Although prostaglandin (PG) F_2α released from the uterus has been shown to cause regression of the bovine corpus luteum (CL), the neuroendocrine, paracrine, and autocrine mechanisms regulating luteolysis and PGF_2α action in the CL are not fully understood. A number of substances produced locally in the CL may be involved in maintaining the equilibrium between luteal development and its regression. The present study was carried out to determine whether noradrenaline (NA) and nitric oxide (NO) regulate the sensitivity of the bovine CL to PGF_2α in vitro and modulate a positive feedback cascade between PGF_2α and luteal oxytocin (OT) in cows. Bovine luteal cells (Days 8–12 of the estrous cycle) cultured in glass tubes were pre-exposed to NA (10⁻⁵ M) or an NO donor (S-nitroso-N-acetylpenicillamine [S-NAP]; 10⁻⁴ M) before stimulation with PGF_2α (10⁻⁶ M). Noradrenaline significantly stimulated the release of progesterone (P₄), OT, PGF_2α, and PGE₂ (P < 0.01); however, S-NAP inhibited P₄ and OT secretion (P < 0.05). Oxytocin secretion and the intracellular level of free Ca² ([Ca² ]_i) were measured as indicators of CL sensitivity to PGF_2α. Prostaglandin F_2α increased both the amount of OT secretion and [Ca² ]_i by approximately two times the amount before (both P < 0.05). The S-NAP amplified the effect of PGF_2α on [Ca² ]_i and OT secretion (both P < 0.001), whereas NA diminished the stimulatory effects of PGF_2α on [Ca² ]_i (P < 0.05). Moreover, PGF_2α did not exert any additionally effects on OT secretion in NA-pretreated cells. The overall results suggest that adrenergic and nitrergic agents play opposite roles in the regulation of bovine CL function. While NA stimulates P₄ and OT secretion, NO may inhibit it in bovine CL. Both NA and NO are likely to stimulate the synthesis of luteal PGs and to modulate the action of PGF_2α. Noradrenaline may be the factor that is responsible for the limited action of PGF_2α on CL and may be involved in the protection of the CL against premature luteolysis. In contrast, NO augments PGF_2α action on CL and it may be involved in the course of luteolysis.