Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA

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Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA. / Klein, A B; Bay, T; Villumsen, I S; Falk-Petersen, C B; Marek, A; Frølund, B; Clausen, R P; Hansen, Hanne Demant; Knudsen, G M; Wellendorph, P.

In: Neurochemistry International, Vol. 100, 11.2016, p. 138-145.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Klein, AB, Bay, T, Villumsen, IS, Falk-Petersen, CB, Marek, A, Frølund, B, Clausen, RP, Hansen, HD, Knudsen, GM & Wellendorph, P 2016, 'Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA' Neurochemistry International, vol. 100, pp. 138-145. https://doi.org/10.1016/j.neuint.2016.09.002

APA

Klein, A. B., Bay, T., Villumsen, I. S., Falk-Petersen, C. B., Marek, A., Frølund, B., ... Wellendorph, P. (2016). Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA. Neurochemistry International, 100, 138-145. https://doi.org/10.1016/j.neuint.2016.09.002

Vancouver

Klein AB, Bay T, Villumsen IS, Falk-Petersen CB, Marek A, Frølund B et al. Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA. Neurochemistry International. 2016 Nov;100:138-145. https://doi.org/10.1016/j.neuint.2016.09.002

Author

Klein, A B ; Bay, T ; Villumsen, I S ; Falk-Petersen, C B ; Marek, A ; Frølund, B ; Clausen, R P ; Hansen, Hanne Demant ; Knudsen, G M ; Wellendorph, P. / Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA. In: Neurochemistry International. 2016 ; Vol. 100. pp. 138-145.

Bibtex

@article{4294497808e0445cb5e411d737ff2405,
title = "Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA",
abstract = "GHB (γ-hydroxybutyric acid) is a compound endogenous to mammalian brain with high structural resemblance to GABA. GHB possesses nanomolar-micromolar affinity for a unique population of binding sites, but the exact nature of these remains elusive. In this study we utilized the highly selective GHB analogue, 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) as a tritiated version ((3)H-HOCPCA) to radioactively label the specific GHB high-affinity binding site and gain further insight into the density, distribution and developmental profile of this protein. We show that, in low nanomolar concentrations, (3)H-HOCPCA displays excellent signal-to-noise ratios using rodent brain autoradiography, which makes it a valuable ligand for anatomical quantification of native GHB binding site levels. Our data confirmed that (3)H-HOCPCA labels only the high-affinity specific GHB binding site, found in high density in cortical and hippocampal regions. The experiments revealed markedly stronger binding at pH 6.0 (Kd 73.8 nM) compared to pH 7.4 (Kd 2312 nM), as previously reported for other GHB radioligands but similar Bmax values. Using (3)H-HOCPCA we analyzed the GHB binding protein profile during mouse brain development. Due to the high sensitivity of this radioligand, we were able to detect low levels of specific binding already at E15 in mouse brain, which increased progressively until adulthood. Collectively, we show that (3)H-HOCPCA is a highly sensitive radioligand, offering advantages over the commonly used radioligand (3)H-NCS-382, and thus a very suitable in vitro tool for qualitative and quantitative autoradiography of the GHB high-affinity site.",
author = "Klein, {A B} and T Bay and Villumsen, {I S} and Falk-Petersen, {C B} and A Marek and B Fr{\o}lund and Clausen, {R P} and Hansen, {Hanne Demant} and Knudsen, {G M} and P Wellendorph",
note = "Copyright {\circledC} 2016 Elsevier Ltd. All rights reserved.",
year = "2016",
month = "11",
doi = "10.1016/j.neuint.2016.09.002",
language = "English",
volume = "100",
pages = "138--145",
journal = "Neurochemistry International",
issn = "0197-0186",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA

AU - Klein, A B

AU - Bay, T

AU - Villumsen, I S

AU - Falk-Petersen, C B

AU - Marek, A

AU - Frølund, B

AU - Clausen, R P

AU - Hansen, Hanne Demant

AU - Knudsen, G M

AU - Wellendorph, P

N1 - Copyright © 2016 Elsevier Ltd. All rights reserved.

PY - 2016/11

Y1 - 2016/11

N2 - GHB (γ-hydroxybutyric acid) is a compound endogenous to mammalian brain with high structural resemblance to GABA. GHB possesses nanomolar-micromolar affinity for a unique population of binding sites, but the exact nature of these remains elusive. In this study we utilized the highly selective GHB analogue, 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) as a tritiated version ((3)H-HOCPCA) to radioactively label the specific GHB high-affinity binding site and gain further insight into the density, distribution and developmental profile of this protein. We show that, in low nanomolar concentrations, (3)H-HOCPCA displays excellent signal-to-noise ratios using rodent brain autoradiography, which makes it a valuable ligand for anatomical quantification of native GHB binding site levels. Our data confirmed that (3)H-HOCPCA labels only the high-affinity specific GHB binding site, found in high density in cortical and hippocampal regions. The experiments revealed markedly stronger binding at pH 6.0 (Kd 73.8 nM) compared to pH 7.4 (Kd 2312 nM), as previously reported for other GHB radioligands but similar Bmax values. Using (3)H-HOCPCA we analyzed the GHB binding protein profile during mouse brain development. Due to the high sensitivity of this radioligand, we were able to detect low levels of specific binding already at E15 in mouse brain, which increased progressively until adulthood. Collectively, we show that (3)H-HOCPCA is a highly sensitive radioligand, offering advantages over the commonly used radioligand (3)H-NCS-382, and thus a very suitable in vitro tool for qualitative and quantitative autoradiography of the GHB high-affinity site.

AB - GHB (γ-hydroxybutyric acid) is a compound endogenous to mammalian brain with high structural resemblance to GABA. GHB possesses nanomolar-micromolar affinity for a unique population of binding sites, but the exact nature of these remains elusive. In this study we utilized the highly selective GHB analogue, 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) as a tritiated version ((3)H-HOCPCA) to radioactively label the specific GHB high-affinity binding site and gain further insight into the density, distribution and developmental profile of this protein. We show that, in low nanomolar concentrations, (3)H-HOCPCA displays excellent signal-to-noise ratios using rodent brain autoradiography, which makes it a valuable ligand for anatomical quantification of native GHB binding site levels. Our data confirmed that (3)H-HOCPCA labels only the high-affinity specific GHB binding site, found in high density in cortical and hippocampal regions. The experiments revealed markedly stronger binding at pH 6.0 (Kd 73.8 nM) compared to pH 7.4 (Kd 2312 nM), as previously reported for other GHB radioligands but similar Bmax values. Using (3)H-HOCPCA we analyzed the GHB binding protein profile during mouse brain development. Due to the high sensitivity of this radioligand, we were able to detect low levels of specific binding already at E15 in mouse brain, which increased progressively until adulthood. Collectively, we show that (3)H-HOCPCA is a highly sensitive radioligand, offering advantages over the commonly used radioligand (3)H-NCS-382, and thus a very suitable in vitro tool for qualitative and quantitative autoradiography of the GHB high-affinity site.

U2 - 10.1016/j.neuint.2016.09.002

DO - 10.1016/j.neuint.2016.09.002

M3 - Journal article

VL - 100

SP - 138

EP - 145

JO - Neurochemistry International

JF - Neurochemistry International

SN - 0197-0186

ER -

ID: 169565554