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Autoradiographic visualization and characterization of [3H]ouabain binding to the Na+,K+-ATPase of rat brain and pineal.

Publication ,  Journal Article
Caspers, ML; Schwartz, RD; Labarca, R; Paul, SM
Published in: Brain Res
April 21, 1987

Ouabain binds to the catalytic subunit of Na+,K+-ATPase and specific [3H]ouabain binding can be used as a measure of the number of active enzyme molecules present in a given tissue. Specific [3H]ouabain binding can be demonstrated in frozen, cryostat sections from rat brain and pineal and these sites have the characteristics of Na+,K+-ATPase. Incubations carried out in the absence of ATP or the presence of excess unlabeled ouabain reduces specific binding by greater than or equal to 98%. The addition of K+ or omission of Mg2+ also result in a decrease in specific binding. Strophanthidin, digoxin and digoxigenin displace [3H]ouabain binding with IC50 values of 0.73, 0.48 and 1.4 microM, respectively. Scatchard analyses of specific [3H]ouabain binding in brain sections shows a single class of non-interacting binding sites with an apparent affinity (Kd) of 339 nM and a maximal binding capacity (Bmax) of 34.9 pmol/mg protein. [3H]Ouabain binding is unevenly distributed throughout the brain with the olfactory nuclei, superior colliculus, dentate gyrus, pontine nuclei and pineal gland having a relatively high density of binding sites. The outer layers (1-3) of the cerebral cortex show more labeling than the inner layers (4-6) and most other brain areas have intermediate levels of [3H]ouabain binding sites, whereas white matter has virtually no specific binding. Computer-assisted densitometry was used to measure changes in specific [3H]ouabain binding after kainic acid injection into the caudate nucleus. An initial increase in [3H]ouabain binding was observed at 1 and 24 h after lesioning and a decrease in [3H]ouabain binding was evident by 9 days after lesioning.(ABSTRACT TRUNCATED AT 250 WORDS)

Duke Scholars

Published In

Brain Res

DOI

ISSN

0006-8993

Publication Date

April 21, 1987

Volume

409

Issue

2

Start / End Page

335 / 342

Location

Netherlands

Related Subject Headings

  • Sodium-Potassium-Exchanging ATPase
  • Rats, Inbred Strains
  • Rats
  • Pineal Gland
  • Ouabain
  • Neurology & Neurosurgery
  • Male
  • Kainic Acid
  • Densitometry
  • Caudate Nucleus
 

Citation

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Caspers, M. L., Schwartz, R. D., Labarca, R., & Paul, S. M. (1987). Autoradiographic visualization and characterization of [3H]ouabain binding to the Na+,K+-ATPase of rat brain and pineal. Brain Res, 409(2), 335–342. https://doi.org/10.1016/0006-8993(87)90719-0
Caspers, M. L., R. D. Schwartz, R. Labarca, and S. M. Paul. “Autoradiographic visualization and characterization of [3H]ouabain binding to the Na+,K+-ATPase of rat brain and pineal.Brain Res 409, no. 2 (April 21, 1987): 335–42. https://doi.org/10.1016/0006-8993(87)90719-0.
Caspers ML, Schwartz RD, Labarca R, Paul SM. Autoradiographic visualization and characterization of [3H]ouabain binding to the Na+,K+-ATPase of rat brain and pineal. Brain Res. 1987 Apr 21;409(2):335–42.
Caspers, M. L., et al. “Autoradiographic visualization and characterization of [3H]ouabain binding to the Na+,K+-ATPase of rat brain and pineal.Brain Res, vol. 409, no. 2, Apr. 1987, pp. 335–42. Pubmed, doi:10.1016/0006-8993(87)90719-0.
Caspers ML, Schwartz RD, Labarca R, Paul SM. Autoradiographic visualization and characterization of [3H]ouabain binding to the Na+,K+-ATPase of rat brain and pineal. Brain Res. 1987 Apr 21;409(2):335–342.
Journal cover image

Published In

Brain Res

DOI

ISSN

0006-8993

Publication Date

April 21, 1987

Volume

409

Issue

2

Start / End Page

335 / 342

Location

Netherlands

Related Subject Headings

  • Sodium-Potassium-Exchanging ATPase
  • Rats, Inbred Strains
  • Rats
  • Pineal Gland
  • Ouabain
  • Neurology & Neurosurgery
  • Male
  • Kainic Acid
  • Densitometry
  • Caudate Nucleus