Localization and characterization of binding sites with high affinity for [3H]ouabain in cerebral cortex of rabbit brain using quantitative autoradiography.

Abstract:

:[3H]Ouabain binding was studied in sections of rabbit somatosensory cortex by quantitative autoradiography and in rabbit brain microsomal membranes using a conventional filtration assay. KD values of 8-12 nM for specific high-affinity binding of [3H]ouabain were found by both methods. High-affinity binding was not uniformly distributed in somatosensory cortex and was localized predominantly to laminae 1, 3, and 4. [3H]Ouabain binding in tissue sections was stimulated by the ligands Mg2+/Pi or Mg2+/ATP/Na+ and was inhibited by K+ (IC50 = 0.7-0.9 mM), N-ethylmaleimide, 5,5'-dithiobis(2-nitrobenzoic acid), and erythrosin B. We conclude that [3H]ouabain is reversibly and specifically bound with high affinity in rabbit brain tissue sections under conditions that favor phosphorylation of Na+,K+-ATPase. Quantitative autoradiography is a powerful tool for assessing the affinity and number of specific ouabain binding sites in brain tissue.

journal_name

J Neurochem

authors

Antonelli MC,Baskin DG,Garland M,Stahl WL

doi

10.1111/j.1471-4159.1989.tb10916.x

subject

Has Abstract

pub_date

1989-01-01 00:00:00

pages

193-200

issue

1

eissn

0022-3042

issn

1471-4159

journal_volume

52

pub_type

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