Fluorescent histochemical localization of neutral endopeptidase-24.11 (enkephalinase) in the rat spinal cord

Stephen Back, C. Gorenstein

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The localization of neutral endopeptidase-24.11 (E.C. 3.4.24.11; enkephalinase) in rat spinal cord was investigated by a novel fluorescent histochemical method. Enkephalinase was localized by using a coupled enzyme assay based upon the sequential cleavage of the synthetic peptide substrate glutaryl-ala-ala-phe-4-methoxy-2-naphthylamide by enkephalinase and exogenous aminopeptidase M. Enzyme distribution was examined in segments from cervical, thoracic, lumbar, and sacral cord. At all spinal cord levels, enkephalinase was localized to discrete regions of the gray matter. The substantia gelatinosa displayed rich enkephalinase staining which overlapped the inner and outer zones of lamina II. A staining pattern similar to that observed in lamina II was observed in the spinal trigeminal nucleus in the medulla. In lamina III the enzyme was associated with small and medium-sized cells. Lamina IV showed staining associated with medium-sized and large cell bodies. The medial boundary of the dorsal gray of laminae IV and V had medium-sized fusiform cells which stained for enkephalinase. In the lateral reticulated areas of lamina V, enkephalinase reaction product was localized to scattered medium-sized and large cells compressed against the white matter of axons bundles. Staining in lamina VI was similar in appearance to lamina V. Enkephalinase reaction product was widely distributed in the ventral horn. Numerous ventral horn motor neurons of varied size and morphology in laminae VIII and XI stained richly for the enzyme. The enzyme was also localized to medium-sized and large cells in lamina X and to cells of the central cervical nucleus. The size and morphology of the cell types associated with the enzyme supported a neuronal association for enkephalinase. The regional distribution of the enzyme overlapped that of enkephalin- and substance-P rich regions of the spinal cord. These findings support a role for enkephalinase in the metabolic regulation of centrally acting neuropeptides.

Original languageEnglish (US)
Pages (from-to)436-450
Number of pages15
JournalJournal of Comparative Neurology
Volume280
Issue number3
StatePublished - 1989
Externally publishedYes

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Neprilysin
Spinal Cord
Substantia Gelatinosa
Enzymes
Staining and Labeling
Spinal Trigeminal Nucleus
Anterior Horn Cells
CD13 Antigens
Enkephalins
Enzyme Assays
Motor Neurons
Horns
Substance P
Neuropeptides
Cell Size
Axons

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Fluorescent histochemical localization of neutral endopeptidase-24.11 (enkephalinase) in the rat spinal cord. / Back, Stephen; Gorenstein, C.

In: Journal of Comparative Neurology, Vol. 280, No. 3, 1989, p. 436-450.

Research output: Contribution to journalArticle

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abstract = "The localization of neutral endopeptidase-24.11 (E.C. 3.4.24.11; enkephalinase) in rat spinal cord was investigated by a novel fluorescent histochemical method. Enkephalinase was localized by using a coupled enzyme assay based upon the sequential cleavage of the synthetic peptide substrate glutaryl-ala-ala-phe-4-methoxy-2-naphthylamide by enkephalinase and exogenous aminopeptidase M. Enzyme distribution was examined in segments from cervical, thoracic, lumbar, and sacral cord. At all spinal cord levels, enkephalinase was localized to discrete regions of the gray matter. The substantia gelatinosa displayed rich enkephalinase staining which overlapped the inner and outer zones of lamina II. A staining pattern similar to that observed in lamina II was observed in the spinal trigeminal nucleus in the medulla. In lamina III the enzyme was associated with small and medium-sized cells. Lamina IV showed staining associated with medium-sized and large cell bodies. The medial boundary of the dorsal gray of laminae IV and V had medium-sized fusiform cells which stained for enkephalinase. In the lateral reticulated areas of lamina V, enkephalinase reaction product was localized to scattered medium-sized and large cells compressed against the white matter of axons bundles. Staining in lamina VI was similar in appearance to lamina V. Enkephalinase reaction product was widely distributed in the ventral horn. Numerous ventral horn motor neurons of varied size and morphology in laminae VIII and XI stained richly for the enzyme. The enzyme was also localized to medium-sized and large cells in lamina X and to cells of the central cervical nucleus. The size and morphology of the cell types associated with the enzyme supported a neuronal association for enkephalinase. The regional distribution of the enzyme overlapped that of enkephalin- and substance-P rich regions of the spinal cord. These findings support a role for enkephalinase in the metabolic regulation of centrally acting neuropeptides.",
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