Faculty Bibliography

How do ensembles of neurons distributed across the hippocampal and entorhinal cortices effectively interact? In the awake-behaving rat, specific subpopulations of hippocampal and entorhinal neurons become entrained into two prominent fast-frequency rhythms (gamma [40-100 Hz], and 200 Hz). These fast rhythms are coupled to slower synchronizing potentials (theta and sharp wave, respectively), are correlated to macroscopic behavioral states, and to some extent are anatomically distinct. These population dynamics allow distributed populations of neurons across the hippocampal and entorhinal cortices to discharge together in time on the order of tens of milliseconds, and thus allow interconnected domains of a distributed neural network to become transiently entraining into synchronized, fast-frequency, population ensembles. We believe that these transient population dynamics allow interconnected domains to 'effectively communicate' and modify their synaptic connectivity

Dendritic morphology and passive cable properties determine many aspects of synaptic integration in complex neurons, together with voltage-dependent membrane conductances. We investigated dendritic properties of CA1 pyramidal neurons intracellularly labeled during in vivo and in vitro physiologic recordings, by using similar intracellular staining and three-dimensional reconstruction techniques. Total dendritic length of the in vivo neurons was similar to that of the in vitro cells. After correction for shrinkage, cell extent in three-dimensional representation was not different between the two groups. Both in vivo and in vitro neurons demonstrated a variable degree of symmetry, with some neurons showing more cylindrical symmetry around the main apical axis, whereas other neurons were more elliptical, with the variation likely due to preparation and preservation conditions. Branch order analysis revealed no difference in the number of branch orders or dendritic complexity. Passive conduction of dendritic signals to the soma in these neurons shows considerable attenuation, particularly with higher frequency signals (such as synaptic potentials compared with steady-state signals), despite a relatively short electrotonic length. Essential aspects of morphometric appearance and complex dendritic integration critical to CA1 pyramidal cell functioning are preserved across neurons defined from the two different hippocampal preparations used in this study

Gamma frequency field oscillations reflect synchronized synaptic potentials in neuronal populations within the approximately 10-40 ms range. The generation of gamma activity in the hippocampus was investigated by intracellular recording from principal cells and basket cells in urethane anaesthetized rats. The recorded neurones were verified by intracellular injection of biocytin. Gamma frequency field oscillations were nested within the slower theta waves. The phase and amplitude of intracellular gamma were voltage dependent with an almost complete phase reversal at Cl- equilibrium potential in pyramidal cells. Basket cells fired at gamma frequency and were phase-locked to the same phase of the gamma oscillation as pyramidal cells. Current-induced depolarization coupled with synaptically induced inhibition resulted in gamma frequency discharge (30-80 Hz) of pyramidal cells without accommodation. These observations suggest that at least part of the gamma frequency field oscillation reflects rhythmic hyperpolarization of principal cells, brought about by the rhythmically discharging basket neurones. Resonant properties of pyramidal cells might facilitate network synchrony in the gamma frequency range

Gamma frequency field oscillations (40-100 Hz) are nested within theta oscillations in the dentate-hilar and CA1-CA3 regions of the hippocampus during exploratory behaviors. These oscillations reflect synchronized synaptic potentials that entrain the discharge of neuronal populations within the approximately 10-25 msec range. Using multisite recordings in freely behaving rats, we examined gamma oscillations within the superficial layers (I-III) of the entorhinal cortex. These oscillations increased in amplitude and regularity in association with entorhinal theta waves. Gamma waves showed an amplitude minimum and reversed in phase near the perisomatic region of layer II, indicating that they represent synchronized synaptic potentials impinging on layer II-III neurons. Theta and gamma oscillations in the entorhinal cortex were coupled with theta and gamma oscillations in the dentate hilar region. The majority of layer II-III neurons discharged irregularly but were phase-related to the negative peak of the local (layer II-III) gamma field oscillation. These findings demonstrate that layer II-III neurons discharge in temporally defined gamma windows (approximately 10-25 msec) coupled to the theta cycle. This transient temporal framework, which emerges in both the entorhinal cortex and the hippocampus, may allow spatially distributed subpopulations to form temporally defined ensembles. We speculate that the theta-gamma pattern in the discharge of these neurons is essential for effective neuronal communication and synaptic plasticity in the perforant pathway

Theta frequency field oscillation reflects synchronized synaptic potentials that entrain the discharge of neuronal populations within the approximately 100-200 ms range. The cellular-synaptic generation of theta activity in the hippocampus was investigated by intracellular recordings from the somata and dendrites of CA1 pyramidal cells in urethane-anesthetized rats. The recorded neurons were verified by intracellular injection of biocytin. Transition from non-theta to theta state was characterized by a large decrease in the input resistance of the neuron (39% in the soma), tonic somatic hyperpolarization and dendritic depolarization. The probability of pyramidal cell discharge, as measured in single cells and from a population of extracellularly recorded units, was highest at or slightly after the negative peak of the field theta recorded from the pyramidal layer. In contrast, cyclic depolarizations in dendrites corresponded to the positive phase of the pyramidal layer field theta (i.e. the hyperpolarizing phase of somatic theta). Current-induced depolarization of the dendrite triggered large amplitude slow spikes (putative Ca2+ spikes) which were phase-locked to the positive phase of field theta. In the absence of background theta, strong dendritic depolarization by current injection led to large amplitude, self-sustained oscillation in the theta frequency range. Depolarization of the neuron resulted in a voltage-dependent phase precession of the action potentials. The voltage-dependent phase-precession was replicated by a two-compartment conductance model. Using an active (bursting) dendritic compartment spike phase advancement of action potentials, relative to the somatic theta rhythm, occurred up to 360 degrees. These data indicate that distal dendritic depolarization of the pyramidal cell by the entorhinal input during theta overlaps in time with somatic hyperpolarization. As a result, most pyramidal cells are either silent or discharge with single spikes on the negative portion of local field theta (i.e., when the somatic region is least polarized). However, strong dendritic excitation may overcome perisomatic inhibition and the large depolarizing theta rhythm in the dendrites may induce spike bursts at an earlier phase of the extracellular theta cycle. The magnitude of dendritic depolarization is reflected by the timing of action potentials within the theta cycle. We hypothesize that the competition between the out-of-phase theta oscillation in the soma and dendrite is responsible for the advancement of spike discharges observed in the behaving animal

In the awake brain, information about the external world reaches the hippocampus via the entorhinal cortex, whereas during sleep the direction of information flow is reversed: population bursts initiated in the hippocampus invade the neocortex. We suggest that neocortico-hippocampal transfer of information and the modification process in neocortical circuitries by the hippocampal output take place in a temporally discontinuous manner associated with theta/gamma oscillations. On the other hand, transfer of the stored representations to neocortical areas is carried by discrete quanta of cooperative neuronal bursts (called sharp wave bursts) initiated in the hippocampus during slow wave sleep. The spatio-temporal participation of principal cells in sharp waves is determined by experience-induced changes in the CA3 recurrent collateral matrix. The co-operative, converging pre-synaptic activity can induce localized fast spikes and associated calcium influx in the apical dendrites of CA1 pyramidal cells, a necessary condition for the induction of synaptic plasticity. In addition, the subcortical effects of hippocampal sharp wave bursts may be critical in the release of various hormones which, in turn, may affect synaptic plasticity. These observations suggest that sleep patterns in the limbic system are essential for the preservation of experience-induced synaptic modifications

Thalamocortical neuronal oscillations underlie various field potentials that are expressed in the neocortex, including sleep spindles and high voltage spike-and-wave patterns (HVSs). The mechanism of extracellular current generation in the neocortex was studied in the anesthetized and awake rat. Field potentials and unit activity were recorded simultaneously along trajectories perpendicular to the cortical layers at spatial intervals of 100 microm by multiple-site recording silicon probes. Current source density (CSD) analysis revealed that the spatial positions of sinks in layers IV, V-VI, and II-III and of the accompanying sources were similar during sleep spindles, HVSs, and thalamic-evoked responses, although their relative strengths and timings differed. The magnitude and relative timing of the multiple pairs of sinks and sources determined the amplitude variability of HVSs and sleep spindles. The presence of temporally shifted dipoles was also supported by the time distribution of unit discharges in different layers. Putative interneurons discharged with repetitive bursts of 300-500 Hz. The spike component of HVSs was associated with fast field oscillations (400-600 Hz 'ripples'). Discharges of pyramidal cells were phase-locked to the ripples. These findings indicate that the major extracellular currents underlying sleep spindles, HVSs, and evoked responses result from activation of intracortical circuitries. We hypothesize that the fast field ripples reflect summed IPSPs in pyramidal cells resulting from the high frequency barrage of interneurons

Recent advances in the physiology of hippocampal interneurons are summarized in this article. These findings suggest that through their interconnectivity inhibitory interneurons can maintain large-scale oscillations at various frequency ranges (theta, gamma, and 200-Hz bands). We suggest that networks of inhibitory interneurons within the forebrain impose coordinated oscillatory 'contexts' for the 'content' carried by networks of principal cells. These oscillating inhibitory networks may provide the precise temporal structure necessary for ensembles of neurons to perform specific functions, such as memory trace formation and retrieval. In addition, synaptic inhibition is shown to reduce the somadendritic backpropagation of sodium spikes and to prevent the occurrence of calcium spikes in dendrites. These observations indicate that interneurons are in an excellent position to control neuronal plasticity and allow synaptic transmission either with or without long-term modification of synaptic strength

The non-competitive N-methyl-D-aspartate antagonist MK-801 has been frequently used to attenuate neurotoxicity mediated by excessive release of glutamate. However, doses of MK-801, effective to prevent cell loss in some areas have been reported to induce pathological changes in retrosplenial cortex [32]. In the present study, we examined the extent of the MK-801-induced damage. Silver staining techniques were used to label damaged neurons, axon terminals and activated microglia. In addition to the retrosplenial cortex, we observed silver-impregnated neurons in the pyriform, and entorhinal cortices, in amygdala in tenia tecti, and in the temporal two thirds of the dentate gyrus. With the exception of the dentate gyrus, signs of early degeneration appeared in the first 4 days in all observed regions. Activated microglia have been found 1 and 3 weeks after the lesion in the same areas. The time course and dose dependence of the damage was also investigated. The distribution of labeled neurons resembled the pattern observed after certain epileptic states. Our data suggest that irreversible cell damage occurred in the affected regions. These findings confirm and extend previous suggestions that, besides its protective effect, MK-801 may lead to neuronal degeneration

The mechanism of afterdischarge termination in the various hippocampal regions was examined in the rat. Stimulation of the perforant path or the commissural system was used to elicit afterdischarges. Combination of multiple site recordings with silicon probes, current source density analysis, and unit recordings in the awake animal allowed for a high spatial resolution of the field events. Interpretation of the field observations was aided by intracellular recordings from anesthetized rats. Irrespective of the evoking conditions, afterdischarges always terminated first in the CA1 region. Termination of the afterdischarge was heralded by a large DC shift initiated in dendritic layers associated with a low amplitude 'afterdischarge termination oscillation' (ATO) at 40 to 80 Hz in the cell body layer. ATOs were also observed in the CA3 region and the dentate gyrus. The DC shift spread at the same velocity (0. 1-0.2 mm/sec) in all directions and could cross the hippocampal fissure. All but 1 of the 25 putative interneurons in the CA1 and dentate regions ceased to fire before the onset of ATO. Intracellularly, ATO and the emerging DC potential were associated with fast depolarizing potentials and firing of pyramidal cells and depolarization block of spike initiation, respectively. Both field ATO and the intracellular depolarization shift were replicated by focal microinjection of potassium. We hypothesize that [K+]o lost by the intensely discharging neurons during the afterdischarge triggers propagating waves of depolarization in the astrocytic network. In turn, astrocytes release potassium, which induces a depolarization block of spike generation in neurons, resulting in 'postictal depression' of the EEG