Three classes of pyramidal neurons in layer V of rat perirhinal cortex

Whole-cell recordings from 140 pyramidal neurons in layer V of rat perirhinal cortex (PR) revealed three distinct firing patterns: regular spiking (RS, 76%), burst spiking (BS, 9%), and late spiking (LS, 14%). LS neurons have not previously been reported in layer V of any cortical region. LS cells in layer V of PR exhibited delays of up to 12 s from onset of a depolarizing current step to spike threshold, followed by sustained firing. In contrast, pyramidal cells in layer V of other cortical regions contain only RS and BS cells. Within PR, the percentage of LS neurons in layer V differs markedly from what we previously observed in layers II/III (50% LS) and VI (90% LS). Morphologically, BS neurons in layer V of PR had thick primary apical dendrites that terminated in a tuft within layer I, whereas RS and LS cells had relatively thin primary apicals that terminated either diffusely or in a layer I tuft. At holding potentials near rest, PR neurons exhibited small (approximately 15 pA), inward, spontaneous postsynaptic currents (PSCs) that were indistinguishable among the three cell types. Currents evoked by minimal stimulation of layer I were about 2.8 times larger than the spontaneous PSCs. Evoked currents had unusually long onset latencies with little variation in latency, consistent with monosynaptic responses evoked by stimulation of unmyelinated fibers. The prevalence of LS cells in combination with the long-latency monosynaptically evoked PSCs suggested that PR is not a region of rapid throughput. This is consistent with anatomical data suggesting that PR is a higher-level association cortex. These data further advance an emerging picture of PR as a cortical region with a unique distribution of cell types different from other cortical regions.     

Cellular properties of principal neurons in the rat entorhinal cortex. I. The lateral entorhinal cortex

The lateral entorhinal cortex (LEC) provides a major cortical input to the hippocampal formation, equaling that of the medial entorhinal cortex (MEC). To understand the functional contributions made by LEC, basic knowledge of individual neurons, in the context of the intrinsic network, is needed. The aim of this study is to compare physiological and morphological properties of principal neurons in different LEC layers in postnatal rats. Using in vitro whole cell current-clamp recordings from up to four post hoc morphologically identified neurons simultaneously, we established that principal neurons show layer specific physiological and morphological properties, similar to those reported previously in adults. Principal neurons in L(ayer) I, LII, and LIII have the majority of their dendrites and axonal collaterals alone in superficial layers. LV contains mainly pyramidal neurons with dendrites and axons extending throughout all layers. A minority of LV and all principal neurons in LVI are neurons with dendrites confined to deep layers and axons in superficial and deep layers. Physiologically, input resistances and time constants of LII neurons are lower and shorter, respectively, than those observed in LV neurons. Fifty-four percent of LII neurons have sag potentials, resonance properties, and rebounds at the offset of hyperpolarizing current injection, whereas LIII and LVI neurons do not have any of these. LV neurons show prominent spike-frequency adaptation and a decrease in spike amplitudes in response to strong depolarization. Despite the well-developed interlaminar communication in LEC, the laminar differences in the biophysical and morphological properties of neurons suggest that their in vivo firing patterns and functions differ, similar to what is known for neurons in different MEC layers.     

Cellular properties of principal neurons in the rat entorhinal cortex. II. The medial entorhinal cortex

Principal neurons in different medial entorhinal cortex (MEC) layers show variations in spatial modulation that stabilize between 15 and 30 days postnatally. These in vivo variations are likely due to differences in intrinsic membrane properties and integrative capacities of neurons. The latter depends on inputs and thus potentially on the morphology of principal neurons. In this comprehensive study, we systematically compared the morphological and physiological characteristics of principal neurons in all MEC layers of newborn rats before and after weaning. We recorded simultaneously from up to four post-hoc morphologically identified MEC principal neurons in vitro. Neurons in L(ayer) I-LIII have dendritic and axonal arbors mainly in superficial layers, and LVI neurons mainly in deep layers. The dendritic and axonal trees of part of LV neurons diverge throughout all layers. Physiological properties of principal neurons differ between layers. In LII, most neurons have a prominent sag potential, resonance and membrane oscillations. Neurons in LIII and LVI fire relatively regular, and lack sag potentials and membrane oscillations. LV neurons show the most prominent spike-frequency adaptation and highest input resistance. The data indicate that adult-like principal neuron types can be differentiated early on during postnatal development. The results of the accompanying paper, in which principal neurons in the lateral entorhinal cortex (LEC) were described (Canto and Witter,2011), revealed that significant differences between LEC and MEC exist mainly in LII neurons. We therefore systematically analyzed changes in LII biophysical properties along the mediolateral axis of MEC and LEC. There is a gradient in properties typical for MEC LII neurons. These properties are most pronounced in medially located neurons and become less apparent in more laterally positioned ones. This gradient continues into LEC, such that in LEC medially positioned neurons share some properties with adjacent MEC cells.     

Electrophysiological and morphological properties of neurons in layer 5 of the rat postrhinal cortex

The postrhinal (POR) cortex of the rat is homologous to the parahippocampal cortex of the primate based on connections and other criteria. POR provides the major visual and visuospatial input to the hippocampal formation, both directly to CA1 and indirectly through connections with the medial entorhinal cortex. Although the cortical and hippocampal connections of the POR cortex are well described, the physiology of POR neurons has not been studied. Here, we examined theelectrical and morphological characteristics of layer 5 neurons from POR cortex of 14‐ to 16‐day‐old rats using an in vitro slice preparation. Neurons were subjectively classified as regular‐spiking (RS), fast‐spiking (FS), or low‐threshold spiking (LTS) based on their electrophysiological properties and similarities with neurons in other regions of neocortex. Cells stained with biocytin included pyramidal cells and interneurons with bitufted or multipolar dendritic patterns. Similarity analysis using only physiological data yielded three clusters that corresponded to FS, LTS, and RS classes. The cluster corresponding to the FS class was composed entirely of multipolar nonpyramidal cells, and the cluster corresponding to the RS class was composed entirely of pyramidal cells. The third cluster, corresponding to the LTS class, was heterogeneous and included both multipolar and bitufted dendritic arbors as well as one pyramidal cell. We did not observe any intrinsically bursting pyramidal cells, which is similar to entorhinal cortex but unlike perirhinal cortex. We conclude that POR includes at least two major classes of neocortical inhibitory interneurons, but has a functionally restricted cohort of pyramidal cells. © 2012 Wiley Periodicals, Inc.     

Morphology and distribution of electrophysiologically defined classes of pyramidal and nonpyramidal neurons in rat ventral subiculum in vitro

Intracellular electrophysiological recordings were made from 210 ventral subicular neurons in rat brain slices. Recordings here classified as burst-firing or nonburst-firing. Eighteen burst-firing neurons were filled with Neurobiotin, and all had pyramidal morphology. Nine of these recordings were made from intrinsically burst-firing (IB) cell bodies, and nine were made from burst-firing dendrites (BD). Twelve nonburst-firing neurons were also filled with Neurobiotin. Eight were regular spiking (RS) and had pyramidal morphology, four were fast spiking (FS) and nonpyramidal. Additional electrophysiological parameters distinguished IB from BD, RS from FS, and RS from IB recordings. The distribution of IB and RS neurons was examined by using 180 recordings. Information from the first series of experiments was used to distinguish between somatic and dendritic recordings. The deep-superficial axis (alveus-hippocampal fissure) was divided into four equal rows. RS neurons accounted for 12%, 28%, 58%, and 50% of presumed somatic recordings in successively more superficial rows. The proximal-distal (CA1-perforant path) axis was divided into five equal columns. RS cells accounted for 52% of presumed somatic impalements in the central column compared with 16% in the most proximal and 10% in the most distal columns. Thus, two electrophysiological classes of pyramidal neuron were localized to particular regions of the ventral subiculum. In the light of existing knowledge of the topography of subicular inputs and outputs, our results are consistent with the hypothesis that the ratio of RS to IB pyramidal neurons will be different in different transhippocampal circuits. J. Comp. Neurol. 380:395–408, 1997. © 1997 Wiley-Liss, Inc.     

Classification of extracellularly recorded neurons by their discharge patterns and their correlates with intracellularly identified neuronal types in the frontal cortex of behaving monkeys

Neurons in the cerebral cortex are not homogeneous. However, neuronal types have been ignored in most previous work studying neuronal processes in behaving monkeys. We propose a new method to identify neuronal types in extracellular recording studies of behaving monkeys. We classified neurons as either bursting or non‐bursting, and then classified the bursting neurons into three types: (i) neurons displaying a burst of many spikes (maximum number of spikes within a burst; NSB max ≥ 8) at a high discharge rate (maximum interspike interval; ISI max < 5 ms); (ii) neurons displaying a burst of fewer spikes (NSB max ≤ 5) at a high discharge rate (ISI max < 5 ms); and (iii) neurons displaying a burst of a few spikes (NSB max ≤ 7) at relatively long ISIs (ISI max > 5 ms). We found that the discharge patterns of the four groups corresponded to those of regular spiking (RS), fast spiking (FS), fast rhythmic bursting (FRB) and intrinsic bursting (IB) neurons demonstrated in intracellular recording studies using in vitro slice preparations, respectively. In addition, we examined correlations with the task events for neurons recorded in the frontal eye field and neuronal interactions for pairs of neurons recorded simultaneously from a single electrode. We found that they were substantially different between RS and FS types. These results suggest that neurons in the frontal cortex of behaving monkeys can be classified into four types based on their discharge patterns, and that these four types contribute differentially to cortical operations.     

Correlation between intrinsic firing patterns and thalamocortical synaptic responses of neurons in mouse barrel cortex.

We used a thalamocortical slice preparation to record both spike trains and synaptically evoked responses from neurons of mouse barrel cortex. Cells were classified as regular spiking (RS), intrinsically bursting (IB), or fast spiking (FS) according to their temporal firing patterns when injected with current. RS cells were further separated into two subtypes, RS1 and RS2 cells, the latter encountered only in the infragranular layers. Synaptic responses were elicited by focal electrical stimuli in the ventrobasal nucleus of the thalamus (VB) while holding the cells at different membrane potentials. Postsynaptic potentials were classified as excitatory (EPSPs) or inhibitory (IPSPs), and their latencies were measured from the onset of the extracellularly recorded fiber volley in layer IV. EPSPs fell into three groups, according to latency. Those in the early cluster had latencies shorter than 1 msec and were coincident with the postsynaptic layer IV population response; they were considered monosynaptic. A second group, with latencies between 1.3 and 2.5 msec, were coincident with all IPSPs and were classified as disynaptic. The rest had latencies longer than 5 msec and were considered polysynaptic. The synaptic order of a cell was correlated with its laminar position and its electrophysiological class. Specifically, monosynaptic responses were restricted to infragranular RS cells and to FS cells, while disynaptic EPSPs were found in supragranular RS cells and in IB cells. Disynaptic IPSPs were found in both deep and superficial layers; in the deep layers they nearly always followed monosynaptic EPSPs, while in the superficial layers they were mostly found in isolation. We conclude that the intrinsic spiking characteristics of a neuron are an important determinant of its position in the cortical circuit and may have a substantial role in determining its response properties.     

Electrophysiological and morphological diversity of neurons from the rat subicular complex in vitro

We combined whole-cell recordings with Neurobiotin labeling to examine the electrophysiological and morphological properties of neurons from the ventral subicular complex in vitro (including the subicular, presubicular, and parasubicular areas). No a priori morphological sampling criteria were used to select cells. Cells were classified as bursting (IB), regular-spiking (RS), and fast-spiking (FS) according to their firing patterns in response to depolarizing current pulses. A number of cells remained unclassified. We found 54% RS, 26% IB, 11% FS, and 9% unclassified cells out of a total of 131 neurons examined. We also found cells showing intrinsic membrane potential oscillations (MPO) (6%), which represented a subgroup of the unclassified cells. We analyzed several electrophysiological parameters and found that RS and IB cells can be subclassified into two separate subgroups. RS cells were subclassified as tonic and adapting, according to the degree of firing adaptation. Both responded with single spikes to orthodromic stimulation. IB cells were subclassified in two subgroups according to their capacity to fire more than one burst, and showed different responses to orthodromic stimulation. We observed that bursting in these two subgroups appeared to involve both Ca2+ and persistent Na+ components. Both IB and RS cells, as well as MPO neurons, were projecting cells. FS cells were morphologically identified as local circuit interneurons. We also analyzed the spatial distribution of these cell types from the vicinity of CA1 to the parasubicular areas. We conclude that, in contrast to the commonly accepted idea of the subicular complex as a bursting structure, there is a wide electrophysiological variability even within a given cellular group.     

Intrinsic connectivity of the rat subiculum: I. Dendritic morphology and patterns of axonal arborization by pyramidal neurons.

The dendritic and axonal morphology of rat subicular neurons was studied in single cells labeled with Neurobiotin. Electrophysiological classification of cells as intrinsic burst firing or regular spiking neurons was correlated with morphologic patterns and cell locations. Every cell had dendritic branches that reached the outer molecular layer, with most cells having branches that reached the hippocampal fissure. All but two pyramidal cells had axon collaterals that entered the deep white matter (alveus). Branching patterns of apical dendrites varied as a function of the cell's soma location along the fissure-alveus axis of the cell layer. The first major dendritic branch point for most cells occurred at the superficial edge of the cell layer giving deep cells long primary apical dendrites and superficial cells short or absent primary apical dendrites. In contrast, basal dendritic arbors were similar across cells regardless of cell position. Apical and basal dendrites of all cells had numerous spines. Superficial and deep cells also differed in axonal collateralization. Deep cells (mostly intrinsically bursting [IB] class) had one or more ascending axon collaterals that typically remained within the region circumscribed by their apical dendrites. Superficial cells (mostly regular spiking [RS] class) tended to have axon collaterals that reached longer distances in the cell layer. Numerous varicosities and axonal extensions were present on axon collaterals in the cell layer and in the apical dendritic region, suggesting intrinsic connectivity. Axonal varicosities and extensions were found on axons that entered presubiculum, entorhinal cortex or CA1, supporting the notion that these were projection cells. Local collaterals were distinctly thinner than collaterals that would leave the subiculum, suggesting little or no myelin on local collaterals and some myelin on efferent fibers. We conclude that both IB and RS classes of subicular principal cells make synaptic contacts in and apical to the cell layer. Based on the patterns of axonal arborization, we suggest that subiculum has at least a crude columnar and laminar architecture, with ascending collaterals of deep cells forming columns and broader axonal arbors of superficial cells serving to distribute activity across multiple columns.     

Presubiculum layer III conveys retrosplenial input to the medial entorhinal cortex

Navigation is mediated by a network of brain areas, and research has focused on the head-direction system in the presubiculum (PrS), the grid cell containing medial entorhinal cortex (EC) (MEC) and place cells in the hippocampus. Less research addressed the interactions of the retrosplenial cortex (RSC) and the navigational system, although it is well established that damage to the RSC leads to navigational deficits. We previously showed that RSC provides a dense input to deep layers of MEC and to superficial layers of PrS. In this study we use confocal microscopical analysis and show that the dense projection from the caudal part of the ventral retrosplenial granular cortex targets neurons in Layer III of PrS, which provide input to superficial layers of MEC. Our high resolution anatomical data indicate that sparsely spiny pyramidal neurons in Layer III of PrS that originate projections to Layer III of MEC are the main target of these retrosplenial projections. Retrosplenial axonal boutons were found to equally contact spines and shafts of basal dendrites in Layer III, but contacts on shafts are more prominent close to the soma, indicating the potential for efficient synaptic transfer. These observations suggest that neurons in Layer III of PrS have an important role in mediating RSC contributions to navigation.     

Synaptic excitability of the burst firing neurons in cat sensorimotor cortex in vitro

We have recently reported that the burst firing neurons are found in layer III as well as in layer V of cat sensorimotor cortex in vitro. In the present study, we examined the synaptic excitability of layer III neurons by white matter stimulation and compared with their firing patterns against the current injections through the recording microelectrodes. The firing patterns of layer III neurons were classified into three main classes as in our previous study, i.e., (1) regular spiking (RS), i.e., the tonic firing that often exhibited spike-frequency adaptation, (2) burst-and-single spiking (BS), i.e., the initial bursting followed by tonic firing, (3) repetitive-bursting (RB), the burst firing that recurred at fast frequency. In RS cells, single action potential was superimposed on the largest EPSPs among all cell types analyzed. BS cells also fired single action potential and never exhibited burst firing synaptically. Only in a part of RB cells, synaptic bursting instead of single action potential was evoked on smaller EPSPs. IPSPs could be observed in about 60% of all the recorded RS and BS cells, however, they were observed in only 10% of the RB cells.     

Morphology and physiology of neurons in the rat perirhinal-lateral amygdala area.

Neuronal structure-function relationships were studied in rat brain slices containing the perirhinal cortex (PR) and immediately adjacent lateral nucleus of the amygdala (ALa). Using video microscopy, whole-cell recordings were made from visually preselected neurons that were labeled with biocytin for subsequent anatomical reconstructions. Most cells were 1 of 4 primary neurophysiological types: fast-spiking (FS), regular-spiking (RS), late-spiking (LS), and burst-spiking (BS). Fast-spiking neurons (small somata) were found throughout PR; RS neurons (stellates and pyramids) were present from layer II/III through VI of PR; BS neurons (large pyramids with thick nonbifurcating apical dendrites) were found in layer Va of PR; and LS neurons (stellates, small pyramids, and cone cells) were encountered in layers II/III and VI of PR. One subpopulation of LS neurons (small pyramids) was found in layer II/III; another (cone cells) was found in clusters spanning layer VI through the lateral portion of ALa. Layer Va also contained large RS pyramidal neurons whose axons were seen traveling in the external capsule, but not entering the ALa. Conversely, the axons of large RS pyramidal neurons in layer Vb typically projected deep into the ALa. The four primary firing patterns were present in ALa, which also contained irregular-spiking, slow-charging, and single-spiking cells. Spontaneous synaptic currents differed markedly among cell types and layers. There was excellent agreement between somatic areas measured from video images of living neurons and somatic areas from the same neurons following fixation. Representative montages, which combined the cellular neuroanatomy and neurophysiology, suggested a circuit-level organization that helps elucidate information processing through the PR-ALa region.     

Developmental regulation of action potential-induced Ca2+ entry in neocortical neurons.

Intracellular calcium plays a critical role in the regulation of membrane excitability, synaptic integration and synaptic plasticity. Using whole-cell recording and calcium imaging, we investigated intracellular calcium dynamics in apical dendrites of layer V pyramidal cells in the immature rat neocortex. Dendritic calcium increases induced by action potentials were only small on postnatal days 4-6 (P4-6), then underwent a gradual enhancement during the second post-natal week and were 2- to 3-fold larger on P16-18 than on P4-6. At P15-18 there were no significant differences in the calcium increases among the three subclasses of layer V pyramidal neurons: adapting regular spiking (RS), non-adapting RS and intrinsically bursting (IB) neurons. Developmental regulation of dendritic calcium dynamics may be crucial for functional maturation of the neocortex.     

Different output properties of perisomatic region‐targeting interneurons in the basal amygdala

The perisomatic region of principal neurons in cortical regions is innervated by three types of GABAergic interneuron, including parvalbumin‐containing basket cells (PVBCs) and axo‐axonic cells (AACs), as well as cholecystokinin and type 1 cannabinoid receptor‐expressing basket cells (CCK/CB1BCs). These perisomatic inhibitory cell types can also be found in the basal nucleus of the amygdala, however, their output properties are largely unknown. Here, we performed whole‐cell recordings in morphologically identified interneurons in slices prepared from transgenic mice, in which the GABAergic cells could be selectively targeted. Investigating the passive and active membrane properties of interneurons located within the basal amygdala revealed that the three interneuron types have distinct single‐cell properties. For instance, the input resistance, spike rate, accommodation in discharge rate, or after‐hyperpolarization width at the half maximal amplitude separated the three interneuron types. Furthermore, we performed paired recordings from interneurons and principal neurons to uncover the basic features of unitary inhibitory postsynaptic currents (uIPSCs). Although we found no difference in the magnitude of responses measured in the principal neurons, the uIPSCs originating from the distinct interneuron types differed in rise time, failure rate, latency, and short‐term dynamics. Moreover, the asynchronous transmitter release induced by a train of action potentials was typical for the output synapses of CCK/CB1BCs. Our results suggest that, despite the similar uIPSC magnitudes originating from the three perisomatic inhibitory cell types, their distinct release properties together with the marked differences in their spiking characteristics may contribute to accomplish specific functions in amygdala network operation.     

Morphological characteristics of layer II projection neurons in the rat medial entorhinal cortex

The entorhinal cortex receives inputs from a variety of neocortical regions. Neurons in layer II of the entorhinal cortex originate one component of the perforant path which conveys this information to the dentate gyrus and hippocampus. The current study extends our previous work on the electroresponsive properties of layer II neurons of the medial entorhinal cortex in which we distinguished two categories of layer II neurons based on their electrophysiological attributes (Alonso and Klink [1993] J Neurophysiol 70:128–143). Here we report on the morphological features of layer II projection neurons, as revealed by in vitro intracellular injection of biocytin. We now report that the two electrophysiologically distinct types of neurons correspond to morphologically distinct types of cells. All neurons (65% of the total cells recorded) that developed sustained, subthreshold, sinusoidal membrane potential oscillations were found to have a stellate appearance. Neurons that did not exhibit oscillatory behavior had either a pyramidal-like (32%) or a horizontal cell morphology (3%). Stellate cells had multiple, thick, primary dendrites. Their widely diverging upper dendritic domain expanded mediolaterally over a distance of around 500 μm close to the pial surface. This mediolateral extent was more than double that of the pyramidal-like cells. Dendrites of stellate cells demonstrated long dendritic appendages, and their dendritic spines had a more complex morphology than those of nonstellates. The stellate cell axons emerged from a primary dendrite and were more than double the thickness (approximately 1.4 μm) of the axons of nonstellate cells. Recurrent axonal collaterization appeared more extensive in axons arising from stellate cells than from pyramidal-like cells. Hippocampus 1997;7:571–583. © 1997 Wiley-Liss, Inc.     

Conserved size and periodicity of pyramidal patches in layer 2 of medial/caudal entorhinal cortex

To understand the structural basis of grid cell activity, we compare medial entorhinal cortex architecture in layer 2 across five mammalian species (Etruscan shrews, mice, rats, Egyptian fruit bats, and humans), bridging ～100 million years of evolutionary diversity. Principal neurons in layer 2 are divided into two distinct cell types, pyramidal and stellate, based on morphology, immunoreactivity, and functional properties. We confirm the existence of patches of calbindin‐positive pyramidal cells across these species, arranged periodically according to analyses techniques like spatial autocorrelation, grid scores, and modifiable areal unit analysis. In rodents, which show sustained theta oscillations in entorhinal cortex, cholinergic innervation targeted calbindin patches. In bats and humans, which only show intermittent entorhinal theta activity, cholinergic innervation avoided calbindin patches. The organization of calbindin‐negative and calbindin‐positive cells showed marked differences in entorhinal subregions of the human brain. Layer 2 of the rodent medial and the human caudal entorhinal cortex were structurally similar in that in both species patches of calbindin‐positive pyramidal cells were superimposed on scattered stellate cells. The number of calbindin‐positive neurons in a patch increased from ～80 in Etruscan shrews to ～800 in humans, only an ～10‐fold over a 20,000‐fold difference in brain size. The relatively constant size of calbindin patches differs from cortical modules such as barrels, which scale with brain size. Thus, selective pressure appears to conserve the distribution of stellate and pyramidal cells, periodic arrangement of calbindin patches, and relatively constant neuron number in calbindin patches in medial/caudal entorhinal cortex. J. Comp. Neurol. 524:783–806, 2016. © 2015 The Authors. The Journal of Comparative Neurology Published by Wiley Periodicals, Inc.     

Size,neurochemistry, and segmental distribution of sensory neurons innervating the rat tibia

Retrograde labeling has been used to identify sensory neurons in the lumbar dorsal root ganglia (DRG) that innervate the rat tibial periosteum, medullary cavity, and trabecular bone. The size, neurochemical profile [isolectin B4 (IB4) binding, substance P (SP), calcitonin gene‐related peptide (CGRP), and NF200 immunoreactivity (‐IR)], and segmental distribution of sensory neurons innervating each of these bony compartments are reported. After injections of fast blue into the periosteum, medullary cavity, and trabecular bone (epiphysis), retrogradely labeled neurons were observed throughout the ipsilateral (but not contralateral) lumbar DRG. They were predominantly small (<800 μm²) or medium‐sized (800–1,800 μm²) neurons. CGRP‐IR and SP‐IR were found in 23% and 16% of the retrogradely labeled neurons, respectively. IB4 binding was observed in 20% and NF200‐IR in 40% of the retrogradely labeled neurons. There were no significant differences in the percentage of neurons labeled with any one of the antisera following injections into each of the three bony compartments. To allow a direct comparison with sensory neurons innervating cutaneous tissues, injections of fast blue were also made into the skin overlying the tibia. The percentage of CGRP‐IR neurons innervating bone was significantly lower than the percentage of CGRP‐IR neurons innervating skin (ANOVA; P < 0.05). No other significant differences in the neurochemical profiles of neurons labeled from bone vs. skin were observed. The findings of the present study show that the periosteum, medullary cavity, and trabecular bone are all innervated by sensory neurons that have size and neurochemical profiles consistent with a role in nociception. J. Comp. Neurol. 517:276–283, 2009. © 2009 Wiley‐Liss, Inc.     

Fast‐spiking interneurons of the rat ventral striatum: temporal coordination of activity with principal cells and responsiveness to reward

Although previous in vitro studies revealed inhibitory synaptic connections of fast‐spiking interneurons to principal cells in the striatum, uncertainty remains about the nature of the behavioural events that correlate with changes in interneuron activity and about the temporal coordination of interneuron firing with spiking of principal cells under natural conditions. Using in vivo tetrode recordings from the ventral striatum in freely moving rats, fast‐spiking neurons were distinguished from putative medium‐sized spiny neurons on the basis of their spike waveforms and rates. Cross‐correlograms of fast‐spiking and putative medium‐sized spiny neuron firing patterns revealed a variety of temporal relationships, including peaks of concurrent firing and transient decrements in medium‐sized spiny neuron spiking around fast‐spiking unit activity. Notably, the onset of these decrements was mostly in advance of the fast‐spiking unit firing. Many of these temporal relationships were dependent on the sleep–wake state. Coordinated activity was also found amongst pairs of the same phenotype, both fast‐spiking units and putative medium‐sized spiny neurons, which was often marked by a broad peak of concurrent firing. When studying fast‐spiking neurons in a reward‐searching task, they generally showed a pre‐reward ramping increment in firing rate but a decrement specifically when the rat received reward. In conclusion, our data indicate that various forms of temporally coordinated activity exist amongst ventral striatal interneurons and principal cells, which cannot be explained by feed‐forward inhibitory circuits alone. Furthermore, firing patterns of ventral striatal fast‐spiking interneurons do not merely correlate with the general arousal state of the animal but display distinct reward‐related changes in firing rate.     

Spatial coding and physiological properties of hippocampal neurons in the Cornu Ammonis subregions

It is well‐established that the feed‐forward connected main hippocampal areas, CA3, CA2, and CA1 work cooperatively during spatial navigation and memory. These areas are similar in terms of the prevalent types of neurons; however, they display different spatial coding and oscillatory dynamics. Understanding the temporal dynamics of these operations requires simultaneous recordings from these regions. However, simultaneous recordings from multiple regions and subregions in behaving animals have become possible only recently. We performed large‐scale silicon probe recordings simultaneously spanning across all layers of CA1, CA2, and CA3 regions in rats during spatial navigation and sleep and compared their behavior‐dependent spiking, oscillatory dynamics and functional connectivity. The accuracy of place cell spatial coding increased progressively from distal to proximal CA1, suddenly dropped in CA2, and increased again from CA3a toward CA3c. These variations can be attributed in part to the different entorhinal inputs to each subregions, and the differences in theta modulation of CA1, CA2, and CA3 neurons. We also found that neurons in the subregions showed differences in theta modulation, phase precession, state‐dependent changes in firing rates and functional connectivity among neurons of these regions. Our results indicate that a combination of intrinsic properties together with distinct intra‐ and extra‐hippocampal inputs may account for the subregion‐specific modulation of spiking dynamics and spatial tuning of neurons during behavior. © 2016 Wiley Periodicals, Inc.     

Presubicular and parasubicular cortical neurons of the rat: Electrophysiological and morphological properties

Intracellular recordings and Neurobiotin-injection were used to examine the electrophysiology and morphology of presubicular and parasubicular cortical neurons in horizontal slices from rat brains. Evoked responses were obtained by stimulation of subicular and entorhinal cortices. Stellate cells were recorded in layers II and V of presubiculum and parasubiculum. Superficial layer cells had spiny dendrites that were found to reach layer I. Deep layer cells had sparsely spiny dendrites or dendrites without spines that did not reach past layer IV. Pyramidal cells were recorded in layers III and V of presubiculum and layers II and V of parasubiculum. Superficial layer cells had spiny dendrites that were found to reach layer I. Deep layer cells had sparsely spiny dendrites or dendrites without spines that could reach layer II. Electrophysiologically, stellate and pyramidal cells were similar to one another, regardless of cell layer, exhibiting repetitive single spiking in response to depolarizing current injection. No cells were found to burst in response to current injection. While there were subtle electrophysiological differences among the cell types, stellate cells were more similar to pyramidal cells from the same or adjacent layers than to other stellate cells from more distant layers. Similarly, pyramidal cells were electrophysiologically more similar to nearby stellate cells than to other distant pyramidal cells. Cells of all layers responded to subicular stimulation with a short latency (<9 ms), excitatory postsynaptic potential. Superficial layer cells responded at short (<9 ms), longer (10–20 ms) and very long latencies (>20 ms) to stimulation of superficial layers of medial entorhinal cortex. Deep layer cells responded at short latencies (<9 ms) to stimulation of deep layers of medial entorhinal cortex. Many cells responded to both subicular and entorhinal inputs. Both pyramidal and stellate cells in the deep layer of pre/parasubiculum could exhibit population bursting behavior in response to stimulation of subiculum or entorhinal cortex. The results define the cellular morphology and basic electrophysiology of presubicular and parasubicular neurons of the rat brain as a step toward understanding the physiology of the retrohippocampal cortices. Hippocampus 7:117–129, 1997. © 1997 Wiley-Liss, Inc.