for the potential secretion of this peptide by pineal cells. Furthermore, the CNP receptor (GC-B) and GC-A (receptor for ANP and BNP) were identified by RT-PCR. Although melatonin secretion was unaffected, natriuretic peptides were able to elevate markedly cGMP production in cultured bovine pinealocytes with a rank order of potency of CNP > BNP = ANP. These findings describe a tissue CNP system in the bovine pineal gland and suggest that CNP may be a local auto- or paracrine modulator of pineal function.