The aim of this work was to characterize the effect of experimental diabetes on neurotransmission in rat vagina. Female Sprague ± Dawley rats were divided into two groups: non-diabetic controls (NDM, n 38) and diabetics (DM, n 38). DM was produced by intraperitoneal injection of streptozotocin. Eight weeks later the animals were killed, the distal part of the vagina was removed, and smooth muscle strips were prepared for functional organ bath experiments and for measurement of nitric oxide synthase (NOS) activity. In DM preparations, the EC 50 value for noradrenaline (NA) was signi®cantly increased (P`0.05) and the maximal contractile response decreased (P 5 0.001). In preparations precontracted with NA, the NO donor SNAP and calcitonin gene-related peptide (CGRP) caused concentration-dependent relaxations, which were signi®cantly decreased (P`0.001) in the DM group. Electrical stimulation of nerves (EFS) caused frequency-dependent contractions, which were signi®cantly lower in DM than in NDM strips (P`0.001). SNAP and CGRP concentration-dependently inhibited EFS evoked contractions in both NDM and DM preparations. The inhibition was signi®cantly lower (P`0.05) in the DM group. In NDM preparations precontracted with NA, EFS evoked frequency-dependent relaxations; such relaxations were inhibited or reduced in DM. Treatment with the NOS inhibitor, L-NOARG 0.1 mM, abolished relaxations in all preparations or produced contraction in DM preparations. Calcium-dependent NOS activity was not signi®cantly different in the DM and NDM groups. However, the DM animals showed a small but signi®cant increase in calcium-independent NOS-activity (P`0.05). Diabetes interferes with adrenergic-, cholinergic-and NANC-neurotransmitter mechanisms in the smooth muscle of the rat vagina. The changes in the nitrergic neurotransmission are not due to reduction in NOS-activity, but seem to be due to interference with later steps in the L-arginineaNOaguanylate cyclaseacGMP system.