1. As it has been shown that oestrogen enhances the cholinergic muscarinic activity in the central nervous system, we studied sex differences in the response to parasympathetic nervous stimulation in the rat heart using in vivo and in vitro preparations. 2. In in situ perfused, innervated hearts, stimulation of bilateral vagus nerves (15 Hz with 1 mumol/L physostigmine) inhibited sympathetic nerve stimulation (5 Hz) induced noradrenaline release to a greater extent in female than in male rats (54 +/- 5 vs 72 +/- 5% of control). Similarly, vagus nerve stimulation at 1-20 Hz reduced heart rate (HR) more in females than males, and this sex difference became more marked in the presence of physostigmine. The chronotropic effect of vagal stimulation was attenuated after ovariectomy but potentiated after castration when compared with sham-operated controls. In contrast, the muscarinic agonist methacholine reduced neural NA release and HR equally well in both sexes. 3. In anaesthetized rats, reduction in HR and mean arterial pressure by vagus nerve stimulation (1-20 Hz) was more pronounced in females than in males after inhibition of acetylcholinesterase with physostigmine. 4. The results indicate that activation of parasympathetic nerve leads to greater presynaptic and postsynaptic effects in female than in male rat hearts, presumably due to a higher level of acetylcholine release following nerve activation.