Recently, motor neurones of the myenteric plexus innervating the muscle layers or the mucosa have been identified in the guinea-pig stomach. We applied the neuronal tracer DiI (1,1'-didodecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorat) onto myenteric ganglia in order to identify populations of interneurones in the myenteric plexus of the guinea-pig stomach. The tracing was combined with the immunohistochemical detection of calbindinD28k (CALB), choline acetyltransferase (ChAT), neuropeptide Y (NPY) and 5-hydroxytryptamine (serotonin) (5-HT) and the results were compared to the neurochemical coding of target specific motor neurones. Long projecting ( approximately 5.4 mm) ChAT/CALB/+/-5-HT-, nitric oxide synthase (NOS)/CALB- and short projecting ( approximately 1.1 mm) ChAT/NPY-neurones were identified as descending interneurones. CALB positive ascending interneurones contained ChAT but rarely 5-HT (code: ChAT/CALB). This study identified ascending and descending interneurones in the gastric myenteric plexus and revealed the neurochemical coding of some of the interneurone populations.