Two human alpha 2-adrenoceptor subtypes alpha 2A-C10 and alpha 2B-C2 expressed in Sf9 cells couple to transduction pathway resulting in opposite effects on cAMP production

Eur J Pharmacol. 1995 Jul 18;290(2):75-83. doi: 10.1016/0922-4106(95)90019-5.

Abstract

The baculovirus expression vector system utilizing the strong polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus (AcNPV) was used for high level expression of the two alpha 2-adrenoceptor subtypes alpha 2A-C10 and alpha 2B-C2 in Spodoptera frugiperda (Sf-9) insect cells. For rapid screening of recombinant viruses the luciferase gene was expressed under the early ETL-promoter (early transcript large) in the same plasmid. Both receptor subtypes showed the same rank order of binding affinity for four agonists tested: dexmedetomidine > l-medetomidine = clonidine > noradrenaline. For the alpha 2A-C10 subtype, these agonists inhibited forskolin stimulated cAMP production through pertussis toxin sensitive G-proteins. In contrast, for the alpha 2B-C2 subtype the agonists stimulated both basal and forskolin stimulated cAMP production.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Cloning, Molecular
  • Cyclic AMP / metabolism*
  • DNA, Recombinant / isolation & purification
  • Genetic Vectors*
  • Humans
  • In Vitro Techniques
  • Luciferases / genetics
  • Plasmids / physiology
  • Radioligand Assay
  • Receptors, Adrenergic, alpha-2 / genetics
  • Receptors, Adrenergic, alpha-2 / metabolism*
  • Spodoptera
  • Transduction, Genetic*

Substances

  • DNA, Recombinant
  • Receptors, Adrenergic, alpha-2
  • Cyclic AMP
  • Luciferases