Neurochemical Research

, Volume 11, Issue 4, pp 599–606 | Cite as

GABA-agonists induce the formation of low-affinity GABA-receptors on cultured cerebellar granule cells via preexisting high affinity GABA receptors

  • Bo Belhage
  • Eddi Meier
  • Arne Schousboe
Original Articles

Abstract

The kinetics of specific GABA-binding to membranes isolated from cerebellar granule cells, cultured for 12 days from dissociated cerebella of 7-day-old rats was studied using [3H]GABA as the ligand. The granule cells were cultured in the presence of the specific GABA receptor agonist 4, 5, 6, 7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP, 150 μM) or THIP plus the antagonist bicuculline methobromide (150 μM of each) or in the absence of the agonist or antagonist. Membranes isolated from granule cells cultured in a medium without the GABA agonist revealed a single binding site for GABA with a binding constant (KD) of 7.9±0.4 nM and aBmax of 3.42±0.08 pmol×mg−1 protein. Membranes from cells cultured in the presence of THIP had two binding sites for GABA withKD-values of 6.8±0.9 nM and 476±311 nM, respectively. The correspondingBmax values were 4.41±0.42 pmol×mg−1 and 5.81±1.20 pmol×mg−1. The effect of culturing the cells in THIP was antagonized by the simultaneous presence of bicuculline in the culture media, i.e. no significant low-affinity binding for GABA was found on the membranes from granule cells cultured in both THIP and bicuculline. TheKD value (14.3±1.4 nM) for the high affinity binding site was, however, slightly increased compared to the non-treated cells. These findings suggest that the ability of THIP to induce formation of low-affinity GABA receptors is mediated by preexisting high-affinity GABA-receptors on the granule cells.

Keywords

Granule Cell Binding Constant High Affinity Binding Simultaneous Presence Gaba Receptor 

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Copyright information

© Plenum Publishing Corporation 1986

Authors and Affiliations

  • Bo Belhage
    • 1
  • Eddi Meier
    • 1
  • Arne Schousboe
    • 1
  1. 1.Department of Biochemistry A Panum InstituteUniversity of CopenhagenCopenhagenDenmark

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