Layers Of Rat Entorhinal Cortex Research Articles

High-voltage-activated Ca 2+ currents show similar patterns of expression in stellate and pyramidal cells from rat entorhinal cortex layer II

High-voltage-activated (HVA) Ca 2+ currents were studied in acutely isolated neurons from rat entorhinal cortex (EC) layer II. Stellate and pyramidal cells, the two main neuronal types of this structure, were visually identified based on morphological criteria. HVA currents were recorded by applying the whole-cell, patch-clamp technique, using 5-mM Ba 2+ as the charge carrier. In both neuronal types, the amplitude of total HVA Ba 2+ currents ( I Bas) showed a significant tendency to increase with postnatal age in the time window considered [postnatal day 15 (P15) to P28–29]. At P20–P29, when I Ba expression reached stable levels, I Ba density per unit of membrane area was not different in stellate versus pyramidal cells. The same was also observed when Ca 2+, instead of Ba 2+, was used as the charge carrier. The pharmacological current subtypes composing total HVA currents were characterized using selective blockers. Again, no significant differences were found between stellate and pyramidal cells with respect to the total–current fractions attributable to specific pharmacological Ca 2+ channel subtypes. In both cell types, ∼52–55% of total I Bas was abolished by the L-type channel blocker, nifedipine (10 μM), ∼23–30% by the N-type channel blocker, ω-conotoxin GVIA (1 μM), ∼22–24% by the P/Q-type channel blocker, ω-agatoxin IVA (100 nM), and ∼11–13% remained unblocked (R-type current) after simultaneous application of L-, N-, and P/Q-type channel blockers. The Ca v 2.3 (α 1E) channel blocker, SNX-482 (100 nM), abolished ∼57–62% of total R-type current. We conclude that HVA Ca 2+ currents are expressed according to similar patterns in the somata and proximal dendrites of stellate and pyramidal cells of rat EC layer II.

Dynamics of rat entorhinal cortex layer II and III cells: characteristics of membrane potential resonance at rest predict oscillation properties near threshold.

Neurones generate intrinsic subthreshold membrane potential oscillations (MPOs) under various physiological and behavioural conditions. These oscillations influence neural responses and coding properties on many levels. On the single-cell level, MPOs modulate the temporal precision of action potentials; they also have a pronounced impact on large-scale cortical activity. Recent studies have described a close association between the MPOs of a given neurone and its electrical resonance properties. Using intracellular sharp microelectrode recordings we examine both dynamical characteristics in layers II and III of the entorhinal cortex (EC). Our data from EC layer II stellate cells show strong membrane potential resonances and oscillations, both in the range of 5-15 Hz. At the resonance maximum, the membrane impedance can be more than twice as large as the input resistance. In EC layer III cells, MPOs could not be elicited, and frequency-resolved impedances decay monotonically with increasing frequency or has only a small peak followed by a subsequent decay. To quantify and compare the resonance and oscillation properties, we use a simple mathematical model that includes stochastic components to capture channel noise. Based on this model we demonstrate that electrical resonance is closely related though not equivalent to the occurrence of sag-potentials and MPOs. MPO frequencies can be predicted from the membrane impedance curve for stellate cells. The model also explains the broad-band nature of the observed MPOs. This underscores the importance of intrinsic noise sources for subthreshold phenomena and rules out a deterministic description of MPOs. In addition, our results show that the two identified cell classes in the superficial EC layers, which are known to target different areas in the hippocampus, also have different preferred frequency ranges and dynamic characteristics. Intrinsic cell properties may thus play a major role for the frequency-dependent information flow in the hippocampal formation.

Long-term synaptic plasticity in deep layer-originated associational projections to superficial layers of rat entorhinal cortex

Superficial layers of the entorhinal cortex (EC) relay the majority of cortical input projections to the hippocampus, whereas deep layers of the EC mediate a large portion of hippocampal output projections back to other cortical areas, suggesting a functional segregation between superficial and deep layers of the EC as input and output structures of the hippocampus, respectively. However, deep layers of the EC send associational projections to superficial layers, suggesting a potential interaction between neocortical input and hippocampus-processed output in superficial layers. This possibility was investigated by examining whether deep to superficial EC projections support long-term synaptic plasticity, and whether they interact with other pathways in superficial layers in rat medial EC slice preparations. Synaptic responses of the deep-to-superficial layer projections were verified based on field potential profiles, paired-pulse facilitation, physical separation between superficial and deep layers, and pharmacological manipulation. Long-term potentiation (LTP) was reliably induced in the deep-to-superficial layer projections by burst stimulations that emulated θ or sharp wave electroencephalogram (EEG),and it was blocked by an N-methyl- d-aspartate receptor antagonist ( d-2-amino-5-phosphonopentanoic acid) and a calcium channel blocker (nifedipine). Prolonged low frequency stimulation induced long-term depression. A weak stimulation of deep layers, which induced a small degree of LTP by itself, generated a much larger degree of LTP when paired with a strong stimulation of superficial layers, indicating that the deep-to-superficial layer projections cooperate with other pathways in the superficial EC to enhance synaptic weights. Our results suggest that neocortical input and hippocampal output information are integrated in superficial layers of the EC.

Increased Persistent Sodium Currents in Rat Entorhinal Cortex Layer V Neurons in a Post–Status Epilepticus Model of Temporal Lobe Epilepsy

Spontaneous seizures in rats emerge several weeks after induction of status epilepticus with pharmacologic treatment or electrical stimulation, providing an animal model for human temporal lobe epilepsy. In this study, we investigated whether status epilepticus caused changes in the function of voltage-gated sodium channels in entorhinal cortex layer V neurons, a cellular group important for the genesis of limbic seizures. We induced status epilepticus in rats, by using lithium-pilocarpine, and then 2-12 weeks later, used whole-cell voltage-clamp to examine voltage-activated sodium currents of acutely dissociated layer V neurons. Transient sodium currents of entorhinal cortex layer V neurons isolated from 9- to 12-week post-status epilepticus rats were similar to currents in age-matched controls; however, low-threshold persistent sodium currents were significantly larger. This increase in persistent activity was not seen 2-3 weeks after pilocarpine treatment; thus it occurred after a delay comparable to the delay in the appearance of spontaneous seizures. Increased persistent currents are expected to accentuate neuronal excitability and thus may contribute to the genesis of spontaneous seizures after status epilepticus.

Variation in Effective Stimulus Patterns for Induction of Long-Term Potentiation Across Different Layers of Rat Entorhinal Cortex

Neuronal activities in superficial (II and III) and deep (V and VI) layers of the entorhinal cortex (EC) are preferentially modulated by theta and sharp wave (SPW) EEG, respectively. We investigated the possibility that distinct EEG patterns represent optimal stimulus patterns for induction of long-term potentiation (LTP) in different layers of the EC. We examined effects of three different stimulation patterns on LTP induction in layers I, II-III, and V of medial EC slices of the rat. The stimulation patterns we used were a single, long high-frequency train (1 sec at 100 Hz, repeated 3x), theta burst stimulation [TBS; 10 bursts (four pulses, 100 Hz) at 5 Hz, repeated 3x], and SPW-like burst stimulation [ SPWBS; three bursts (20 pulses, 200 Hz) at 2 Hz, repeated 6x]. Similar degrees of LTP were induced by the three stimulation patterns in layer I. In layers II--III and layer V, however, the largest degrees of LTP were induced by TBS and SPWBS, respectively. These results suggest that burst stimulation constructed to mimic naturally occurring patterns of neuronal activity in the corresponding layer is optimal for LTP induction in layers II-III and layer V of the EC. The differences may play important roles in shaping hippocampal-neocortical interactions in encoding and retrieval of memory.

Activation of presynaptic group III metabotropic glutamate receptors depresses spontaneous inhibition in layer V of the rat entorhinal cortex

Whole cell voltage clamp recording was used to investigate neurotransmitter release onto neurones in deep and superficial layers of rat entorhinal cortex in vitro. Activation of metabotropic glutamate receptors with the agonist (1 S,3 R,4 S)-1-aminocyclopentane-1,2,4-tricarboxylic acid depressed spontaneous release of the inhibitory neurotransmitter GABA in layer V, but not in layer II. Depression of transmitter release did not persist in the presence of the sodium channel blocker tetrodotoxin. It seems likely that activation of presynaptic glutamate heteroreceptors inhibits action potential dependent release of neurotransmitter via a direct action at the presynaptic terminal. We confirmed that depression of inhibitory neurotransmission in layer V was mediated by group III metabotropic glutamate receptors using a specific group III antagonist, ( RS)-cyclopropyl-4-phosphonophenylglycine. Application of the antagonist alone did not alter the frequency of spontaneous neurotransmitter release, suggesting that the metabotropic glutamate receptor is not tonically active. In layer V of the entorhinal cortex, activation of presynaptic metabotropic glutamate receptors enhances spontaneous glutamate release, and inhibits spontaneous release of GABA. These effects may combine to increase random action potential firing in this layer, thereby reducing its capacity for synchrony generation. Our results are consistent with an anticonvulsant action for group III metabotropic glutamate receptors in the entorhinal cortex.

Persistent sodium channel activity mediates subthreshold membrane potential oscillations and low-threshold spikes in rat entorhinal cortex layer V neurons

Entorhinal cortex layer V occupies a critical position in temporal lobe circuitry since, on the one hand, it serves as the main conduit for the flow of information out of the hippocampal formation back to the neocortex and, on the other, it closes a hippocampal–entorhinal loop by projecting upon the superficial cell layers that give rise to the perforant path. Recent in vitro electrophysiological studies have shown that rat entorhinal cortex layer V cells are endowed with the ability to generate subthreshold oscillations and all-or-none, low-threshold depolarizing potentials. In the present study, by applying current-clamp, voltage-clamp and single-channel recording techniques in rat slices and dissociated neurons, we investigated whether entorhinal cortex layer V cells express a persistent sodium current and sustained sodium channel activity to evaluate the contribution of this activity to the subthreshold behavior of the cells. Sharp-electrode recording in slices demonstrated that layer V cells display tetrodotoxin-sensitive inward rectification in the depolarizing direction, suggesting that a persistent sodium current is present in the cells. Subthreshold oscillations and low-threshold regenerative events were also abolished by tetrodotoxin, suggesting that their generation also requires the activation of such a low-threshold sodium current. The presence of a persistent sodium current was confirmed in whole-cell voltage-clamp experiments, which revealed that its activation “threshold” was negative by about 10 mV to that of the transient sodium current. Furthermore, stationary noise analysis and cell-attached, patch-clamp recordings indicated that whole-cell persistent sodium currents were mediated by persistent sodium channel activity, consisting of relatively high-conductance (∼18 pS) sustained openings. The presence of a persistent sodium current in entorhinal cortex layer V cells can cause the generation of oscillatory behavior, bursting activity and sustained discharge; this might be implicated in the encoding of memories in which the entorhinal cortex participates but, under pathological situations, may also contribute to epileptogenesis and neurodegeneration.

Slow voltage-dependent inactivation of a sustained sodium current in stellate cells of rat entorhinal cortex layer II.

Slow voltage-dependent inactivation of a sustained sodium current in stellate cells of rat entorhinal cortex layer II.

NMDA receptor 2C subunit is selectively decreased by MK-801 in the entorhinal cortex

Administration of the non-competitive NMDA receptor antagonist MK-801 (5.methyl-10,11-dihydro-5 H-dibenzo[1, d]cyclohepten-5-10-imine) produces paradoxicl neurotoxicity in limbic cortical regions which incluide the entorhinal cortex. The expression of NMDAR-2C but not -2A, -2B, or -2D subunits was significantly decreased in rat entorhinal cortex layer III following MK-801 administration. These results suggest an important role for the NMDAR-2C subunit in the response to MK-801-induced neurotoxicity in brain regions highly vulnerable to injury.