Types Of Interneurons Research Articles

Major Contribution of Somatostatin-Expressing Interneurons and Cannabinoid Receptors to Increased GABA Synaptic Activity in the Striatum of Huntington's Disease Mice.

Huntington’s disease (HD) is a heritable neurological disorder that affects cognitive and motor performance in patients carrying the mutated huntingtin (HTT) gene. In mouse models of HD, previous reports showed a significant increase in spontaneous GABAA receptor-mediated synaptic activity in striatal spiny projection neurons (SPNs). In this study, using optogenetics and slice electrophysiology, we examined the contribution of γ-aminobutyric acid (GABA)-ergic parvalbumin (PV)- and somatostatin (SOM)-expressing interneurons to the increase in GABA neurotransmission using the Q175 (heterozygote) mouse model of HD. Patch clamp recordings in voltage-clamp mode were performed on SPNs from brain slices of presymptomatic (2 months) and symptomatic (8 and 12 months) Q175 mice and wildtype (WT) littermates. While inhibitory postsynaptic currents (IPSCs) evoked in SPNs following optical activation of PV- and SOM-expressing interneurons differed in amplitude, no genotype-dependent differences were observed at all ages from both interneuron types; however, responses evoked by either type were found to have faster kinetics in symptomatic mice. Since SOM-expressing interneurons are constitutively active in striatal brain slices, we then examined the effects of acutely silencing these neurons in symptomatic mice with enhanced Natronomonas pharaonis halorhodopsin (eNpHR). Optically silencing SOM-expressing interneurons resulted in a greater decrease in the frequency of spontaneous IPSCs (sIPSCs) in a subset of SPNs from Q175 mice compared to WTs, suggesting that SOM-expressing interneurons are the main contributors to the overall increased GABA synaptic activity in HD SPNs. Additionally, the effects of activating GABAB and cannabinoid (CB1) receptors were investigated to determine whether these receptors were involved in modulating interneuron-specific GABA synaptic transmission and if this modulation differed in HD mice. When selectively activating PV- and SOM-expressing interneurons in the presence of the CB1 receptor agonist WIN-55,212, the magnitudes of the evoked IPSCs in SPNs decreased for both interneuron types although this change was less prominent in symptomatic Q175 SPNs during SOM-expressing interneuron activation. Overall, these findings show that dysfunction of SOM-expressing interneurons contributes to the increased GABA synaptic activity found in HD mouse models and that dysregulation of the endocannabinoid system may contribute to this effect.

Reciprocal Inhibitory Glomerular Circuits Contribute to Excitation-Inhibition Balance in the Mouse Olfactory Bulb.

The major inhibitory interneurons in olfactory bulb (OB) glomeruli are periglomerular cells (PGCs) and short axon cells (SACs). PGCs and SACs provide feedforward inhibition to all classes of projection neurons, but inhibition between PGCs and SACs is not well understood. We crossed Cre and GFP transgenic mice and used virally-delivered optogenetic constructs to selectively activate either SACs or GAD65cre-ChR2-positive PGCs while recording from identified GAD65cre-ChR2-positive PGCs or SACs, respectively, to investigate inhibitory interactions between these two interneuron types. We show that GAD65cre-ChR2-positive PGCs robustly inhibit SACs and SACs strongly inhibit PGCs. SACs form the interglomerular circuit, which inhibits PGCs in distant glomeruli. Activation of GAD65cre-ChR2-positive PGCs monosynaptically inhibit mitral cells (MCs), which complements recent findings that SACs directly inhibit MCs. Thus, both classes of glomerular inhibitory neurons inhibit each other, as well as OB output neurons. We further show that olfactory nerve input to one glomerulus engages the interglomerular circuit and inhibits PGCs in distant glomeruli. Sensory activation of the interglomerular circuit directly inhibits output neurons in other glomeruli and by inhibiting intraglomerular PGCs, may potentially disinhibit output neurons in other glomeruli. The nature and context of odorant stimuli may determine whether inhibition or excitation prevails so that odors are represented in part by patterns of active and inactive glomeruli.

Examination of morphological and synaptic features of calbindin-immunoreactive neurons in deep layers of the rat olfactory bulb with correlative laser and volume electron microscopy.

The olfactory bulb (OB) contains various interneuron types that play key roles in processing olfactory information via synaptic contacts. Many previous studies have reported synaptic connections of heterogeneous interneurons in superficial OB layers. In contrast, few studies have examined synaptic connections in deep layers because of the lack of a selective marker for intrinsic neurons located in the deeper layers, including the mitral cell layer, internal plexiform layer (IPL) and granule cell layer. However, neural circuits in the deep layers are likely to have a strong effect on the output of the OB because of the cellular composition of these regions. Here, we analyzed the calbindin-immunoreactive neurons in the IPL, one of the clearly neurochemically defined interneuron types in the deep layers, using multiple immunolabeling and confocal laser scanning microscopy combined with electron microscopic three-dimensional serial-section reconstruction, enabling correlated laser and volume electron microscopy (EM). Despite a resemblance to the morphological features of deep short axon cells, IPL calbindin-immunoreactive (IPL-CB-ir) neurons lacked axons. Furthermore, multiple immunolabeling for plural neurochemicals indicated that IPL-CB-ir neurons differed from any interneuron types reported previously. We identified symmetrical synapses formed by IPL-CB-ir neurons on granule cells (GCs) using correlated laser and volume EM. These synapses might inhibit GCs and thus disinhibit mitral and tufted cells. Our present findings indicate, for the first time, that IPL-CB-ir neurons are involved in regulating the activities of projection neurons, further suggesting their involvement in synaptic circuitry for output from the deeper layers of the OB, which has not previously been clarified.

Phase relations of theta oscillations in a computer model of the hippocampal CA1 field: Key role of Schaffer collaterals

The hippocampal theta rhythm (4–12 Hz) is one of the most important electrophysiological processes in the hippocampus, it participates in cognitive hippocampal functions, such as navigation in space, novelty detection, and declarative memory. We use neural network modeling to study the mechanism of theta rhythm emergence in the CA1 microcircuitry. Our model of the CA1 field includes biophysical representation of major cell types related to the theta rhythm emergence: excitatory pyramidal cells and two types of inhibitory interneurons, PV+ basket cells and oriens lacunosum–moleculare (OLM) cells. The main inputs to the CA1 cells come from the entorhinal cortex via perforant pathway, the CA3 field via Schaffer collaterals, and the medial septum via fimbria–fornix. By computer simulations we investigated the influence of each input, intrinsic parameters of neurons, and connections between neurons on phase coupling between the theta rhythm and the firing of pyramidal, PV+ basket and OLM cells in the CA1. We found that the input from the CA3 field via Schaffercollaterals plays a major role in the formation of phase relations that have been observed in experiments in vivo. The direct input from the medial septum participates in the formation of proper phase relations, but it is not crucial for the production of the theta rhythm in CA1 neural populations.

Genetic Single Neuron Anatomy Reveals Fine Granularity of Cortical Axo-Axonic Cells

Parsing diverse nerve cells into biological types isnecessary for understanding neural circuit organization. Morphology is an intuitive criterion for neuronal classification and a proxy of connectivity, but morphological diversity and variability often preclude resolving the granularity of neuron types. Combining genetic labeling with high-resolution, large-volume light microscopy, we established a single neuron anatomy platform that resolves, registers, and quantifies complete neuron morphologies in the mouse brain. We discovered that cortical axo-axonic cells (AACs), a cardinal GABAergic interneuron type that controls pyramidal neuron (PyN) spiking at axon initial segments, consist of multiple subtypes distinguished by highly laminar-specific soma position and dendritic and axonal arborization patterns. Whereas the laminar arrangements of AAC dendrites reflect differential recruitment by input streams, the laminar distribution and local geometry of AAC axons enable differential innervation of PyNensembles. This platform will facilitate genetically targeted, high-resolution, and scalable single neuron anatomy in the mouse brain.

Molecular Classification and Comparative Taxonomics of Foveal and Peripheral Cells in Primate Retina

High-acuity vision in primates, including humans, is mediated by a small central retinal region called the fovea. As more accessible organisms lack a fovea, its specialized function and its dysfunction in ocular diseases remain poorly understood. We used 165,000 single-cell RNA-seq profiles to generate comprehensive cellular taxonomies of macaque fovea and peripheral retina. More than 80% of >60 cell types match between the two regions but exhibit substantial differences in proportions and gene expression, some of which we relate to functional differences. Comparison of macaque retinal types with those of mice reveals that interneuron types are tightly conserved. In contrast, projection neuron types and programs diverge, despite exhibiting conserved transcription factor codes. Key macaque types are conserved in humans, allowing mapping of cell-type and region-specific expression of >190 genes associated with 7 human retinal diseases. Our work provides a framework for comparative single-cell analysis across tissue regions and species.

Two Groups of eGFP-Expressing Neurons with Distinct Characteristics in the Neocortex of GIN Mice

GIN (GFP-expressing inhibitory interneuron) transgenic mice are believed to express the enhanced GFP (eGFP) in a subset of somatostatin (SST)-expressing interneurons in the neocortex and have been widely used in the study on SST interneurons. Previous studies showed that eGFP+ neurons in the neocortex are distributed in the layer II-IV and upper layer V (cortical eGFP neurons) and contain SST. In this study, we reported a new group of eGFP+ neurons in GIN mice at early postnatal ages, which was located in the deep layer of the lateral neocortex as clusters (cluster eGFP neurons). Cluster eGFP neurons were noticeable at birth but disappeared within two months, in contrast to cortical eGFP neurons that started to appear around postnatal day 3 to 5 and existed through life. Cluster eGFP neurons were not immunoreactive for SST antibodies, contrary to cortical eGFP neurons. They were also not immunolabeled by parvalbumin, a marker for another major type of interneurons, and Ca2+/calmodulin-dependent kinases II, a commonly used marker for excitatory neurons. Firing rate, afterhyperpolarization, and excitatory synaptic activity significantly enhanced in cortical eGFP neurons during postnatal development, but these properties remained mostly unchanged in cluster eGFP neurons. Short-term plasticity of the excitatory synapse showed robust facilitation in cortical eGFP neurons but depression in cluster eGFP neurons. These results implied that eGFP might also be expressed in other types of cortical neurons in addition to SST-containing interneurons in GIN mice at early postnatal ages.

Intersectional monosynaptic tracing for dissecting subtype-specific organization of GABAergic interneuron inputs.

Functionally and anatomically distinct cortical substructures such as areas or layers contain different principal neuron (PN) subtypes that generate output signals representing particular information. Various types of cortical inhibitory interneurons (INs) differentially but coordinately regulate PN activity. Despite a potential determinant for functional specialization of PN subtypes, the spatial organization of IN subtypes that innervate defined PN subtypes remains unknown. Here we develop a genetic strategy combining a recombinase-based intersectional labeling method and rabies viral monosynaptic tracing, which enables subtype-specific visualization of cortical IN ensembles sending inputs to defined PN subtypes. Our approach reveals not only cardinal but also underrepresented connections between broad, non-overlapping IN subtypes and PNs. Furthermore, we demonstrate that distinct PN subtypes defined by areal or laminar positions display different organization of input IN subtypes. Our genetic strategy will facilitate understanding of the wiring and developmental principles of cortical inhibitory circuits at unparalleled levels.

Photoreceptor projection from a four-tiered retina to four distinct regions of the first optic ganglion in a jumping spider.

Jumping spiders have four pairs of eyes (ocelli) of which only the principal eyes (PEs) are used to detect features of objects. Photoreceptors in the retina of the PEs form four layers (PL1-4) and terminate in the first optic ganglion (FOG). Here, we focus on Hasarius adansoni because it has unique depth vision besides color vision and its FOG appears to contribute to the initial processing of these visual modalities. We first investigated the neuroanatomical organization of the FOG. The three-dimensional structure of the FOG revealed by synapsin immunostaining is horseshoe-shaped and consists of four terminal zones (TZ1-4). Then, we traced single photoreceptors through serial sections and found that green-sensitive receptors of PL1 and 2 terminate in TZ1 and 2, respectively, by keeping retinotopic organization. In contrast to TZ1 and 2, TZ3 receives terminals of ultraviolet-sensitive receptors from lateral regions of both PL3 and 4, while photoreceptors of the medial region of PL3 and 4 terminate in TZ4. We further studied details of photoreceptor terminals and the branching pattern of interneurons in the FOG in Golgi stained preparations. Photoreceptors have long lateral processes in each terminal zone. Some photoreceptors terminating in TZ3 have branches innervating TZ1, indicating that TZ1 receives different spectral information. A type of interneuron connects TZ1 and 2, while others have branches within a single terminal zone or in the entire FOG. These results suggest that TZ1 and 2 contribute to color, shape, and depth vision, while TZ3 and 4 have specific roles for UV vision.

Expression and Roles of the Immunoglobulin Superfamily Recognition Molecule Sidekick1 in Mouse Retina.

Processes of >100 types of interneurons (bipolar and amacrine cells) and projection neurons (retinal ganglion cells, RGCs) form specific and stereotyped patterns of connections in the inner plexiform layer (IPL) of the mouse retina. Four closely related homophilic immunoglobulin superfamily recognition molecules (Sidekick [Sdk] 1, Sdk 2, Dscam, and DscamL1) have been shown to play roles in patterning neuronal arbors and connections in chick retina, and all but Sdk1 have been shown to play related roles in mice. Here, we compare patterns of Sdk1 and Sdk2 expression in mouse retina and use genetic methods to assess roles of Sdk1. In adult retina, 3 neuronal types express sdk1 but not sdk2 at detectable levels, 5 express sdk2 but not sdk1 and 3 express both. Patterns of gene expression and protein localization at or near synapses are established during the first postnatal week. Dendrites of amacrine cells and RGCs that express sdk1 but not sdk2 arborize in the same narrow stratum in the center of the IPL. In the absence of Sdk1, this laminar restriction is degraded. Overexpression of sdk1 in developing cells that normally express sdk2 reorients their dendrites to resemble those of endogenously Sdk1-positive cells, indicating that Sdk1 plays an instructive role in patterning the IPL. Sdk1 fails to affect arbors when introduced after they are mature, suggesting that it is required to form but not maintain laminar restrictions. The effect of ectopically expressed sdk1 requires the presence of endogenous Sdk1, suggesting that the effect requires homophilic interactions among Sdk1-positive neurites. Together with previous results on Sdk2, Dscam, DscamL1, as well as the related Contactins, our results support the idea that an elaborate immunoglobulin superfamily code plays a prominent role in establishing neural circuits in the retina by means of tightly regulated cell type-specific expression and homophilically restricted intercellular interactions.

LHX1-a multifunctional regulator in preoptic area-derived interneuron development.

LHX1-a multifunctional regulator in preoptic area-derived interneuron development.

Synaptic inhibition tunes contrast computation in the retina.

Inhibition shapes activity and signal processing in neural networks through numerous mechanisms mediated by many different cell types. Here, we examined how one type of GABAergic interneuron in the retina, the A17 amacrine cell, influences visual information processing. Our results suggest that A17s, which make reciprocal feedback inhibitory synapses onto rod bipolar cell (RBC) synaptic terminals, extend the luminance range over which RBC synapses compute temporal contrast and enhance the reliability of contrast signals over this range. Inhibition from other amacrine cells does not influence these computational features. Although A17-mediated feedback is mediated by both GABAA and GABAC receptors, the latter plays the primary role in extending the range of contrast computation. These results identify specific functions for an inhibitory interneuron subtype, as well as specific synaptic receptors, in a behaviorally relevant neural computation.

Pathway-Specific Drive of Cerebellar Golgi Cells Reveals Integrative Rules of Cortical Inhibition.

Cerebellar granule cells (GrCs) constitute over half of all neurons in the vertebrate brain and are proposed to decorrelate convergent mossy fiber (MF) inputs in service of learning. Interneurons within the GrC layer, Golgi cells (GoCs), are the primary inhibitors of this vast population and therefore play a major role in influencing the computations performed within the layer. Despite this central function for GoCs, few studies have directly examined how GoCs integrate inputs from specific afferents, which vary in density to regulate GrC population activity. We used a variety of methods in mice of either sex to study feedforward inhibition recruited by identified MFs, focusing on features that would influence integration by GrCs. Comprehensive 3D reconstruction and quantification of GoC axonal boutons revealed tightly clustered boutons that focus feedforward inhibition in the neighborhood of GoC somata. Acute whole-cell patch-clamp recordings from GrCs in brain slices showed that, despite high GoC bouton density, fast phasic inhibition was very sparse relative to slow spillover mediated inhibition. Dynamic-clamp simulating inhibition combined with optogenetic MF activation at moderate rates supported a predominant role of slow spillover mediated inhibition in reducing GrC activity. Whole-cell recordings from GoCs revealed a role for the density of active MFs in preferentially driving them. Thus, our data provide empirical confirmation of predicted rules by which MFs activate GoCs to regulate GrC activity levels.SIGNIFICANCE STATEMENT A unifying framework in neural circuit analysis is identifying circuit motifs that subserve common computations. Wide-field inhibitory interneurons globally inhibit neighbors and have been studied extensively in the insect olfactory system and proposed to serve pattern separation functions. Cerebellar Golgi cells (GoCs), a type of mammalian wide-field inhibitory interneuron observed in the granule cell layer, are well suited to perform normalization or pattern separation functions, but the relationship between spatial characteristics of input patterns to GoC-mediated inhibition has received limited attention. This study provides unprecedented quantitative structural details of GoCs and identifies a role for population input activity levels in recruiting inhibition using in vitro electrophysiology and optogenetics.

Not just uninhibited: Interneurons and seizure onset

Optogenetic study in vivo demonstrates that different types of interneurons show consistently elevated but distinct firing patterns in the period preceding seizure onset in a rodent model of epilepsy.

Morphological Properties of the Two Types of Caudate Interneurons: Kohonen Self-Organizing Maps and Correlation-Comparison Analysis.

Our previous study found that caudate and putaminal interneurons are morphologically very different, and that accordingly they could be divided in two separate clusters. In addition, it also demonstrated, as a collateral result, that the caudate cluster itself consists of two clusters of morphologically different interneurons. Hence, the objective of this study is a morphological description and subtle differing of morphologies of these two types of caudate interneurons, i.e., an investigation of those morphological traits which characterize them uniquely, and which would distinguish them. Binary two-dimensional images of caudate interneurons, taken from deceased adult human subjects, were analyzed by using 46 parameters, describing the morphology of interneurons. The parameters can be divided in the following classes: size (surface) of a neuron, neuronal shape, length of neuronal morphological compartments, dendritic branching, morphological organization, and complexity. The morphological determination of caudate interneurons was performed in a step-wise manner. The first step was the assignment of each individual neuron to an adequate cluster where it belonged according to morphological criteria. This was done by using the trained artificial neural network, Kohonen self-organizing map. After the clusters were formed, the analysis is further continued by the precise, feature-wise determination of morphological differences found between clusters of caudate interneurons and then finished by defining correlation-based, mutual, inter-parametric relations for each of the clusters. The first was performed by using single-factor analysis, and the second by correlation-comparison analysis. Single-factor analysis showed significance for 34 parameters (morphological features) that distinguish between the clusters. Correlation-comparison analysis extended the results of single-factor analysis by demonstrating significance for 198 inter-parametric correlation pairs that represent 19.13% of mismatched correlations of the first kind among the total number of correlations. This represents a significant inter-cluster separation zone. In addition, the analysis extracted one correlation of the second kind, namely, the DO-MDCBO, very highly significant (p<0.001), positive (r=0.45) in the cluster I, while negative (r=-0.13), also significant (p<0.05) in the cluster II. The two clusters of caudate interneurons were found to be significantly morphologically different. These differences, albeit not strong as the caudate-putaminal differences, are more numerous with respect to significant morphological properties defining them. They probably underlie, influence, and modulate different neurofunctional behavior of the two types of interneurons, which need to be further investigated by future studies.

Astrocytes integrate and drive action potential firing in inhibitory subnetworks

Many brain functions depend on the ability of neural networks to temporally integrate transient inputs to produce sustained discharges. This can occur through cell-autonomous mechanisms in individual neurons and through reverberating activity in recurrently connected neural networks. We report a third mechanism involving temporal integration of neural activity by a network of astrocytes. Previously, we showed that some types of interneurons can generate long-lasting trains of action potentials (barrage firing) following repeated depolarizing stimuli. Here we show that calcium signaling in an astrocytic network correlates with barrage firing; that active depolarization of astrocyte networks by chemical or optogenetic stimulation enhances; and that chelating internal calcium, inhibiting release from internal stores, or inhibiting GABA transporters or metabotropic glutamate receptors inhibits barrage firing. Thus, networks of astrocytes influence the spatiotemporal dynamics of neural networks by directly integrating neural activity and driving barrages of action potentials in some populations of inhibitory interneurons.

The Epigenetic Factor CBP Is Required for the Differentiation and Function of Medial Ganglionic Eminence-Derived Interneurons

The development of inhibitory circuits depends on the action of a network of transcription factors and epigenetic regulators that are critical for interneuron specification and differentiation. Although the identity of many of these transcription factors is well established, much less is known about the specific contribution of the chromatin-modifying enzymes that sculpt the interneuron epigenome. Here, we generated a mouse model in which the lysine acetyltransferase CBP is specifically removed from neural progenitors at the median ganglionic eminence (MGE), the structure where the most abundant types of cortical interneurons are born. Ablation of CBP interfered with the development of MGE-derived interneurons in both sexes, causing a reduction in the number of functionally mature interneurons in the adult forebrain. Genetic fate mapping experiments not only demonstrated that CBP ablation impacts on different interneuron classes, but also unveiled a compensatory increment of interneurons that escaped recombination and cushion the excitatory-inhibitory imbalance. Consistent with having a reduced number of interneurons, CBP-deficient mice exhibited a high incidence of spontaneous epileptic seizures, and alterations in brain rhythms and enhanced low gamma activity during status epilepticus. These perturbations led to abnormal behavior including hyperlocomotion, increased anxiety and cognitive impairments. Overall, our study demonstrates that CBP is essential for interneuron development and the proper functioning of inhibitory circuitry in vivo.

Synapse Type-Dependent Expression of Calcium-Permeable AMPA Receptors.

Calcium-permeable (CP) AMPA-type glutamate receptors (AMPARs) are known to mediate synaptic plasticity in several different interneuron (IN) types. Recent evidence suggests that CP-AMPARs are synapse-specifically expressed at excitatory connections onto a subset of IN types in hippocampus and neocortex. For example, CP-AMPARs are found at connections from pyramidal cells (PCs) to basket cells (BCs), but not to Martinotti cells (MCs). This synapse type-specific expression of CP-AMPARs suggests that synaptic dynamics as well as learning rules are differentially implemented in local circuits and has important implications not just in health but also in disease states such as epilepsy.

Lateral inhibition by Martinotti interneurons is facilitated by cholinergic inputs in human and mouse neocortex

A variety of inhibitory pathways encompassing different interneuron types shape activity of neocortical pyramidal neurons. While basket cells (BCs) mediate fast lateral inhibition between pyramidal neurons, Somatostatin-positive Martinotti cells (MCs) mediate a delayed form of lateral inhibition. Neocortical circuits are under control of acetylcholine, which is crucial for cortical function and cognition. Acetylcholine modulates MC firing, however, precisely how cholinergic inputs affect cortical lateral inhibition is not known. Here, we find that cholinergic inputs selectively augment and speed up lateral inhibition between pyramidal neurons mediated by MCs, but not by BCs. Optogenetically activated cholinergic inputs depolarize MCs through activation of ß2 subunit-containing nicotinic AChRs, not muscarinic AChRs, without affecting glutamatergic inputs to MCs. We find that these mechanisms are conserved in human neocortex. Cholinergic inputs thus enable cortical pyramidal neurons to recruit more MCs, and can thereby dynamically highlight specific circuit motifs, favoring MC-mediated pathways over BC-mediated pathways.

Inhibitory Connectivity Dominates the Fan Cell Network in Layer II of Lateral Entorhinal Cortex.

Fan cells in layer II of the lateral entorhinal cortex (LEC) form a main component of the projection to the dentate gyrus, CA3 and CA2 of the hippocampal formation. This projection has a counterpart originating from stellate cells in layer II of the medial entorhinal cortex (MEC). Available evidence suggests that the two pathways carry different information, exemplified by a difference in spatial tuning of cells in LEC and MEC. The grid cell, a prominent position-modulated cell type present in MEC, has been postulated to derive its characteristic hexagonal firing pattern from dominant disynaptic inhibitory connections between hippocampal-projecting stellate cells. Given that grid cells have not been described in LEC, we aim to describe the local synaptic connectivity of fan cells, to explore whether the network architecture is similar to that of the MEC stellate cell. Using a combination of in vitro multicell electrophysiological and optogenetic approaches in acute slices from rodents of either sex, we show that excitatory connectivity between fan cells is very sparse. Fan cells connect preferentially with two distinct types of inhibitory interneurons, suggesting disynaptic inhibitory coupling as the main form of communication among fan cells. These principles are similar to those reported for stellate cells in MEC, indicating an overall comparable local circuit architecture of the main hippocampal-projecting cell types in the lateral and medial entorhinal cortex.SIGNIFICANCE STATEMENT Our data provide the first description of the synaptic microcircuit of hippocampal-projecting layer II cells in the lateral entorhinal cortex. We show that these cells make infrequent monosynaptic connections with each other, and that they preferentially communicate through a disynaptic inhibitory network. This is similar to the microcircuit of hippocampal-projecting stellate cells in layer II of the medial entorhinal cortex, but dissimilar to the connectivity observed in layer 2 of neocortex. In medial entorhinal cortex, the observed network structure has been proposed to underlie the firing pattern of grid cells. This opens the possibility that layer II cells in lateral entorhinal cortex exhibit regular firing patterns in an unexplored domain.