title: Ultracytochemical localization of adenylate cyclase and guanylate cyclase in crushed peripheral nerves.
abstract: Cellular and subcellular distribution of adenylate cyclase (AC) and guanylate cyclase (GC) activities in crushed peripheral nerves during regeneration were studied at the electron microscope level. In unlesioned nerves, no AC reaction product could be evidenced, whereas GC was detectable on the plasma membranes of Schwann cells, myelinated and nonmyelinated fibers, and within nonmyelinated axons. At 24 hours after the crush, AC reaction product was found within axonal segments proximal to the zone of the crush in association with mitochondria. At this stage, macrophage-like cells, which probably are transformed Schwann cells, polymorphonuclear leucocytes, and endothelial cells displaying an intense AC reaction product could be detected. On the other hand, at 24 hours after the crush, GC was no longer detectable, except on occasional unlesioned nerve fibers. At 48 hours after the lesion, AC reaction product was no longer detectable within axons, and all AC positivity was associated with plasma membranes of non-neuronal cells, including transformed Schwann cells, occasional macrophages, polymorphonuclear leucocytes, fibroblasts, and elongated cells. As to GC, images similar to those obtained at 24 hours were observed until 48 hours after the crush. From the 7th to the 28th postlesion day, AC activity was localized exclusively to the plasma membranes of fibroblasts and elongated cells. Transformed Schwann cells were no longer detectable, whereas normal Schwann cells and regenerating axons could be seen, and these showed no AC reaction product in analogy to the absence of AC reaction product of unlesioned nerves. During the same period, GC again was detectable on regenerating fibers with the same subcellular localization as that of unlesioned nerves. The present results strongly suggest that starting from the second postcrush day, cells invading the lesioned zone and transformed Schwann cells, all taking part in the formation of the new perineurial tissue, display a high AC activity, which should be taken into account when measuring cyclic adenosine monophosphate (cAMP) levels under these conditions. Also, our data suggest that GC is involved primarily in regeneration processes that occur in crushed peripheral nerves. Thus, the pattern of AC distribution in peripheral unlesioned and lesioned nerves appears to be exactly the opposite of the GC localization examined under similar experimental conditions insofar as nervous fibers are concerned.
authors.affiliation: Department of Experimental Medicine, University of Perugia, Italy.
authors.forename: M G
authors.initials: MG
authors.lastname: Rambotti
authors.forename: A
authors.initials: A
authors.lastname: Spreca
authors.forename: M
authors.initials: M
authors.lastname: Rende
authors.forename: R
authors.initials: R
authors.lastname: Donato
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001369:Axons / Q000201:enzymology / Q000648:ultrastructure; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D006162:Guanylate Cyclase / Q000378:metabolism*; D006651:Histocytochemistry; D008854:Microscopy, Electron; D009410:Nerve Degeneration; D009416:Nerve Regeneration; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values; D012584:Sciatic Nerve / Q000201:enzymology* / Q000502:physiology / Q000648:ultrastructure
chemical_list: D000262:Adenylyl Cyclases; D006162:Guanylate Cyclase