Role of hilar mossy cells in the CA3-dentate gyrus network during sharp wave-ripple activity in vitro

Der Gyrus dentatus (DG) des Hippokampus wird als Eingangsstation für Informationen aus dem entorhinalen Kortex betrachtet. In das DG-Netzwerk sind zwei exzitatorische Zelltypen eingebettet: Körnerzellen, die Signale von dem entorhinalen Kortex empfangen, und Hilus-Mooszellen (MCs), die Signale von Körnerzellen als auch von feedback-Projektionen von CA3-Pyramidenzellen (PCs) empfangen. Postsynaptische Ziele von MC-Projektionen umfassen DG Körnerzellen und verschiedene Interneurone in der selben und in der kontralateralen Hemisphäre des Gehirns. Die Rolle von MCs während rhythmischer Populationsaktivität, und insbesondere während Sharp-Wave / Ripple-Komplexen (SWRs), ist bisher weitgehend unerforscht. SWRs sind prominente Ereignisse im Hippocampus während des Tiefschlafs (Slow wave sleep) und des ruhigen Wachzustandes, und sie sind an der Gedächtniskonsolidierung beteiligt. In der vorliegenden Arbeit, untersuchen wir mithilfe eines in-vitro-Modells von SWRs, inwieweit Mooszellen an SWRs in CA3 beteiligt sind. Mit CA3-Feldpotential-Ableitungen und gleichzeitigen ‚cell-attached‘ Messungen von einzelnen MCs konnten wir beobachten, dass ein wesentlicher Anteil von MCs (47%) während der SWRs in das aktive neuronale Netzwerk rekrutiert werden. Darüber hinaus fanden wir in MCs SWR-assoziierte synaptische Aktivität, bei denen sowohl die exzitatorischen als auch die inhibitorischen Komponenten phasenkohärent und verzögert zur Ripple Oszillation in CA3 auftreten. Simultane Patch-clamp Messungen von CA3-Pyramidenzellen und MCs zeigten längere exzitatorische und inhibitorische Latenzzeiten bei MCs, was die Hypothese einer von CA3 ausgehenden Feedback-Rekrutierung unterstützt. Unsere Daten zeigen zusätzlich, dass das Verhältnis exzitatorischer zu inhibitorischer Aktivität in MCs höher ist als in CA3-Pyramidenzellen, wodurch die MCs mit höherer Wahrscheinlichkeit während SWRs überschwellig aktiviert werden. Schließlich zeigen wir, dass ein signifikanter Anteil (66%) der getesteten Körnerzellen SWR-assoziierte exzitatorische Signale erhalten, im Vergleich zu MCs zeitlich verzögert, was auf eine indirekte Aktivierung von Körnerzellen durch CA3 PCs über MCs hinweist. Zusammengefasst zeigen unsere Daten die aktive Beteiligung von Mooszellen an SWRs und deuten auf eine funktionelle Bedeutung als Schaltstelle für das CA3- Gyrus dentatus Netzwerk in diesem wichtigen physiologischen Netzwerkzustand hin.The dentate gyrus (DG) is considered as the hippocampal input gate for the information arriving from the entorhinal cortex. Embedded into the DG network are two excitatory cell types –granule cells (GCs), which receive inputs from the entorhinal cortex, and hilar mossy cells (MCs), which receive input from GCs and feedback projections from CA3 pyramidal cells (PCs). The postsynaptic targets of MC projections are the GCs and hilar interneurons in both ipsilateral and contralateral hemispheres of the brain. The role of MCs during rhythmic population activity, and in particular during sharp-wave/ripple complexes (SWRs), has remained largely unexplored. SWRs are prominent field events in the hippocampus during slow wave sleep and quiet wakefulness, and are involved in memory consolidation and future planning. In this study, we sought to understand whether MCs participate during CA3 SWRs using an in vitro model of SWRs. With simultaneous CA3 field potential– and cell-attached recordings from MCs, we observed that a significant fraction of MCs (47%) are recruited into the active neuronal network during SWRs. Moreover, MCs receive pronounced, compound, ripple-associated synaptic input where both excitatory and inhibitory components are phase-coherent with and delayed to the CA3 ripple. Simultaneous patch recordings from CA3 pyramidal neurons and MCs revealed longer excitatory and inhibitory latencies in MCs, supporting a feedback recruitment from CA3. Our data also show that the excitatory to inhibitory charge transfer (E/I) ratio in MCs is higher than in the CA3 PCs, making the MCs more likely to spike during SWRs. Finally, we demonstrate that a significant fraction (66%) of tested GCs receive SWR-associated excitatory inputs that are delayed compared to MCs, indicating an indirect activation of GCs by CA3 PCs via MCs. Together, our data suggest the involvement of mossy cells during SWRs and their importance as a relay for CA3-dentate gyrus networks in this important physiological network state

Conditional Spike Transmission Mediated by Electrical Coupling Ensures Millisecond Precision-Correlated Activity among Interneurons In Vivo

Many GABAergic interneurons are electrically coupled and in vitro can display correlated activity with millisecond precision. However, the mechanisms underlying correlated activity between interneurons in vivo are unknown. Using dual patch-clamp recordings in vivo, we reveal that in the presence of spontaneous background synaptic activity, electrically coupled cerebellar Golgi cells exhibit robust millisecond precision-correlated activity which is enhanced by sensory stimulation. This precisely correlated activity results from the cooperative action of two mechanisms. First, electrical coupling ensures slow subthreshold membrane potential correlations by equalizing membrane potential fluctuations, such that coupled neurons tend to approach action potential threshold together. Second, fast spike-triggered spikelets transmitted through gap junctions conditionally trigger postjunctional spikes, depending on both neurons being close to threshold. Electrical coupling therefore controls the temporal precision and degree of both spontaneous and sensory-evoked correlated activity between interneurons, by the cooperative effects of shared synaptic depolarization and spikelet transmission

The role of the dopamine D4 receptor in modulating state-dependent gamma oscillations

Rhythmic oscillations in neuronal activity display variations in amplitude (power) over a range of frequencies. Attention and cognitive performance correlate with increases in cortical gamma oscillations (40-70Hz) that are generated by the coordinated firing of glutamatergic pyramidal neurons and GABAergic interneurons, and are modulated by dopamine. In the medial prefrontal cortex (mPFC) of rats, gamma power increases during treadmill walking, or after administration of an acute subanesthetic dose of the NMDA receptor antagonist ketamine. Ketamine is also used to mimic symptoms of schizophrenia, including cognitive deficits, in healthy humans and rodents. Additionally, the ability of a drug to modify ketamine-induced gamma power has been proposed to predict its pro-cognitive therapeutic efficacy. However, the mechanism underlying ketamine-induced gamma oscillations is poorly understood. We hypothesized that gamma oscillations induced by walking and ketamine would be generated by a shared mechanism in the mPFC and one of its major sources of innervation, the mediodorsal thalamus (MD). Recordings from chronically implanted electrodes in rats showed that both treadmill walking and ketamine increased gamma power, firing rates, and spike-gamma LFP correlations in the mPFC. By contrast, in the MD, treadmill walking increased all three measures, but ketamine decreased firing rates and spike-gamma LFP correlations while increasing gamma power. Therefore, walking- and ketamine-induced gamma oscillations may arise from a shared circuit in the mPFC, but different circuits in the MD. Recent work in normal animals suggests that dopamine D4 receptors (D4Rs) synergize with the neuregulin/ErbB4 signaling pathway to modulate gamma oscillations and cognitive performance. Consequently, we hypothesized that drugs targeting the D4Rs and ErbB receptors would show pro-cognitive potential by reducing ketamine-induced gamma oscillations in mPFC. However, when injected before ketamine, neither the D4R agonist nor antagonist altered ketamine’s effects on gamma power or firing rates in the mPFC, but the pan-ErbB antagonist potentiated ketamine’s increase in gamma power, and prevented ketamine from increasing firing rates. This indicates that D4Rs and ErbB receptors influence gamma power via distinct mechanisms that interact with NMDA receptor antagonism differently. Our results highlight the value of using ketamine-induced changes in gamma power as a means of testing novel pharmaceutical agents

IST Austria Thesis

Distinguishing between similar experiences is achieved by the brain in a process called pattern separation. In the hippocampus, pattern separation reduces the interference of memories and increases the storage capacity by decorrelating similar inputs patterns of neuronal activity into non-overlapping output firing patterns. Winners-take-all (WTA) mechanism is a theoretical model for pattern separation in which a "winner" cell suppresses the activity of the neighboring neurons through feedback inhibition. However, if the network properties of the dentate gyrus support WTA as a biologically conceivable model remains unknown. Here, we showed that the connectivity rules of PV+interneurons and their synaptic properties are optimizedfor efficient pattern separation. We found using multiple whole-cell in vitrorecordings that PV+interneurons mainly connect to granule cells (GC) through lateral inhibition, a form of feedback inhibition in which a GC inhibits other GCs but not itself through the activation of PV+interneurons. Thus, lateral inhibition between GC–PV+interneurons was ~10 times more abundant than recurrent connections. Furthermore, the GC–PV+interneuron connectivity was more spatially confined but less abundant than PV+interneurons–GC connectivity, leading to an asymmetrical distribution of excitatory and inhibitory connectivity. Our network model of the dentate gyrus with incorporated real connectivity rules efficiently decorrelates neuronal activity patterns using WTA as the primary mechanism. This process relied on lateral inhibition, fast-signaling properties of PV+interneurons and the asymmetrical distribution of excitatory and inhibitory connectivity. Finally, we found that silencing the activity of PV+interneurons in vivoleads to acute deficits in discrimination between similar environments, suggesting that PV+interneuron networks are necessary for behavioral relevant computations. Our results demonstrate that PV+interneurons possess unique connectivity and fast signaling properties that confer to the dentate gyrus network properties that allow the emergence of pattern separation. Thus, our results contribute to the knowledge of how specific forms of network organization underlie sophisticated types of information processing

Focal Augmentation of Somatostatin Interneuron Function and Subsequent Circuit Effects in Developmentally Malformed, Epileptogenic Cortex

Drug-resistant epilepsy (DRE) is a common clinical sequela of developmental cortical malformations such as polymicrogyria. Unfortunately, much remains unknown about the aberrant GABA-mediated circuit alterations that underlie DRE\u27s onset and persistence in this context. To address this knowledge gap, we utilized the transcranial freeze lesion model in optogenetic mice lines (Somatostatin (SST)-Cre or Parvalbumin (PV)-Cre x floxed channelrhodopsin-2) to dissect features of the SST, PV, and pyramidal neuron microcircuit that are potentially associated with DRE. Investigations took place within developmental microgyria’s known pathological substrate, the adjoined and epileptogenic paramicrogyral region (PMR). As well, microcircuit relationships within the previously unexplored range of normal-appearing cortex beyond PMR’s terminus were also interrogated. We previously demonstrated SST interneuron output enhancement onto postsynaptic layer V pyramidal neurons of PMR. Dissertation studies elaborated on this SST-interneuron mediated effect through the utilization of ex vivo slice electrophysiology in conjunction with selective optical activation of either SST or PV interneurons. An ostensible mechanism was identified in the form of a novel structural schematic for SST interneurons of PMR whereby they exhibit wider reaching, within-layer arborization of axons within this pathological substrate. Also, within PMR, SST interneuron output was not enhanced onto postsynaptic layer V PV interneurons, indicating targeting specificity of the SST to pyramidal neuron effect. Moving beyond PMR, past its terminus, SST interneuron output onto layer V pyramidal cells was found to be equivalent to controls, indicating effect focality. Finally, a novel disinhibitory relationship was demonstrated beyond PMR’s terminus, wherein PV interneurons exhibited output enhancement onto postsynaptic layer V SST interneurons. This indicates a putative in vivo mechanism for the PMR-focality of the SST to pyramidal neuron output enhancement scheme. These novel discoveries will provide the field with more context as to the role SST and PV interneurons potentially play in the emergence and/or modulation of drug-resistant epilepsy in and outside the terminus of PMR

Regulation of circuit organization and function through inhibitory synaptic plasticity

Diverse inhibitory neurons in the mammalian brain shape circuit connectivity and dynamics through mechanisms of synaptic plasticity. Inhibitory plasticity can establish excitation/inhibition (E/I) balance, control neuronal firing, and affect local calcium concentration, hence regulating neuronal activity at the network, single neuron, and dendritic level. Computational models can synthesize multiple experimental results and provide insight into how inhibitory plasticity controls circuit dynamics and sculpts connectivity by identifying phenomenological learning rules amenable to mathematical analysis. We highlight recent studies on the role of inhibitory plasticity in modulating excitatory plasticity, forming structured networks underlying memory formation and recall, and implementing adaptive phenomena and novelty detection. We conclude with experimental and modeling progress on the role of interneuron-specific plasticity in circuit computation and context-dependent learning

Cognitive Impairment and Aberrant Plasticity in the Kindling Model of Epilepsy

Epilepsy is a neurological disorder that affects approximately 1% of the population worldwide. Although motor seizures are the best known feature of epilepsy, many patients also experience severe interictal (between-seizure) behavioral and cognitive comorbidities that have a greater negative influence on quality of life than seizure control or frequency. To study the characteristics of these interictal comorbidities and the neural mechanisms that underlie them, I use the kindling model of epilepsy. Kindling refers to the brief electrical stimulation of a discrete brain site that produces a gradual and permanent increase in the severity of elicited seizure activity. The repeated seizures associated with kindling induce robust structural and functional plasticity that appears to be primarily aberrant. Importantly, the aberrant plasticity evoked by repeated seizures is thought to contribute to the pathophysiology of epilepsy and its associated behavioral and cognitive comorbidities. Unfortunately, the relationship between aberrant plasticity and cognition dysfunction following repeated seizures remains poorly understood. The aim of this dissertation is to gain a better understanding of the effects of repeated convulsions on aberrant neural plasticity and interictal behavior. In Chapter 2, I will examine the effect of short- and long-term amygdala kindling on amygdala- and hippocampal-dependent forms of operant fear conditioning. To evaluate whether kindling alters neural circuits important in memory, I will analyze post-mortem measures of neural activity following the retrieval of fearful memories. In Chapter 3, I will evaluate whether deficits in operant fear learning and memory are a general consequence of convulsions induced by kindling stimulations or whether these deficits occur following kindling of specific brain regions. To evaluate whether aberrant plasticity following kindling of different brain regions contributes to learning and memory deficits, I will make post-mortem examinations of the inhibitory neurotransmitter neuropeptide Y and its Y2 receptor. In Chapter 4, I will investigate the relationship between hippocampal neurogenesis and cognition. Specifically, I will determine whether kindling of different brain regions induces an aberrant form of hippocampal neurogenesis that contributes to cognitive dysfunction. In Chapter 5, I will investigate whether kindling of different brain regions alters different subpopulations of hippocampal GABAergic interneurons, in terms of number and morphological features. Finally, Chapter 6 will provide preliminary evidence that the cognitive impairments associated with kindling can be ameliorated through intrahippocampal infusions of recombinant reelin. The collection of studies in this dissertation improves our understanding of the relationship between aberrant plasticity and cognitive impairments associated with repeated convulsions