α4βδ GABA receptors are high-affinity targets for γ-hydroxybutyric acid (GHB)

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

α4βδ GABA receptors are high-affinity targets for γ-hydroxybutyric acid (GHB). / Absalom, N.; Karim, N.; Eghorn, L.F.; Villumsen, I.S.; Bay, T.; Bräuner-Osborne, H.; Frølund, B.; Clausen, R.P.; Olsen, J.V.; Knudsen, G.M.; Chebib, M.; Wellendorph, P.

In: Proceedings of the National Academy of Sciences USA (PNAS), Vol. 109, No. 33, 14.08.2012, p. 13404-13409.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Absalom, N, Karim, N, Eghorn, LF, Villumsen, IS, Bay, T, Bräuner-Osborne, H, Frølund, B, Clausen, RP, Olsen, JV, Knudsen, GM, Chebib, M & Wellendorph, P 2012, 'α4βδ GABA receptors are high-affinity targets for γ-hydroxybutyric acid (GHB)', Proceedings of the National Academy of Sciences USA (PNAS), vol. 109, no. 33, pp. 13404-13409. https://doi.org/10.1073/pnas.1204376109

APA

Absalom, N., Karim, N., Eghorn, L. F., Villumsen, I. S., Bay, T., Bräuner-Osborne, H., ... Wellendorph, P. (2012). α4βδ GABA receptors are high-affinity targets for γ-hydroxybutyric acid (GHB). Proceedings of the National Academy of Sciences USA (PNAS), 109(33), 13404-13409. https://doi.org/10.1073/pnas.1204376109

Vancouver

Absalom N, Karim N, Eghorn LF, Villumsen IS, Bay T, Bräuner-Osborne H et al. α4βδ GABA receptors are high-affinity targets for γ-hydroxybutyric acid (GHB). Proceedings of the National Academy of Sciences USA (PNAS). 2012 Aug 14;109(33):13404-13409. https://doi.org/10.1073/pnas.1204376109

Author

Absalom, N. ; Karim, N. ; Eghorn, L.F. ; Villumsen, I.S. ; Bay, T. ; Bräuner-Osborne, H. ; Frølund, B. ; Clausen, R.P. ; Olsen, J.V. ; Knudsen, G.M. ; Chebib, M. ; Wellendorph, P. / α4βδ GABA receptors are high-affinity targets for γ-hydroxybutyric acid (GHB). In: Proceedings of the National Academy of Sciences USA (PNAS). 2012 ; Vol. 109, No. 33. pp. 13404-13409.

Bibtex

@article{c4a1008177a8493a8945ed53b91e639a,
title = "α4βδ GABA receptors are high-affinity targets for γ-hydroxybutyric acid (GHB)",
abstract = "γ-Hydroxybutyric acid (GHB) binding to brain-specific high-affinity sites is well-established and proposed to explain both physiological and pharmacological actions. However, the mechanistic links between these lines of data are unknown. To identify molecular targets for specific GHB high-affinity binding, we undertook photolinking studies combined with proteomic analyses and identified several GABA receptor subunits as possible candidates. A subsequent functional screening of various recombinant GABA receptors in Xenopus laevis oocytes using the two-electrode voltage clamp technique showed GHB to be a partial agonist at αβδ- but not αβγ-receptors, proving that the δ-subunit is essential for potency and efficacy. GHB showed preference for α4 over α(1,2,6)-subunits and preferably activated α4β1δ (EC = 140 nM) over α4β(2/3)δ (EC = 8.41/1.03 mM). Introduction of a mutation, α4F71L, in α4β 1(δ)-receptors completely abolished GHB but not GABA function, indicating nonidentical binding sites. Radioligand binding studies using the specific GHB radioligand [ H](E,RS)-(6,7,8,9-tetrahydro-5-hydroxy-5H- benzocyclohept-6-ylidene)acetic acid showed a 39{\%} reduction (P = 0.0056) in the number of binding sites in α4 KO brain tissue compared with WT controls, corroborating the direct involvement of the α4-subunit in high-affinity GHB binding. Our data link specific GHB forebrain binding sites with α4-containing GABA receptors and postulate a role for extrasynaptic α4β-containing GABA receptors in GHB pharmacology and physiology. This finding will aid in elucidating the molecular mechanisms behind the proposed function of GHB as a neurotransmitter and its unique therapeutic effects in narcolepsy and alcoholism.",
author = "N. Absalom and N. Karim and L.F. Eghorn and I.S. Villumsen and T. Bay and H. Br{\"a}uner-Osborne and B. Fr{\o}lund and R.P. Clausen and J.V. Olsen and G.M. Knudsen and M. Chebib and P. Wellendorph",
note = "Keywords: gamma-hydroxybutyric acid receptor, gamma-hydroxybutyric acid high-affinity binding sites, alpha4-subunit knockout, photoaffinity ligand",
year = "2012",
month = "8",
day = "14",
doi = "10.1073/pnas.1204376109",
language = "English",
volume = "109",
pages = "13404--13409",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "The National Academy of Sciences of the United States of America",
number = "33",

}

RIS

TY - JOUR

T1 - α4βδ GABA receptors are high-affinity targets for γ-hydroxybutyric acid (GHB)

AU - Absalom, N.

AU - Karim, N.

AU - Eghorn, L.F.

AU - Villumsen, I.S.

AU - Bay, T.

AU - Bräuner-Osborne, H.

AU - Frølund, B.

AU - Clausen, R.P.

AU - Olsen, J.V.

AU - Knudsen, G.M.

AU - Chebib, M.

AU - Wellendorph, P.

N1 - Keywords: gamma-hydroxybutyric acid receptor, gamma-hydroxybutyric acid high-affinity binding sites, alpha4-subunit knockout, photoaffinity ligand

PY - 2012/8/14

Y1 - 2012/8/14

N2 - γ-Hydroxybutyric acid (GHB) binding to brain-specific high-affinity sites is well-established and proposed to explain both physiological and pharmacological actions. However, the mechanistic links between these lines of data are unknown. To identify molecular targets for specific GHB high-affinity binding, we undertook photolinking studies combined with proteomic analyses and identified several GABA receptor subunits as possible candidates. A subsequent functional screening of various recombinant GABA receptors in Xenopus laevis oocytes using the two-electrode voltage clamp technique showed GHB to be a partial agonist at αβδ- but not αβγ-receptors, proving that the δ-subunit is essential for potency and efficacy. GHB showed preference for α4 over α(1,2,6)-subunits and preferably activated α4β1δ (EC = 140 nM) over α4β(2/3)δ (EC = 8.41/1.03 mM). Introduction of a mutation, α4F71L, in α4β 1(δ)-receptors completely abolished GHB but not GABA function, indicating nonidentical binding sites. Radioligand binding studies using the specific GHB radioligand [ H](E,RS)-(6,7,8,9-tetrahydro-5-hydroxy-5H- benzocyclohept-6-ylidene)acetic acid showed a 39% reduction (P = 0.0056) in the number of binding sites in α4 KO brain tissue compared with WT controls, corroborating the direct involvement of the α4-subunit in high-affinity GHB binding. Our data link specific GHB forebrain binding sites with α4-containing GABA receptors and postulate a role for extrasynaptic α4β-containing GABA receptors in GHB pharmacology and physiology. This finding will aid in elucidating the molecular mechanisms behind the proposed function of GHB as a neurotransmitter and its unique therapeutic effects in narcolepsy and alcoholism.

AB - γ-Hydroxybutyric acid (GHB) binding to brain-specific high-affinity sites is well-established and proposed to explain both physiological and pharmacological actions. However, the mechanistic links between these lines of data are unknown. To identify molecular targets for specific GHB high-affinity binding, we undertook photolinking studies combined with proteomic analyses and identified several GABA receptor subunits as possible candidates. A subsequent functional screening of various recombinant GABA receptors in Xenopus laevis oocytes using the two-electrode voltage clamp technique showed GHB to be a partial agonist at αβδ- but not αβγ-receptors, proving that the δ-subunit is essential for potency and efficacy. GHB showed preference for α4 over α(1,2,6)-subunits and preferably activated α4β1δ (EC = 140 nM) over α4β(2/3)δ (EC = 8.41/1.03 mM). Introduction of a mutation, α4F71L, in α4β 1(δ)-receptors completely abolished GHB but not GABA function, indicating nonidentical binding sites. Radioligand binding studies using the specific GHB radioligand [ H](E,RS)-(6,7,8,9-tetrahydro-5-hydroxy-5H- benzocyclohept-6-ylidene)acetic acid showed a 39% reduction (P = 0.0056) in the number of binding sites in α4 KO brain tissue compared with WT controls, corroborating the direct involvement of the α4-subunit in high-affinity GHB binding. Our data link specific GHB forebrain binding sites with α4-containing GABA receptors and postulate a role for extrasynaptic α4β-containing GABA receptors in GHB pharmacology and physiology. This finding will aid in elucidating the molecular mechanisms behind the proposed function of GHB as a neurotransmitter and its unique therapeutic effects in narcolepsy and alcoholism.

UR - http://www.scopus.com/inward/record.url?scp=84865165962&partnerID=8YFLogxK

U2 - 10.1073/pnas.1204376109

DO - 10.1073/pnas.1204376109

M3 - Journal article

VL - 109

SP - 13404

EP - 13409

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 33

ER -

ID: 38405483