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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F16%3A00469234" target="_blank" >RIV/61388963:_____/16:00469234 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.neuint.2016.09.002" target="_blank" >http://dx.doi.org/10.1016/j.neuint.2016.09.002</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.neuint.2016.09.002" target="_blank" >10.1016/j.neuint.2016.09.002</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

GHB (gamma-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 (H-3-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, 3H-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 3H-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 (K-d 73.8 nM) compared to pH 7.4 (Kd 2312 nM), as previously reported for other GHB radioligands but similar B-max values. Using H-3-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 H-3-HOCPCA is a highly sensitive radioligand, offering advantages over the commonly used radioligand H-3-NCS-382, and thus a very suitable in vitro tool for qualitative and quantitative autoradiography of the GHB high-affinity site.

Název v anglickém jazyce

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

Popis výsledku anglicky

GHB (gamma-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 (H-3-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, 3H-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 3H-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 (K-d 73.8 nM) compared to pH 7.4 (Kd 2312 nM), as previously reported for other GHB radioligands but similar B-max values. Using H-3-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 H-3-HOCPCA is a highly sensitive radioligand, offering advantages over the commonly used radioligand H-3-NCS-382, and thus a very suitable in vitro tool for qualitative and quantitative autoradiography of the GHB high-affinity site.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

—

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2016

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Neurochemistry International

ISSN

0197-0186

e-ISSN

—

Svazek periodika

100

Číslo periodika v rámci svazku

Nov

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

8

Strana od-do

138-145

Kód UT WoS článku

000388780300016

EID výsledku v databázi Scopus

2-s2.0-84988521805