Quantitative expression and localization of GABAB receptor protein subunits in hippocampi from patients with refractory temporal lobe epilepsy

SHEILABI, Mariam, BATTACHARYYA, Dev, CAETANO, Laura, THOM, Maria, REUBER, Markus, DUNCAN, John S and PRINCIVALLE, Alessandra (2017). Quantitative expression and localization of GABAB receptor protein subunits in hippocampi from patients with refractory temporal lobe epilepsy. Neuropharmacology. (In Press)

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Link to published version:: 10.1016/j.neuropharm.2017.08.001

Abstract

This study investigates GABAB protein expression and mRNA levels in three types of specimens. Two types of specimens from patients with temporal lobe epilepsy (TLE), secondary to hippocampal sclerosis, sclerotic hippocampal samples (TLE-HS), and tissue from the structurally preserved non-spiking ipsilateral superior temporal gyrus (TLE-STG) removed from the same patient during epilepsy surgery; and third specimen is hippocampal tissue from individuals with no history of epilepsy (post-mortem controls, PMC). mRNA expression of GABAB subunits was quantified in TLE-HS, TLE-STG and PMC specimens by qRT-PCR. Qualitative and quantitative Western blot (WB) and immunohistochemistry techniques were employed to quantify and localize GABAB proteins subunits. qRT-PCR data demonstrated an overall decrease of both GABAB1 isoforms in TLE-HS compared to TLE-STG. These results were mirrored by the WB findings. GABAB2 mRNA and protein were significantly reduced in TLE-HS samples compared to TLE-STG; however they appeared to be upregulated in TLE-HS compared to the PMC samples. Immunohistochemistry (IHC) showed that GABAB proteins were widely distributed in PMC and TLE-HS hippocampal sections with regional differences in the intensity of the signal. The higher expression of mature GABAB protein in TLE-HS than PMC is in agreement with previous studies. However, these findings could be due to post-mortem changes in PMC specimens. The TLE-STG samples examined here represent a better 'control' tissue compared to TLE-HS samples characterized by lower than expected GABAB expression. This interpretation provides a better explanation for previous functional studies suggesting reduced inhibition in TLE-HS tissue due to attenuated GABAB currents. [Abstract copyright: Copyright © 2017. Published by Elsevier Ltd.]

Item Type: Article
Additional Information: ** From PubMed via Jisc Publications Router. ** History: ** received: 16-06-2017 ** revised: 01-08-2017 ** accepted: 02-08-2017
Uncontrolled Keywords: GABA(B) qRT-PCR, Hippocampal sclerosis, Human temporal lobe epilepsy, Immunohistochemistry, Quantitative Western blot
Research Institute, Centre or Group: Biomolecular Sciences Research Centre
Identification Number: 10.1016/j.neuropharm.2017.08.001
SWORD Depositor: Jill Hazard
Depositing User: Jill Hazard
Date Deposited: 18 Aug 2017 08:37
Last Modified: 19 Aug 2017 11:01
URI: http://shura.shu.ac.uk/id/eprint/16563

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