Pharmacological characterisation of a cell line expressing GABA B1b and GABA B2 receptor subunits

Biochem Pharmacol. 2003 Apr 1;65(7):1103-13. doi: 10.1016/s0006-2952(02)01658-1.

Abstract

The gamma-aminobutyric acid (GABA(B)) receptor has been shown to be a heterodimer consisting of two receptor subunits, GABA(B1) and GABA(B2). We have stably co-expressed these two subunits in a CHO cell line, characterised its pharmacology and compared it to the native receptor in rat brain membranes. Radioligand binding using [3H]CGP54626A demonstrated a similar rank order of potency between recombinant and native receptors: CGP62349>CGP54626A>SCH 50911>3-aminopropylphosphinicacid(3-APPA)>GABA>baclofen>saclofen>phaclofen. However, differences were observed in the affinity of agonists, which were higher at the native receptor, suggesting that in the recombinant system a large number of the receptors were in the low agonist affinity state. In contrast, [35S]GTPgammaS binding studies did not show any differences between recombinant and native receptors with the full agonists GABA and 3-APPA. Measurement of cAMP accumulation in the cells revealed a degree of endogenous coupling of the receptors to G-proteins. This is most likely to be due to the high expression levels of receptors (B(max)=22.5+/-2.5pmol/mg protein) in this experimental system. There was no evidence of GABA(B2) receptors, when expressed alone, binding [3H]CGP54626A, [3H]GABA, [3H]3-APPA nor of GABA having any effect on basal [35S]GTPgammaS binding or cAMP levels.

MeSH terms

  • Animals
  • Binding Sites
  • CHO Cells
  • Cell Line
  • Cricetinae
  • Cyclic AMP / metabolism
  • Guanosine 5'-O-(3-Thiotriphosphate) / metabolism
  • Radioligand Assay
  • Receptors, GABA / drug effects
  • Receptors, GABA / metabolism*
  • Receptors, GABA-B / drug effects
  • Receptors, GABA-B / metabolism*
  • Sulfur Radioisotopes

Substances

  • Receptors, GABA
  • Receptors, GABA-B
  • Sulfur Radioisotopes
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Cyclic AMP