Frequency potentiation in the medial cortex of young turtle brains in vitro.

Turtle brains have a relatively primitive cortex. Glutamate receptors in the cortex of turtles include N-methyl-D-aspartate (NMDA) and DL-alpha-amino-3-hydroxy-5-methylisoxazole-propionic acid (AMPA). Our aim was to determine whether the medial cortex in turtles, like the cortex and hippocampus in mammals exhibits frequency potentiation, a non-lasting form of synaptic plasticity, and, if so, to identify the involved receptors. Our results indicate that (1) the medial cortex exhibits this phenomenon with septal stimulation at 2 Hz, the frequency with maximum power spectral density in the electrocorticogram of turtles, showing an increase in both the excitatory postsynaptic potential and the evoked potential amplitudes; (2) the frequency potentiation of the medial cortex in turtles is mediated by AMPA type glutamate receptors; (3) the dynamics of frequency potentiation development in turtles show a number of differences from that in mammals. In summary, the cortex in this group of reptiles exhibits a functional trait of the cortex in mammals that is related to learning and memory; this trait, frequency potentiation, may have appeared as an independent specialization in both groups.

Homo- and heterosynaptic long-term potentiation in the medial cortex of the turtle brain in vitro.

Long-term potentiation of excitatory synaptic transmission was studied in an in vitro turtle brain that manifests spontaneous electrocorticogram activity. We show that in the turtle medial cortex, there is evidence of homosynaptic long-term potentiation in the septum-medial cortex pathway. This potentiation has two components, one dependent on NMDA receptor activation and the other independent of this receptor and suppressed by nifedipine, an antagonist of voltage-dependent Ca2+ channels (VDCCs), as occurs in the CA1 of rat hippocampus. Heterosynaptic long-term potentiation was also found in the medial cortex, as a tetanus in the septum also increased the cortico medial-cortico medial response. The intracellular response of pyramidal cells showed that the medial cortex EPSP increased its amplitude paripassu with an increase in the response of evoked field potential after tetanic stimulation in the septum.

Membrane and circuit properties of lateral septum neurons: relationships with hippocampal rhythms.

The lateral septum receives the most important afferents from the hippocampus, has been proposed to contribute to theta (theta) rhythm generation. Our aim was to study the membrane and circuital properties of lateral septum neurons and their relationship with hippocampal rhythms. Extra- and intracellular recordings (n=81) were obtained in urethane-anesthetized rats. Two neuronal populations were found, one of them with hippocampal theta; rhythm dependence (theta-D, 68%), and the other group independent of hippocampal theta; rhythm (theta-I, 32%). Other differences were spontaneous firing rate (theta-D=13.20+/-2.09, theta-I=6.99+/-1.18; p<0.005) with a bursting pattern in the theta-D group and single discharges in the theta-I group. Intracellular recordings showed higher synaptic activity in theta-D than in theta-I neurons. Both groups showed fast spikes while only theta-D neurons had high-threshold and low-threshold slow spikes. theta-D neurons had theta-oscillations in their membrane potential. Stimulation of the contralateral lateral septum resets the hippocampal theta rhythm and the theta rhythm recorded intracellularly in the lateral septum neurons. Some neurons (10.6%) showed rhythmic oscillations lasting a few seconds, at a higher frequency than those of the simultaneously recorded hippocampal EEG. This high frequency appeared spontaneously or could be evoked by stimulations of the fornix and reticularis pontis oralis nucleus (RPO). Homologous high frequency oscillations appeared in the simultaneously recorded hippocampal EEG, synchronized with the neuronal activity, during fornix stimulation. RPO stimulation evoked increments (57%) or decrements (43%) in the firing rate of lateral septum neurons. Thus, they could participate in different septal networks to modulate the theta rhythm. The marked functional relationship between lateral septum neurons and hippocampal theta rhythm supports the proposal that lateral septum represents a feedback system for the improvement of medial septum theta rhythm. The hypothalamic projections could be a way to introduce theta and higher rhythms into this structure that control many biological rhythms.

Changes in hippocampal cell discharge patterns and theta rhythm spectral properties as a function of walking velocity in the guinea pig.

Dorsal hippocampal theta rhythm (theta) and extracellular unit activity from CA1 pyramidal layer were recorded in awake guinea pigs, both during standing and during walking on a conveyor belt at increasing speeds. Amplitude, frequency and rhythmicity of theta increased linearly with the movement speed. In this preparation we found the same three types of unit discharge patterns that have been described in anesthetized rats in the presence of spontaneous or induced hippocampal theta: type 1, rhythmic at theta frequency and phase-locked with theta; type 2, discharging non-rhythmically but phase-locked with theta; and type 3, discharging at random. Furthermore, all units modified their firing pattern when the animals walked, either by increasing their rhythmicity and/or phase-locking with theta or by increasing their firing frequency. During walking, some type 3 units changed into type 2 or type 1, type 2 units changed to type 1, and type I increased their rhythmicity. Consequently, the unit discharge rhythmicity and phase-locking with theta increased with the speed of movement. The mean rate of neuronal discharges increased linearly as a function of walking speed. In this paper we show that the progressive spectral theta changes determined by the intensity of movement are concomitant with the increase in rhythmicity of hippocampal cells. Moreover, the firing rate of these cells, and the amplitude, frequency and rhythmicity of theta, increased linearly as a function of walking speed, suggesting that neuronal excitation may be basically responsible for these changes in theta properties.

Nucleus basalis of Meynert cell responses in awake monkeys.

Extracellular cell activity was recorded in the intermediate and posterior subdivisions of the nucleus basalis of Meynert (NBM) of awake monkeys to determine cell characteristics and the motor and sensory participation. Animals were trained to move a lever by elbow flexion-extensions to receive a reward. Cell activity was recorded when the animal was at rest and executing the task. The electromyogram of the upper limb, contralateral to the recording site, was recorded simultaneously with NBM neuron activity. The effect of visual, auditory, and tactile stimuli were also studied after performance of the learned task. A moderate number of cells responded to the reward (16%), while a higher percentage of them was associated with unexpected, unspecific stimuli (22%). Firing rates correlated positively with limb movement (30%). Visual (34%) and auditory (15%) responses were also found. No NBM cell responded to tactile stimulation. Considering these findings and the anatomical projections over the cortex, the NBM role ion complex integrative processes in discussed.

In vivo intracellular recordings of medial septal and diagonal band of Broca neurons: relationships with theta rhythm.

Transmembrane potentials from medial septal and diagonal band of Broca (MS-DBB) neurons and hippocampal field activity were recorded in curarized and urethanized rats. MS-DBB cells were studied during large amplitude irregular activity and during hippocampal theta rhythm, elicited by either sensory (i.e. stroking the fur on the animal's back) or electrical stimulation of the reticularis pontis oralis nucleus (RPO). Three types of cells were described according to their firing pattern and the characteristics of their "intracellular theta" rhythm. Type A neurons displayed continuous rhythmic oscillations in the membrane potential (Vm) of approximately 17 mV. These oscillations generated rhythmic high-frequency spike trains which were phase-locked with hippocampal theta rhythm. Type A cells revealed intracellular theta rhythm even in the absence of hippocampal theta rhythm, suggesting that the activity of this type of cell was the most important in hippocampal theta genesis. Type B cells were characterized by marked postspike after hyperpolarization and intracellular theta oscillations of smaller amplitude than in type A cells. Type C cells revealed a postspike afterdepolarization and a lower amplitude, intracellular theta rhythm only in the presence of hippocampal theta rhythm. Type C neurons could fire slow spikes at depolarizing (46% of cells) or hyperpolarizing (15% of cells) Vms. Type B and C cells were intracellularly stained with Lucifer yellow. Although type B and C neurons revealed dissimilar electrophysiological properties, they had comparable morphological shapes. RPO electrical stimulation generated hippocampal theta rhythm and intracellular theta rhythm in types A and B cells but not in type C cells, and increased the spike rate in type C neurons. Electrical stimulation of the fornix only evoked synaptic responses in type B and C neurons, with antidromic responses being elicited in 12% of type C cells. These results indicate that probably most of the type A rhythmic cells did not receive direct hippocampal feedback and that at least some type C cells were projecting neurons. The present findings demonstrate that theta rhythm oscillations in the Vm of MS-DBB neurons elicit different rhythmic discharge patterns.

Polymodal sensory and motor convergence in substantia nigra neurons of the awake monkey.

Responses of substantia nigra (SN) pars compacta (SNc) and reticulata (SNr) neurons to sensory stimulation and movement were investigated in awake Macaca fascicularis. Concern was assigned to polysensory and sensory/motor convergence on the same cell, which were found in 12% and 9% of the neurons, respectively. Others were modality specific (35%) or unresponsive (44%). The convergence lends support to the notion that SN plays an important role in sensory-motor integration.

Hippocampal EEG theta power density is similar during slow-wave sleep and paradoxical sleep. A long-term study in rats.

Rat hippocampal EEG and motor activity were studied during 15 days in relation to the vigilance state and to the light-dark cycle with a computerized system. During slow-wave sleep (SS) the hippocampal EEG has an outstanding mean power density in the theta band, similar to the large values present during paradoxical sleep. A circadian modulation was found for motor activity in SS and wakefulness (i.e. day-sleep is more restless, day-wakefulness is more quiet than night), and for EEG mean theta power in SS (i.e. less during day-sleep than night-sleep). These data underline the importance of analyzing the dark period when studying nocturnal animals.

Hippocampal and entorhinal glucose metabolism in relation to cholinergic theta rhythm.

Hippocampal and entorhinal cortex glucose metabolism were studied by 14C-2-deoxyglucose (2-DG) autoradiography in anesthetized rats with and without continuous theta rhythm (theta). 2-Deoxyglucose changes in specific cytoarchitectonic regions were precisely assessed by n innovative approach. In the absence of theta there were areas with a higher glucose metabolism corresponding to neuropile regions at CA3, dentate gyrus, and subiculum, while the cellular layers always showed lower values. In the presence of theta, provoked by intraventricular injections of anticholinesterases (i.e., physostigmine) or curarimimetics (i.e., d-tubocurarine), 2-DG uptake showed two opposite significant changes in relation to controls: a) it increased in the outer zone of the molecular layer (inner blade) of the dentate gyrus, and in the stratum lacunosum-moleculare of CA3, suggesting an increase in perforant path input during theta rhythm; b) it decreased in the hilar dentate region. This noteworthy decrease in metabolic activity probably reflects an hilar inhibition by local circuits during theta rhythm generation.

ERP components related to stimulus selection processes.

Event-related potential (ERP) components associated with target stimulus selection in a double discrimination visual task were studied. The experimental paradigm consisted in the presentation of low intensity stimuli that varied according to two physical features: geometrical form (squares and circles) and location (a spot in different positions inside the stimulus). Subjects performed 3 tasks on these stimuli: control task in which they looked passively at the stimuli, and 2 discrimination tasks, in which they had to respond to a certain stimulus (a specific conjunction of form and spot location). The early components (P1 and N1) obtained in the control and discrimination tasks were associated with sensory analysis of simple stimulus features. Relevance of a particular feature modified the latency and/or the area of these components. The longer-latency components (N2 and P3) were elicited only in the discrimination tasks. N2 was associated with target stimulus selection because its area was significantly larger for target stimuli and because its "offset" latency correlated with choice reaction time. Results are discussed and contrasted with various models of target selection.

Activity in monkey substantia nigra neurons related to a simple learned movement.

Single cell activity was recorded in the pars compacta (SNc) and pars reticulata (SNr) of the substantia nigra (SN) in 4 unanesthetized Macaca fascicularis to determine the motor role of the nucleus. Animals were trained to perform a simple task that involved moving a lever by elbow flexion-extensions, in the horizontal plane using the hand contralateral to the recording site. Two monkeys learnt to execute the task on both sides. Electromyograms (EMG) of limb muscles were recorded simultaneously with SN neurons. Discharge rate modulation related to specific movement phases was present in 35% of the neurons. A significant positive correlation of the discharge rate with movement velocity and amplitude was found in SNc and SNr neurons. Some SNr cells discharged in anticipation of the EMG, suggesting a participation of the nucleus in the preparation of movement. The activity of SNr neurons was also related to movement of the left and right upper limb. In conclusion, the SN seems to play an important role in the control of specific motor mechanisms, probably modulating movement velocity, amplitude and direction, with little participation of somatosensory feedback. The involvement of the SNr in the coordination of bilateral arm activity is discussed.

Frequency potentiation in granule cells in vivo at theta frequency perforant path stimulation.

The effect of frequency potentiation on the postsynaptic potential in granule cells was studied stimulating the perforant path in curarized and urethanized rats. At stimulation frequencies between 2.0-5.0 Hz, synaptic efficacy in eliciting an orthodromic action potential increased despite the hyperpolarization of the transmembrane potential. The excitatory postsynaptic potential (EPSP) slope and duration also increased while the inhibitory postsynaptic potential (IPSP) was reduced. Stimulation frequencies greater than 5.0 Hz produced similar changes in the transmembrane potential and EPSP-IPSP sequence, but they did not increase synaptic efficacy. The frequency potentiation at frequencies into the theta band suggest that this potentiation participates in theta rhythm genesis in this structure which, in turn, suggests that the fascia dentata could work as a band-pass filter. In 12.5% of cases postpotentiation was also observed.

Electrocorticograms of hippocampal and dorsal cortex of two reptiles: comparison with possible mammalian homologs.

To compare the ongoing electrical activity in possibly homologous structures of reptiles and mammals, the electrographic activity (micro-EEG) from major parts of the cortex of unanesthetized turtles (Pseudemys) and geckos (Gekko) was recorded with and without acute and chronic stimuli, physostigmine and atropine. Electrodes were placed in the medial cortex (MC) and in the dorsal cortex (DC), the possible homologs of the mammalian hippocampus and transitional or/and isocortex, respectively. The resting corticograms (1-50 Hz) are different in the two cortical areas. Both are wide-band; power falls steadily with frequency above a single maximum about 2 Hz. The MC has a nonrhythmic, low-voltage activity with occasional superimposed large sharp waves (LSWs), generally biphasic, 100-300 microV and lasting 0.25-0.75 s. The DC has smaller amplitudes (ca. 3-6 dB) at all frequencies and fewer LSWs. Reptilian LSWs are reminiscent of mammalian hippocampal sharp waves or spikes, a correlate of decreased arousal. The immobility-related rhythmic slow activity (theta), so characteristic of the hippocampus in a number of mammals, was not found in the cortex of either species of reptile under a variety of conditions. We cannot exclude the possibility of movement-related theta waves. Physostigmine injection does not produce theta, although it acts like an arousing stimulus, producing a disappearance of the LSWs and a substantial increase in the amplitude of the frequencies 12-24 Hz; these changes were more obvious in the DC. Atropine reversed the effects of physostigmine. Theta may represent a trait of the more highly differentiated hippocampal field of mammals. The condition represented by these reptiles, in which the EEG differs between parts of the pallium but without theta or reciprocal changes in the MC and DC, may be an earlier evolutionary stage. A distinctive reptilian EEG is not recognizable in Pseudemys and Gekko, but a number of differences from the EEG in familiar mammals are shared by these two neurologically quite different reptiles.

Relationships of nucleus reticularis pontis oralis neuronal discharge with sensory and carbachol evoked hippocampal theta rhythm.

The activity of 72 neurons recorded in the reticularis pontis oralis nucleus (RPO) was examined in anesthetized and curarized rats during hippocampal theta (theta) rhythm elicited by either sensory stimulation or carbachol microinjections. During hippocampal theta rhythm evoked by sensory stimulation, 63.9% of RPO neurons increased their discharge rate while the firing rate decreased in 20.8%. In all cases, the RPO neurons maintained a non-rhythmic discharge pattern. In 44% of the neurons the discharges tended to occur on the positive wave of the theta rhythm. Similar firing patterns were seen in 18 RPO neurons recorded during theta rhythm elicited by both, sensory stimulation and a carbachol microinjection; this effect was blocked by atropine. These results indicate that the RPO region contributes to the generation of hippocampal theta rhythm with a tonic and nonrhythmic outflow through a cholinergic system which may be muscarinic.

Electroencephalogram in vitro and cortical transmembrane potentials in the turtle Chrysemys d'orbigny.

An experimental stable model of an in vitro turtle brain (Chrysemys d'orbigny) was developed in order to compare electrographic activity (EEG) with transmembrane potentials. Two preparations were used: a whole intact hemisphere and a whole open hemisphere. The latter permitted easier impalement of cortical neurons through the ependymal surface. The EEG characteristics were similar to those described in turtles in vivo. The EEG was nonrhythmic (rhythmicity coefficient less than 0.40). The power spectrum presented a high energy band between 1 and 3 Hz, decreasing progressively towards the higher frequencies. Total power of the EEG was one order of magnitude greater than the system noise. Random large amplitude sharp waves (22-300 microV, 500-1,900 ms) were recorded spontaneously. Hypoxia produced an increase in frequency and amplitude of the large sharp waves, without modification of either EEG background activity or membrane potentials. Physostigmine provoked the disappearance of the large sharp waves, an effect reversed by atropine. The addition of TTX to the medium provoked the abolition of the EEG, although spikes and plateaus determined by Ca2+ conductances persisted. The power spectra band of maximum relative potency was 0.8-2.5 Hz for both EEG and slow membrane potentials.

Synaptic contributions to theta rhythm genesis in rat CA1-CA3 hippocampal pyramidal neurons in vivo.

The effects of intracellular Cl- diffusion and hyperpolarizing current pulses on inhibitory postsynaptic potentials (IPSPs) and the transmembrane theta rhythm of CA1-CA3 pyramidal neurons were tested in urethanized and curarized rats. Cl- diffusion and hyperpolarizing currents decreased the amplitude of IPSPs evoked by fornix stimulation without modifying the theta rhythm amplitude and phase. The membrane conductance was typically 22-46% higher at the positive than negative intracellular theta peaks. Results indicate that in curarized rats excitatory postsynaptic potentials were the main components of intracellular theta without an important participation of IPSPs in theta rhythm genesis.

Slow intrinsic spikes recorded in vivo in rat CA1-CA3 hippocampal pyramidal neurons.

The membrane potential of 45 CA1-CA3 hippocampal pyramidal cells was recorded in curarized and urethanized rats. Two slow spike types were observed together with the usual Na+ type action potential. Slow spikes were termed HTS and LTS because they were essentially identical to the high and low threshold Ca2+ spikes observed in vitro and probably represent the same kinds of activities. LTS and HTS (22 and 29 cells, respectively) were triggered at potentials greater than or equal to 65 mV and less than or equal to 55 mV, had mean durations of 23.7 and 90.4 ms and mean amplitudes of 11.5 and 39.3 mV, respectively, and fired an overriding burst of the action potentials. LTS and HTS were sometimes present in the same neuron (n = 16). Depolarizing pulses triggered rhythmic HTS at rates that increased with depolarization up to a 5 impulses/s maximum. The same limit was found with imposed membrane potential sine currents at frequencies within the theta rhythm range. With spontaneous or imposed hyperpolarizations LTS were evoked by depolarizing, at the break of hyperpolarizing pulses, or spontaneously. They were also evoked at the depolarizing, or recovery, slopes of inhibitory postsynaptic potentials. HTS and LTS supply pyramidal neurons with different firing patterns which enhance the system's integrative possibilities. Evidence is provided that theta is not exclusively generated by network properties, since rhythmic HTS may participate.

Nocturnal growth hormone surges in type 1 diabetes mellitus are both sleep- and glycemia-dependent: assessment under continuous sleep monitoring.

Our work has studied the relationship between nocturnal growth hormone (GH) surges, sleep and glycemia in seven conventionally treated type 1 diabetic patients under continuous sleep monitoring and the results were compared to those found in five age-matched healthy controls. On the experimental day, sleep was monitored from 24.00 to 07.00. Blood glucose levels and GH were assayed in both groups. As a group the diabetics had nocturnal GH responses higher than those in controls. However, the sleep-related GH release is not abnormally high in patients who maintain strict normoglycemia. Early-night hypoglycemia and/or rapidly decreasing blood glucose concentrations enhance sleep-related GH secretion in diabetics, whereas hypoglycemia not associated with slow-wave sleep (SWS) causes a moderate increase in GH. Late-onset nocturnal hypoglycemia is not potent enough to stimulate GH. It is proposed that in diabetics sleep-related GH production is probably not abnormally elevated within a wide range of stable glucose levels, but when these thresholds are crossed or when there is a rapid decrease in blood glucose, then GH secretion is inversely related to the changing blood glucose. Therefore, our study supports the conclusion that sleep-related GH secretion is finely modulated by the actual glycemic fluctuations in diabetic patients.

In vivo electrophysiological analysis of lucifer yellow-coupled hippocampal pyramids.

Small transient all-or-none depolarizations (also termed in the literature fast prepotentials, spikelets, pseudospikes, d-spikes, or short latency depolarizations) and their association with lucifer yellow (LY) dye-coupling were analyzed in CA1-CA3 hippocampal pyramidal cells in urethane anesthetized rats. It was found that (a) 15 of the 24 LY-injected pyramidal neurons (63%) showed dye-coupling; (b) spontaneous, anti- and orthodromically evoked spikelets (3-7 ms in duration; 3- to 12-mV peak) were recorded in 40 of 95 cells (42%); (c) there was a significantly higher probability of dye-coupled neurons with spikelets and of uncoupled ones without spikelets; (d) spikelet waveform and amplitude were unaffected by spontaneous or imposed polarizations; (e) large hyperpolarizations could reduce the rate and even prevent spikelets; and (f) spikelets could precede or follow spikes, the latter were more frequent with large depolarizations. Electrophysiological findings, and the association of dye-coupling and spikelets, suggest strongly that at least some spikelets are coupling potentials. This implies that pyramidal cells may be electronically coupled under physiological conditions.

In vivo intracellular analysis of rat dentate granule cells.

In vivo intracellular recordings and dentate EEG were obtained in urethanized-curarized rats. Granule cells (GCs) were identified by antidromic activation as well as by intracellular staining with Lucifer yellow (LY). GCs fired spikes which, in 43.6% of the cases, had brief post-hyperpolarization. Slow spikes were recorded at hyperpolarized levels during the rebound of hyperpolarizing pulses or during inhibitory postsynaptic potentials (IPSPs). Medial septal nucleus or perforant pathway stimulation evoked an excitatory postsynaptic potential (EPSP)-IPSP sequence. During theta (theta) rhythm, the membrane potential of 80.3% of the GCs showed rhythmic sine-like waves of up to 15 mV at a theta frequency which were phase-locked with extracellular theta. GCs were classified into 3 types: type 1 (67.1%) showed intracellular theta and rhythmic firing; type 2 (13.2%) revealed intracellular theta and random firing, but spikes tended to occur at a preferred phase of the dentate theta; and type 3 (19.7%) had neither intracellular theta nor rhythmic firing. Intracellular theta amplitude was wider during injection of the hyperpolarizing current and narrower during depolarizing ones, indicating that rhythmic EPSPs contribute to theta genesis. Intracellular theta was unaffected by Cl- or Cs+ diffusion, suggesting that IPSP is not essential to theta genesis.