Characterization of opioid receptors in cultured neurons

Pierre J J Vaysse, R. Suzanne Zukin, Kay L. Fields, John A. Kessler

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The appearance of μ, δ-, and κ-opioid receptors was examined in primary cultures of embryonic rat brain. Membranes prepared from striatal, hippocampal, and hypothalamic neurons grown in dissociated cell culture each exhibited high-affinity opioid binding sites as determined by equilibrium binding of the universal opioid ligand (-)[3]bremazocine. The highest density of binding sites (per mg of protein) was found in membranes prepared from cultured striatal neurons (Bmax = 210 ± 40 fmol/mg protein); this density is approximately two-thirds that of adult striatal membranes. By contrast, membranes of cultured cerebellar neurons and cultured astrocytes were devoid of opioid binding sites. The opioid receptor types expressed in cultured striatal neurons were characterized by equilibrium binding of highly selective radioligands. Scatchard analysis of binding of the μ-specific ligand [3H]D-Ala2,N-Me-Phe4,Gly-ol 5-enkephalin to embryonic striatal cell membranes revealed an apparent single class of sites with an affinity (KD) of 0.4 ± 0.1 nM and a density (Bmax) of 160 ± 20 fmol/mg of protein. Specific binding of (-)-[3H]bremazocine under conditions in which μ- and δ-receptor binding was suppressed (κ-receptor labeling conditions) occurred to an apparent single class of sites (KD = 2 ± 1 nM/; Bmax = 40 ± 15 fmol/mg of protein). There was no detectable binding of the selective δ-ligand [3H]D-Pen22,D-Pen5-enkephalin. Thus, cultured striatal neurons expressed μ- and κ-receptor sites at densities comparable to those found in vivo for embryonic rat brain, but not δ-receptors.

Original languageEnglish (US)
Pages (from-to)624-631
Number of pages8
JournalJournal of Neurochemistry
Volume55
Issue number2
StatePublished - 1990

Fingerprint

Corpus Striatum
Opioid Receptors
Neurons
Opioid Analgesics
Membranes
Enkephalins
Binding Sites
Ligands
Rats
Brain
Proteins
Cell membranes
Cell culture
Labeling
Astrocytes
Cell Culture Techniques
Cell Membrane

Keywords

  • Astroglia
  • Cell culture
  • Neuron
  • Opioid receptors
  • Primary culture

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Vaysse, P. J. J., Zukin, R. S., Fields, K. L., & Kessler, J. A. (1990). Characterization of opioid receptors in cultured neurons. Journal of Neurochemistry, 55(2), 624-631.

Characterization of opioid receptors in cultured neurons. / Vaysse, Pierre J J; Zukin, R. Suzanne; Fields, Kay L.; Kessler, John A.

In: Journal of Neurochemistry, Vol. 55, No. 2, 1990, p. 624-631.

Research output: Contribution to journalArticle

Vaysse, PJJ, Zukin, RS, Fields, KL & Kessler, JA 1990, 'Characterization of opioid receptors in cultured neurons', Journal of Neurochemistry, vol. 55, no. 2, pp. 624-631.
Vaysse, Pierre J J ; Zukin, R. Suzanne ; Fields, Kay L. ; Kessler, John A. / Characterization of opioid receptors in cultured neurons. In: Journal of Neurochemistry. 1990 ; Vol. 55, No. 2. pp. 624-631.
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