[3H]Ryanodine binding sites in rat brain demonstrated by membrane binding and autoradiography

Rodolfo A. Padua, Weihua Wan, James I. Nagy, Jonathan D. Geiger

Research output: Contribution to journalArticle

40 Scopus citations

Abstract

High affinity [3H]ryanodine binding sites were characterized in P1 (crude nuclear), P2 (mitochondrial/synaptosomal) and P3 (microsomal) subcellular fractions of rat brain. Binding in each of the fractions was highest at 37°C and pH 8-9, optimal in the presence of 100 μM Ca2+, 550 μM ATP and 1.0 M KCl, and increased linearly as a function of protein. Saturation analyses revealed a single class of binding sites with mean KD values (nM) of 8.9, 1.6 and 5.7 and Bmax values (fmol/mg protein) of 122, 69 and 106 for the P1, P2 and P3 fractions, respectively. The levels of [3H]ryanodine binding in P1 and P2 fractions of 4 brain regions were fairly uniform while those in P3 fractions were 5-fold greater in cerebral cortex than in the other areas examined. By autoradiography, a high concentration of [3H]ryanodine binding sites was seen in the dentate gyrus and CA3 subregions of the hippocampus. The results suggest that [3H]ryanodine binding sites, perhaps similar to [3H]ryanodine receptors in muscle, are associated with various subcellular structures and are heterogeneously distributed in the CNS.

Original languageEnglish (US)
Pages (from-to)135-140
Number of pages6
JournalBrain Research
Volume542
Issue number1
DOIs
StatePublished - Feb 22 1991

Keywords

  • Autoradiography
  • Calcium channel
  • Membrane binding
  • Ryanodine receptor
  • Subcellular fraction

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

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