A parvalbumin-containing, axosomatic synaptic network in the rat medial septum: relevance to rhythmogenesis

The medial septal diagonal band complex (MS/DB), made up of cholinergic and GABAergic neurons, plays an important role in the generation of the hippocampal theta rhythm. A GABAergic neuron type in the MS/DB that has fast spiking properties was shown previously to contain parvalbumin immunoreactivity and to form axosomatic connections with unidentified somata. The aim in the current study was to determine the neurochemical identities of these target neurons. In slices and in perfused-fixed brain, staining for parvalbumin immunoreactivity first of all revealed the presence of two types of parvalbumin-positive somata in the MS/DB: medially located neurons with parvalbumin-positive basket-like terminals on them, and more laterally located neurons with fewer parvalbumin-positive contacts on them. In MS/DB slices, the terminals of fast spiking neurons filled with biocytin correspondingly made either numerous contacts that surrounded the parvalbumin-positive cell body in basket-like formation, or 1-5 contacts on a localized patch of the soma. These contacts were shown by electron microscopy to form synaptic junctions. No terminals of biocytin-filled fast spiking neurons were observed on cholinergic neurons, and dual staining in perfused-fixed brain did not reveal the presence of parvalbumin-containing terminals on cholinergic somata. Our results suggest therefore that there are two subtypes of parvalbumin-containing neuron in the MS/DB, and that these are interconnected via axosomatic synapses. The contrasting topographical organization of the two types of parvalbumin-containing neuron suggests that they may receive different types of afferent input, but this will require substantiation in future studies. We propose that generation of rhythmic activity in the MS/DB is controlled by contrasting contributions from two types of parvalbumin-positive neuron, and that the role of the cholinergic neuron is modulatory.     

Saccadic omnipause and burst neurons in monkey and human are ensheathed by perineuronal nets but differ in their expression of calcium-binding proteins

The extracellular matrix of the brain contains large aggregates of chondroitin sulfate proteoglycans (CSPG), which form lattice-like cell coatings around distinct neuron populations and are termed perineuronal nets. The function of perineuronal nets is not fully understood, but they are often found around neurons containing the calcium-binding protein parvalbumin, suggesting a function in primarily highly active neurons. In the present paper the distribution of perineuronal nets was studied in two functional cell groups of the primate oculomotor system with well-known firing properties: 1) the saccadic omnipause neurons in the nucleus raphe interpositus (RIP) exhibit a high tonic firing rate, which is only interrupted during saccades; they are inhibitory and use glycine as a transmitter; and 2) premotor burst neurons for vertical saccades in the rostral interstitial nucleus of the medial longitudinal fascicle (RiMLF) fire with high-frequency bursts during saccades; they are excitatory and use glutamate and/or aspartate as a transmitter. In the macaque monkey, both cell populations were identified by their parvalbumin immunoreactivity and were studied for the presence of perineuronal nets using CSPG antibodies or lectin binding with Wisteria floribunda agglutinin. In addition, the expression of another calcium-binding protein, calretinin, was studied in both cell groups. Double- and triple-immunofluorescence methods revealed that both omnipause and burst neurons are selectively ensheathed with strongly labeled perineuronal nets. Calretinin was coexpressed in at least 70% of the saccadic burst neurons, but not in the omnipause neurons. Parallel staining of human tissue revealed strongly labeled perineuronal nets around the saccadic omnipause and burst neurons, in corresponding brainstem regions, which specifically highlighted these neurons within the poorly structured reticular formation. These findings support the hypothesis that perineuronal nets may provide a specialized microenvironment for highly active neurons to maintain their fast-spiking activity and are not related to the transmitter or the postsynaptic action of the ensheathed neurons.     

Morphology of local axon collaterals of electrophysiologically characterised neurons in the rat medial septal/ diagonal band complex

Neurons in the medial septal/diagonal band complex (MS/DB) in vivo exhibit rhythmic burst-firing activity that is phase-locked with the hippocampal theta rhythm. The aim was to assess the morphology of local axon collaterals of electrophysiologically identified MS/DB neurons using intracellular recording and biocytin injection in vitro. Cells were classified according to previous criteria into slow-firing, fast-spiking, regular-spiking, and burst-firing neurons; previous work has suggested that the slow-firing neurons are cholinergic and that the other types are GABAergic. A novel finding was the existence of two types of burst-firing neuron. Type I burst-firing neurons had significantly longer duration after hyperpolarisation potentials when held at -60 mV, and at -75 mV, type I neurons exhibited a low-threshold spike with more rapid activation and inactivation kinetics than those of type II neurons. We have, also for the first time, described the main features of the local axon collaterals of the five neuron types. All filled neurons possessed a main axon that gave forth 1-12 local primary axon collaterals. All electrophysiological types, except for the type I burst-firing neuron, had a main axon that coursed toward the fornix. Myelination of the main axon was a prominent feature of all but the slow-firing neurons. Branching of the primary axon collaterals of the fast-spiking and type I burst-firing neurons was more extensive than that of the other cell types, with those of the slow-firing neurons exhibiting the least branching. All cell types possessed axon collaterals of the en passant type, and some in addition had twiglike or basketlike axon terminals. All cell types made synapses on distal dendrites; a proportion of the fast-spiking and burst-firing cells in addition had basketlike terminals that made synaptic contacts on proximal dendrites and on somata. Two morphological types of somata were postsynaptic to the basket cells: large (20-30-microm) oval cells with dark cytoplasm, and large oval cells with paler cytoplasm, often with an apical dendrite. The presence of lamellar bodies in the large dark neurons suggests that they may be cholinergic neurons, because previous work has localised these structures in some neurons that stain for choline acetyltransferase. Our work suggests therefore that there may be GABAergic neurons in the MS/DB that form basket synaptic contacts on at least two types of target cell, possibly cholinergic and GABAergic neurons, which means that the basket cells could play a key role in the generation of rhythmic activity in the MS/DB.     

Morphology and distribution of electrophysiologically defined classes of pyramidal and nonpyramidal neurons in rat ventral subiculum in vitro

Intracellular electrophysiological recordings were made from 210 ventral subicular neurons in rat brain slices. Recordings here classified as burst-firing or nonburst-firing. Eighteen burst-firing neurons were filled with Neurobiotin, and all had pyramidal morphology. Nine of these recordings were made from intrinsically burst-firing (IB) cell bodies, and nine were made from burst-firing dendrites (BD). Twelve nonburst-firing neurons were also filled with Neurobiotin. Eight were regular spiking (RS) and had pyramidal morphology, four were fast spiking (FS) and nonpyramidal. Additional electrophysiological parameters distinguished IB from BD, RS from FS, and RS from IB recordings. The distribution of IB and RS neurons was examined by using 180 recordings. Information from the first series of experiments was used to distinguish between somatic and dendritic recordings. The deep-superficial axis (alveus-hippocampal fissure) was divided into four equal rows. RS neurons accounted for 12%, 28%, 58%, and 50% of presumed somatic recordings in successively more superficial rows. The proximal-distal (CA1-perforant path) axis was divided into five equal columns. RS cells accounted for 52% of presumed somatic impalements in the central column compared with 16% in the most proximal and 10% in the most distal columns. Thus, two electrophysiological classes of pyramidal neuron were localized to particular regions of the ventral subiculum. In the light of existing knowledge of the topography of subicular inputs and outputs, our results are consistent with the hypothesis that the ratio of RS to IB pyramidal neurons will be different in different transhippocampal circuits. J. Comp. Neurol. 380:395–408, 1997. © 1997 Wiley-Liss, Inc.     

Distribution of parvalbumin-containing neurons and lectin-binding perineuronal nets in the rat basal forebrain

In sections of rat brain treated forWisteria floribunda agglutinin (WFA) labelling the occurrence of parvalbumin (PARV)-, calbindin (CALB)- or choline acetyltransferase (ChAT) immunoreactivity was analyzed in the basal forebrain using dual-peroxidase and double-fluorescence methods. Only PARV-immunoreactive (-ir) neurons were surrounded by WFA-labelled, i.e.N-acetylgalactosamine-containing, perineuronal lattice-like structures known as perineuronal nets. The distribution of these nets and PARV-ir cells in the rat basal forebrain was documented to obtain detailed data on their co-existence. A remarkable diversity distribution of both markers was observed, as PARV-ir neurons are only associated with nets in the medial septal nucleus, the nuclei of the diagonal band and the magnocellular preoptic nucleus, but not in the ventral pallidum or the substantia innominata/nucleus basalis complex. These differences in the neuronal microenvironment may reflect system-related specializations of neurons within the basal forebrain nuclei.     

Characterisation of hyperpolarization-activated currents (I(h)) in the medial septum/diagonal band complex in the mouse

Hyperpolarization-activated cyclic nucleotide gated (HCN) channel subunits are distributed widely, but selectively, in the central nervous system, and underlie hyperpolarization-activated currents (I(h)) that contribute to rhythmicity in a variety of neurons. This study investigates, using current and voltage-clamp techniques in brain slices from young mice, the properties of I(h) currents in medial septum/diagonal band (MS/DB) neurons. Subsets of neurons in this complex, including GABAergic and cholinergic neurons, innervate the hippocampal formation, and play a role in modulating hippocampal theta rhythm. In support of a potential role for I(h) in regulating MS/DB firing properties and consequently hippocampal neuron rhythmicity, I(h) currents were present in around 60% of midline MS/DB complex neurons. The I(h) currents were sensitive to the selective blocker ZD7288 (10 microM). The I(h) current had a time constant of activation of around 220 ms (at -130 mV), and tail current analysis revealed a half-activation voltage of -98 mV. Notably, the amplitude and kinetics of I(h) currents in MS/DB neurons were insensitive to the cAMP membrane permeable analogue 8-bromo-cAMP (1 mM), and application of muscarine (100 microM). Immunofluoresence using antibodies against HCN1, 2 and 4 channel subunits revealed that all three HCN subunits are expressed in neurons in the MS/DB, including neurons that express the calcium binding protein parvalbumin (marker of fast spiking GABAergic septo-hippocampal projection neurons). The results demonstrate, for the first time, that specific HCN channel subunits are likely to be coexpressed in subsets of MS/DB neurons, and that the resultant I(h) currents show both similarities, and differences, to previously described I(h) currents in other CNS neurons.     

The two faces of perineuronal nets

Perineuronal nets are extracellular structures enwrapping the soma and proximal dendrites of some neurons known to be parvalbumin immunoreactive. The composition of the nets is not completely known, but it can change between different neurons. We studied the heterogeneous composition of a specific component of perineuronal nets, the signaling molecule Janusin (or Tenascin R), by means of a double immunofluorescence using lectin from Wisteria floribunda as a general marker for perineuronal nets and an antibody against Janusin. The presence of two kinds of perineuronal nets, one rich in Janusin (the majority) and a second one devoid of this glycoprotein, indicates differential roles of these neurons, as well as differences in their afferents, or a difference in their functional state.     

Complementary Distribution of Calbindin D-28k and Parvalbumin in the Basal Forebrain and Midbrain of the Squirrel Monkey

The distribution of cell bodies expressing either calbindin D-28k or parvalbumin immunoreactivity in the basal forebrain and midbrain of squirrel monkeys (Saimiri sciureus) was studied on contiguous sections incubated with monoclonal antibodies raised against calbindin or parvalbumin. In the nucleus accumbens, medium-sized calbindin-positive neurons formed two cell bridges joining the ventral part of the striatum to the olfactory tubercle, whereas medium-sized parvalbumin-positive cells in the same area were much less numerous and more uniformly distributed. The medial and dorsal septal nuclei contained a small number of elongated calbindin-positive neurons and only a few parvalbumin-immunoreactive cells. In the nucleus of the diagonal band of Broca, calbindin and parvalbumin were found to label two distinct but closely intermingled neuronal populations. In the striatum, medium-sized calbindin-immunoreactive cells occurred in very large numbers and appeared to be confined to the extrastriosomal matrix. Medium-sized, parvalbumin-immunoreactive neurons were also present in the striatum but they were less numerous than the calbindin-positive cells. The calbindin-positive neurons in the dorsal portion of the striatum were less intensely stained than those in the ventral portion, whereas this pattern did not occur for neurons expressing parvalbumin immunoreactivity. At the pallidal level, neurons in both segments were devoid of calbindin but displayed a very strong parvalbumin immunoreactivity. Most of the large neurons of the nucleus basalis of Meynert were strongly calbindin-immunoreactive and many of them invaded dorsally the medullary laminae of the pallidal complex. The neurons of the subthalamic nucleus were markedly enriched with parvalbumin but displayed only light calbindin staining. In the substantia nigra/ventral tegmental area complex, calbindin-immunoreactive cells abounded in the ventral tegmental area and in the dorsal tier of the pars compacta of the substantia nigra, but were absent in the ventral tier of the pars compacta and in the entire pars reticulata of the substantia nigra. In contrast, numerous parvalbumin-immunoreactive neurons occurred in the pars reticulata and pars lateralis, but none were found in the pars compacta and ventral tegmental area. These findings reveal that the patterns of calbindin and parvalbumin distribution in primate basal forebrain and midbrain are strikingly complementary, suggesting a synergistic role for these calcium-binding proteins in basal forebrain and midbrain function.     

Muscarinic Modulation of Intrinsic Burst Firing in Rat Hippocampal Neurons

Intracellular recordings in rat hippocampal slices were used to examine how exogenous and endogenous cholinergic agonists modulate the firing pattern of intrinsically burst-firing pyramidal cells. About 24% of CA1 pyramidal cells generated all-or-none, high-frequency bursts of fast action potentials in response to intracellular injection of long positive current pulses. Application of carbachol (5 μM) converted burst firing in these neurons into regular trains of independent spikes. Acetylcholine (5 μM) exerted a similar effect, provided acetylcholine esterase activity was blocked with neostigmine (2 μM). Atropine (1 μM) reversed this cholinergic effect, indicating its mediation by muscarinic receptors. Cholinergic agonists also caused mild neuronal depolarization but the block of intrinsic burst firing was independent of this effect. Repetitive stimulation of cholinergic fibres in the presence of neostigmine (2 μM) evoked a slow cholinergic excitatory postsynaptic potential (EPSP) lasting about a minute. During the slow EPSP, burst firing could not be evoked by depolarizing pulses and the neurons fired in regular mode. These effects were prevented by pretreatment with atropine (1 μM). Exogenously applied cholinergic agonists and endogenously released acetylcholine also reduced spike frequency accommodation and suppressed the long-duration afterhyperpolarization in burst-firing pyramidal cells in an atropine-sensitive manner. A membrane-permeable cAMP analogue (8-bromo-cAMP; 1 mM) also reduced frequency accommodation and blocked the long-duration afterhyperpolarization, but did not affect intrinsic burst firing at all. Taken together, the data show that muscarinic receptor stimulation transforms the stereotyped, phasic response of burst-firing neurons into stimulus-graded, tonic discharge.     

Input and frequency-specific entrainment of postsynaptic firing by IPSPs of perisomatic or dendritic origin

Correlated activity of cortical neurons underlies cognitive processes. Networks of several distinct classes of gamma-aminobutyric acid (GABA)ergic interneurons are capable of synchronizing cortical neurons at behaviourally relevant frequencies. Here we show that perisomatic and dendritic GABAergic inputs provided by two classes of GABAergic cells, fast spiking and bitufted interneurons, respectively, entrain the timing of postsynaptic spikes differentially in both pyramidal cells and interneurons at beta and gamma frequencies. Entrainment of pyramidal as well as regular spiking non-pyramidal cells was input site and inhibitory postsynaptic potential frequency dependent. Gamma frequency input from fast spiking cells entrained pyramidal cells on the positive phase of an intrinsic cellular theta oscillation, whereas input from bitufted cells was most effective in gamma frequency entrainment on the negative phase of the theta oscillation. The discharge of regular spiking interneurons was phased at gamma frequency by dendritic input from bitufted cells, but not by perisomatic input from fast spiking cells. Action potentials in fast spiking GABAergic neurons were phased at gamma frequency by both other fast spiking and bitufted cells, regardless of whether the presynaptic GABAergic input was at gamma or beta frequency. The interaction of cell type-specific intrinsic properties and location-selective GABAergic inputs could result in a spatio-temporally regulated synchronization and gating of cortical spike propagation in the network.     

Neuronal correlates of vestibulo-ocular reflex adaptation in the alert guinea-pig

The spiking behaviour of 66 second-order vestibular neurons was studied in alert, chronically prepared guinea-pigs during the horizontal vestibulo-ocular reflex (VOR). Among the 66 studied neurons, seven were held long enough (> 1 h) to compare their spiking behaviour before and after a training procedure inducing a decrease in the gain of the VOR. When tested in darkness following adaptation, five of them showed a significant decrease of their sensitivity to head rotation. However, the resting discharge of these five neurons remained unchanged. This suggests that VOR adaptation is mediated not only by changes in synaptic efficacities but also by modifications in the spike generator which transforms synaptic inputs into a pattern of action potentials.     

Respiratory cycle entrainment of septal neurons mediates the fast coupling of sniffing rate and hippocampal theta rhythm

Memory for odour information may result from temporal coupling between the olfactory and hippocampal systems. Respiration defines the frequency of olfactory perception, but how the respiratory rate affects hippocampal oscillations remains poorly understood. The afferent connectivity of the medial septum/diagonal band of Broca complex (MS/DB) proposes this region as a crossroads between respiratory and limbic pathways. Here we investigate if the firing rates of septal neurons integrate respiratory rate signals. We demonstrate that approximately 50% of MS/DB neurons are temporally correlated with sniffing frequency. Moreover, a group of slow‐spiking septal neurons are phase‐locked to the sniffing cycle. We show that inter‐burst intervals of MS/DB theta cells relate to the sniff rate. Intranasal odour infusion evokes sniff phase preference for the activity of fast‐spiking MS/DB neurons. Concurrently, the infusion augments the correlation between sniffing and limbic theta oscillations. During periods of sniffing–theta correlation, CA1 place cells fired preferentially during the inhalation phase, suggesting the theta cycle as a coherent time frame for central olfactory processing. Furthermore, injection of the GABAergic agonist muscimol into medial septum induces a parallel decrease of sniffing and theta frequencies. Our findings provide experimental evidence that MS/DB does not merely generate theta rhythm, but actively integrates sensorimotor stimuli that reflect sniffing rate. Such integration may provide temporal oscillatory synchronisation of MS/DB‐innervated limbic structures with the sniffing cycle.     

Burst-firing activity of presumed 5-HT neurones of the rat dorsal raphe nucleus: electrophysiological analysis by antidromic stimulation

We recently reported raphe neurones which frequently fired spikes in short bursts. However, the action potentials were broad and the neurones fired in a slow and regular pattern, suggesting they were an unusual type of 5-hydroxytryptamine (5-HT) neurone. In the present study, we investigated whether these putative burst-firing 5-HT neurones project to the forebrain and whether all spikes fired in bursts propagate along the axon. In anaesthetised rats, electrical stimulation of the medial forebrain bundle evoked antidromic spikes in both burst-firing neurones and in single-spiking, classical 5-HT neurones recorded in the dorsal raphe nucleus. Although the antidromic spike latency of the single-spiking and burst-firing neurones showed a clear overlap, burst-firing neurones had a significantly shorter latency than single-spiking neurones. For both burst-firing neurones and classical 5-HT neurones, antidromic spikes made collisions with spontaneously occurring spikes. Furthermore, in all burst-firing neurones tested, first, second and third order spikes in a burst could be made to collide with an antidromic spike. Interestingly, in a small number of burst-firing neurones, antidromic stimulation evoked spike doublets, similar to those recorded spontaneously. From these data we conclude that burst-tiring neurones in the dorsal raphe nucleus project to the forebrain, and each spike generated by the burst propagates along the axon and could thereby release transmitter (5-HT).     

Apamin increases NMDA-induced burst-firing of rat mesencephalic dopamine neurons

Intracellular recordings made in vitro from rat midbrain dopamine neurons showed that apamin (100 nM) did not alter the regular spontaneous firing of the neurons, but it increased the occurrence of bursts of action potentials in N-methyl-d-aspartate. Apamin appeared to facilitate burst-firing induced by NMDA because, by blocking an outward calcium-activated potassium current, it increased the depolarizing action of NMDA.     

Fast oscillations trigger bursts of action potentials in neocortical neurons in vitro: a quasi-white-noise analysis study

PURPOSE: Recent evidence supports the importance of action potential bursts in physiological neural coding, as well as in pathological epileptogenesis. To better understand the temporal dynamics of neuronal input currents that trigger burst firing, we characterized spectral patterns of stimulation current that generate bursts of action potentials from regularly spiking neocortical neurons in vitro. METHODS: Sharp microelectrodes were used for intracellular recording and stimulation of cortical neurons in rat brain slices. Quasi-white-noise (0-2 kHz) and "chirp" sine wave currents of decreasing wavelength were applied to represent a broad spectrum of stimulation frequencies. Action potential-related averaging of the stimulation current variations preceding bursting was used to characterize stimulation current patterns more likely to result in a burst rather than a single-spike response. RESULTS: Bursts of action potentials were most reliably generated by a preceding series of > or = 2 positive current transients at 164+/-37 Hz of the quasi-white-noise, and to sine wave currents with frequencies greater than 90 Hz. The intraburst action potential rate was linearly related to the frequency of the input sine wave current. CONCLUSIONS: This study demonstrates that regularly spiking cortical neurons in vitro burst in response to fast oscillations of input currents. In the presence of positive cortical feedback loops, encoding input frequency in the intraburst action potential rate may be safer than producing a high-frequency regular output spike train. This leads to the experimentally testable and therapeutically important hypothesis that burst firing could be an antiepileptogenic and/or anti-ictogenic mechanism.     

Wisteria floribunda agglutinin-labelled nets surround parvalbumin-containing neurons.

Net-like structures surrounding several types of neurones contain glycoconjugates which are detectable by lectins specific for N-acetylgalactosamine. Wisteria floribunda agglutinin (WFA) was introduced as a further marker for the visualization of such perineuronal nets, which were also revealed in regions of the rat brain where these structures could not be clearly demonstrated using other lectins. The WFA-labelled perineuronal nets resembled in detail those which could be visualized using Vicia villosa agglutinin, colloidal iron or hyaluronectin as markers. Furthermore, WFA-stained perineuronal net components appeared to be similar to proteoglycan-immunoreactive structures. Dual-peroxidase experiments and fluorescence double labelling demonstrated that WFA-binding structures frequently ensheath GABAergic neurons containing the calcium-binding protein parvalbumin in the areas investigated.     

Morphology and electrophysiological properties of immunocytochemically identified rat dopamine neurons recorded in vitro

In vitro intracellular recordings were made from neurons in the rat midbrain slice. Two neuronal types could be distinguished in dopamine-containing (DA) midbrain regions based on electrophysiological criteria. One neuron type exhibited short duration action potentials (less than 1.5 msec), could fire at high frequencies (greater than 10 Hz), and exhibited either phasic or burst firing patterns. This neuron did not exhibit tyrosine hydroxylase immunoreactivity. A second neuronal type exhibited a unique set of electrophysiological properties, which included (1) a spontaneous pacemaker-like depolarizing potential, (2) a highly regular firing pattern, (3) long duration (greater than 2 msec) action potentials, and (4) a high (i.e., depolarized) spike threshold. This neuron was consistently double labeled using intracellular staining and immunocytochemical localization of the catecholamine-specific enzyme tyrosine hydroxylase, and thus represented the DA neuronal type. Midbrain DA neurons stained with Lucifer yellow could be separated into 3 classes based on their location and morphology: (1) fusiform neurons with laterally projecting dendrites in the dorsal substantia nigra zona compacta region, (2) multipolar cells with laterally and ventrally projecting dendrites in the ventral substantia nigra zona compacta, and (3) neurons with fusiform and multipolar somata and radially projecting dendrites in the ventral tegmental area. The dendrites also exhibited spine-like protrusions and ended with specialized forked processes. Spontaneously firing DA cells recorded in vitro had a number of distinguishing electrophysiological characteristics in common with those of DA neurons recorded in vivo, such as the presence of a slow depolarizing potential driving spike activity and a characteristic depolarized spike threshold (approximately-36 mV). However, in contrast to that found in vivo, the DA cells characterized here exhibited substantially higher input resistances and fired spontaneously in a very regular pacemaker pattern. Burst firing was not observed. Spike activity was apparently dependent on 4 depolarizing events: (1) a voltage-dependent TTX-sensitive slow depolarization, (2) a cobalt-sensitive low threshold depolarization that was activated during the rebound from brief membrane hyperpolarizations, (3) high threshold dendritic calcium spikes which gave rise to the spike afterhyperpolarization, and (4) a high threshold initial segment sodium spike. These depolarizations were modulated by several processes, including a 4-aminopyridine-insensitive delayed repolarization, an instantaneous and time-dependent anomalous rectifier, and an afterhyperpolarization. Although low threshold depolarizations and rebound action potentials could be triggered by the membrane repolarization following small membrane hyperpolarizations, comparatively larger hyperpolarizations attenuated this rebound activation, thereby suppressing anodal break excitation.(ABSTRACT TRUNCATED AT 400 WORDS)     

Electrophysiology of the Neurons in the Area of the Enkephalinergic Magnocellular Dorsal Nucleus of the Guinea-pig Hypothalamus, Studied by Intracellular and Whole-cell Recordings

The electrophysiological characteristics of 103 hypothalamic neurons in the area of the guinea-pig enkephalinergic magnocellular dorsal nucleus were studied in a thick slice preparation with sharp microelectrodes (63 neurons) and patch pipettes for whole-cell recordings (40 neurons). Of the sampled cells, 79.6% displayed tetrodotoxin-resistant, calcium-dependent slow-depolarizing potentials when the membrane potential was hyperpolarized to ?70 mV (type I neurons). Half of them showed robust slow depolarizing potentials, generating bursts of fast action potentials. In the remaining neurons, the slow-depolarizing potentials did not cause burst-firing action potentials but triggered single action potentials. The other class of neurons (20.4% of the sample: type II neurons) did not exhibit calcium-dependent slow-depolarizing potentials. Resting potential, input resistance and the membrane time constant did not distinguish among the two classes of neurons. Current-voltage relationships were heterogeneous. A transient outward rectification was observed in the two classes. This was not totally blocked by 2 mM 4-aminopyridine but was abolished when using perfusion with cobalt instead of calcium. Input resistance and the time constant were higher when measured in the whole-cell mode but the other electrical parameters and the sampling of the recorded neurons were strikingly similar between the two methods of recording. Intracellular staining of 22 neurons retrogradely labelled from the lateral septum allowed confirmation of their location within the magnocellular dorsal nucleus. The study indicates that the electrical properties of these neurons did not differ from those of neurons found throughout the area explored. It also indicates the presence of distinct electrophysiological types of cells in the magnocellular dorsal nucleus, although the nucleus is composed of a single type of enkephalinergic neuron. It provides a basis for the study of the regulation of activity of the neurons at the origin of an enkephalinergic tractus which is involved in neuroendocrine, psychoneuroendocrine and immune processes.     

Distinct classes of pyramidal cells exhibit mutually exclusive firing patterns in hippocampal area CA3b

It is thought that CA3 pyramidal neurons communicate mainly through bursts of spikes rather than so-called trains of regular firing action potentials. Reports of both burst firing and nonburst firing CA3 cells suggest that they may fire with more than one output pattern. With the use of whole-cell recording methods we studied the firing properties of rat hippocampal pyramidal neurons in vitro within the CA3b subregion and found three distinct types of firing patterns. Approximately 37% of cells were regular firing where spikes generated by minimal current injection (rheobase) were elicited with a short latency and with stronger current intensities trains of spikes exhibited spike frequency adaptation (SFA). Another 46% of neurons exhibited a delayed onset at rheobase with a weakly-adapting firing pattern upon stronger stimulation. The remaining 17% of cells showed a burst-firing pattern, though only elicited in response to strong current injection and spontaneous bursts were never observed. Control experiments indicated that the distinct firing patterns were not due to our particular slicing methods or recording techniques. Finally, computer modeling was used to identify how relative differences in K+ conductances, specifically K(C), K(M), and K(D), between cells contribute to the different characteristics of the three types of firing patterns observed experimentally.