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Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies. / Charlotte Amy Hunt-Jones

Swansea University Author: Charlotte Amy Hunt-Jones

Abstract

Epilepsy affects over 450,000 people in the UK and there are over 50 epilepsy phenotypes; genetic generalised epilepsy (GGE) account for up to 30% of seizure types. It is established that GGE and other neurological disorders are, in some cases, caused by channelopathies within post-synaptic inhibito...

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Published: 2015
Institution: Swansea University
Degree level: Doctoral
Degree name: Ph.D
URI: https://cronfa.swan.ac.uk/Record/cronfa43036
first_indexed 2018-08-02T18:56:08Z
last_indexed 2019-10-21T16:48:54Z
id cronfa43036
recordtype RisThesis
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spelling 2018-08-29T14:53:53.4948224 v2 43036 2018-08-02 Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies. 9321a53ff7b0f65901a979f9b451be3d NULL Charlotte Amy Hunt-Jones Charlotte Amy Hunt-Jones true true 2018-08-02 Epilepsy affects over 450,000 people in the UK and there are over 50 epilepsy phenotypes; genetic generalised epilepsy (GGE) account for up to 30% of seizure types. It is established that GGE and other neurological disorders are, in some cases, caused by channelopathies within post-synaptic inhibitory neurotransmitter systems such as GAB A (epilepsy) and Glycine (hyperekplexia). GAB A is the primary inhibitory neurotransmitter in the brain and is synthesised from glutamate by GAD65 and 67, and is released from the pre-synaptic nerve terminal into the synaptic cleft, where it binds to post-synaptic GABA receptors and initiate neuroinhibition. This inhibition is removed by post-synaptic GABA transporters (GAT1 and GAT3) that uptake GABA back into the cell for re-packaging in presynaptic vesicles or breakdown by transamination. Abnormalities in this system have been linked to diseases including anxiety, psychosis, Parkinsons’s Disease and epilepsy. GABAergic animal models have demonstrated a tendency to seizure, including GABA transporter and enzyme models in relation to epilepsy.Given the above, the aim of this study was to identify GGE causing variants in four GABAergic genes. GGE patient samples (n=101) were recruited from 3 global centres and screened for variations in GAT1, GAT3, GAD65, GAD67 using high-throughput LightScanner analysis and bi-directional Sanger sequencing. Control population studies («=480) were carried out and analysis of online databases to determine the frequency of variants. Twenty novel or very rare variants were identified in 48 patient samples representing a detection rate of 6.8%, where a clustering of phenotypes included a predisposition towards absence seizures. The biological consequences of these variants were predicted using three online predictive programmes, multiple phylogenetic alignments and 3D structural modelling. Mutation expression constructs were prepared and expression levels were validated by immunocytochemistry. Functional characterisation of these variants will hopefully improve genetic diagnosis in GGE and determine causality of GABAergic absence seizures. E-Thesis Epliepsy 31 12 2015 2015-12-31 COLLEGE NANME Swansea University Medical School COLLEGE CODE Swansea University Doctoral Ph.D 2018-08-29T14:53:53.4948224 2018-08-02T16:24:31.1318014 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Charlotte Amy Hunt-Jones NULL 1 0043036-02082018162541.pdf 10821426.pdf 2018-08-02T16:25:41.6300000 Output 28431577 application/pdf E-Thesis true 2018-08-02T16:25:41.6300000 false
title Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies.
spellingShingle Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies.
Charlotte Amy Hunt-Jones
title_short Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies.
title_full Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies.
title_fullStr Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies.
title_full_unstemmed Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies.
title_sort Mutation analysis of GABAergic neuroinhibitory genes in childhood genetic generalised epilepsies.
author_id_str_mv 9321a53ff7b0f65901a979f9b451be3d
author_id_fullname_str_mv 9321a53ff7b0f65901a979f9b451be3d_***_Charlotte Amy Hunt-Jones
author Charlotte Amy Hunt-Jones
author2 Charlotte Amy Hunt-Jones
format E-Thesis
publishDate 2015
institution Swansea University
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 1
active_str 0
description Epilepsy affects over 450,000 people in the UK and there are over 50 epilepsy phenotypes; genetic generalised epilepsy (GGE) account for up to 30% of seizure types. It is established that GGE and other neurological disorders are, in some cases, caused by channelopathies within post-synaptic inhibitory neurotransmitter systems such as GAB A (epilepsy) and Glycine (hyperekplexia). GAB A is the primary inhibitory neurotransmitter in the brain and is synthesised from glutamate by GAD65 and 67, and is released from the pre-synaptic nerve terminal into the synaptic cleft, where it binds to post-synaptic GABA receptors and initiate neuroinhibition. This inhibition is removed by post-synaptic GABA transporters (GAT1 and GAT3) that uptake GABA back into the cell for re-packaging in presynaptic vesicles or breakdown by transamination. Abnormalities in this system have been linked to diseases including anxiety, psychosis, Parkinsons’s Disease and epilepsy. GABAergic animal models have demonstrated a tendency to seizure, including GABA transporter and enzyme models in relation to epilepsy.Given the above, the aim of this study was to identify GGE causing variants in four GABAergic genes. GGE patient samples (n=101) were recruited from 3 global centres and screened for variations in GAT1, GAT3, GAD65, GAD67 using high-throughput LightScanner analysis and bi-directional Sanger sequencing. Control population studies («=480) were carried out and analysis of online databases to determine the frequency of variants. Twenty novel or very rare variants were identified in 48 patient samples representing a detection rate of 6.8%, where a clustering of phenotypes included a predisposition towards absence seizures. The biological consequences of these variants were predicted using three online predictive programmes, multiple phylogenetic alignments and 3D structural modelling. Mutation expression constructs were prepared and expression levels were validated by immunocytochemistry. Functional characterisation of these variants will hopefully improve genetic diagnosis in GGE and determine causality of GABAergic absence seizures.
published_date 2015-12-31T04:46:43Z
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score 11.310851

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