Interneuron Types and Their Circuits in the Basolateral Amygdala

The basolateral amygdala (BLA) is a cortical structure based on its cell types, connectivity features, and developmental characteristics. This part of the amygdala is considered to be the main entry site of processed and multisensory information delivered via cortical and thalamic afferents. Although GABAergic inhibitory cells in the BLA comprise only 20% of the entire neuronal population, they provide essential control over proper network operation. Previous studies have uncovered that GABAergic cells in the basolateral amygdala are as diverse as those present in other cortical regions, including the hippocampus and neocortex. To understand the role of inhibitory cells in various amygdala functions, we need to reveal the connectivity and input-output features of the different types of GABAergic cells. Here, I review the recent achievements in uncovering the diversity of GABAergic cells in the basolateral amygdala with a specific focus on the microcircuit organization of these inhibitory cells

A hippokampális éleshullámok kialakulásának hálózati mechanizmusai. = Circuit mechanisms underlying the generation of hippocampal sharp wave/ripple oscillations

A hippokampális EEG egyik jellemző aktivitásmintázata az ún. éleshullám, mely fontos szerepet játszik egyes kognitív folyamatokban, mint pl. a memórianyomok bevésésében. Az in vivo kutatások az éleshullámok számos sajátságát feltárták, de a háttérben álló pontos sejtszintű mechanizmusok mindmáig tisztázatlanok. Célunk volt, hogy egy in vitro modell segítségével felderítsük, miként járulnak hozzá a különböző sejttípusok ezen hálózati események kialakulásához. Vizsgálataink feltárták, hogy az éleshullámok a hippokampusz CA3 régiójában spontán keletkeznek a piramissejtek szporadikus aktivitásának a következtében. A rekurrens kollaterálisokon keresztül jelentős aktivitási szintet ér el a piramissejtek populációs aktivitása, amelynek a parvalbumin tartalmú gátlósejtek kisülése vet véget, ami mint éleshullám jelentkezik a lokális mezőpotenciálban. Ez a mechanizmus hasonló az általunk korábban feltárt gamma oszcillációk sejtszintű folyamataihoz. Kísérleteinkben továbbá tisztáztuk, hogy a CA3 régió neuronhálózata az éleshullám-aktivitási állapotból a gamma oszcillációba azáltal tud váltani, ha az idegsejtek serkenthetőbbek lesznek, ill. a köztük lévő szinaptikus kapcsolatok legyengülnek. Az aktivitási mintázatok közti ’átkapcsolást’ pl. az acetilkolin receptorainak az ingerlése eredményezheti. A hálózati oszcillációk sejtszintű mechanizmusainak az azonosítása elősegítheti a patológiás aktivitások, mint pl. az epilepsziás rohamok kialakulási körülményeinek a megértését. | The hippocampal EEG is often decorated with so-called sharp wave-ripple (SWR) activities that were shown to play an important role in memory consolidation. In vivo studies have revealed several features of these synchronous events, yet the underlying mechanisms are still unknown. Our aim was to uncover the contribution of distinct neuron types to SWRs using an in vitro model. We found that SWRs in the CA3 region of the hippocampal slices were emerged as a consequence of sporadic activity of pyramidal cells. Through the recurrent collaterals the activity level in the pyramidal cell population reaches a threshold, when parvalbumin containing inhibitory cells are recruited. The firing of these GABAergic cells terminates the population burst. The high frequency discharge of inhibitory cells results in a deflection in local field potential detected as a SWR. These synaptic mechanisms resemble those that were uncovered for the generation of gamma oscillations. In addition, we revealed that in CA3 the SWR activity state can be transformed into gamma oscillations by increasing the excitability within the network and, in parallel, by decreasing the synaptic strengths. Such changes in network parameters can be achieved e.g. by activation of acetylcholine receptors. Revealing the cellular mechanisms underlying the synchronous activities my help to understand the changes in neuronal networks that can produces pathological activities, including epileptiform events

A különböző gátlósejttípusok hozzájárulása a hippokampális éleshullámok kialakulásához. = Contribution by distinct types of GABAergic interneuron to hippocampal sharp wave/ripple oscillations.

A hippokampusz neuronhálózatában spontán keletkeznek az éleshullámok, amelyek kulcsszerepet játszanak a memóriafolyamatokban. Egy in vitro modellt használva feltérképeztük az egyes idegsejttípusok bemeneti és kimeneti tulajdonságait az éleshullámok alatt. Az találtuk, hogy a legaktívabb gátlósejtek parvalbumint tartalmaztak, míg a piramissejtek többsége nem tüzelt. Meghatároztuk, hogy az éleshullámok alatti tüzelési aktivitás korrelált a serkentő szinaptikus bemenettel. Farmakológiai kísérletekkel kiderítettük, hogy a parvalbumin tartalmú gátlósejtek nagyfrekvenciás kisüléséből eredő periszomatikus gátló áramok generálják a lokális mezőpotenciálban mérhető éleshullámokat. Hasonlóan, ezek a gátlósejtek felelősek a gamma oszcillációk létrehozásáért is a hippokampális agyszeletekben. Ezen túlmenően megállapítottuk, hogy a kolinerg receptorok aktivációja, amely növeli a serkenthetőséget, de csökkenti a szinaptikus kommunikáció hatékonyságát, képes a hippokampusz alapműködését, az éleshullám-aktivitást átkapcsolni gamma oszcillációvá. Az eredményeink azt mutatják, hogy az éber állatra jellemző hálózati aktivitásokat, az éleshullámokat és a gamma oszcillációt ugyan az a hippokampális neuronhálózat generálja, amely a piramissejtek és a parvalbumin tartamú gátlósejtekből áll. | Sharp wave/ripple oscillations (SPW-Rs), that play a crucial role in memory formation, are spontaneously emerging synchronous network events in the hippocampal circuitry. Using an in vitro model, we uncovered that the input-output properties of distinct types of neurons during SPW-Rs. We found that the most active GABAergic cells were parvalbumin containing interneurons, while the vast majority of pyramidal cells was silent. Our analysis revealed that in all cell types the firing during SPW-Rs was driven by excitatory synaptic input. Pharmacological manipulations uncovered that perisomatic inhibitory currents predominantly originated from the high frequency discharge of parvalbumin containing interneurons generate the majority of the field potential that is seen as a sharp wave. Similarly, these GABAergic cells were found to generate the gamma oscillations in hippocampal slices as well. In addition, we elucidated that by cholinergic receptor activation, which increases the excitability, but reduces the efficiency of synaptic communication, the default mode of the hippocampal operation, the SPW-R state can be readily switched to gamma oscillation. Our results propose that the behaviorally relevant network activities, SPW-Rs and gamma oscillations are generated by the same neuronal circuitries in the hippocampus, comprised of pyramidal cells and parvalbumin containing interneurons

Feedforward Inhibition Underlies the Propagation of Cholinergically Induced Gamma Oscillations from Hippocampal CA3 to CA1.

Gamma frequency (30-80 Hz) oscillations are implicated in memory processing. Such rhythmic activity can be generated intrinsically in the CA3 region of the hippocampus from where it can propagate to the CA1 area. To uncover the synaptic mechanisms underlying the intrahippocampal spread of gamma oscillations, we recorded local field potentials, as well as action potentials and synaptic currents in anatomically identified CA1 and CA3 neurons during carbachol-induced gamma oscillations in mouse hippocampal slices. The firing of the vast majority of CA1 neurons and all CA3 neurons was phase-coupled to the oscillations recorded in the stratum pyramidale of the CA1 region. The predominant synaptic input to CA1 interneurons was excitatory, and their discharge followed the firing of CA3 pyramidal cells at a latency indicative of monosynaptic connections. Correlation analysis of the input-output characteristics of the neurons and local pharmacological block of inhibition both agree with a model in which glutamatergic CA3 input controls the firing of CA1 interneurons, with local pyramidal cell activity having a minimal role. The firing of phase-coupled CA1 pyramidal cells was controlled principally by their inhibitory inputs, which dominated over excitation. Our results indicate that the synchronous firing of CA3 pyramidal cells rhythmically recruits CA1 interneurons and that this feedforward inhibition generates the oscillatory activity in CA1. These findings identify distinct synaptic mechanisms underlying the generation of gamma frequency oscillations in neighboring hippocampal subregions

Mechanisms of sharp wave initiation and ripple generation

Replay of neuronal activity during hippocampal sharp wave-ripples (SWRs) is essential in memory formation. To understand the mechanisms underlying the initiation of irregularly occurring SWRs and the generation of periodic ripples, we selectively manipulated different components of the CA3 network in mouse hippocampal slices. We recorded EPSCs and IPSCs to examine the buildup of neuronal activity preceding SWRs and analyzed the distribution of time intervals between subsequent SWR events. Our results suggest that SWRs are initiated through a combined refractory and stochastic mechanism. SWRs initiate when firing in a set of spontaneously active pyramidal cells triggers a gradual, exponential buildup of activity in the recurrent CA3 network. We showed that this tonic excitatory envelope drives reciprocally connected parvalbumin-positive basket cells, which start ripple-frequency spiking that is phase-locked through reciprocal inhibition. The synchronized GABAA receptor-mediated currents give rise to a major component of the ripple-frequency oscillation in the local field potential and organize the phase-locked spiking of pyramidal cells. Optogenetic stimulation of parvalbumin-positive cells evoked full SWRs and EPSC sequences in pyramidal cells. Even with excitation blocked, tonic driving of parvalbumin-positive cells evoked ripple oscillations. Conversely, optogenetic silencing of parvalbumin-positive cells interrupted the SWRs or inhibited their occurrence. Local drug applications and modeling experiments confirmed that the activity of parvalbumin-positive perisomatic inhibitory neurons is both necessary and sufficient for ripple-frequency current and rhythm generation. These interneurons are thus essential in organizing pyramidal cell activity not only during gamma oscillation, but, in a different configuration, during SWRs

Különböző típusú GABAerg interneuronok szerepe a hippokampális gamma oszcillációkban = The role of distinct types of GABAergic interneurons in hippocampal network oscillations at gamma frequency

A kérgi neuronhálózatokban megfigyelt gamma (30-100 Hz) oszcillációk alapvető szerepet játszanak olyan kognitív folyamatokban, mint pl. a szenzoros információfeldolgozás. Funkciójuk megértéséhez ismernünk kell a neuronhálózatokat alkotó serkentő és gátlósejtek viselkedését ill. szerepét az oszcillációk kialakításában. Pályázatunk célja a hippokampális ideghálózatok szinkronizált működését kialakító sejtszintű mechanizmusok felderítése volt. In vitro farmakológiailag indukált oszcillációk során vizualizált patch-clamp méréstechnika segítségével megállapítottuk, hogy a hippokampusz CA3 régiójában keletkező gamma oszcillációkat a gyorsan tüzelő kosársejtek és a piramissejtek időben összehangolt kisülése generálja szinaptikus visszacsatolás révén. A hippokampusz CA1 régiójába a gamma oszcilláció előrecsatoló gátlással terjed át a CA3 régióból. Mindkét régióban a gátlósejtek oszcillációhoz viszonyított fáziskapcsolt tüzelését a rájuk érkező szinaptikus serkentés, míg a piramissejtek kisülését a szinaptikus gátlás határozta meg. Kifejlesztettünk egy szabadalmi bejelentéssel védett szeletkamrát in vitro mérésekhez, melyben az agyszeletek oxigénellátása megközelíti az in vivo körülményeket. Az eredményeinknek klinikai vonatkozása is elképzelhető, hiszen az epilepszia tünetcsoportban tapasztalt hiperszinkonitás kialakulásában is kulcsfontosságú szerepet játszhatnak a gyorsan tüzelő kosársejtek, amely gátlósejtek működésének célzott szabályozása egy potenciális gyógyszercélpont lehet. | Cortical network oscillations at gamma (30-100 Hz) frequencies were suggested to be linked to several cognitive tasks including sensory processing. To understand the role of oscillations in neuronal operation, the behavior and the function of different neuronal types during oscillatory activities need to be revealed. The aim of our project was to uncover the basic cellular mechanisms generating synchronous network activities in hippocampal neuronal circuitries. The combination of visualized patch-clamp recordings with pharmacologically-induced in vitro oscillations allowed us to determine that in CA3 hippocampal region the precisely timed discharge of fast spiking basket cells and pyramidal cells could generate the gamma oscillations via a synaptic feed-back loop. The gamma oscillation emerged intrinsically in CA3 propagates to CA1 via feed-forward inhibition. In both regions, the phase-coupled firing of inhibitory cells was controlled by synaptic excitation, whereas the discharge of pyramidal cells was primarily determined by synaptic inhibition. For in vitro recordings we developed a new type of slice chamber protected by a patent, where the oxygen supply of brain slices approaches the in vivo circumstances. Our results also have clinical relevance implying the pivotal role of fast spiking basket cells in hypersynchrony during epileptic discharges, therefore the modulation of the fast spiking basket cell operation might be a novel target for drug development

Presynaptic Calcium Channel Inhibition Underlies CB1 Cannabinoid Receptor-Mediated Suppression of GABA Release.

CB1 cannabinoid receptors (CB1) are located at axon terminals and effectively control synaptic communication and thereby circuit operation widespread in the CNS. Although it is partially uncovered how CB1 activation leads to the reduction of synaptic excitation, the mechanisms of the decrease of GABA release upon activation of these cannabinoid receptors remain elusive. To determine the mechanisms underlying the suppression of synaptic transmission by CB1 at GABAergic synapses, we recorded unitary IPSCs (uIPSCs) at cholecystokinin-expressing interneuron-pyramidal cell connections and imaged presynaptic [Ca(2+)] transients in mouse hippocampal slices. Our results reveal a power function with an exponent of 2.2 between the amplitude of uIPSCs and intrabouton [Ca(2+)]. Altering CB1 function by either increasing endocannabinoid production or removing its tonic activity allowed us to demonstrate that CB1 controls GABA release by inhibiting Ca(2+) entry into presynaptic axon terminals via N-type (Cav2.2) Ca(2+) channels. These results provide evidence for modulation of intrabouton Ca(2+) influx into GABAergic axon terminals by CB1, leading to the effective suppression of synaptic inhibition

DAG-sensitive and Ca(2+) permeable TRPC6 channels are expressed in dentate granule cells and interneurons in the hippocampal formation

Members of the transient receptor potential (TRP) cation channel family play important roles in several neuronal functions. To understand the precise role of these channels in information processing, their presence on neuronal elements must be revealed. In this study, we investigated the localization of TRPC6 channels in the adult hippocampal formation. Immunostainings with a specific antibody, which was validated in Trpc6 knockout mice, showed that in the dentate gyrus, TRPC6 channels are strongly expressed in granule cells. Immunogold staining revealing the subcellular localization of TRPC6 channels clarified that these proteins were predominantly present on the membrane surface of the dendritic shafts of dentate granule cells, and also in their axons, often associated with intracellular membrane cisternae. In addition, TRPC6 channels could be observed in the dendrites of some interneurons. Double immunofluorescent staining showed that TRPC6 channels were present in the dendrites of hilar interneurons and hippocampal interneurons with horizontal dendrites in the stratum oriens expressing mGlu1a receptors, whereas parvalbumin immunoreactivity was revealed in TRPC6-expressing dendrites with radial appearance in the stratum radiatum. Electron microscopy showed that the immunogold particles depicting TRPC6 channels were located on the surface membranes of the interneuron dendrites. Our results suggest that TRPC6 channels are in a key position to alter the information entry into the trisynaptic loop of the hippocampal formation from the entorhinal cortex, and to control the function of both feed-forward and feed-back inhibitory circuits in this brain region

Strategically Positioned Inhibitory Synapses of Axo-axonic Cells Potently Control Principal Neuron Spiking in the Basolateral Amygdala.

Axo-axonic cells (AACs) in cortical regions selectively innervate the axon initial segments (AISs) of principal cells (PCs), where the action potentials are generated. These GABAergic interneurons can alter the activity of PCs, but how the efficacy of spike control correlates with the number of output synapses remains unclear. Moreover, the relationship between the spatial distribution of GABAergic synapses and the action potential initiation site along the AISs is not well defined. Using paired recordings obtained in the mouse basolateral amygdala, we found that AACs powerfully inhibited or delayed the timing of PC spiking by 30 ms, if AAC output preceded PC spiking with no more than 80 ms. By correlating the number of synapses and the probability of spiking, we revealed that larger numbers of presynaptic AAC boutons giving rise to larger postsynaptic responses provided more effective inhibition of PC spiking. At least 10-12 AAC synapses, which could originate from 2-3 AACs on average, were necessary to veto the PC firing under our recording conditions. Furthermore, we determined that the threshold for the action potential generation along PC axons is the lowest between 20 and 40 mum from soma, which axonal segment received the highest density of GABAergic inputs. Single AACs preferentially innervated this narrow portion of the AIS where action potentials were generated with the highest likelihood, regardless of the number of synapses forming a given connection. Our results uncovered a fine organization of AAC innervation maximizing their inhibitory efficacy by strategically positioning synapses along the AISs