two-photon-calcium-imaging-in-neuronal-dendrites-in-brain-slices

Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices

Calcium (Ca2+) imaging is a powerful tool to investigate the spatiotemporal dynamics of intracellular Ca2+ signals in neuronal dendrites. Ca2+ ...

Université Laval

This study tested the hypothesis that early functional alterations in neuronal synchrony in the partially deafferented cortex may lead to spontaneously occurring electrographic seizures. In vivo experiments with partial deafferentation of cat suprasylvian gyrus after extensive undercut of the white matter were conducted using multi-site EEG, extracellular unit and intracellular recordings. The amplitudes of EEG waves were much higher in the areas surrounding deafferented cortical fields as compared with control and with undercut cortex. In 40% of animals with undercut cortex, paroxysmal activity occurred 2-3 h after the undercut and was initiated in the relatively intact cortex, adjacent to the more disconnected one. The seizures that followed the undercut consisted of spike-wave/polyspike-wave complexes and fast runs, resembling the electrographic patterns of some clinical epileptic syndromes. An increased local synchrony in the relatively intact cortex evolved into paroxysmal activity that ultimately spread to the deafferented cortex. The electrographic seizures were found only in animals that showed a propagation of the slow sleep-like oscillation in control conditions. The increase of long-range synchrony within a given seizure was associated with seizure termination. These results indicate that alterations in neuronal synchrony following neuronal trauma can be a critical factor triggering electrographic seizures.

Partial cortical deafferentation promotes development of paroxysmal activity.

Cortical trauma can lead to development of electrographic paroxysmal activities. Current views of trauma-induced epileptogenesis suggest that chronic neuronal hyperexcitability and extensive morphological reorganization of the traumatized cortex are required for the generation of electrographic seizures. However, the mechanisms responsible for the initiation of electrographic seizures shortly after cortical injury are poorly understood. Here we show that, in the experimental model of partially deafferented (undercut) cortex, an increase in intrinsic and synaptic excitability of neurons in areas adjacent to the undercut cortex is sufficient for the generation of electrographic paroxysmal activity within few hours after partial cortical deafferentation. Locally increased and spatially restricted neuronal excitability arose from the increased incidence of intrinsically bursting neurons, enhanced intrinsic and synaptic neuronal responsiveness, and slight disinhibition. These mechanisms only operate in neurons located in the vicinity of partially deafferented sites because, after the cortical injury, partially deafferented neurons are mostly silent and hypoexcitable. Our results suggest that trauma-induced electrographic seizures first arise in cortical fields that are closest to the site of injury and such seizures do not require long-term neuronal reorganization.

Hyperexcitability of intact neurons underlies acute development of trauma-related electrographic seizures in cats in vivo.

Activity-dependent regulation of synaptic inputs in neurons is controlled by highly compartmentalized and dynamic dendritic calcium signaling. Among multiple Ca(2+) mechanisms operating in neuronal dendrites, voltage-sensitive Ca(2+) channels (VSCCs) represent a major source of Ca(2+) influx; however, their use-dependent implication, regulation, and function in different types of central neurons remain widely unknown. Using two-photon microscopy to probe Ca(2+) signaling in dendrites of hippocampal oriens/alveus interneurons, we found that intense synaptic activity or local activation of mGluR5 induced long-lasting potentiation of action potential evoked Ca(2+) transients. This potentiation of dendritic Ca(2+) signaling required mGluR5-induced intracellular Ca(2+) release and PKC activation and was expressed as a selective compartmentalized potentiation of L-type VSCCs. Thus, in addition to mGluR1a-dependent synaptic plasticity, hippocampal interneurons in the feedback inhibitory circuit demonstrate a novel form of mGluR5-induced dendritic plasticity. Given an implication of L-type VSCCs in the induction of Hebbian LTP at interneuron excitatory synapses, their activity-dependent regulation may represent a powerful mechanism for regulating synaptic plasticity.

Activity-dependent compartmentalized regulation of dendritic Ca2+ signaling in hippocampal interneurons.

In most central neurons, action potentials (APs), generated in the initial axon segment, propagate back into dendrites and trigger considerable Ca(2+) entry via activation of voltage-sensitive calcium channels (VSCCs). Despite the similarity in its underlying mechanisms, however, AP-evoked dendritic Ca(2+) signalling often demonstrates a cell type-specific profile that is determined by the neuron dendritic properties. Using two-photon Ca(2+) imaging in combination with patch-clamp whole-cell recordings,we found that in distinct types of hippocampal inhibitory interneurons Ca(2+) transients evoked by backpropagating APs not only were shaped by the interneuron-specific properties of dendritic Ca(2+) handling but also involved specific Ca(2+) mechanisms that were regulated dynamically by distinct activity patterns. In dendrites of regularly spiking basket cells, AP-evoked Ca(2+) rises were of large amplitude and fast kinetics; however, they decreased with membrane hyperpolarization or following high-frequency firing episodes. In contrast, AP-evoked Ca(2+) elevations in dendrites of Schaffer collateral-associated cells exhibited significantly smaller amplitude and slower kinetics, but increased with membrane hyperpolarization. These cell type-specific properties of AP-evoked dendritic Ca(2+) signalling were determined by distinct endogenous buffer capacities of the interneurons examined and by specific types of VSCCs recruited by APs during different patterns of activity. Furthermore, AP-evoked Ca(2+) transients summated efficiently during theta-like bursting and were associated with the induction of long-term potentiation at inhibitory synapses onto both types of interneurons. Therefore, the cell type-specific profile of AP-evoked dendritic Ca(2+) signalling is shaped in an activity-dependent manner, such that the same pattern of hippocampal activity can be differentially translated into dendritic Ca(2+) signals in different cell types. However, Cell type-specific differences in Ca(2+) signals can be 'smoothed out' by changes in neuronal activity, providing a means for common, cell-type-independent forms of synaptic plasticity.

Cell type-specific and activity-dependent dynamics of action potential-evoked Ca2+ signals in dendrites of hippocampal inhibitory interneurons.

Stratum oriens-lacunosum moleculare interneurons (O-LM INs) represent the major element of the hippocampal feedback inhibitory circuit, which provides inhibition to the distal dendritic sites of CA1 pyramidal neurons. Although the intrinsic conductance profile and the properties of glutamatergic transmission to O-LM INs have become a subject of intense investigation, far less is known about the properties of the inhibitory synapses formed onto these cells. Here, we used whole-cell patch-clamp recordings in acute mouse hippocampal slices to study the properties and plasticity of GABAergic inhibitory synapses onto O-LM INs. Surprisingly, we found that the kinetics of inhibitory postsynaptic currents (IPSCs) were slower in mature synapses (P26-40) due to the synaptic incorporation of the α5 subunit of the GABA(A) receptor (a5-GABA(A)R). Moreover, this age-dependent synaptic expression of a5-GABA(A)Rs was directly associated with the emergence of long-term potentiation at IN inhibitory synapses. Finally, the slower time course of IPSCs observed in O-LM INs of mature animals had a profound effect on IN excitability by significantly delaying its spike firing. Our data suggest that GABAergic synapses onto O-LM INs undergo significant modifications during postnatal maturation. The developmental switch in IPSC properties and plasticity is controlled by the synaptic incorporation of the a5-GABA(A)R subunit and may represent a potential mechanism for the age-dependent modifications in the inhibitory control of the hippocampal feedback inhibitory circuit.

Age-dependent remodelling of inhibitory synapses onto hippocampal CA1 oriens-lacunosum moleculare interneurons.

Calcium (Ca(2+) ) is a major second messenger in the regulation of different forms of synaptic and intrinsic plasticity. Tightly organized in space and time, postsynaptic Ca(2+) transients trigger the activation of many distinct Ca(2+) signaling cascades, providing a means for a highly specific signal transduction and plasticity induction. High-resolution two-photon microscopy combined with highly sensitive synthetic Ca(2+) indicators in brain slices allowed for the quantification and analysis of postsynaptic Ca(2+) dynamics in great detail. Much of our current knowledge about postsynaptic Ca(2+) mechanisms is derived from studying Ca(2+) transients in the dendrites and spines of pyramidal neurons. However, postsynaptic Ca(2+) dynamics differ considerably among different cell types. In particular, distinct rules of postsynaptic Ca(2+) signaling and, accordingly, of Ca(2+) -dependent plasticity operate in GABAergic interneurons. Here, I review recent progress in understanding the complex organization of postsynaptic Ca(2+) signaling and its relevance to several forms of long-term potentiation at excitatory synapses in cortical GABAergic interneurons.

Dendritic calcium mechanisms and long-term potentiation in cortical inhibitory interneurons.

Information processing within neural circuits depends largely on the dynamic interactions between the principal cells and inhibitory interneurons. It is further determined by the efficacy of synaptic transmission between individual circuit elements, which is in turn tightly regulated by changes in network activity to allow for numerous adaptations to occur at a single synapse. Intracellular calcium (Ca2+) is a crucial factor in the regulation of synaptic efficacy in neuronal networks. Evidence from high-resolution imaging studies has revealed the intricacies of how Ca2+ signalling is organised in the dendrites of different cell types. Inhibitory interneurons exhibit a variety of postsynaptic Ca2+ mechanisms, which are recruited by distinct activity patterns and are responsible for the formation of functionally segregated dendritic Ca2+ microdomains. Furthermore, postsynaptic Ca2+ signals in these cells not only contribute to the induction of synaptic plasticity but also may themselves undergo different forms of plastic modifications, depending on the activity level. This compartmentalised regulation of postsynaptic Ca2+ signalling may have a significant impact on the induction of synaptic plasticity and on single-interneuron and network computations.

Functional compartmentalisation and regulation of postsynaptic Ca2+ transients in inhibitory interneurons.

Communication between neurons is achieved by rapid signal transduction via highly specialized structural elements known as synaptic contacts. In addition, numerous extrasynaptic mechanisms provide a flexible platform for the local regulation of synaptic signals. For example, peri- and extra-synaptic signaling through the group I metabotropic glutamate receptors (mGluRs) can be involved in the highly compartmentalized regulation of dendritic ion conductances, the induction of input-specific synaptic plasticity, and the local release of retrograde messengers. Therefore, extrasynaptic mechanisms appear to play a key role in the local tuning of dendritic computations. Here, we review recent findings on the role of group I mGluRs in the dendritic signaling of inhibitory interneurons. We propose that group I mGluRs provide a dual-mode signaling device that integrates different patterns of neural activity. By implementing distinct forms of intrinsic and synaptic regulation, group I mGluRs may be responsible for the local fine-tuning of dendritic function.

Dendritic Signaling in Inhibitory Interneurons: Local Tuning via Group I Metabotropic Glutamate Receptors.

Information processing within neuronal networks is determined by a dynamic partnership between principal neurons and local circuit inhibitory interneurons. The population of GABAergic interneurons is extremely heterogeneous and comprises, in many brain regions, cells with divergent morphological and physiological properties, distinct molecular expression profiles, and highly specialized functions. GABAergic interneurons have been studied extensively during the past two decades, especially in the hippocampus, which is a relatively simple cortical structure. Different types of hippocampal inhibitory interneurons control spike initiation [e.g., axo-axonic and basket cells (BCs)] and synaptic integration (e.g., bistratified and oriens-lacunosum moleculare interneurons) within pyramidal neurons and synchronize local network activity, providing a means for functional segregation of neuronal ensembles and proper routing of hippocampal information. Thus, it is thought that, at least in the hippocampus, GABAergic inhibitory interneurons represent critical regulating elements at all stages of information processing, from synaptic integration and spike generation to large-scale network activity. However, this raises an important question: if inhibitory interneurons are fundamental for network computations, what are the mechanisms that control the activity of the interneurons themselves? Given the essential role of synaptic inhibition in the regulation of neuronal activity, it would be logical to expect that specific inhibitory mechanisms have evolved to control the operation of interneurons. Here, we review the mechanisms of synaptic inhibition of interneurons and discuss their role in the operation of hippocampal inhibitory circuits.

Inhibitory control of hippocampal inhibitory neurons.

Postsynaptic calcium (Ca2+) nonlinearities allow neuronal coincidence detection and site-specific plasticity. Whether such events exist in dendrites of interneurons and play a role in regulation of synaptic efficacy remains unknown. Here, we used a combination of whole-cell patch-clamp recordings and two-photon Ca2+ imaging to reveal Ca2+ nonlinearities associated with synaptic integration in dendrites of mouse hippocampal CA1 fast-spiking interneurons. Local stimulation of distal dendritic branches within stratum oriens/alveus elicited fast Ca2+ transients, which showed a steep sigmoidal relationship to stimulus intensity. Supralinear Ca2+ events required Ca2+ entry through AMPA receptors with a subsequent Ca2+ release from internal stores. To investigate the functional significance of supralinear Ca2+ signals, we examined activity-dependent fluctuations in transmission efficacy triggered by Ca2+ signals of different amplitudes at excitatory synapses of interneurons. Subthreshold theta-burst stimulation (TBS) produced small amplitude postsynaptic Ca2+ transients and triggered long-term potentiation. In contrast, the suprathreshold TBS, which was associated with the generation of supralinear Ca2+ events, triggered long-term depression. Blocking group I/II metabotropic glutamate receptors (mGluRs) during suprathreshold TBS resulted in a slight reduction of supralinear Ca2+ events and induction of short-term depression. In contrast, blocking internal stores and supralinear Ca2+ signals during suprathreshold TBS switched the direction of plasticity from depression back to potentiation. These data reveal a novel type of supralinear Ca2+ events at synapses lacking the GluA2 AMPA subtype of glutamate receptors and demonstrate a general mechanism by which Ca2+ -permeable AMPA receptors, together with internal stores and mGluRs, control the direction of plasticity at interneuron excitatory synapses.

Dendritic calcium nonlinearities switch the direction of synaptic plasticity in fast-spiking interneurons.

In cortical networks, different types of inhibitory interneurons control the activity of glutamatergic principal cells and GABAergic interneurons. Principal neurons represent the major postsynaptic target of most interneurons; however, a population of interneurons that is dedicated to the selective innervation of GABAergic cells exists in the CA1 area of the hippocampus. The physiological properties of these cells and their functional relevance for network computations remain unknown. Here, we used a combination of dual simultaneous patch-clamp recordings and targeted optogenetic stimulation in acute mouse hippocampal slices to examine how one class of interneuron-specific (IS) cells controls the activity of its GABAergic targets. We found that type 3 IS (IS3) cells that coexpress the vasoactive intestinal polypeptide (VIP) and calretinin contact several distinct types of interneurons within the hippocampal CA1 stratum oriens/alveus (O/A), with preferential innervation of oriens-lacunosum moleculare cells (OLMs) through dendritic synapses. In contrast, VIP-positive basket cells provided perisomatic inhibition to CA1 pyramidal neurons with the asynchronous GABA release and were not connected with O/A interneurons. Furthermore, unitary IPSCs recorded at IS3-OLM synapses had a small amplitude and low release probability but summated efficiently during high-frequency firing of IS3 interneurons. Moreover, the synchronous generation of a single spike in several IS cells that converged onto a single OLM controlled the firing rate and timing of OLM interneurons. Therefore, dendritic inhibition originating from IS cells is needed for the flexible activity-dependent recruitment of OLM interneurons for feedback inhibition.

Dendritic inhibition provided by interneuron-specific cells controls the firing rate and timing of the hippocampal feedback inhibitory circuitry.

It has been recognized for some time that different subtypes of cortical inhibitory interneurons innervate specific dendritic domains of principal cells and release GABA at particular times during behaviorally relevant network oscillations. However, the lack of basic information on how the activity of interneurons can be controlled by GABA released in particular behavioral states has hindered our understanding of the rules that govern the spatio-temporal organization and function of dendritic inhibition. Similar to principal cells, any given interneuron may receive several functionally distinct inhibitory inputs that target its specific subcellular domains. We recently found that local circuitry of the so-called interneuron-specific (IS) interneurons is responsible for dendritic inhibition of different subtypes of hippocampal interneurons with a great impact on cell output. Here, we will review the properties and the specificity of connections of IS interneurons in the CA1 hippocampus and neocortex, and discuss their possible role in the activity-dependent regulation of dendritic inhibition received by pyramidal neurons.

Coordination of dendritic inhibition through local disinhibitory circuits.

Status epilepticus (SE) is associated with complex reorganization of hippocampal circuits involving a significant loss of specific subtypes of GABAergic interneurons. While adaptive circuit plasticity may increase the chances for recruitment of surviving interneurons, the underlying mechanisms remain largely unknown. We studied the alterations in the inhibitory tone received by the hippocampal CA1 oriens/alveus (O/A) interneurons from the vasoactive intestinal peptide (VIP)- and calretinin (CR)-expressing interneurons using the pilocarpine-induced status epilepticus (SE) model of epilepsy. Our data showed that, while the overall density of the VIP/CR-co-expressing interneurons remained preserved, the number of axonal boutons made by these cells within the CA1 O/A was significantly lower after SE. Furthermore, VIP/CR interneurons exhibited significant alterations in their dendritic morphology and passive membrane properties. Subsequently, while all O/A interneuron types, including oriens-lacunosum moleculare (OLM), bistratified (Bis) and basket cells, exhibited decrease in spontaneous inhibitory drive, Bis and basket cells showed a smaller amplitude of light-evoked IPSCs mediated by the selective activation of VIP-positive interneurons. These data point to the target cell-specific changes in the inhibitory tone provided by the VIP cells to O/A interneurons following SE. Given that basket, Bis and OLM cells coordinate different subcellular domains of pyramidal neurons, significant disinhibition of basket and Bis cells along with a previously reported loss of the OLMs may result in a redistribution of inhibition converging onto pyramidal neurons, with a direct impact onto their recruitment to epileptiform network activity and seizure propagation.

Article	Year
Partial cortical deafferentation promotes development of paroxysmal activity. Cerebral cortex (New York, N.Y. : 1991)\| PubMed ID: 12853375	2003
Hyperexcitability of intact neurons underlies acute development of trauma-related electrographic seizures in cats in vivo. The European journal of neuroscience\| PubMed ID: 12911745	2003
Activity-dependent compartmentalized regulation of dendritic Ca2+ signaling in hippocampal interneurons. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 19357290	2009
Cell type-specific and activity-dependent dynamics of action potential-evoked Ca2+ signals in dendrites of hippocampal inhibitory interneurons. The Journal of physiology\| PubMed ID: 21486769	2011
Age-dependent remodelling of inhibitory synapses onto hippocampal CA1 oriens-lacunosum moleculare interneurons. The Journal of physiology\| PubMed ID: 21825029	2011
Dendritic calcium mechanisms and long-term potentiation in cortical inhibitory interneurons. The European journal of neuroscience\| PubMed ID: 22304664	2012
Functional compartmentalisation and regulation of postsynaptic Ca2+ transients in inhibitory interneurons. Cell calcium\| PubMed ID: 22656961	2012
Dendritic Signaling in Inhibitory Interneurons: Local Tuning via Group I Metabotropic Glutamate Receptors. Frontiers in physiology\| PubMed ID: 22934015	2012
Inhibitory control of hippocampal inhibitory neurons. Frontiers in neuroscience\| PubMed ID: 23162426	2012
Dendritic calcium nonlinearities switch the direction of synaptic plasticity in fast-spiking interneurons. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 24623765	2014
Dendritic inhibition provided by interneuron-specific cells controls the firing rate and timing of the hippocampal feedback inhibitory circuitry. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 24671999	2014
Coordination of dendritic inhibition through local disinhibitory circuits. Frontiers in synaptic neuroscience\| PubMed ID: 25767448	2015
Target-specific alterations in the VIP inhibitory drive to hippocampal GABAergic cells after status epilepticus. Experimental neurology\| PubMed ID: 28315308	2017

Lisa Topolnik

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