Localisation of immunoreactive factor VIII, nitric oxide synthase, substance P, endothelin-1 and 5-hydroxytryptamine...

This pre-embedding electron-immunocytochemical study investigated the localisation of endothelial (type III) and neuronal (type I) isoforms of nitric oxide synthase, substance P, endothelin-1 and 5-hydroxytryptamine in the human middle cerebral artery taken up to 40 h postmortem. To ¿recover' from the anoxic period some of the vessels were incubated in oxygenated Krebs solution prior to the immunoprocedure. At this long postmortem time, immunoreactivity to type III and type I nitric oxide synthase, substance P, endothelin-1 and 5-hydroxytryptamine was found in a subpopulation of intact cells present in the vessel intima; immunoreactivity to type I nitric oxide synthase was also observed in a subpopulation of adventitial perivascular nerve fibres. Cultures of the cells from the intima of the postmortem vessels showed that the cells were proliferating and positive immunoreactivity to factor VII identified them as endothelial cells. The results therefore indicate that even after up to 40 h postmortem, endothelium of human middle cerebral artery is immunoreactive for a number of vasoactive agents and perivascular nerve fibres show nitric oxide synthase immunoreactivity.