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Activation of dentate hilar neurons by stimulation of the fimbria in rat hippocampal slices

Scharfman, H E
It is has been shown that the major afferent input to the dentate gyrus, the perforant path, excites dentate hilar neurons. However, little is known about the other inputs to hilar cells. Therefore, we examined the responses of hilar neurons to stimulation of the fimbria. We positioned our stimulating electrodes so that granule cells were not excited antidromically by fimbria stimulation, although action potentials were easily triggered in area CA3b and CA3c pyramidal cells by such stimulation. In these experiments, fimbria stimulation evoked responses from every hilar cell tested, including examples of both of the major cell types, the spiny hilar 'mossy' cells (n = 15) and the relatively aspiny, 'fast-spiking' cells (putative interneurons, n = 5). Hilar cell responses consisted primarily of EPSPs that could trigger action potentials, but small IPSPs were also evoked in some cases, particularly in the fast-spiking cells. Excitation was blocked by an antagonist of the AMPA/kainate receptor subtype of excitatory amino acid receptors, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 5 microM, n = 5), whereas the cholinergic antagonist atropine (10 microM) had no effect (n = 4). When sequential intracellular recordings were made from hilar cells and area CA3 pyramidal cells in the same slice, hilar cell EPSPs began after action potentials of CA3b pyramidal cells, and stimulus strengths required to evoke hilar cell EPSPs were above threshold for area CA3b pyramidal cells.(ABSTRACT TRUNCATED AT 250 WORDS)
PMCID:3281807
PMID: 8105429
ISSN: 0304-3940
CID: 73504

Electron microscopy of intracellularly labeled neurons in the hippocampal slice preparation

Kunkel, D D; Scharfman, H E; Schmiege, D L; Schwartzkroin, P A
We have assessed the properties of three intracellular markers, horseradish peroxidase, biocytin/Neurobiotin, and Lucifer Yellow, and have compared their usefulness as neuronal markers for light and electron microscopic visualization. Neurons in the acute slice preparation of rat hippocampus were filled with one of these markers, and the marker was converted to an optical and electron-dense reaction product. Dimethylsulfoxide (DMSO) greatly facilitated penetration of recognition reagents while preserving membrane integrity. The markers were compared with respect to injection parameters, mobility and recognition, stability and visibility, and ultrastructural clarity. Horseradish peroxidase (HRP)-labeled neurons, recognized histochemically with diaminobenzedine (DAB), were easily visualized by the density of the DAB reaction product; however, the electron density was often so great as to obscure ultrastructural details. Biocytin (BC)-/Neurobiotin (NB)-labeled neurons were recognized by avidin-HRP, followed by histochemical localization of HRP with DAB. The optically dense reaction product gave complete visualization of the soma and processes at the light microscopic level. The electron density was homogeneously distributed throughout the cell, so that ultrastructural features were easily identified. Lucifer Yellow (LY), a fluorescent marker, was converted to an optical and electron-dense reaction product via immunocytochemical staining with a rabbit anti-LY antibody, followed by goat anti-rabbit IgG-HRP and DAB histochemical localization. Similar to BC/NB, the reaction product was evenly dispersed, providing good light microscopic and ultrastructural clarity. Under our experimental conditions, BC/NB and LY were superior markers that could be used routinely to label neurons, and give excellent visualization not only at the light but also at the electron microscopic level
PMID: 7679592
ISSN: 1059-910x
CID: 73498

Spiny neurons of area CA3c in rat hippocampal slices have similar electrophysiological characteristics and synaptic responses despite morphological variation

Scharfman, H E
Area CA3c is an area of morphologically diverse neurons. In addition to the presence of interneurons and pyramidal cells that are similar to those found in other subfields of area CA3, many neurons of area CA3c are different. They do not resemble interneurons, since they bear numerous spines, yet they also differ substantially from pyramidal cells in their morphology. To determine if the variants of area CA3c spiny cells are distinct physiologically as well as morphologically, intracellular recordings were made to record the electrophysiological properties of area CA3c cells in rat hippocampal slices, and each cell was identified morphologically following intracellular dye injection. The results show that the spiny cells, regardless of their often extensive morphological variation, have relatively uniform, pyramidal-like electrophysiological properties. The aspiny cells are quite different from the spiny cells morphologically (i.e., in their paucity or complete lack of spines), and are also extremely different electrophysiologically, exhibiting features of 'fast-spiking' cells. Thus, spiny cells in area CA3c correspond to cells with pyramidal-like electrophysiology, and the aspiny cells in area CA3c correspond to cells with interneuronal physiological properties. This correlation between structure and function appears to be a rule that pertains to each of the subfields of the hippocampus
PMID: 8364685
ISSN: 1050-9631
CID: 73506

Blockade of excitation reveals inhibition of dentate spiny hilar neurons recorded in rat hippocampal slices

Scharfman, H E
1. Extracellular and intracellular recordings in rat hippocampal slices were used to compare the synaptic responses to perforant path stimulation of granule cells of the dentate gyrus, spiny 'mossy' cells of the hilus, and area CA3c pyramidal cells of hippocampus. Specifically, we asked whether aspects of the local circuitry could explain the relative vulnerability of spiny hilar neurons to various insults to the hippocampus. 2. Spiny hilar cells demonstrated a surprising lack of inhibition after perforant path activation, despite robust paired-pulse inhibition and inhibitory postsynaptic potentials (IPSPs) in adjacent granule cells and area CA3c pyramidal cells in response to the same stimulus in the same slice. However, when the slice was perfused with excitatory amino acid antagonists [6-cyano-7-nitro-quinoxaline-2,3-dione (CNQX), or CNQX with 2-amino-5-phosphonovaleric acid (APV)], IPSPs could be observed in spiny hilar cells in response to perforant path stimulation. 3. The IPSPs evoked in spiny hilar cells in the presence of CNQX were similar in their reversal potentials and bicuculline sensitivity to IPSPs recorded in dentate granule cells or hippocampal pyramidal cells in the absence of CNQX. 4. These results demonstrate that, at least in slices, perforant path stimulation of spiny hilar cells is primarily excitatory and, when excitation is blocked, underlying inhibition can be revealed. This contrasts to the situation for dentate and hippocampal principal cells, which are ordinarily dominated by inhibition, and only when inhibition is compromised can the full extent of excitation be appreciated.(ABSTRACT TRUNCATED AT 250 WORDS)
PMCID:3286008
PMID: 1359025
ISSN: 0022-3077
CID: 73436

Differentiation of rat dentate neurons by morphology and electrophysiology in hippocampal slices: granule cells, spiny hilar cells and aspiny 'fast-spiking' cells

Scharfman, H E
Intracellular recording and intracellular dye injection of single cells in the dentate region of rat hippocampal slices have been used to understand the different types of cells in the dentate and their possible functional organization. On the basis of combined electrophysiological and morphological data, the cells that have been sampled fall into three distinct groups: the granule cells, the spiny cells located in the hilus (the 'mossy' cell being the prototype), and the aspiny, 'fast-spiking' cells located throughout the region (many of which are likely to be GABAergic interneurons). Although there is some variability within each group, this variability is minor compared to the large differences between groups. To clarify these groups, each one is described first morphologically, at the level of the light microscope and histochemically, and then the three groups are described electrophysiologically, in terms of intrinsic electrophysiological characteristics, synaptic responses to perforant path stimulation, and possible roles in dentate circuitry. It is proposed that this apparent organization of neurons into three major classes be used as a starting point in our evolving understanding of the functional organization of the dentate region, and, in particular, the hilus. In addition, the possibility is raised that area CA3c cells of the hippocampus could be included in the dentate region as a fourth group. Together with the hilar cells, area CA3c could have the obviously important role of integrating the dentate circuitry with that of the hippocampus proper
PMCID:3281805
PMID: 1361334
ISSN: 0922-9833
CID: 73437

Dentate hilar cells with dendrites in the molecular layer have lower thresholds for synaptic activation by perforant path than granule cells

Scharfman, H E
Neurons in the dentate hilus or area CA3c of rat hippocampal slices were recorded intracellularly with electrodes containing the fluorescent dye Lucifer yellow. Stimulation of perforant path fibers in the molecular layer of the fascia dentata strongly excited most hilar neurons, with a much lower threshold for action potential generation than granule cells and area CA3c pyramidal cells that were recorded in the same area of the slice. Examination of dye-filled hilar neurons with a confocal microscope showed that hilar cells with a low threshold were morphologically heterogeneous: some were spiny 'mossy' cells, and others were aspiny interneurons. However, all hilar cells with low thresholds possessed dendrites that penetrated the granule cell layer and passed into the molecular layer, often reaching the outer molecular layer. The few hilar cells that had a threshold similar to, or greater than, granule cells did not possess visible dendrites in the molecular layer. The results suggest that the circuitry of the dentate region allows for (1) excitation of both granule cells and hilar cells by perforant path stimuli, and (2) strong excitation of most hilar cells when most granule cells are subthreshold for action potential generation. Given that hilar neurons project to many different sites in the ipsilateral and contralateral fascia dentata (Blackstad, 1956; Zimmer, 1971; Swanson et al., 1978; Laurberg and Sorensen, 1981), it is quite likely that hilar neurons are involved in the processing of information passing from entorhinal cortex to hippocampus
PMID: 2045880
ISSN: 0270-6474
CID: 73479

Comparisons of neocortex and hippocampus [Letter]

Scharfman, H E
PMID: 1699324
ISSN: 0166-2236
CID: 73466

N-methyl-D-aspartate receptors contribute to excitatory postsynaptic potentials of cat lateral geniculate neurons recorded in thalamic slices

Scharfman, H E; Lu, S M; Guido, W; Adams, P R; Sherman, S M
Neurons of the cat's dorsal lateral geniculate nucleus were recorded intracellularly to study the contribution of N-methyl-D-aspartate (NMDA) receptors to excitatory postsynaptic potentials (EPSPs) and low-threshold calcium spikes. EPSPs were evoked by stimulation of retinogeniculate axons in the optic tract and/or corticogeniculate axons in the optic radiations; EPSPs from both sources were similar. These EPSPs had one or two components, and the second component had several characteristics of NMDA receptor-mediated events. For example, EPSP amplitude decreased when neurons were hyperpolarized and increased when stimulus frequency was increased; these EPSPs could also be blocked reversibly by application of the selective NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (APV). We also studied the influence of NMDA receptors on low-threshold calcium spikes, which are large, voltage- and calcium-dependent depolarizations that are often accompanied by high-frequency action potential discharge. APV blocked synaptically activated low-threshold calcium spikes, but APV had no effect on low-threshold calcium spikes that were elicited by current injection. Therefore, APV does not appear to have a direct effect on the T-type calcium channel that is involved in generation of low-threshold calcium spikes. The voltage and frequency dependence of the NMDA receptor-mediated component of the EPSPs, as well as its ability to trigger low-threshold calcium spikes, provide for complex signal processing in the lateral geniculate nucleus
PMCID:54153
PMID: 1972275
ISSN: 0027-8424
CID: 73477

Similarities in circuitry between Ammon's horn and dentate gyrus: local interactions and parallel processing

Schwartzkroin, P A; Scharfman, H E; Sloviter, R S
We present a 'model' of hippocampal information processing based on a review of recent data regarding the local circuitry of Ammon's horn and the dentate gyrus. We have been struck by the parallels in cell type and connectivity in Ammon's horn and the dentate gyrus, and have focused on similarities between CA3 pyramidal cells and mossy cells. Important conclusions of our analysis include the following: (1) The idea of serial processing of afferent information, from one hippocampal subregion to the next, is inadequate and based on an over-simplification of circuitry; information processing undoubtedly occurs over parallel, as well as serial, pathways. (2) Local circuitry within a given hippocampal subregion gives rise predominantly to feedforward inhibition; recurrent inhibition is present, but less potent. (3) There are multiple populations of local circuit neurons, each of which has a specific function, characteristic interconnections, and special cell properties. It is misleading to categorize these cells into a single category of inhibitory interneuron
PMID: 1975454
ISSN: 0079-6123
CID: 73478

Synaptic connections of dentate granule cells and hilar neurons: results of paired intracellular recordings and intracellular horseradish peroxidase injections

Scharfman, H E; Kunkel, D D; Schwartzkroin, P A
Simultaneous intracellular recordings were made in the dentate gyrus of rat hippocampal slices, from pairs of the following cell types: granule cells, interneurons located in the granule cell layer, hilar interneurons, and spiny hilar 'mossy cells'. Granule cells were found to have strong excitatory effects on mossy cells and interneurons. Interneurons inhibited granule cells as well as other interneurons. No synaptic connections from mossy cells onto other cell types were found, within the confines of the slice, using intracellular recording methods. However, at the ultrastructural level, axon terminals of horseradish peroxidase-filled mossy cells were found making synaptic contacts in the hilus on dendrites of interneurons. These studies provide the first step towards determining the functional interactions of the various cell types in the fascia dentata
PMID: 2247219
ISSN: 0306-4522
CID: 73480