Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2006. / Härtill 4 uppsatser.

While the causes of myriad medical and infectious illnesses have been identified, the etiologies of neuropsychiatric illnesses remain elusive. This is due to two major obstacles. First, the risk for neuropsychiatric disorders, such as schizophrenia, is determined by both genetic and environmental factors. Second, numerous genes influence susceptibility for these illnesses. Genome-wide association studies have identified at least 108 genomic loci for schizophrenia, and more are expected to be published shortly. In addition, numerous biological processes contribute to the neuropathology underlying schizophrenia. These include immune dysfunction, synaptic and myelination deficits, vascular abnormalities, growth factor disruption, and N-methyl-D-aspartate receptor (NMDAR) hypofunction. However, the field of psychiatric genetics lacks a unifying model to explain how environment may interact with numerous genes to influence these various biological processes and cause schizophrenia. Here we describe a biological cascade of proteins that are activated in response to environmental stimuli such as stress, a schizophrenia risk factor. The central proteins in this pathway are critical mediators of memory formation and a particular form of hippocampal synaptic plasticity, long-term depression (LTD). Each of these proteins is also implicated in schizophrenia risk. In fact, the pathway includes four genes that map to the 108 loci associated with schizophrenia: GRIN2A, nuclear factor of activated T-cells (NFATc3), early growth response 1 (EGR1) and NGFI-A Binding Protein 2 (NAB2); each of which contains the "Index single nucleotide polymorphism (SNP)" (most SNP) at its respective locus. Environmental stimuli activate this biological pathway in neurons, resulting in induction of EGR immediate early genes: EGR1, EGR3 and NAB2. We hypothesize that dysfunction in any of the genes in this pathway disrupts the normal activation of Egrs in response to stress. This may result in insufficient electrophysiologic, immunologic, and neuroprotective, processes that these genes normally mediate. Continued adverse environmental experiences, over time, may thereby result in neuropathology that gives rise to the symptoms of schizophrenia. By combining multiple genes associated with schizophrenia susceptibility, in a functional cascade triggered by neuronal activity, the proposed biological pathway provides an explanation for both the polygenic and environmental influences that determine the complex etiology of this mental illness.

No description available.

The ventral tegmental area (VTA) is an important component of the mesolimbic dopamine circuit and processes reward and motivational behaviors. In response to drug exposure, synaptic connections of this circuit can be rewired via synaptic plasticity—a phenomenon thought be responsible for the pathology of addiction. While much is known about dopamine neuron plasticity, less is known regarding plasticity exhibited by VTA GABA cells, specifically inhibitory inputs from outside the VTA. Expanding on the work of Bocklisch et al. (2013), we investigated the plasticity of inhibitory inputs to VTA GABA neurons. Using whole cell electrophysiology in GAD67 GFP mice, we observed that these VTA GABA cells can experience either long-term potentiation (LTP) or long-term depression (LTD) in response to a 5 Hz stimulus. While neither the LTP nor LTD appear to be mediated by the cannabinoid-1 receptor (CB1), the nitric oxide synthase (NOS) pathway, or the dopamine-2 (D2) receptor, the LTP is blocked by APV, an NMDA receptor antagonist, and the LTD is blocked by CGP 54626, an antagonist of the GABAB receptor. Additionally, µ-opioid and adenosine-1 receptors modulated plasticity at this synapse, but chronic morphine administration (10mg/kg) did not block the observed LTP or LTD. Furthermore, we used an optogenetic approach in VGAT-Cre mice to target inhibitory inputs from the lateral hypothalamus (LH) to the VTA. An optical stimulus (5 Hz) caused these inputs to depress, which has not been previously described and may be behaviorally important in reward processing. These novel findings increase our understanding of VTA neural circuitry, ultimately increasing our capacity to better comprehend and treat the pathology of addiction. Additionally, changes in synaptic strength in hippocampal CA1 pyramidal cells are thought to be responsible for the acquisition and retention of short-term memory. Feedforward stratum radiatum interneurons of many subtypes experience LTD, short-term depression (STD), or lack of plasticity, but it is not known whether plasticity correlates with specific interneuron subtypes. Using whole cell electrophysiology and qPCR, we characterized the plasticity expressed by hippocampal interneurons in correlation with their mRNA expression patterns to determine cell subtype. We also assessed the expression of endocannabinoid (eCB) biosynthetic enzymes as well as metabotropic glutamate receptor subunits known to mediate plasticity. Cells exhibiting LTD tended to express mRNA for at least one of the eCB biosynthetic enzymes and the metabotropic glutamate receptor subunit mGluR5. mGluR5 was not expressed by cells exhibiting STD or no plasticity. Cells that exhibited short-term depression tended to express mRNA for at least one of the eCB biosynthetic enzymes, but not mGluR5. This suggests that stratum radiatum interneuron plasticity can be predicted based on mGluR expression, and that these different types of plasticity may have some importance in hippocampal function.

Endocannabinoids (eCBs) are lipophilic signals that are produced by postsynaptic neurons in an activity-dependent manner, and signal in a retrograde fashion to modulate neurotransmitter release. As such, eCBs are highly involved in synaptic plasticity, a process that strengthens or weakens synapses. eCB-mediated synaptic plasticity is involved in many brain processes including learning, short-term memory, and adaptive reward, which are processed in the hippocampus and ventral tegmental area (VTA), respectively. However, the expression of eCB biosynthetic enzyme mRNA within hippocampal and VTA neurons, as well as the relationship between these mRNA species and the occurrence of synaptic plasticity, remains unclear. The goal of these studies was to demonstrate the expression pattern of eCB biosynthetic enzyme mRNA within hippocampal and VTA neurons, and to describe the relationship between synaptic plasticity and mRNA expression. Using whole-cell electrophysiology and real-time quantitative PCR, I tested hippocampal and VTA neurons for the presence of eCB biosynthetic enzyme mRNA and described the relationship between these enzymes and synaptic plasticity. The data presented herein demonstrate the importance of eCB signaling within the hippocampus and VTA and the expression patterns of eCB biosynthetic machinery within several neuron types. These data provide evidence that eCB signaling plays a critical role in learning, short-term memory, and adaptive reward.

Recent evidence indicates that the mature neocortex retains a higher degree of plasticity than traditionally assumed. Up- and down-regulation of synaptic strength, long-term potentiation (LTP) and long-term depression (LTD), is thought to be the primary mechanism mediating experience-dependent plasticity of cortical networks. The present thesis investigate factors that regulate adult cortical plasticity, focusing on the role of neuromodulators, recent sensory experience, and different anatomical divisions of the cortex in influencing synaptic strength. First, I investigated the role of the neuromodulator histamine in gating plasticity in the primary visual cortex (V1) of urethane anesthetized adult rats. Histamine applied locally in V1 produced an enhancement of LTP elicited by theta burst stimulation (TBS) of dorsal lateral geniculate nucleus (dLGN) and allowed a sub-threshold TBS to produce stable LTP. Second, the impact of visual deprivation on LTP in V1 was assessed. Animals that received 2 and 5 hr dark exposure showed greater potentiation of field potentials when stimulated though retinal light flashes or weak TBS of the dLGN, which failed to induce LTP in control animals kept in continuous light. Third, I performed a detailed characterization of LTP induced by different TBS protocols, recording in either the monocular or binocular segment of both V1 hemispheres (i.e., ipsi- and contralateral to the stimulated dLGN). Stronger, NMDA receptor-independent LTP was found in the contralateral V1. Interestingly, weak TBS induced LTD that was NMDA receptor-dependent in the ipsilateral V1. Furthermore, a lower LTP induction threshold was observed in the binocular than the monocular segment of ipsilateral V1. Lastly, I investigated cholinergic modulation of sensory-induced activity in the barrel cortex. Basal forebrain stimulation enhanced multi-unit activity elicited by whisker deflection, an effect that was more pronounced for weaker response driven by a secondary whisker than principal whisker deflection. This thesis demonstrates that neocortical plasticity consists of multiple forms of synaptic modification. Adult cortical plasticity is greatly influenced by preceding activity of the synapse by various neuromodulator systems, and by anatomical subdivisions within primary sensory cortex fields. Together, these mechanisms may facilitate the detection, amplification, and storage of inputs to primary sensory fields of the neocortex. / Thesis (Ph.D, Psychology) -- Queen's University, 2010-04-29 14:02:30.742

Many molecules and the complex interactions between them underlie plasticity in the cerebellum. However, the exact relationship between cerebellar plasticity and the different signalling cascades remains unclear. Calcium-calmodulin dependent protein kinase II (CaMKII) regulates many forms of synaptic plasticity, but very little is known about its function during plasticity induction in the cerebellum. The aim of this thesis is to contribute to a better understanding of the molecular mechanisms that regulate the induction of synaptic plasticity in cerebellar Purkinje cells (PCs). The focus of the thesis is to investigate the role of CaMKII isoforms in the bidirectional modulation of plasticity induction at parallel fibre (PF)-PC synapses. For this investigation, computational models that represent the CaMKII activation and the signalling network that mediates plasticity induction at these synapses were constructed. The model of CaMKII activation by calcium-calmodulin developed by Dupont et al (2003) replicates the experiments by De Koninck and Schulman (1998). Both theoretical and experimental studies have argued that the phosphorylation and activation of CaMKII depends on the frequency of calcium oscillations. Using a simplified version of the Dupont model, it was demonstrated that the CaMKII phosphorylation is mostly determined by the average calcium-calmodulin concentration, and therefore depends only indirectly on the actual frequency of calcium oscillations. I have shown that a pulsed application of calcium-calmodulin is, in fact, not required at all. These findings strongly indicate that the activation of CaMKII depends on the average calcium-calmodulin concentration and not on the oscillation frequency per se as asserted in those studies. This thesis also presents the first model of AMPA receptor phosphorylation that simulates the induction of long-term depression (LTD) and potentiation (LTP) at the PF-PC synapse. The results of computer simulations of a simple mathematical model suggest that the balance of CaMKII-mediated phosphorylation and protein phosphatase 2B (PP2B)-mediated dephosphorylation of AMPA receptors determines whether LTD or LTP occurs in cerebellar PCs. This model replicates the experimental observations by Van Woerden et al (2009) that indicate that CaMKII controls the direction of plasticity at PF-PC synapses. My computer simulations support Van Woerden et al’s original suggestion that filamentous actin binding can enable CaMKII to regulate bidirectional plasticity at these synapses. The computational models of intracellular signalling constructed in this thesis advance the understanding of the mechanisms of synaptic plasticity induction in the cerebellum. These simple models are significant tools for future research by the scientific community.

Long-term potentiation (LTP) and depression (LTD) are activity dependent long-lasting changes in synaptic efficacy and have been proposed as mechanisms for learning and memory. Although the exact relationship of LTP and LTD to memory is not known, they do share some properties and mechanisms that relate to memory, such as the strengthening and weakening of synapses. LTP and LTD have been studied extensively in hippocampal brain-slice preparations, due to its relatively organized structure, ease of induction, and its critical function in memory storage. Less work has been done in the neocortex despite the belief that it is heavily involved in the storage of long-term memories. Activity dependent plasticity has also been demonstrated in the basal ganglia in vivo and in vitro, but the results have been somewhat inconsistent. The experiments presented in this thesis explore a novel form of neural plasticity in two excitatory pathways (corticostriatal and thalamocortical) of the basal ganglia motor loop in the intact brain in awake, freely behaving rats. In thalamocortical slice preparations, simultaneous presynaptic stimulation and postsynaptic depolarization can induce L TP in animals prior to the critical period. However the results presented in this thesis show that applied stimulation to the thalamocortical pathway failed to produce either LTP or LTD in the awake freely moving animal.Corticostriatal LTD has been shown in slice preparations following direct tetanic stimulation of the striatum. In the current experiment, cortical stimulation failed to induce LTD although there was an observable decrease in the evoked potential following low-frequency stimulation. Corticostriatal L TP has been shown to depend on the type of stimulation applied. High-frequency and theta burst stimulation produced long-lasting changes in response amplitude in the corticostriatal pathway, with theta burst stimulation appearing to be the more effective stimulation protocol for inducing LTP in both the early and late components. Paired stimulation of the substantia nigra pars compacta and cortex indicated a modulatory action of dopamine on corticostriatal synaptic plasticity. Pairing led to a stable increase in the amplitude of LTP of both early and late components. We also report that a temporal relationship exists in the striatum with respect to the release of nigral dopamine and cortical glutamate. Simultaneous stimulation produced a more robust L TP compared to the two other conditions in which there was an applied stimulation delay to either the corticostriatal or nigrostriatal pathway. The results demonstrate the mechanistic differences, not only between the thalamocortical and corticostriatal pathways, but also slice and anesthetized preparations. The results also emphasize the need for further study on mechanisms of L TP and LTD in the various excitatory and inhibitory pathways of the basal ganglia motor loop. / Thesis / Doctor of Philosophy (PhD)

Cannabinoids have been shown to alter synaptic plasticity but the mechanism by which this occurs at hippocampal CA3-CA1 synapses in vivo is not yet known. Utilizing in vivo electrophysiological recordings of field excitatory postsynaptic potentials (fEPSP) on anesthetized rats and mice as well as three lines of conditional knockout mouse models, the objective was to show a two-part mechanistic breakdown of cannabinoid-evoked CA3-CA1 long-term depression (LTD) in its induction as well as early and later-phase expression stages. It was determined that this cannabinoid-induced in vivo LTD requires cannabinoid type-1 receptors (CB1Rs) on astrocytes, but not CB1Rs on glutamatergic or GABAergic neuronal axons/terminals. Pharmacological testing determined that cannabinoid-induced in vivo LTD also requires activation of NMDA receptors (NMDAR) and subsequent postsynaptic endocytosis of AMPA receptors (AMPAR). There exists a clear role for NR2B-containing NMDARs in a persistent, transitory form, potentially related to prolonged or delayed glutamate release (possibly as a result of the astrocytic network). A key determination of the expression phase is the involvement of new protein synthesis (using translation and transcription inhibitors) – further evidence of the long-term action of the synaptic plasticity from a single cannabinoid dose.

Cannabinoids have been shown to alter synaptic plasticity but the mechanism by which this occurs at hippocampal CA3-CA1 synapses in vivo is not yet known. Utilizing in vivo electrophysiological recordings of field excitatory postsynaptic potentials (fEPSP) on anesthetized rats and mice as well as three lines of conditional knockout mouse models, the objective was to show a two-part mechanistic breakdown of cannabinoid-evoked CA3-CA1 long-term depression (LTD) in its induction as well as early and later-phase expression stages. It was determined that this cannabinoid-induced in vivo LTD requires cannabinoid type-1 receptors (CB1Rs) on astrocytes, but not CB1Rs on glutamatergic or GABAergic neuronal axons/terminals. Pharmacological testing determined that cannabinoid-induced in vivo LTD also requires activation of NMDA receptors (NMDAR) and subsequent postsynaptic endocytosis of AMPA receptors (AMPAR). There exists a clear role for NR2B-containing NMDARs in a persistent, transitory form, potentially related to prolonged or delayed glutamate release (possibly as a result of the astrocytic network). A key determination of the expression phase is the involvement of new protein synthesis (using translation and transcription inhibitors) – further evidence of the long-term action of the synaptic plasticity from a single cannabinoid dose.