Postnatal development and kinetics of [3H]gaboxadol binding in rat brain: in vitro homogenate binding and quantitative autoradiography

Brain Res. 2007 Sep 19:1170:39-47. doi: 10.1016/j.brainres.2007.07.031. Epub 2007 Jul 20.

Abstract

The postnatal development of the binding of the GABA(A) receptor agonist [(3)H]gaboxadol in rat brain was investigated. Using brain tissue from rats obtained at postnatal days 1, 10, 25, and >25 (adult), the binding of [(3)H]gaboxadol and the benzodiazepine [(3)H]flunitrazepam to GABA(A) receptors was compared in homogenate binding assays and quantitative receptor autoradiography. Kinetic and equilibrium data obtained in homogenate binding studies revealed two different [(3)H]gaboxadol affinities. A kinetically derived K(D) of 3.7 nM in adult cerebellum, calculated from the association and dissociation rate constants k(on) (1.45 x 10(8) M(-1) min(-1)) and k(off) (0.54 min(-1)) was contrasted by an equilibrium K(D) of 38.6 nM, obtained by homologous competition experiments. Quantitative analysis of autoradiographic data revealed an increase in specific [(3)H]gaboxadol binding sites during brain development, which resembles the anatomical and temporal pattern of the postnatal expression of the extrasynaptic delta subunit of GABA(A) receptors. In conclusion, by the radioligand binding data obtained on native tissue, binding of gaboxadol to GABA(A) receptors located outside the synaptic junctions could be postulated.

MeSH terms

  • Aging / physiology*
  • Animals
  • Animals, Newborn
  • Autoradiography / methods
  • Binding, Competitive / drug effects
  • Binding, Competitive / physiology
  • Brain / drug effects*
  • Brain / growth & development*
  • Brain / metabolism
  • Brain Chemistry / physiology*
  • Flunitrazepam / metabolism
  • Flunitrazepam / pharmacokinetics
  • GABA Agonists / metabolism
  • GABA Agonists / pharmacokinetics
  • GABA Modulators / metabolism
  • GABA Modulators / pharmacokinetics
  • Isoxazoles / metabolism*
  • Isoxazoles / pharmacokinetics*
  • Kinetics
  • Radioligand Assay / methods
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, GABA-A / drug effects
  • Receptors, GABA-A / metabolism
  • Subcellular Fractions
  • gamma-Aminobutyric Acid / metabolism

Substances

  • GABA Agonists
  • GABA Modulators
  • Isoxazoles
  • Receptors, GABA-A
  • gamma-Aminobutyric Acid
  • Flunitrazepam
  • gaboxadol