Numerous ganglia or single neurones immunoreactive to protein gene-product 9.5 (PGP) were demonstrated in the chicken ureter. Ganglia were observed in the main nerve trunks accompanying the ureter (400-2,000 cells), in the adventitia (1-45 cells; density; 79 +/- 12 ganglia/cm2; mean +/- S.E.M.), in the circular muscle (1-9 cells; 76 +/- 10 ganglia/cm2) and in the longitudinal muscle (1-8 cells; 232 +/- 41 ganglia/cm2). Most of the PGP-positive neurones in the nerve trunk ganglia (approximately 66%) and in the smooth muscle layers (85%) were encircled by a dense plexus of varicose nerve fibres containing both substance P (SP) and calcitonin gene-related peptide (CGRP). SP-positive somata were rarely observed. Immunogold electron microscopy revealed that SP- and CGRP-immunoreactivity were colocalised in the same dense core vesicles. A strong reduction of SP-positive nerve fibres was observed in organ cultures of the ureter, indicating their extrinsic origin. The fibres might originate from the dorsal root ganglia, where SP and CGRP were colocalised in 20-30% of the neurones. The sensitivity of ureteric neurones to SP and CGRP was investigated in recordings obtained from mechanosensitive nerve fibres with cell bodies located in or adjacent to the ureter (U-G units). The majority (71%) of the U-G units was excited by local application of SP in a dose-dependent manner. The SP-sensitive U-G neurones had higher mechanical thresholds (29 +/- 5 mmHg) as opposed to the SP-insensitive ones (10 +/- 3 mmHg). Repeated applications of high doses of SP to the U-G units resulted in desensitisation and reduced the response to mechanical stimuli. None of the U-G units responded to local application of CGRP, but all U-G units were excited by acetylcholine. The data support the hypothesis that SP-containing primary afferents are involved in the modulation of the activity of ureteric neurons in the chicken.