Home LiteratureArticle Details
PMID: 6297646 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characteristics of GABAB receptor binding sites on rat whole brain synaptic membranes.

British journal of pharmacology ·Vol. 78 ·No. 1 ·1983-01-00 ·Pages 191-206

Bowery NG, Hill DR, Hudson AL

Abstract

1 Saturable binding of (+/-)-[3H]-baclofen and [3H]-gamma- aminobutyric acid ([3H]-GABA) to rat brain crude synaptic membranes has been examined by means of a centrifugation assay. 2 The binding of [3H]-baclofen could be detected in fresh or previously frozen tissue and was dependent on the presence of physiological concentrations of Ca2+ or Mg2+ although a lower affinity Na+ -dependent component could also be observed. Both components probably reflect binding to receptor recognition sites. 3 The saturable portion of bound [3H]-baclofen formed 20.3 +/- 6.9% of total bound ligand. This could be displaced by GABA (IC50 = 0.04 microM), (-)-baclofen (0.04 microM) and to a much lesser extent by (+)-baclofen (33 microM). Isoguvacine, piperidine-4-sulphonic acid and bicuculline methobromide were inactive (up to 100 microM) and muscimol was only weakly active (IC50 = 12.3 microM). 4 Saturable binding of [3H]-GABA increased on adding CaCl2 or MgSO4 (up to 2.5 mM and 5.0 mM respectively) to the Tris-HCl incubation solution. This binding (GABAB site binding) was additional to the bicuculline-sensitive binding of GABA (GABAA site binding) and could be completely displaced by (-)-baclofen (IC50 = 0.13 microM). 5 Increasing the Ca2+ concentration (0 to 2.5 mM) increased the binding capacity of the membranes without changing their affinity for the ligand. 6 The binding of [3H]-GABA to GABAB sites could be demonstrated in fresh as well as previously frozen membranes with a doubling of the affinity being produced by freezing. Further incubation with the non-ionic detergent Triton-X-100 (0.05% v/v) reduced the binding capacity by 50%. 7 The pharmacological profile of displacers of [3H]-GABA from GABAB sites correlated well with that for [3H]-baclofen displacement. A correlation with data previously obtained in isolated preparations of rat atria and mouse vas deferens was also apparent. 8 It is concluded that [3H]-baclofen or [3H]-GABA are both ligands for the same bicuculline-insensitive, divalent cation-dependent binding sites in the rat brain.

MeSH Terms
Animals Baclofen/pharmacology Bicuculline/pharmacology Brain/metabolism In Vitro Techniques Kinetics Male Nerve Tissue Proteins/metabolism Octoxynol Polyethylene Glycols/pharmacology Rats Rats, Inbred Strains Receptors, Cell Surface/metabolism Receptors, GABA-A Synaptic Membranes/metabolism
Chemicals
Nerve Tissue Proteins Receptors, Cell Surface Receptors, GABA-A Polyethylene Glycols Octoxynol Baclofen Bicuculline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Bowery N G
Hill D R
Hudson A L
References (56)
56 references, click to expand
  1. The uptake of [3H]GABA by slices of rat cerebral cortex.
    J Neurochem. 1968 Oct;15(10):1141-9 PMID: 5711127
  2. GABA, bicuculline and central inhibition.
    Nature. 1970 Jun 27;226(5252):1222-4 PMID: 4393081
  3. Normalization in the fitting of data by iterative methods. Application to tracer kinetics and enzyme kinetics.
    Biochem J. 1973 Jul;134(3):729-36 PMID: 4749273
  4. Central effects of beta-(para-chlorophenyl)-gamma-aminobutyric acid.
    Brain Res. 1974 Apr 26;70(3):493-9 PMID: 4362456
  5. The action of beta-phenyl-GABA derivatives on neurones of the cat cerebral cortex.
    Brain Res. 1974 Apr 26;70(3):501-5 PMID: 4821062
  6. Amino acid transmitters in the mammalian central nervous system.
    Ergeb Physiol. 1974;69(0):97-188 PMID: 4151806
  7. The effects of Lioresal on synaptic activity in the isolated spinal cord.
    Neurology. 1974 Oct;24(10):957-63 PMID: 4370458
  8. Depolarizing actions of gamma-aminobutyric acid and related compounds on rat superior cervical ganglia in vitro.
    Br J Pharmacol. 1974 Feb;50(2):205-18 PMID: 4154116
  9. Gamma-aminobutyric acid binding to receptor sites in the rat central nervous system.
    Proc Natl Acad Sci U S A. 1974 Dec;71(12):4802-7 PMID: 4155072
  10. Properties of gamma-aminobutyric acid (GABA) receptor binding in rat brain synaptic membrane fractions.
    Brain Res. 1975 Dec 12;100(1):81-97 PMID: 241459
  11. Influences ions, enzymes, and detergents on gamma-aminobutyric acid-receptor binding in synaptic membranes of rat brain.
    Mol Pharmacol. 1977 May;13(3):442-53 PMID: 876033
  12. A new class of GABA agonist.
    Nature. 1977 Jul 7;268(5615):53-5 PMID: 196200
  13. The action of baclofen on neurons of the substantia nigra and of the ventral tegmental area.
    Brain Res. 1977 Oct 14;134(3):577-80 PMID: 902118
  14. Muscimol binding in rat brain: association with synaptic GABA receptors.
    Brain Res. 1978 Jun 9;148(1):153-62 PMID: 207386
  15. Use of 3H-muscimol for GABA receptor studies.
    Nature. 1978 Jun 1;273(5661):392-4 PMID: 207993
  16. GABAergic modulation of benzodiazepine binding site sensitivity.
    Nature. 1978 Jul 27;274(5669):383-5 PMID: 27722
  17. Is GABA release modulated by presynaptic receptors?
    Nature. 1978 Aug 31;274(5674):904-5 PMID: 210397
  18. Action of baclofen on mammalian synaptic transmission.
    Neuroscience. 1978;3(6):495-515 PMID: 211462
  19. Purification of an endogenous protein inhibitor of the high affinity binding of gamma-aminobutyric acid to synaptic membranes of rat brain.
    Proc Natl Acad Sci U S A. 1978 Aug;75(8):4024-8 PMID: 279020
  20. The biological activity of d- and l-baclofen (Lioresal).
    Eur J Pharmacol. 1978 Nov 1;52(1):133-6 PMID: 214308
  21. Action of beta-(4-chlorophenyl-GABA) on uptake and metabolism of GABA in different subcellular fractions of rat brain.
    Experientia. 1978 Sep 15;34(9):1137-8 PMID: 720504
  22. Baclofen: effects on amino acid release and metabolism in slices of guinea pig cerebral cortex.
    J Neurochem. 1979 Jan;32(1):103-9 PMID: 759561
  23. Central depressant action of baclofen [proceedings].
    J Physiol. 1978 Nov;284:131P PMID: 731481
  24. Inhibition of Na+-independent [3H]GABA binding to synaptic membranes of rat brain by beta-substituted GABA derivatives.
    J Neurochem. 1979 Mar;32(3):1123-5 PMID: 430047
  25. Is stimulation of benzodiazepine receptor binding mediated by a novel GABA receptor?
    Nature. 1979 Apr 19;278(5706):748-9 PMID: 219374
  26. gamma-Aminobutyric acid receptors in cerebellar membranes of rat brain after a treatment with Triton X-100.
    J Neurochem. 1979 May;32(5):1379-86 PMID: 438810
  27. Characterization of [3H]muscimol binding to mouse brain membranes.
    Biochem Pharmacol. 1979 Apr 1;28(7):1123-8 PMID: 444270
  28. gamma-Aminobutyric acid reduces the evoked release of [3H]-noradrenaline from sympathetic nerve terminals [proceedings].
    Br J Pharmacol. 1979 May;66(1):108P PMID: 454903
  29. Partial agonists for brain GABA/benzodiazepine receptor complex.
    Nature. 1979 Jul 26;280(5720):331-3 PMID: 460407
  30. Delta-aminolaevulinic acid is a potent agonist for GABA autoreceptors.
    Nature. 1979 Aug 9;280(5722):514-5 PMID: 223065
  31. Baclofen and muscimol: behavioural and neurochemical sequelae of unilateral intranigral administration and effects on 3H-GABA receptor binding.
    Naunyn Schmiedebergs Arch Pharmacol. 1979 Apr;306(3):275-80 PMID: 471079
  32. Biochemical evidence for two classes of GABA receptors in rat brain.
    Brain Res. 1979 Aug 31;172(3):566-71 PMID: 224975
  33. Pharmacological characterization of different types of GABA and glutamate receptors in vertebrates and invertebrates.
    Prog Neurobiol. 1979;13(2):117-235 PMID: 227014
  34. (-)Baclofen decreases neurotransmitter release in the mammalian CNS by an action at a novel GABA receptor.
    Nature. 1980 Jan 3;283(5742):92-4 PMID: 6243177
  35. Piperidine-4-sulphonic acid, a new specific GABA agonist.
    J Neurochem. 1980 Mar;34(3):756-9 PMID: 7354350
  36. The binding of 3H-isoguvacine to mouse brain synaptic membranes.
    Life Sci. 1980 Apr 14;26(15):1239-45 PMID: 7392797
  37. Presence of a low molecular weight endogenous inhibitor on 3H-muscimol binding in synaptic membranes.
    Nature. 1980 Jun 26;285(5767):670-3 PMID: 6248791
  38. GABA binding in mammalian brain: inhibition by endogenous GABA.
    Life Sci. 1980 Sep 15;27(11):1001-11 PMID: 7432094
  39. 3H-baclofen and 3H-GABA bind to bicuculline-insensitive GABA B sites in rat brain.
    Nature. 1981 Mar 12;290(5802):149-52 PMID: 6259535
  40. Selective depression of synaptic excitation in cat spinal neurones by baclofen: an iontophoretic study.
    Br J Pharmacol. 1981 Feb;72(2):373-84 PMID: 7214103
  41. GABA receptor pharmacology. Functional considerations.
    Biochem Pharmacol. 1981 May 1;30(9):907-13 PMID: 6112992
  42. Bicuculline-insensitive GABA receptors on peripheral autonomic nerve terminals.
    Eur J Pharmacol. 1981 Apr 24;71(1):53-70 PMID: 6263651
  43. The effects of 4-aminopyridine on the isolated vas deferens and its effects on the inhibitory properties of adenosine, morphine, noradrenaline and gamma-aminobutyric acid.
    Br J Pharmacol. 1981 Jul;73(3):791-6 PMID: 7248668
  44. Selective effects of (-)-baclofen on spinal synaptic transmission in the cat.
    Exp Brain Res. 1981;42(2):158-70 PMID: 6266854
  45. Characterization of the binding of the GABA agonist [3H]piperidine-4-sulphonic acid to bovine brain synaptic membranes.
    J Neurochem. 1981 Aug;37(2):401-9 PMID: 6267201
  46. Two types of gamma-aminobutyric acid receptor on embryonic sensory neurones.
    Br J Pharmacol. 1981 Nov;74(3):579-85 PMID: 6271320
  47. Autoradiographic localization of GABAB receptors in rat cerebellum.
    Nature. 1981 Dec 10;294(5841):584-7 PMID: 6273750
  48. Endogenous GABA determines the characteristics of [3H]GABA-binding.
    Eur J Pharmacol. 1981 Oct 22;75(2-3):83-92 PMID: 6274665
  49. The binding of the GABA agonist [3H]THIP to rat brain synaptic membranes.
    J Neurochem. 1982 Apr;38(4):1123-9 PMID: 6278083
  50. gamma-Aminobutyric acid (GABA) receptor stimulation. II. Specificity of progabide (SL 76002) and SL 75102 for the GABA receptor.
    J Pharmacol Exp Ther. 1982 Mar;220(3):672-7 PMID: 6121050
  51. Two actions of gamma-aminobutyric acid on the responses of the isolated basilar artery from the rabbit.
    Br J Pharmacol. 1982 Jan;75(1):177-81 PMID: 7074280
  52. gamma-Aminobutyric acid and postganglionic sympathetic transmission in the pulmonary artery of the rabbit.
    J Auton Pharmacol. 1980 Nov;1(1):45-51 PMID: 6281282
  53. Evidence that SL75102 is an agonist at GABAb as well as GABAa receptors.
    Neuropharmacology. 1982 May;21(5):391-5 PMID: 6287331
  54. Presynaptic gamma-aminobutyric acid receptors in the rat anococcygeus muscle and their antagonism by 5-aminovaleric acid.
    Br J Pharmacol. 1982 Sep;77(1):163-8 PMID: 6289954
  55. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  56. The isolation of nerve endings from brain: an electron-microscopic study of cell fragments derived by homogenization and centrifugation.
    J Anat. 1962 Jan;96:79-88 PMID: 13901297
Article Info
Journal
British journal of pharmacology
Abbr.
Br J Pharmacol
ISSN
0007-1188
DOI
10.1111/j.1476-5381.1983.tb09380.x
Published
1983-01-00
Pages
191-206
Language
English
Region
England
NLM ID
7502536
PMCID
PMC2044790
Subset
IM
External Links
PubMed source DOI: 10.1111/j.1476-5381.1983.tb09380.x PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2