Pharmacological similarities between native brain and heterologously expressed α4β2 nicotinic receptors

Navid Shafaee, McCann Houng, Anthony Truong, Nareerat Viseshakul, Antonio Figl, Sumandeep Sandhu, John R. Forsayeth, Linda P. Dwoskin, Peter A. Crooks, Bruce N. Cohen

Research output: Contribution to journalArticlepeer-review

26 Scopus citations


1. We studied the pharmacological properties of native rat brain and heterologously expressed rat α4β2 nicotinic receptors immunoprecipitated onto a fixed substrate with the anti-α4 antibody mAb 299. 2. Immunodepletion with the anti-β2 antibody mAb 270 showed that 89% of the mAb-299-precipitated rat brain receptors contained β2. 3. The association and dissociation rate constants for 30 pM ± [3H]-epibatidine binding to α4β2 receptors expressed in oocytes were 0.02 ± 0.01 and 0.03 ± 0.01 min-1 (± standard error, degrees of freedom = 7-8) at 20-23°C. 4. The Hill coefficients for ± [3H] epibatidine binding to the native brain, α4β2 receptors expressed in oocytes, and α4β2 receptors expressed in CV-1 cells (using recombinant adenovirus) were 0.69-0.70 suggesting a heterogeneous receptor population. Fits of the ± [3H]-epibatidine concentration-binding data to a two-site model gave K(D)s of 8-30 and 560-1.200 pM. The high-affinity sites comprised 73-74% of the native brain and oocyte α4β2 receptor population, 85% of the CV-1 α4β2 receptor population. 5. The expression of rat α4β2 receptors in CV-1 cells using vaccinia viral infection-transfection resulted in a more homogeneous receptor population (Hill coefficient of 1.0 ± 0.2). Fits of the ± [3H]-epibatidine binding data to a single-site model gave a K(D) of 40 ± 3 pM. 6. DHβE (IC50 = 260-470 nM) and the novel nicotine analogue NDNI (IC50 = 7-10 μM) inhibited 30 pM ± [3H]-epibatidine binding to the native brain and heterologously expressed α4β2 receptors equally well. 7. The results show that α4β2-containing nicotinic receptors in the rat brain and heterologously expressed rat α4β2 receptors have similar affinities for ± [3H]-epibatidine, DHβE, and NDNI.

Original languageEnglish
Pages (from-to)1291-1299
Number of pages9
JournalBritish Journal of Pharmacology
Issue number6
StatePublished - 1999


  • Dihydro-β-erythroidine
  • Epibatidine binding
  • Immunoprecipitation
  • Neuronal nicotinic acetylcholine receptors
  • Xenopus oocytes
  • mAb 299 antibody
  • α4
  • β2

ASJC Scopus subject areas

  • Pharmacology


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