Electrophysiologic properties of red nucleus neurons in the rat brain slices]

Intracellular recordings from the red nucleus (RN) neurons were made in experiments on the rat brain slices. Passive membrane properties (input resistance and membrane time constant) of the RN neurons were evaluated. Phenomena of potential-dependent rebound depolarization and time-dependent inward rectification were revealed by means of passing hyperpolarizing current pulses through the recorded cells. Injections of depolarizing currents caused repetitive firing of neurons with frequencies directly depending on the intensity of injected currents. Repetitive firing was also characterized by a fast frequency adaptation during injections of currents of different intensities. Stimulation of a region of slices presumably corresponding to the decussation of brachium conjunctivum evoked mainly monosynaptic EPSPs with a "fast"-rise time in the RN neurons, which suggests activation of the synaptic input from the cerebellar nucleus interpositus. Stimulation of the same region sometimes evoked EPSP-IPSP mixtures or pure IPSPs in the RN neurons.     

In vitro electrophysiology of rat subicular bursting neurons

Intracellular recordings were used to study the electrophysiological properties of rat subicular neurons in a brain slice preparation in vitro. Cells were classified as bursting neurons (n = 102) based on the firing pattern induced by depolarizing current pulses. The bursting response recorded at resting membrane potential (−66.1 ± 6.2 mV, mean ± SD n = 94) was made up of a cluster of fast action potentials riding on a slow depolarization and was followed by an afterhyperpolarization. Tonic firing occurred at a membrane potential of approximately −55 mV. A burst also occurred upon termination of a hyperpolarizing current pulse. Tetrodotoxin (TTX, 1 μM) blocked the burst and decreased or abolished the underlying slow depolarization. These effects were not induced by the concomitant application of the Ca²⁺ channel blockers Co²⁺ (2 mM) and Cd²⁺ (1 mM). Subicular bursting neurons displayed voltage- and time-dependent inward rectifications of the membrane during depolarizing and hyperpolarizing current pulses. The inward rectification in the depolarizing direction was abolished by TTX, while that in the hyperpolarizing direction was blocked by extracellular Cs⁺ (3 mM), but not modified by Ba²⁺ (0.5–1 mM), TTX, or Co²⁺ and Cd²⁺. Tetraethylammonium (10 mM)-sensitive, outward rectification became apparent in the presence of TTX. These results suggest that neurons in the rat subiculum can display voltage-dependent bursts of action potentials as well as membrane rectification in the depolarizing and hyperpolarizing directions. These results also indicate that activation of a voltage-gated Na⁺ conductance may be instrumental in the initiation of bursting activity. Hippocampus 7:48–57, 1997. © 1997 Wiley-Liss, Inc.     

Electrophysiological properties and their modulation by norepinephrine in the ambiguus neurons of the guinea pig

Electrophysiological properties of guinea pig ambiguus (AMB) neurons were studied in a brainstem slice preparation. During subthreshold depolarization AMB neurons displayed an early slow depolarization and a late outward rectification both of which were blocked by replacing Ca²⁺ with Co²⁺ in the extracellular solution. AMB neurons showed hyperpolarizing inward rectification which was blocked by extracellular Cs⁺ and is likely caused by the activation of Ih. In 58% (n = 49) of AMB neurons spike firing was restricted to the early phase of a long-lasting depolarizing current injection (phasic firing). The remaining AMB neurons showed repetitive firing throughout the depolarization (tonic firing). A Ca²⁺-mediated K⁺ current (I_K(Ca)) caused an afterhyperpolarization that followed both single and repetitive spike firing. I_K(Ca) also controlled the firing pattern in both types of firing, especially in the phasic firing. Norepinephrine (NE) blocked both the hyperpolarizing inward rectification and the Ca²⁺-dependent AHP. These effects of NE were antagonized by propranolol. It is proposed that the blockade of I_K(Ca) and Ih contribute to the improvement of the ‘signal-to-noise ratio’ by NE in AMB neurons.     

Changes during the postnatal development in physiological and anatomical characteristics of rat motoneurons studied in vitro

The postnatal maturation of rat brainstem (oculomotor and hypoglossal nuclei) and spinal motoneurons, based on data collected from in vitro studies, is reviewed here. Membrane input resistance diminishes with age, but to a greater extent for hypoglossal than for oculomotor motoneurons. The time constant of the membrane diminishes with age in a similar fashion for both oculomotor and hypoglossal motoneurons. The current required to reach threshold (rheobase) decreases in oculomotor motoneurons, in contrast with the increase observed in hypoglossal motoneurons. The depolarization voltage required to generate an action potential also diminishes in oculomotor motoneurons, whereas it remains constant in hypoglossal motoneurons. A membrane potential rectification (sag) appears in response to negative current steps, hyperpolarizing brainstem motoneurons more than 20 mV relative to the rest. This membrane response is more frequent in adult motoneurons. The durations of the action potential and its medium afterhyperpolarization (mAHP) decrease with postnatal development in all motoneurons studied, although the shortening of mAHP is more evident in oculomotor motoneurons. A rise in firing rate for all motoneurons with age is universal; this trend is also more pronounced in oculomotor motoneurons. Developing motoneurons exhibit a postinhibitory rebound depolarization that is capable of triggering an action potential or a short burst of spikes. This phenomenon is voltage-dependent and requires less of a membrane hyperpolarization to elicit an action potential in adult than in neonatal cells. In all developing brainstem and spinal motoneurons, the adult somal size is reached within the newborn period, although their dendrites continue to elongate. In summary, input resistance, time constant, and durations of action potential and mAHP decrease, while the frequency of sag and postinhibitory rebound, as well as the motoneuron firing rate and dendritic length, increase with postnatal age. These trends are universal to all the motoneuronal populations studied; however, the extent of these changes differs for each motoneuronal pool. A further distinction is evident in the inconsistent age-dependent change in rheobase and depolarization voltage for the two brainstem motoneuron nuclei.     

Synaptic and intrinsic properties of neurons of origin of the perforant path in layer II of the rat entorhinal cortex in vitro

Layer II of the entorhinal cortex (EC) provides the first step in the hippocampal trisynaptic loop via the perforant path projection to the dentate gyrus. While a great deal is known about this projection and the properties of the dentate granule cells, much less information is available concerning the properties of and synaptic inputs to the cells of origin of the pathway in layer II. The present experiments have employed a slice preparation of the rat EC to study the intrinsic membrane properties and synaptic organization of layer II neurons. Two types of neurons could be identified electrophysiologically. The majority were designated type I and displayed a pronounced time-dependent inward rectification in the hyperpolarizing direction. Type II displayed little evidence of this characteristic. However, morphological examination suggested that both types were spiny stellate neurons projecting via the perforant path. Synaptic responses of both types displayed evidence of excitatory inputs mediated by both N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptors. In general, however, at low frequencies the responses were dominated by inhibitory inputs mediated by both GABA_A and GABA_B receptors. At higher frequencies the bias was shifted much more toward excitation. The contribution of synaptic and intrinsic properties of layer II neurons to the processing capabilities of the EC is discussed. © 1994 Wiley-Liss, Inc.     

Morphological and electrophysiological properties of lateral entorhinal cortex layers II and III principal neurons

The intrinsic electrophysiology and morphology of neurons from layers II and III of the lateral entorhinal cortex (EC) was investigated in a rat brain slice preparation by intracellular recording and biocytin labeling. Morphologically, we distinguished three groups of layer II principal neurons. The most numerous group included cells with multiple radiating dendrites that spread over layers II and I in a fan-like fashion. While morphologically "fan" neurons were similar to the "stellate" cells of the medial EC, electrophysiologically the fan cells lacked the persistent rhythmic subthreshold oscillations and the very pronounced time-dependent inward rectification typical of the stellate cells. The second group consisted of pyramidal cells that manifested regular spike firing and had a more negative resting potential and a longer spike duration than the fan cells. In the third group we included all those neurons that had diverse multipolar appearances distinct from the fan cells. Neurons in this group had electrophysiological profiles intermediate between those of the fan and pyramidal cells. All neurons recorded in layer III were pyramidal in shape with a basal dendritic tree that could extend into layer V and an axon that could also give off collaterals into layer V. Electrophysiologically, layer III pyramidal cells were very similar to those of layer II. On the basis of these and other data we suggest that in different EC regions layer II neurons may be conducting more input-dependent specialized processing, while cells from layer III may perform a more global or generalized function.     

Inward Rectification (Ih) in Immunocytochemically-ldentified Vasopressin and Oxytocin Neurons of Guinea-Pig Supraoptic Nucleus

Intracellular recordings of magnocellular neurons from the supraoptic nucleus of guinea-pigs were made with KCI/K citrate- and biocytin-filled electrodes. Fifty of 99 cells exhibited a time-dependent inward rectification (TDR). The TDR was activated during hyperpolarizing current pulses to membrane potentials more hyperpolarized than −75 mV. In voltage-clamp recordings, an inward current appeared at voltage steps more hyperpolarized than −75 mV, with properties similar to the slow inward rectifier (I_h) described in other tissues. The I_h was blocked by 2 mM CsCI. BaCI₂ (100 to 500 μM) did not block the I_h. Immunocytochemical identification of the recorded cells revealed that both vasopressin (AVP)- and oxytocin (OT)- containing neurons exhibited an I_h.     

Mechanisms concerned in the direct effect of isoflurane on rat hippocampal and human neocortical neurons

The effect of isoflurane on postsynaptic neurons was studied by intracellular recordings from rat hippocampus and human neocortex in vitro. Isoflurane caused a hyperpolarization of the cell membrane. The hyperpolarization was reversed (although incompletely in some neurons) by increasing the membrane potential. The reversal potential was -80 +/- 12 mV (mean +/- S.D.) or 12 +/- 6 mV negative to the resting membrane potential. Potassium channel blockers reduced the isoflurane-induced hyperpolarization, while chloride loading was without effect. The transient depolarization preceding the hyperpolarization in some of the neurons was not reversed by hyperpolarization. The action potential was prolonged by 19 +/- 3% due to a slower rate of rise. The rise time was almost doubled. Firing threshold was increased by 4 +/- 3 mV (relative to the reference electrode). Subthreshold inward rectification was reduced or abolished. Some cells showed subthreshold outward rectification during isoflurane administration. These results suggest that isoflurane depressed neuronal excitability by (1) hyperpolarizing the cell membrane, at least partly by an increase in potassium conductance, (2) slowing the rate of rise of the action potential, presumably due to interference with the fast sodium channel, (3) decreasing subthreshold inward rectification and (4) increasing firing threshold.     

Electrical properties of facial motoneurons in brainstem slices from guinea pig

Electrical properties of guinea pig facial motoneurons (FMNs) were studied in a brainstem slice preparation. FMNs were identified histologically and by antidromic activation. They displayed time-varying responses and inward rectification during both subthreshold depolarization and hyperpolarization. The depolarizing rectification was caused by a persistent Na+ current (INaP); the Cs+-sensitive hyperpolarizing response had a different mechanism. Hyperpolarizing prepulses caused a 4-aminopyridine-sensitive delay of spike initiation. An evoked spike was followed by a fast- and a medium-duration hyperpolarization (the fAHP and mAHP, respectively). Blockade of Ca2+ influx abolished the mAHP without affecting spike duration, whereas spikes were prolonged and the fAHP was abolished by TEA or 4-AP. Adequate depolarization evoked tonic repetitive firing characterized by a steep F-I slope and fast adaptation. Abolition of the mAHP was associated with reduced fast adaptation and increased F-I slope, whereas the mAHP was enhanced and firing rate was slowed after TEA application. Three outward ionic currents were identified during voltage clamp: a rapidly inactivating current, a slowly inactivating current and a slow persistent Ca2+-mediated current (IK(Ca]. We conclude that spike repolarization and the fAHP are governed mainly by fast voltage-dependent currents, whereas progressive activation of IK(Ca) causes fast adaptation and, together with INaP, regulates firing rate.     

Electrophysiological properties of the rat area postrema neurons displaying both the transient outward current and the hyperpolarization-activated inward current

We found coexistence of the transient outward potassium current (I(TO)) and the hyperpolarization-activated inward current (I(H)) in 26 of 82 area postrema neurons tested using the whole-cell patch-clamp technique in rat brain slices. Cells displaying both the I(TO) and the I(H) typically showed "voltage sag" and "rebound potentials" in response to hyperpolarizing current injection and repetitive firing with strong adaptation was seen with depolarizing current injection. When cells were held at membrane potentials more negative than the resting level (e.g., -85mV), the afterhyperpolarization was enhanced. Voltage clamp recordings were performed to examine the characteristics of I(TO) and I(H) in and the contributions of these currents to the electroresponsiveness of area postrema cells. We show, in this study, the voltage-dependent properties of I(H) and I(TO), and how these currents modulate the intrinsic membrane properties of area postrema cells. We discuss the functional significance of the specific subset of area postrema neurons whose cells have both I(H) and I(TO) channels.     

Intracellular study of rat entopeduncular nucleus neurons in an in vitro slice preparation: electrical membrane properties

Electrical properties of rat entopeduncular nucleus (EP) neurons were studied in vitro using slice preparations. Of 108 EP neurons recorded, 104 were classified into two types based on their membrane properties. Type I neurons (n = 86) possessed: (1) a strong, time-dependent anomalous rectification that was sensitive to Cs⁺; (2) a weak spike adaptation; and (3) a strong rebound excitation with a low threshold Ca-spike and fast spikes. Many Type I neurons displayed spontaneous repetitive firing. Some of them generated spontaneous Ca-dependent plateau potentials with fast spikes upon application of tetraethylammonium bromide. Type II neurons (n = 18) had: (1) no apparent rectification; (2) a strong spike adaptation; and (3) a ramp-shaped repolarization, similar to the A-current, at the offset of a hyperpolarizing pulse. Features common to both types included: (1) a similar range of the input resistance; (2) capability of generating high threshold Ca-spike; and (3) generation of postactive hyperpolarizations (i.e. Ca-activated K-conductance). The great majority (Type I) of rat EP neurons share similar electrical properties. A minority of neurons (Type II) behave differently from Type I neurons and share similar properties among themselves.     

Negative slope conductance due to a persistent subthreshold sodium current in cat neocortical neurons in vitro

The voltage dependent ionic currents of large layer V neurons of cat sensory/motor cortex were examined in an in vitro slice preparation using a single-microelectrode voltage clamp. These cells exhibit a persistent inward current in a voltage range below spike threshold. This inward current is responsible for the increase of input resistance upon depolarization seen in these cells in response to a constant current pulse and is activated at the same voltages traversed by the membrane potential between spikes during rhythmic firing. The inward current appears to be a persistent sodium current, since it is unaffected by extracellular Ba2+ or Co2+ but is blocked by extracellular TTX or intracellular QX314.     

Hyperpolarization-activated (Ih) current in mouse vestibular primary neurons

The presence of a hyperpolarization-activated inward current (Ih) was investigated in mouse vestibular primary neurons using the whole-cell patch-clamp technique. In current-clamp configuration, injection of hyperpolarizing currents induced variations of membrane voltage with prominent time-dependent rectification increasing with current amplitudes. This effect was abolished by 2 mM Cs+ or 100 microM ZD7288. In voltage-clamp configuration, hyperpolarization pulses from -60 mV to -140 mV triggered a slow activating and non inactivating inward current that was sensitive to the two blockers, but insensitive to 5 mM Ba2+. Changing Na+ and K+ concentrations demonstrated that Ih current is carried by both these monovalent cations. This is the first demonstration of a Ih current in vestibular primary neurons.     

Voltage-dependent nonlinearities in the membrane of locust nonspiking local interneurons, and their significance for synaptic integration

Some aspects of the electronic and active membrane properties of nonspiking local interneurons were studied in isolated locust thoracic ganglia, using the switched current- and voltage-clamp techniques in neuropilar recordings. The average transmembrane potential (Vr) of the interneurons was -58 +/- 6mV (n = 85), and the input resistance (in the linear region of the current-voltage curve) was 16.5 +/- 8 M omega (n = 19, range 8 to 32 M omega). The membrane and equalizing time constants were estimated from charging curves evoked by the injection of low density hyperpolarizing current pulses from about -80 mV, i.e., from voltages in the linear region of the I-V curve. The curves yielded 2 time constants (tau m and tau l) whose average values were 33.2 +/- 9 msec and 3.3 +/- 1 msec (n = 18), respectively. The mean specific membrane resistance is therefore about 33 k omega.cm2, assuming that the membrane capacitance is ca. 1 microF.cm-2. An outward rectification was always observed upon depolarization from potentials more negative than Vr and was accompanied by a decrease in input resistance and membrane time constant. The "resting" membrane, for example, had a time constant of 26.4 +/- 8 msec (n = 31). This outward rectification was due to at least 2 conductances with different inactivation kinetics, similar to the transient "A" and "delayed-rectifier" potassium conductances. No inward rectification was ever observed upon injection of hyperpolarizing current. In about 60% of the recordings, an active and TTX-resistant depolarizing process could be evoked by rapid depolarization around Vr. The voltage-dependent properties of the membrane of the nonspiking local interneurons had dramatic effects on the shape and time course of natural or evoked unitary PSPs. The half-width of EPSPs, for example, decreased by a factor of 7.5 if the membrane potential was shifted from -93 to -50 mV. When the membrane potential of an interneuron was altered with a triangular current waveform, the reduction of tonically occurring IPSPs depended more on the sign and rate of the induced voltage change than on the absolute transmembrane potential. For 2 identical instantaneous values of membrane potential, for example, the reduction of the PSPs was greater during the depolarizing phase than during the hyperpolarizing phase of the current waveform. The possible nature of the active membrane conductances underlying the nonlinear electrical behavior of the membrane is discussed, together with their functional significance for local circuit synaptic integration.     

Electrophysiology of regular firing cells in the rat perirhinal cortex.

The electrophysiological properties of neurons in the rat perirhinal cortex were analyzed with intracellular recordings in an in vitro slice preparation. Cells included in this study (n = 59) had resting membrane potential (RMP) = -73.9 +/- 8.5 mV (mean +/- SD), action potential amplitude = 95.5 +/- 10.4 mV, input resistance = 36.1 +/- v 15.7 M omega, and time constant = 13.9 +/- 3.4 ms. When filled with neurobiotin (n = 27) they displayed a pyramidal shape with an apical dendrite and extensive basal dendritic tree. Injection of intracellular current pulses revealed: 1) a tetrodotoxin (TTX, 1 microM)-sensitive, inward rectification in the depolarizing direction (n = 6), and 2) a time- and voltage-dependent hyperpolarizing sag that was blocked by extracellular Cs+ (3 mM, n = 5) application. Prolonged (up to 3 s) depolarizing pulses made perirhinal cells discharge regular firing of fast action potentials that diminished over time in frequency and reached a steady level (i.e., adapted). Repetitive firing was followed by an afterhyperpolarization that was decreased, along with firing adaptation, by the Ca(2+)-channel blocker Co2+ (2 mM, n = 6). Action potential broadening became evident during repetitive firing. This behavior, which was more pronounced when larger pulses of depolarizing current were injected (and thus when repetitive firing attained higher rates), was markedly decreased by Co2+ application. Subthreshold membrane oscillations at 5-12 Hz became apparent when cells were depolarized by 10-20 mV from RMP, and action potential clusters appeared with further depolarization. Application of glutamatergic and GABAA receptor antagonists (n = 4), CO2+ (n = 6), or Cs+ (n = 5) did not prevent the occurrence of these oscillations that were abolished by TTX (n = 6). Our results show that pyramidal-like neurons in the perirhinal cortex are regular firing cells with electrophysiological features resembling those of other cortical pyramidal elements. The ability to generate subthreshold membrane oscillations may play a role in synaptic plasticity and thus in the mnemonic processes typical of this limbic structure.     

The physiological and morphological characteristics of interneurons caudal to the trigeminal motor nucleus in rats

In this study we have characterized the membrane properties and morphology of interneurons which lie between the caudal pole of the trigeminal motor nucleus and the rostral border of the facial motor nucleus. Previous studies suggest that many of these interneurons may participate in the genesis of rhythmical jaw movements. Saggital brainstem slices were taken from rats aged 5-8 days. Interneurons lying caudal to the trigeminal motor nucleus were visualized using near-infrared differential interference contrast (DIC) microscopy, and were recorded from using patch pipettes filled with a K-gluconate- and biocytin-based solution. The 127 neurons recorded could be categorized into three subtypes on the basis of their responses to injection of depolarizing current pulses, namely tonic firing (type I), burst firing (type II) and spike-adaptive (type III) neurons. Type I interneurons had a higher input resistance and a lower rheobase than type II neurons. All three neuron subtypes showed 'sag' of the voltage response to injection of large-amplitude hyperpolarizing current pulses, and, in addition, also showed rectification of the voltage response to injection of depolarizing current pulses, with type II neurons showing significantly greater rectification than type I neurons. The axonal arborizations were reconstructed for 44 of 63 neurons labelled with tracer. Neurons of each subtype were found to issue axon collaterals terminating in the brainstem nuclei, including the parvocellular reticular nucleus (PCRt), the trigeminal motor nucleus (Vmot), the supratrigeminal nucleus or the trigeminal mesencephalic nucleus. Twenty-five of the 43 neurons issued collaterals which terminated in the Vmot and the other brainstem nuclei. When viewed under 100x magnification, the collaterals of some interneurons were seen to give off varicosities and end-terminations which passed close to the somata of unidentified neurons in the trigeminal motor nucleus and in the area close to the interneuron soma itself. This suggests that the interneurons may make synaptic contacts both on motoneurons and also on nearby interneurons. These results provide data on the membrane properties of trigeminal interneurons and evidence for their synaptic connections both with nearby interneurons and also with motoneurons. Thus, the interneurons examined could play roles in the shaping, and possibly also in the generation, of rhythmical signals to trigeminal motoneurons.     

Intracellular labeling study of neurons in the superficial part of the magnocellular layer of the medullary dorsal horn of the rat

Morphology and electrical membrane properties of neurons in the superficial part of the magnocellular layer of the rat medullary dorsal horn (MDH: caudal subnucleus of the spinal trigeminal nucleus) were examined by using horizontal slice preparations. Intracellular recording and biocytin-injection combined with histochemical and immunohistochemical staining were done. Twenty-four neurons were examined successfully and classified into projection neurons (PNs) and intrinsic neurons (INs). The PNs were further divided into type I PNs (I-PNs) and type II PNs (II-PNs). The I-PNs sent axons to the medullary reticular formation; the II-PNs sent axons to the interpolar subnucleus of the spinal trigeminal nucleus but had no axons extending to the medullary reticular formation. The INs that sent no axons to the brain regions outside the MDH were also divided into small INs with spiny dendrites (INSSs) and large INs with aspiny dendrites (INLAs). The dendritic fields of the PNs extended to laminae I and II of the MDH and occasionally further to the spinal tract of the trigeminal nerve, whereas those of the INs were confined within the magnocellular layer of the MDH. The axonal branches of each IN formed a dense axonal mesh around the cell body of the parent neuron. Although the main bodies of the axonal fields of the INs were located in the magnocellular layer, some axonal branches extended to laminae I and II of the MDH. Immunoreactivity for NK1 receptor (substance P receptor) was found in approximately half of the PNs but not in the INs. Although no strong correlation was found between morphology and electrical membrane properties, there were some differences in electrical properties among the morphologically classified neuron groups, e.g., hyperpolarizing sag was observed in some PNs but not in the Ins; inward rectification was observed in some of the INSSs and INLAs but not in the PNs; the slow ramp depolarization and the slow afterdepolarization were observed in all INSSs examined but not in the PNs or INLAs.     

Heterogeneity in the Membrane Current Pattern of Identified Glial Cells in the Hippocampal Slice

Glial cells, acutely isolated or in tissue culture, have previously been shown to express a variety of voltage-gated channels. To resolve the question whether such channels are also expressed by glial cells in their normal cellular environment, we have applied the patch-clamp technique to study glial cells in hippocampal slices of 10–12-day-old mice. Based on the membrane current pattern, we distinguished four glial cell types. One was characterized by passive, symmetrical K⁺ currents activated in depolarizing and hyperpolarizing directions. A second population showed a similar current pattern, but with a marked decay of the current during the 50-ms voltage jumps. In a third population, the decaying passive currents were superimposed with a delayed rectifier outward current and, in some cases, with a slow inward current activated by depolarization. The fourth population expressed delayed rectifying outward currents, an inward rectifier K⁺ current and fast inward currents activated by depolarization. To unequivocally identify the glial cells we combined electrophysiological and ultrastructural characterizations. Therefore, cells were filled with the fluorescent dye lucifer yellow during characterization of their membrane currents, the fluorescence of the dye was used to convert diaminobenzidine to an electron-dense material, and subsequently slices were inspected in the electron microscope. Recordings were obtained from cells in the stratum radiatum and were identified as glial by their size, the characteristic chromatin distribution, and the lack of synaptic membrane specializations.     

Morphologic features and electrical membrane properties of projection neurons in the marginal layer of the medullary dorsal horn of the rat

Possible correspondence between morphologic features and electrical membrane properties of projection neurons in lamina I [the marginal zone (MZ)] of the caudal subnucleus of the spinal trigeminal nucleus [the medullary dorsal horn (MDH)] was examined by using intracellular recordings and biocytin-injections combined with histochemical and immunohistochemical staining techniques. The experiments were done in horizontal slice preparations of the rat brain. Thirteen MZ neurons were recorded stably and stained successfully. These neurons were confirmed to send their axons to the brain regions outside the MDH by camera lucida reconstruction. They were divided into two types on the basis of branching patterns of their axons within the MDH: Type I projection (P-I) neurons (n = 7 neurons) had main axons that rarely emitted axon collaterals within the MDH, whereas type II projection (P-II) neurons (n = 6 neurons) had main axons that emitted many axon collaterals within laminae I, II (substantia gelatinosa), and III (magnocellular part) of the MDH and also to the spinal tract of the trigeminal nerve; these axon collaterals usually constituted a dense mesh of axonal processes within laminae I and II of the MDH, especially in lamina II. About half of the neurons of each type showed immunoreactivity for the neurokinin-1 receptor. Resting membrane potentials were significantly more positive in P-I neurons than in P-II neurons. The P-II neurons had higher input resistance, a longer membrane time constant, and a higher threshold for spike than P-I neurons. In response to weak, long depolarizing current pulses, P-II neurons often showed slow ramp depolarization; the same neurons exhibited delayed repolarization to the resting potential (slow after depolarization) after the offset of the long depolarizing current pulses. Neither the slow-ramp depolarization nor the slow after depolarization was observed in P-I neurons. Slow return to resting membrane potential after offset of hyperpolarizing current pulses also was observed frequently in P-II neurons but not in P-I neurons. The results indicate that P-II neurons differ in their membrane properties compared with P-I neurons, and P-II neurons may be involved in the local circuit mechanism within the MDH more deeply than P-I neurons.     

Morphological and electrophysiological characteristics of layer V neurons of the rat medial entorhinal cortex

This study aimed to characterize the morphological and electrophysiological properties of neurons in layer V of the entorhinal cortex in the rat brain. Using the in vitro slice preparation and sharp electrode techniques, we recorded from layer V neurons located in the medial entorhinal cortex. Recorded cells were also labeled with biocytin. Based on morphological criteria, layer V of the entorhinal cortex is comprised of three categories of neurons: pyramidal cells, horizontal cells, and polymorphic cells. Horizontal cells could be easily distinguished from the pyramidal cells because the bulk of their dendritic plexus extended horizontally within layer V. Polymorphic cells vary in size and shape. Interestingly, they typically do not have apical dendrites, and some of them have dendrites that extend into the subiculum. Based on electrophysiological criteria alone, it was not possible to unequivocally distinguish the morphological cell types because they were somewhat heterogeneous with respect to several parameters including inward rectification, spike-frequency adaptation, and intrinsic oscillations. Nevertheless, although most horizontal cells displayed time-dependent inward rectification, most pyramidal cells displayed fast inward rectification exclusively. None of the entorhinal cortex layer V cells displayed oscillatory activity like that of neocortical layer V "bursting" cells, although neurons from all groups displayed rhythmic subthreshold membrane potential oscillations. In summary, we have found that layer V of the rat medial entorhinal cortex consists of three morphologically distinct neuronal subtypes that cannot be clearly distinguished from each other by traditional electrophysiological measures.