eprintid: 1569998 rev_number: 31 eprint_status: archive userid: 608 dir: disk0/01/56/99/98 datestamp: 2017-08-13 01:49:31 lastmod: 2021-09-29 22:13:36 status_changed: 2017-10-24 14:01:20 type: article metadata_visibility: show creators_name: Sheilabi, MA creators_name: Battacharyya, D creators_name: Caetano, L creators_name: Thom, M creators_name: Reuber, M creators_name: Duncan, JS creators_name: Princivalle, AP title: Quantitative expression and localization of GABAB receptor protein subunits in hippocampi from patients with refractory temporal lobe epilepsy ispublished: pub divisions: UCL divisions: B02 divisions: C07 divisions: D07 divisions: F81 keywords: GABA(B) qRT-PCR, Hippocampal sclerosis, Human temporal lobe epilepsy, Immunohistochemistry, Quantitative Western blot note: © 2017 Published by Elsevier Ltd. This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. 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 characterised 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. date: 2018-07-01 date_type: published official_url: http://doi.org/10.1016/j.neuropharm.2017.08.001 oa_status: green full_text_type: other language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1410514 doi: 10.1016/j.neuropharm.2017.08.001 pii: S0028-3908(17)30368-4 lyricists_name: Duncan, John lyricists_name: Thom, Maria lyricists_id: JSDUN52 lyricists_id: MTHOM26 actors_name: Thom, Maria actors_name: Laslett, David actors_id: MTHOM26 actors_id: DLASL34 actors_role: owner actors_role: impersonator full_text_status: public publication: Neuropharmacology volume: 136 number: A pagerange: 117-128 event_location: England issn: 1873-7064 citation: Sheilabi, MA; Battacharyya, D; Caetano, L; Thom, M; Reuber, M; Duncan, JS; Princivalle, AP; (2018) Quantitative expression and localization of GABAB receptor protein subunits in hippocampi from patients with refractory temporal lobe epilepsy. Neuropharmacology , 136 (A) pp. 117-128. 10.1016/j.neuropharm.2017.08.001 <https://doi.org/10.1016/j.neuropharm.2017.08.001>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/1569998/1/Thom_Revised%20GABAB_TLE%20V7%20MR-AP%20V2.pdf