An immunohistochemical study of endopeptidase-24.11 ("enkephalinase") in the pig nervous system

Abstract

Endopeptidase-24.11, a plasma membrane ectoenzyme with the ability to hydrolyse a variety of neuropeptides, has been localized in the pig nervous system by an immunoperoxidase technique. The endopeptidase was mapped in cryostat sections of the fore and mid-brain to the following structures: caudate-putamen, globus pallidus, olfactory tubercle, nucleus interpeduncularis and substantia nigra. Endopeptidase-24.11-like immunoreactivity was also found in the pia mater, choroid plexus and ependymal lining of the central canal. In the spinal cord, weak staining was observed in the dorsal horn, but strong staining was found in the dorsal root ganglia and nerve roots. Within the central nervous system, endopeptidase immunoreactivity was confined to gray matter and within the positive areas of the striatum densely staining areas, corresponding to striosomes, were discernible. These well-defined structures were exploited in serial sections to examine the alignment of the enzyme-rich patches of neuropil with correspondingly strong staining for other antigens. A consistent match was observed with a monoclonal antibody to neurofilament protein, but there was a poor correlation with a polyclonal antibody to glial fibrillary acidic protein. Substance P-like and [Leu]enkephalin-like immunoreactivity were also studied in sections adjacent to those stained for the endopeptidase. Good matching between enzyme-rich and peptide-rich areas was observed, but some enkephalin-rich areas did not align with enzyme staining and indeed endopeptidase-rich areas were not necessarily matched with areas rich in either peptide. These findings suggest a neuronal rather than an astrocytic location for endopeptidase-24.11 in the CNS and lend support to the view that it plays a central role in neuropeptide metabolism at membrane surfaces. In the peripheral nervous system, the endopeptidase was located in Schwann cell membranes surrounding dorsal root ganglion cells and nerve fibres, while in the pituitary the main concentration was in the adenohypophysis, where only a proportion of the endocrine cells were found to be immunoreactive.