Membrane peptidases in the peripheral nervous system of the pig: their localization by immunohistochemistry at light and electron microscopic levels.
The presence and cellular localization of five membrane peptidases has been investigated in peripheral nerves, including those of the autonomic nervous system, in the pig. Endopeptidase-24.11 ("enkephalinase") peptidyl dipeptidase A, aminopeptidase N, aminopeptidase W and dipeptidyl peptidase IV were studied by both enzymic assays of membranes prepared from samples of nerve and by immunoperoxidase histochemistry at light and in two cases, endopeptidase-24.11 and aminopeptidase W, at electron microscopic levels. All five peptidases could be quantified by enzymic assay, though the activities were about 1% of those in renal microvilli and less than those of choroid plexus membranes. Endopeptidase-24.11 was associated with Schwann cell membranes in all types of nerve examined, including major nerves containing predominantly myelinated fibres as well as autonomic nerves, such as the vagus and splenic nerves and the sympathetic chain, staining being observed in membranes associated with myelinated and unmyelinated fibres. The Schwann cell location of endopeptidase-24.11 was confirmed by correlation with immunostaining for glial fibrillary acidic protein and by electron microscopy. This peptidase is known to have a wide repertoire of susceptible substrates among neuropeptides which was here shown to include vasoactive intestinal polypeptide (Km 268 microM, kcat 568 min-1), one of a number of neuropeptides present in peripheral nerve fibres. Three of the peptidases, peptidyl dipeptidase A, aminopeptidase N and dipeptidyl peptidase IV, were associated with microvessels of peripheral nerves. Aminopeptidase N was also observed in connective tissue elements, including the perineurium. Aminopeptidase W was unique among the five peptidases in having a neuronal localization. This was observed in unmyelinated and myelinated nerves and was supported by comparison with the pattern of staining observed for neurofilament protein and by electron microscopic immunoperoxidase staining. This observation was unexpected since aminopeptidase W has not been detected as a neuronal marker in the brain. Some possible roles for the membrane peptidases in peripheral nerves are discussed.
Barnes
K
K
Department of Biochemistry, University of Leeds, U.K.
Bourne
A
A
Cook
P A
PA
Turner
A J
AJ
Kenny
A J
AJ
D000818:Animals; D001917:Brachial Plexus / Q000201:enzymology; D020689:Exopeptidases; D005267:Femoral Nerve / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D008565:Membrane Proteins / Q000032:analysis*; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009479:Neuropeptides / Q000378:metabolism; D010447:Peptide Hydrolases / Q000032:analysis*; D010525:Peripheral Nerves / Q000201:enzymology* / Q000648:ultrastructure; D013552:Swine / Q000378:metabolism*
D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D020689:Exopeptidases; D010447:Peptide Hydrolases