Localization of sodium/potassium adenosine triphosphatase in multiple cell types of the murine nervous system with antibodies raised against the enzyme from kidney

• Published in: The Journal of neuroscience Vol. 5; no. 10; pp. 2581 - 2596
• Main Authors: Ariyasu, RG, Nichol, JA, Ellisman, MH
• Format: Journal Article
• Language: English
• Published: Washington, DC Soc Neuroscience 01.10.1985; Society for Neuroscience
• Subjects: Animals; Antibodies - immunology; Biochemistry and metabolism; Biological and medical sciences; Central nervous system; Fundamental and applied biological sciences. Psychology; Histocytochemistry; Immunochemistry; Immunologic Techniques; Kidney - enzymology; Mice; Mice, Inbred Strains; nervous system; Nervous System - cytology; Nervous System - enzymology; Neurons - classification; Neurons - enzymology; Rats; Rats, Inbred Strains; sodium; Sodium-Potassium-Exchanging ATPase - immunology; Sodium-Potassium-Exchanging ATPase - isolation & purification; Sodium-Potassium-Exchanging ATPase - metabolism; Tissue Distribution; Vertebrates: nervous system and sense organs; Vertebrata; Mammalia; Transmission electron microscopy; Rat; Enzyme; Ultrastructure; Rodentia; Na,K-ATPase; Nervous system; Cytochemistry
• ISSN: 0270-6474; 1529-2401
• DOI: 10.1523/jneurosci.05-10-02581.1985

This report describes the development and characterization of a battery of highly specific antibodies to sodium/potassium (Na+ + K+)-ATPase and their use in localizing this enzyme in nervous tissue. The immunolabeling characteristics of polyclonal antibodies and monoclonal antibodies (Schenk, D. B., and H. L. Leffert (1983) Proc. Natl. Acad. Sci. U. S. A. 80: 5281-5285) raised against rat renal (Na+ + K+)-ATPase were compared. The interspecies cross-reactivity of the polyclonal anti-rat antibodies was examined by determining their binding to purified rat, eel, or dog enzyme. The immunostaining characteristics of the IgG fraction of the polyclonal antibody preparations, their affinity-purified derivatives, and the monoclonal antibodies were compared. The results obtained with each of these were similar, providing information about where focal concentrations of the enzyme exist within nervous tissue. The IgG fraction of the polyclonal antibody preparations provided the most sensitive probe, facilitating localization of the (Na+ + K+)-ATPase in the tissue sections from various regions of the nervous system. (Na+ + K+)-ATPase-like immunoreactivity was observed along the plasmalemma of alpha-motor neurons and at the nodal axolemma of myelinated axons from the central or peripheral nervous system. It was determined that the absence of labeling for the enzyme along the paranodal or internodal regions of the axolemma was not an artifact due to a limited accessibility of antibody to these regions. Some central nervous system glial cells demonstrated abundant amounts of plasmalemmal and intracellular (Na+ + K+)-ATPase-like immunoreactivity. These cells were identified as astroglia by positive labeling of cells in serial sections for glial fibrillary acid protein immunoreactivity in the soma and radial processes in optic nerve, or velous processes in the cerebellum. Astrocyte processes overlying the nodal axolemma also stained positively for the enzyme. (Na+ + K+)-ATPase-like immunoreactivity was not observed in association with oligodendroglia cell bodies or their processes forming myelin sheaths. In contrast, the plasmalemma of myelinating Schwann cells showed greatest immunoreactivity in the region of the node of Ranvier. Although a focal concentration of immunoreactivity was observed along node- and paranode-associated regions of Schwann cells, a lower level of uniform staining was noted along the entire Schwann cell surface membrane.