Microvessels have been isolated from goat cerebral cortex and caudate nucleus. The purity of the preparations was assessed by light microscopy and by the high enrichment in the marker enzymes alkaline phosphatase and gamma-glutamyltransferase. Choline acetyltransferase activity was detected in the vascular fractions, being significantly higher in capillaries than in larger vessels. Acetylcholinesterase (AChE)-containing fibers were visualized in vessels of different caliber. Vessels with diameters larger than 70-90 microns showed a network pattern of fibers similar to that of pial arteries. In small vessels (10-70 microns) longitudinal or helical fibers were observed with occasional side-branches that surround the vessel. No AChE staining was visualized in isolated capillaries under light microscopy. This study shows that isolated intracerebral microvessels are suitable preparations for histochemical studies of perivascular nerves. Taken together, the biochemical and histological results are in accordance with a cholinergic innervation of the goat intracerebral vasculature.