Regular Spiking Non-pyramidal Cells Research Articles

Cannabinoid type 2 receptors mediate a cell type-specific self-inhibition in cortical neurons

Endogenous cannabinoids are diffusible lipid ligands of the main cannabinoid receptors type 1 and 2 (CB1R and CB2R). In the central nervous system endocannabinoids are produced in an activity-dependent manner and have been identified as retrograde modulators of synaptic transmission. Additionally, some neurons display a cell-autonomous slow self-inhibition (SSI) mediated by endocannabinoids. In these neurons, repetitive action potential firing triggers the production of endocannabinoids, which induce a long-lasting hyperpolarization of the membrane potential, rendering the cells less excitable. Different endocannabinoid receptors and effector mechanisms have been described underlying SSI in different cell types and brain areas. Here, we investigate SSI in neurons of layer 2/3 in the somatosensory cortex. High-frequency bursts of action potentials induced SSI in pyramidal cells (PC) and regular spiking non-pyramidal cells (RSNPC), but not in fast-spiking interneurons (FS). In RSNPCs the hyperpolarization was accompanied by a change in input resistance due to the activation of G protein-coupled inward-rectifying K+ (GIRK) channels. A CB2R-specific agonist induced the long-lasting hyperpolarization, whereas preincubation with a CB2R-specific inverse agonist suppressed SSI. Additionally, using cannabinoid receptor knockout mice, we found that SSI was still intact in CB1R-deficient but abolished in CB2R-deficient mice. Taken together, we describe an additional SSI mechanism in which the activity-induced release of endocannabinoids activates GIRK channels via CB2Rs. These findings expand our knowledge about cell type-specific differential neuronal cannabinoid receptor signaling and suggest CB2R-selective compounds as potential therapeutic approaches.

Whisker experience modulates long‐term depression in neocortical γ‐aminobutyric acidergic interneurons in barrel cortex

Long-term plasticity is believed to be an important mechanism that allows neural circuits to be modulated in an use-dependent manner. However, evidence regarding the role of sensory experience in modulating long-term plasticity of glutamatergic synapses in neocortical GABAergic interneurons is unavailable. Here, we focused on regular-spiking nonpyramidal (RSNP) interneurons and examined spike-pairing-induced plasticity of glutamatergic synapses and its modulation by sensory experiences ex vivo. In our previous studies, RSNP interneurons do not show experience-dependent plasticity of intrinsic properties or alteration of thalamocortical (TC) synapses and exhibit robust modulation by mGluRs. Here we report a spike-pairing-induced long-term depression (spLTD) of glutamatergic synapses in RSNP interneurons of barrels cortex. Using paired recording and thalamic-induced responses, we found that the spLTD was specific for intracortical but not TC synapses. The spLTD was mediated via presynaptic mGluRs. The spLTD was not modulated by chronic or acute administration of NMDAR antagonists but was enhanced by sensory deprivation (via whisker trimming) during a postnatal sensitive period ex vivo. The synapses specific spLTD to intracortical glutamatergic synapses in RSNP cells and their modulation by sensory deprivation may contribute to sensory-dependent remodeling of cortical circuits and redistribution circuit activity along the whisker-related columns.

Subtype-Specific Dendritic Ca2+Dynamics of Inhibitory Interneurons in the Rat Visual Cortex

The Ca(2+) increase in dendrites that is evoked by the backpropagation of somatic action potentials (APs) is involved in the activity-dependent modulation of dendritic and synaptic functions that are location dependent. In the present study, we investigated dendritic Ca(2+) dynamics evoked by backpropagating APs (bAPs) in four subtypes of inhibitory interneurons classified by their spiking patterns: fast spiking (FS), late spiking (LS), burst spiking (BS), and regular-spiking nonpyramidal (RSNP) cells. Cluster analysis, single-cell RT-PCR, and immunohistochemistry confirmed the least-overlapping nature of the grouped cell populations. Somatic APs evoked dendritic Ca(2+) transients in all subtypes of inhibitory interneurons with different spatial profiles along the tree: constantly linear in FS and LS cells, increasing to a plateau in BS cells and bell-shaped in RSNP cells. The increases in bAP-evoked dendritic Ca(2+) transients brought about by the blocking of A-type K(+) channels were similar in whole dendritic trees of each subtype of inhibitory interneurons. However, in RSNP cells, the increases in the distal dendrites were larger than those in the proximal dendrites. On cholinergic activation, nicotinic inhibition of bAP-evoked dendritic Ca(2+) transients was observed only in BS cells expressing cholecystokinin and vasoactive intestinal peptide mRNAs, with no muscarinic modulation in all subtypes of inhibitory interneurons. Cell subtype-specific differential spatial profiles and their modulation in bAP-evoked dendritic Ca(2+) transients might be important for the domain-specific modulation of segregated inputs in inhibitory interneurons and differential control between the excitatory and inhibitory networks in the visual cortex.

Experience-Dependent Intrinsic Plasticity in Interneurons of Barrel Cortex Layer IV

It is unclear whether intrinsic excitabilities of specific interneurons are modulated by sensory experiences. Here, I examined the intrinsic excitabilities of interneurons in "sensory-spared" and "sensory-deprived" cortices of GAD67-GFP mice. The results showed that whisker trimming, begun at postnatal day 7 for 3 wk, induced significant changes in intrinsic and firing properties of fast-spiking (FS) but not regular spiking nonpyramidal (RSNP) cells. Firing threshold, spike frequency, spike adaptation index, and input resistance of FS cells were significantly altered by sensory deprivation such that FS cells became less excitable. An up-regulation of IA currents in FS cells appeared to be responsible. Along with changes in the intrinsic properties of FS cells, whisker trimming also induced a robust reduction in the number of vesicular glutamate transporter 2 positive varicosities and parvalbumin expression and the strength of thalamocortical (TC) excitatory postsynaptic currents in FS cells in the "sensory-deprived barrels." The probability of spike induction by TC stimulus was reduced by 30% and the spike jitter was increased in sensory-deprived FS cells. These results suggest that the FS networks are selectively inhibited by sensory deprivation. The concurrent changes of intrinsic properties and expression of parvalbumin in FS but not RSNP cells with TC synapses support a contribution from the TC pathway and glutamate to sensory-induced activity-dependent intrinsic plasticity of inhibitory networks in barrel cortex.

Attractor dynamics in a modular network model of neocortex

Starting from the hypothesis that the mammalian neocortex to a first approximation functions as an associative memory of the attractor network type, we formulate a quantitative computational model of neocortical layers 2/3. The model employs biophysically detailed multi-compartmental model neurons with conductance based synapses and includes pyramidal cells and two types of inhibitory interneurons, i.e., regular spiking non-pyramidal cells and basket cells. The simulated network has a minicolumnar as well as a hypercolumnar modular structure and we propose that minicolumns rather than single cells are the basic computational units in neocortex. The minicolumns are represented in full scale and synaptic input to the different types of model neurons is carefully matched to reproduce experimentally measured values and to allow a quantitative reproduction of single cell recordings. Several key phenomena seen experimentally in vitro and in vivo appear as emergent features of this model. It exhibits a robust and fast attractor dynamics with pattern completion and pattern rivalry and it suggests an explanation for the so-called attentional blink phenomenon. During assembly dynamics, the model faithfully reproduces several features of local UP states, as they have been experimentally observed in vitro, as well as oscillatory behavior similar to that observed in the neocortex.

Gap-Junctional Coupling between Neurogliaform Cells and Various Interneuron Types in the Neocortex

Electrical synapses contribute to the generation of synchronous activity in neuronal networks. Several types of cortical GABAergic neurons acting via postsynaptic GABA(A) receptors also form electrical synapses with interneurons of the same class, suggesting that synchronization through gap junctions could be limited to homogenous interneuron populations. Neurogliaform cells elicit combined GABA(A) and GABA(B) receptor-mediated postsynaptic responses in cortical pyramidal cells, but it is not clear whether neurogliaform cells are involved in networks linked by electrical coupling. We recorded from pairs, triplets, and quadruplets of cortical neurons in layers 2 and 3 of rat somatosensory cortex (postnatal day 20-35). Neurogliaform cells eliciting slow IPSPs on pyramidal cells also triggered divergent electrical coupling potentials on interneurons. Neurogliaform cells were electrically coupled to other neurogliaform cells, basket cells, regular-spiking nonpyramidal cells, to an axoaxonic cell, and to various unclassified interneurons showing diverse firing patterns and morphology. Electrical interactions were mediated by one or two electron microscopically verified gap junctions linking the somatodendritic domain of the coupled cells. Our results suggest that neurogliaform cells have a unique position in the cortical circuit. Apart from eliciting combined GABA(A) and GABA(B) receptor-mediated inhibition on pyramidal cells, neurogliaform cells establish electrical synapses and link multiple networks formed by gap junctions restricted to a particular class of interneuron. Widespread electrical connections might enable neurogliaform cells to monitor the activity of different interneurons acting on GABA(A) receptors at various regions of target cells.

Input and frequency‐specific entrainment of postsynaptic firing by IPSPs of perisomatic or dendritic origin

Correlated activity of cortical neurons underlies cognitive processes. Networks of several distinct classes of gamma-aminobutyric acid (GABA)ergic interneurons are capable of synchronizing cortical neurons at behaviourally relevant frequencies. Here we show that perisomatic and dendritic GABAergic inputs provided by two classes of GABAergic cells, fast spiking and bitufted interneurons, respectively, entrain the timing of postsynaptic spikes differentially in both pyramidal cells and interneurons at beta and gamma frequencies. Entrainment of pyramidal as well as regular spiking non-pyramidal cells was input site and inhibitory postsynaptic potential frequency dependent. Gamma frequency input from fast spiking cells entrained pyramidal cells on the positive phase of an intrinsic cellular theta oscillation, whereas input from bitufted cells was most effective in gamma frequency entrainment on the negative phase of the theta oscillation. The discharge of regular spiking interneurons was phased at gamma frequency by dendritic input from bitufted cells, but not by perisomatic input from fast spiking cells. Action potentials in fast spiking GABAergic neurons were phased at gamma frequency by both other fast spiking and bitufted cells, regardless of whether the presynaptic GABAergic input was at gamma or beta frequency. The interaction of cell type-specific intrinsic properties and location-selective GABAergic inputs could result in a spatio-temporally regulated synchronization and gating of cortical spike propagation in the network.

β and γ Frequency Synchronization by Dendritic GABAergic Synapses and Gap Junctions in a Network of Cortical Interneurons

Distinct interneuron populations innervate perisomatic and dendritic regions of cortical cells. Perisomatically terminating GABAergic inputs are effective in timing postsynaptic action potentials, and basket cells synchronize each other via gap junctions combined with neighboring GABAergic synapses. The function of dendritic GABAergic synapses in cortical rhythmicity, and their interaction with electrical synapses is not understood. Using multiple whole-cell recordings in layers 2-3 of rat somatosensory cortex combined with light and electron microscopic determination of sites of interaction, we studied the interactions between regular spiking nonpyramidal cells (RSNPCs). Random samples of unlabeled postsynaptic targets showed that RSNPCs placed GABAergic synapses onto dendritic spines (53 +/- 12%) and shafts (45 +/- 10%) and occasionally somata (2 +/- 4%). GABAergic interactions between RSNPCs were mediated by 4 +/- 2 axodendritic synapses and phased postsynaptic activity at beta frequency but were ineffective in phasing at gamma rhythm. Electrical interactions of RSNPCs were transmitted via two to eight gap junctions between dendritic shafts and/or spines. Elicited at beta and gamma frequencies, gap junctional potentials timed postsynaptic spikes with a phase lag, however strong electrical coupling could synchronize presynaptic and postsynaptic activity. Combined unitary GABAergic and gap junctional connections of moderate strength produced beta and gamma frequency synchronization of the coupled RSNPCs. Our results provide evidence that dendritic GABAergic and/or gap junctional mechanisms effectively transmit suprathreshold information in a population of interneurons at behaviorally relevant frequencies. A coherent network of GABAergic cells targeting the dendrites could provide a pathway for rhythmic activity spatially segregated from perisomatic mechanisms of synchronization.

Trans-synaptically induced bursts in regular spiking non-pyramidal cells in deep layers of the cat motor cortex.

In deep layers of the cat motor cortex, we have investigated the properties of neurons displaying trans-synaptically induced bursts. In in vivo experiments, extracellularly recorded burst neurons were separated into two subtypes based on their dependence on stimulation sites, the medullary pyramid or the ventrolateral (VL) thalamic nucleus, from which bursts of 10-20 spikes were triggered. The spike amplitude attenuation and frequency adaptation during a burst were more prominent in pyramid-dependent burst neurons than in VL-dependent burst neurons. Intracellular recordings in in vivo experiments revealed that pyramid-dependent bursts emerged from a long-lasting depolarization, while each spike during a VL-dependent burst was narrow in half-width and was followed by a fast AHP, similar to fast spiking neurons. In in vitro slice experiments, intracellular recordings were obtained from neurons that displayed a burst of attenuated spikes emerging from a long-lasting depolarization, and were also obtained from fast spiking neurons. They were morphologically recovered to be multipolar cells with sparsely spiny dendrites and local axonal networks, suggesting that they are inhibitory interneurons. The multipolar neurons displaying bursts of attenuated spikes may mediate the recurrent inhibition of pyramidal tract cells.

Neurochemical features and synaptic connections of large physiologically-identified GABAergic cells in the rat frontal cortex

Physiological and morphological properties of large non-pyramidal cells immunoreactive for cholecystokinin, parvalbumin or somatostatin were investigated in vitro in the frontal cortex of 18–22-day-old rats. These three peptides were expressed in separate populations including large cells. Cholecystokinin cells and parvalbumin cells made boutons apposed to other cell bodies, but differed in their firing patterns in response to depolarizing current pulses. Parvalbumin cells belonged to fast-spiking cells. Parvalbumin fast-spiking cells also included chandelier cells. In contrast, cholecystokinin cells were found to be regular-spiking non-pyramidal cells or burst-spiking non-pyramidal cells with bursting activity from hyperpolarized potentials (two or more spikes on slow depolarizing humps). Large somatostatin cells belonged to the regular-spiking non-pyramidal category and featured wide or ascending axonal arbors (wide arbor cells and Martinotti cells) which did not seem to be apposed to the somata so frequently as large cholecystokinin and parvalbumin cells. For electron microscopic observations, another population of eight immunohistochemically-uncharacterized non-pyramidal cells were selected: (i) five fast spiking cells including one chandelier cell which are supposed to contain parvalbumin, and (ii) three large regular-spiking non-pyramidal cells with terminals apposed to somata, which are not considered to include somatostatin cells, but some of which may belong to cholecystokinin cells. The fast-spiking cells other than a chandelier cell and the large regular-spiking non-pyramidal cells made GABA-positive synapses on somata (4% and 12% of the synapses in two small to medium fast-spiking cells, 22% and 35% of the synapses in two large fast-spiking cells, and 10%, 18% and 37% of the synapses in three large regular-spiking non-pyramidal cells). A few terminals of the fast-spiking and regular-spiking non-pyramidal cells innervated GABAergic cells. About 30% of the fast-spiking cell terminals innervated spines, but few of the regular-spiking non-pyramidal cell terminals did. A fast-spiking chandelier cell made GABA-positive synapses on GABA-negative axon initial segments. These results suggest that large GABAergic cells are heterogeneous in neuroactive substances, firing patterns and synaptic connections, and that cortical cells receive heterogeneous GABAergic somatic inputs.

Molecular and Physiological Diversity of Cortical Nonpyramidal Cells

The physiological and molecular features of nonpyramidal cells were investigated in acute slices of sensory-motor cortex using whole-cell recordings combined with single-cell RT-PCR to detect simultaneously the mRNAs of three calcium binding proteins (calbindin D28k, parvalbumin, and calretinin) and four neuropeptides (neuropeptide Y, vasoactive intestinal polypeptide, somatostatin, and cholecystokinin). In the 97 neurons analyzed, all expressed mRNAs of at least one calcium binding protein, and the majority (n = 73) contained mRNAs of at least one neuropeptide. Three groups of nonpyramidal cells were defined according to their firing pattern. (1) Fast spiking cells (n = 34) displayed tonic discharges of fast action potentials with no accommodation. They expressed parvalbumin (n = 30) and/or calbindin (n = 19) mRNAs, and half of them also contained transcripts of at least one of the four neuropeptides. (2) Regular spiking nonpyramidal cells (n = 48) displayed a firing behavior characterized by a marked accommodation and presented a large diversity of expression patterns of the seven biochemical markers. (3) Finally, a small population of vertically oriented bipolar cells, termed irregular spiking cells (n = 15), fired bursts of action potentials at an irregular frequency. They consistently co-expressed calretinin and vasoactive intestinal polypeptide. Additional investigations of these cells showed that they also co-expressed glutamic acid decarboxylase and choline acetyl transferase. Our results indicate that neocortical nonpyramidal neurons display a large diversity in their firing properties and biochemical patterns of co-expression and that both characteristics could be correlated to define discrete subpopulations.

Depolarization-induced slow potentiation of depolarizing afterpotential.

Effects of strong membrane depolarization on the excitability of regular spiking (RS) non-pyramidal cells in layer VI of the cat motor cortex were examined in in vitro slice preparations. RS non-pyramidal cells that expressed depolarizing afterpotential (DAP) following spikes displayed either of the two distinct responses: immediate and successive potentiation of DAP in association with repetition of firing, or slow enhancement of DAP after repetitive injections of strong depolarizing current pulses leading to burst firing. In those neurones that displayed the immediate potentiation of DAP, slow potentiation was never induced. This difference in changes of DAP was consistent with morphological differences. DAP-lacking RS non-pyramidal cells displayed neither immediate nor slow changes in excitability.

Physiological and morphological identification of somatostatin- or vasoactive intestinal polypeptide-containing cells among GABAergic cell subtypes in rat frontal cortex

Physiological and morphological characteristics of GABAergic nonpyramidal cells in frontal cortex of young rats identified immunohistochemically as containing somatostatin or vasoactive intestinal polypeptide (VIP) were studied in vitro by whole-cell recording and biocytin injection. We have found that most somatostatin- or VIP-containing neurons were different from two other types of GABAergic cells, the parvalbumin-containing fast-spiking cells and the late-spiking cells (neurogliaform cells). In response to injected currents, somatostatin- or VIP-containing nonpyramidal cells showed either bursts of a few spikes on a slow-depolarizing hump, burst-spiking nonpyramidal cells, or single spikes only on depolarization, regular-spiking nonpyramidal cells. Morphologically, both somatostatin- and VIP-containing cells had vertical axonal arbors terminating in symmetrical synapses that were immunoreactive for GABA in electron micrographs. Somatostatin cells included neurons with main ascending axons sending collaterals into layer I (Martinotti cells in deep layers). Some of the Martinotti cells in layer V also contained calbindin D 28k. VIP cells included neurons the main descending axons of which had more descending than ascending collaterals (bipolar cells and double bouquet cells). Two other morphological forms of the VIP cells were those with short descending axons with collaterals bearing multiple boutons on other cell bodies (small basket cells) or with short ascending main axons with collaterals forming arcades (arcade cells). Some of these neurons also contained calretinin. From these results, it appears that the GABAergic neurons controlling circuits in the neocortical layers may be characterized further based on whether they contain somatostatin or VIP.

Physiological subgroups of nonpyramidal cells with specific morphological characteristics in layer II/III of rat frontal cortex

Physiological and morphological properties of nonpyramidal cells in layer II/III of frontal cortex of young rats were studied in vitro by whole-cell recording and intracellular staining with biocytin. Layer II/III nonpyramidal cells could be divided into four subgroups by their firing patterns in response to depolarizing current pulses and their patterns of dendritic and axonal arborizations. (1) Fast-spiking (FS) cells (35% of the total sample) showed an abrupt episode of nonadapting repetitive discharges of shorter-duration action potentials. FS cells had local and horizontal axonal arbors which did not enter layer I. This type of FS cell was immunoreactive for parvalbumin and included some basket cells. Three chandelier cells were identified as FS cells, although one chandelier cell was not identified as an FS cell. (2) Late-spiking (LS) cells (11%) exhibited slowly developing ramp depolarizations near threshold. LS cells were neurogliaform cells. (3) Low-threshold spike (LTS) cells (5%) had prominent low-threshold spikes when stimulated from hyperpolarizations. The main axons of LTS cells ascended, and the collaterals entered into layer I. (4) The remaining cells [regular-spiking nonpyramidal (RSNP) cells] (49%) could not be categorized into the above three subgroups. In some RSNP cells depolarizing pulses from hyperpolarizations induced fast depolarizing notches with small amplitude. RSNP cells had vertically elongated axonal fields, extending from layer I to V, sometimes to layer VI. This subgroup included double bouquet cells and bipolar cells. Each subgroup with a different firing mode may differentially contribute to neocortical laminar and columnar circuitry.