Characterization of the binding of radioligands to the N-methyl-D-aspartate, phencyclidine, and glycine receptors in buffy coat membranes

Susan M. Jones, Lawrence D. Snell, Kenneth M. Johnson

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Rapid vacuum filtration assays were used to quantitate radioligand binding to phencyclidine (PCP), N-methyl-D-aspartate (NMDA), and strychnine-insensitive glycine receptors in a simple buffy coat preparation of rat cortical membranes. KD and Bmax values for [3H]glycine binding were very similar to those previously reported by workers who used centrifugation for the separation of free and bound [3H]glycine. We also found that this preparation had a high percentage of NMDA-displaceable L-[3H]glutamate binding sites, which demonstrated a pharmacology very similar to that previously observed in more purified synaptic plasma membranes. Hill analysis of the displacement curves indicated that glutamate bound to a single class of sites, but that NMDA and NMDA antagonists may interact with this site in a negatively cooperative fashion. This preparation was also found to be suitable for the study of NMDA and glycine receptor regulation of the associated ion channel, as these effectors, alone and in combination, increased the affinity with which [3H]TCP bound to the PCP receptor believed to be located within the ion channel. Thus, the ability to measure radioligand binding to these three sites in the same simple membrane preparation should greatly facilitate the study of the interaction between them.

Original languageEnglish (US)
Pages (from-to)161-168
Number of pages8
JournalJournal of Pharmacological Methods
Volume21
Issue number2
DOIs
StatePublished - 1989

Fingerprint

Phencyclidine Receptors
Glycine Receptors
N-Methylaspartate
N-Methyl-D-Aspartate Receptors
Membranes
Ion Channels
Glycine
Glutamic Acid
Sarcosine
Phencyclidine
Synaptic Membranes
Centrifugation
Cell membranes
Vacuum
Rats
Assays
Binding Sites
Cell Membrane
Pharmacology

Keywords

  • Binding
  • Glycine
  • N-methyl-D-aspartate
  • Phencyclidine

ASJC Scopus subject areas

  • Pharmacology

Cite this

Characterization of the binding of radioligands to the N-methyl-D-aspartate, phencyclidine, and glycine receptors in buffy coat membranes. / Jones, Susan M.; Snell, Lawrence D.; Johnson, Kenneth M.

In: Journal of Pharmacological Methods, Vol. 21, No. 2, 1989, p. 161-168.

Research output: Contribution to journalArticle

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