Costas Papatheodoropoulos

Decreased ability of rat temporal hippocampal CA1 region to produce long-term potentiation

Tetanic stimulation of Schaffer collaterals in the CA1 region of transverse slices, taken from the septal (dorsal) part of young rat hippocampus, produced N-Methyl-D-aspartate-dependent long-term potentiation (LTP) of the rising slope of excitatory postsynaptic potential (mean 38%). Under identical conditions of stimulation (100 Hz, 1 s) slices taken from the temporal (ventral) third of hippocampus presented a substantially reduced ability for LTP (mean 5%). The defect appeared to lie with the induction rather than the maintenance phase of LTP. These results suggest that a significant functional differentiation at the local synaptic plasticity level occurs between the two poles of hippocampus, which together with the substantial differences in their extrinsic connections, may help explain the reported differential participation of neurons in these parts of hippocampus during animal memory tests.

Differential expression of NMDA and AMPA receptor subunits in rat dorsal and ventral hippocampus.

Several studies have demonstrated anatomical and functional segregation along the dorsoventral axis of the hippocampus. This study examined the possible differences in the AMPA and NMDA receptor subunit composition and receptor binding parameters between dorsal and ventral hippocampus, since several evidence suggest diversification of NMDA receptor-dependent processes between the two hippocampal poles. Three sets of rat dorsal and ventral hippocampus slices were prepared: 1) transverse slices for examining a) the expression of the AMPA (GluRA, GluRB, GluRC) and NMDA (NR1, NR2A, NR2B) subunits mRNA using in situ hybridization, b) the protein expression of NR2A and NR2B subunits using Western blotting, and c) by using quantitative autoradiography, c(1)) the specific binding of the AMPA receptor agonist [(3)H]AMPA and c(2)) the specific binding of the NMDA receptor antagonist [(3)H]MK-801, 2) longitudinal slices containing only the cornus ammonis 1 (CA1) region for performing [(3)H]MK-801 saturation experiments and 3) transverse slices for electrophysiological measures of NMDA receptor-mediated excitatory postsynaptic potentials. Ventral compared with dorsal hippocampus showed for NMDA receptors: 1) lower levels of mRNA and protein expression for NR2A and NR2B subunits in CA1 with the ratio of NR2A /NR2B differing between the two poles and 2) lower levels of [(3)H]MK-801 binding in the ventral hippocampus, with the lowest value observed in CA1, apparently resulting from a decreased receptor density since the B(max) value was lower in ventral hippocampus. For the AMPA receptors CA1 our results showed in ventral hippocampus compared with dorsal hippocampus: 1) lower levels of mRNA expression for GluRA, GluRB and GluRC subunits, which were more pronounced in CA1 and in dentate gyrus region and 2) lower levels of [(3)H]AMPA binding. Intracellular recordings obtained from pyramidal neurons in CA1 showed longer NMDA receptor-mediated excitatory postsynaptic potentials in ventral hippocampus compared with dorsal hippocampus. In conclusion, the differences in the subunit mRNA and protein expression of NMDA and AMPA receptors as well as the lower density of their binding sites observed in ventral hippocampus compared with dorsal hippocampus suggest that the glutamatergic function differs between the two hippocampal poles. Consistently, the lower value of the ratio NR2A/NR2B seen in the ventral part would imply that the ventral hippocampus NMDA receptor subtype is functionally different than the dorsal hippocampus subtype, as supported by our intracellular recordings. This could be related to the lower ability of ventral hippocampus for long-term synaptic plasticity and to the higher involvement of the NMDA receptors in the epileptiform discharges, observed in ventral hippocampus compared with dorsal hippocampus.

Dorsal-ventral differentiation of short-term synaptic plasticity in rat CA1 hippocampal region

Two forms of short-term synaptic plasticity (STP), paired-pulse facilitation (PPF) and frequency potentiation (FP) of CA1 field excitatory postsynaptic potentials (EPSP) to afferent stimulation were compared in slices taken from the dorsal and ventral parts of rat hippocampus. While dorsal slices showed significant PPF at all interpulse intervals (20-1400 ms, 80% at 40 ms), PPF in ventral slices was substantially weaker at intervals shorter than 100 ms (19% at 40 ms) and nil at longer intervals. While dorsal slices showed substantial FP at frequencies 1-40 Hz and frequency depression at 50-100 Hz, ventral slices showed only a much smaller potentiation at 1 Hz and substantial depression at 20-100 Hz. Decreasing [Ca(2+)](o) from 2 to 1 and 0.5 mM substantially reduced the baseline EPSPs in both groups of slices but its effect on PPF was greater in ventral slices. On the contrary when [Ca(2+)](o) was increased to 5 mM only dorsal slices showed an enhancement of baseline EPSP. It is concluded that ventral hippocampus CA1 area has a specific deficit in STP, which is related to the important presynaptic role of calcium and is consistent with a relatively higher transmitter release probability.

Enhanced neuronal plasticity and elevated endogenous sAPPα levels in mice over-expressing MMP9

J. Neurochem. (2012) 121, 239–251.

Spontaneous GABAA-dependent synchronous periodic activity in adult rat ventral hippocampal slices

The present study shows that adult rat transverse slices from the ventral hippocampus perfused with standard medium persistently generate spontaneous synchronous field potentials. In CA1 st. pyramidale this regular ventral hippocampus spontaneous synchronous activity (VHSSA) was positive with mean amplitude 0.18 +/- 0.02 mV (n=80 slices) and occurred every 0.48 +/- 0.02 s. Simultaneous intracellular recordings from CA1 pyramidal neurons demonstrated that concomitant hyperpolarizations invariably occurred in association to this field activity and could thus constitute its electrical generators. These hyperpolarizations, had mean amplitude 2.7 +/- 0.6 mV, duration at half amplitude 44.8 +/- 6.6 ms, they reversed at -72.6 +/- 1.5 mV (n=10 cells), they effectively suspended the depolarization-induced tonic neuronal firing of all ten pyramidal neurons and they were reversibly abolished, together with field potentials, by the GABA(A) receptor antagonist bicuculline (5 microM, n=4). VHSSA was also dependent on fast glutamatergic transmission, since it was blocked by the antagonist of AMPA/kainate receptors 6-Cyano-7-nitroquinoxaline-2,3-dione disodium (10 microM, n=3). We propose that, under standard in vitro conditions, synchronous GABA(A)-mediated hyperpolarizing potentials are spontaneously generated in pyramidal neurons presumably resulting from the phasic quasi-rhythmic discharge of a local interneuronal network of ventral hippocampus.

Differential expression of γ-aminobutyric acid-A receptor subunits in rat dorsal and ventral hippocampus

Recent data demonstrate weaker γ‐aminobutyric acid (GABA)‐ergic inhibition in ventral (VH) compared with dorsal (DH) hippocampus. Therefore, we examined possible differences regarding the GABAA receptors between VH and DH as follows: 1) the expression of the GABAA receptor subunits (α1/2/4/5, β1/2/3, γ2, δ) mRNA and protein and 2) the quantitative distribution and kinetic parameters of [3H]muscimol (GABAA receptor agonist) binding. VH compared with DH showed: 1) lower levels for α1, β2, γ2 but higher levels for α2 and β1 subunits in CA1, CA2, and CA3, the differences being more pronounced in CA1 region; in the CA1 region, the mRNA levels of α5 were higher, whereas those of α4 subunit were slightly lower; in dentate gyrus, the mRNA levels of α4, β3, and δ subunits were significantly lower, presumably suggesting a lower expression of the α4/β3/δ receptor subtype; and 2) lower levels of [3H]muscimol binding, with the lowest value observed in CA1, apparently resulting from weaker binding affinity, insofar as the KD values were higher in VH, whereas the Bmax values were similar between DH and VH. The differences in the subunit expression and the lower affinity of GABAA receptor binding observed predominantly in the CA1 region of VH suggest that the α1/β2/γ2 GABAA receptor subtype dominates in DH, and the α2/β1/γ2 subtype prevails in VH. This could underlie the lower GABAA‐mediated inhibition observed in VH and, to some extent, explain 1) the higher liability of VH for epileptic activity and 2) the differential involvement of DH and VH in cognitive and emotional processes.

Spontaneous, low frequency (∼2–3 Hz) field activity generated in rat ventral hippocampal slices perfused with normal medium

Abstract This study demonstrates that transverse slices taken from the ventral hippocampus of adult rats perfused with a medium of normal ionic composition sustain spontaneous periodic field potentials due to the synchronous activity of a population of neurons. This ventral hippocampus spontaneous synchronous activity (VHSSA) in CA1 stratum pyramidale consisted of positive potentials (∼0.12 mV, 55 ms) occurring at a frequency of 2.8 ± 0.2 Hz for hours without interruption. VHSSA was most frequently observed in slices taken 1–3 mm from the ventral end of hippocampus, and was absent in slices taken from tissue more than 4.5 mm away from it. Stimulation of Schaffer collaterals primed the appearance of potentials, which were similar to VHSSA and clearly distinguishable from excitatory postsynaptic potentials. In view of the known relative proneness of ventral hippocampus to epilepsy, we perfused ventral slices with high-[K + ] o medium (8 mM). Albeit reduced in amplitude, VHSSA persisted during the high-[K + ] o induced interictal-like epileptiform activity. We could not document any temporal relationship between the two phenomena. Low concentrations of the antagonist of γ-amino-butyric acid receptors, type A, bicuculline (2–3 μM), which enhanced the high-[K + ] o induced epileptiform activity, reversibly blocked the VHSSA. We conclude that under standard in vitro conditions small circuits in the ventral hippocampus are most often and for long periods of time engaged in synchronous quasi-rhythmic low-frequency activity, generated locally by mechanisms substantially differing from those supporting epileptiform discharges.

Weaker synaptic inhibition in CA1 region of ventral compared to dorsal rat hippocampal slices.

Extracellular and intracellular recordings were made from slices taken from the dorsal (DH) and ventral (VH) part of rat hippocampus. Using paired-pulse stimulation of Schaffer collaterals, at different interpulse intervals (IPIs), and records of the population spike (PS) we found that the strength and duration of paired-pulse inhibition was much weaker in VH compared to DH slices: at the IPI of 10 ms the decrease of PS in VH (40%) was significantly smaller compared to that in DH slices (76%), while at 20 ms the decrease of PS in DH slices (60%) corresponded to facilitation in VH slices. Moreover, the amplitude and duration of intracellularly recorded fast inhibitory postsynaptic potentials (fast-IPSPs) were found significantly smaller in VH (5.2+/-0.6 mV, 54.8+/-5.8 ms) than in DH (11.2+/-1.1 mV, 105+/-10 ms) neurons. The smaller and shorter fast-IPSP recorded in VH neurons may at least in part explain the results in paired-pulse inhibition. The demonstrated weaker inhibition may underlie the higher propensity of the ventral hippocampus for epileptiform activity.

Age-related Changes in Excitability and Recurrent Inhibition in the Rat CA1 Hippocampal Region

In hippocampal slices from male Wistar rats aged 1–34 months, we recorded the synaptic field potential responses of the CA1 neurons to stimulation of Schaffer collaterals. Eight electrophysiological indexes were extracted from input/output curves and compared in 11 age groups from 1 to 30 months. Neuronal excitability presented a U‐shaped curve of development with a minimum at ˜7–8 months of age. There was a significant continuous increase in neuronal excitability, i.e. a decrease in excitatory postsynaptic potential (EPSP) producing both the threshold and half‐maximal population spike from middle age (8–10 months) to senescence (30 months). Synaptic efficiency also increased in old rats to reach a maximum during senescence, i.e. both the current for threshold EPSP and that for half‐maximal EPSP reached a minimum in senescence, although the earlier developmental patterns of these two indexes were non‐linear. The duration of the field EPSP elicited with maximal stimulation presented an abrupt decay after the first month. Aged animals presented a relatively small maximal population spike. Recurrent inhibition was most prominent on neuronal excitability rather than synaptic strength. Measured as the percentage change in the half‐maximal EPSP and half‐maximal population spike, recurrent inhibition was found to decrease during the first 7–10 months of life and remained small in later development.

GABA-mediated synchronization in the human neocortex: elevations in extracellular potassium and presynaptic mechanisms

Field potential and extracellular [K(+)] ([K(+)](o)) recordings were made in the human neocortex in an in vitro slice preparation to study the synchronous activity that occurs in the presence of 4-aminopyridine (50 microM) and ionotropic excitatory amino acid receptor antagonists. Under these experimental conditions, negative or negative-positive field potentials accompanied by rises in [K(+)](o) (up to 4.1 mM from a baseline of 3.25 mM) occurred spontaneously at intervals of 3-27 s. Both field potentials and [K(+)](o) elevations were largest at approximately 1000 microm from the pia. Similar events were induced by neocortical electrical stimuli. Application of medium containing low [Ca(2+)]/high [Mg(2+)] (n=3 slices), antagonism of the GABA(A) receptor (n=7) or mu-opioid receptor activation (n=4) abolished these events. Hence, they represented network, GABA-mediated potentials mainly reflecting the activation of type A receptors following GABA release from interneurons. The GABA(B) receptor agonist baclofen (10-100 microM, n=11) reduced and abolished the GABA-mediated potentials (ID(50)=18 microM). Baclofen effects were antagonized by the GABA(B) receptor antagonist CGP 35348 (0.1-1 mM, n=6; ID(50)=0.19 mM). CGP 38345 application to control medium increased the amplitude of the GABA-mediated potentials and the concomitant [K(+)](o) rises without modifying their rate of occurrence. The GABA-mediated potentials were not influenced by the broad-spectrum metabotropic glutamate agonist (+/-)-1-aminocyclopentane-trans-1,3-dicarboxylic acid (100 microM, n=10), but decreased in rate with the group I receptor agonist (S)-3,5-dihydroxyphenylglycine (10-100 microM, n=9). Our data indicate that human neocortical networks challenged with 4-aminopyridine generate glutamatergic-independent, GABA-mediated potentials that are modulated by mu-opioid and GABA(B) receptors presumably located on interneuron terminals. These events are associated with [K(+)](o) elevations that may contribute to interneuron synchronization in the absence of ionotropic excitatory synaptic transmission.