Synapse Dysfunction Induced by Autism Spectrum Disorder Associated Shank3 Mutations
type: CreativeWork
description: Autism spectrum disorders (ASD) comprise a range of neurodevelopmental disorders characterised by deficits in social interaction and communication, and by repetitive behaviours. Recent research has identified ASD-associated mutations in important synaptic proteins, and this has prompted investigations into understanding the roles of synapse dysfunction in the pathogenesis of ASD. Shank3 is a postsynaptic scaffolding protein that is essential to regulating both the function and structure of excitatory synapses and Shank3 genetic mutations have been described in people affected with ASD. However, currently little is known about the effects of ASD-associated mutations in Shank3 on synapse structure, function and plasticity. The work in this thesis has examined the structural and physiological synaptic changes that are induced by alterations in Shank3 expression levels or by the expression of ASD-associated mutations in Shank3. In these experiments, dissociated rat hippocampal neurons grown in vitro were transiently transfected with Shank3 wild-type (WT) to increase normal Shank3 levels, Shank3 short hairpin ribonucleic acid (shRNA) to down-regulate Shank3 expression, or mutant forms of Shank3 that have been identified in people with ASDs including R87C and R375C point mutations, and the InsG frameshift mutation. The first part of this thesis aimed to investigate the changes that occur in γ- aminobutyric acid (GABA)-ergic inhibitory synapses induced by up- or down-regulation of Shank3 expression or expression of ASD-associated mutations in Shank3. We specifically investigated changes in the expression level and density of GABAergic pre- and postsynaptic proteins by immunocytochemistry, and examined inhibitory synaptic transmission using whole cell patch clamp electrophysiology. To directly compare the results observed in GABAergic inhibitory synapses to glutamatergic excitatory synapses, the density of excitatory synapses and the properties of excitatory synaptic transm…
abstract: Autism spectrum disorders (ASD) comprise a range of neurodevelopmental disorders characterised by deficits in social interaction and communication, and by repetitive behaviours. Recent research has identified ASD-associated mutations in important synaptic proteins, and this has prompted investigations into understanding the roles of synapse dysfunction in the pathogenesis of ASD. Shank3 is a postsynaptic scaffolding protein that is essential to regulating both the function and structure of excitatory synapses and Shank3 genetic mutations have been described in people affected with ASD. However, currently little is known about the effects of ASD-associated mutations in Shank3 on synapse structure, function and plasticity. The work in this thesis has examined the structural and physiological synaptic changes that are induced by alterations in Shank3 expression levels or by the expression of ASD-associated mutations in Shank3. In these experiments, dissociated rat hippocampal neurons grown in vitro were transiently transfected with Shank3 wild-type (WT) to increase normal Shank3 levels, Shank3 short hairpin ribonucleic acid (shRNA) to down-regulate Shank3 expression, or mutant forms of Shank3 that have been identified in people with ASDs including R87C and R375C point mutations, and the InsG frameshift mutation. The first part of this thesis aimed to investigate the changes that occur in γ- aminobutyric acid (GABA)-ergic inhibitory synapses induced by up- or down-regulation of Shank3 expression or expression of ASD-associated mutations in Shank3. We specifically investigated changes in the expression level and density of GABAergic pre- and postsynaptic proteins by immunocytochemistry, and examined inhibitory synaptic transmission using whole cell patch clamp electrophysiology. To directly compare the results observed in GABAergic inhibitory synapses to glutamatergic excitatory synapses, the density of excitatory synapses and the properties of excitatory synaptic transm…
author: Lee, Kevin
license: https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm
date published: 2015
date modified: 2026-07-24T01:06:48Z
provider: University of Auckland
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: ResearchSpace@Auckland

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property ID: Handle
value: https://hdl.handle.net/2292/26586