Electrophysiological evidence for axonal branching of ambiguous laryngeal motoneurons

Laryngeal motoneurons in the nucleus ambiguous (NA) were identified antidromically by stimulation of the ipsilateral superior laryngeal nerve (S) and/or the recurrent laryngeal nerve (R). In some NA motoneurons, antidromic spikes elicited by both S and R stimulation collided with the spontaneously occurring discharges. In the same neuron, spikes evoked antidromically by stimulation of one laryngeal nerve always collided with antidromic spikes elicited by stimulation of the other laryngeal nerve. Of 105 NA neurons activated by S and R stimulation, 36 neurons satisfied the criteria, and were classified as NA neurons with branching axons (branching NA (B-NA) neurons). Those neurons activated by either S or R stimulation but not both were classified as NA neurons without branching axons (unbranched NA (UB-NA) neurons). Mean antidromic latencies of B-NA neurons were 0.79 +/- 0.20 ms to S stimulation and 1.91 +/- 0.45 ms to R stimulation and those values for UB-NA neurons were 0.84 +/- 0.17 ms to S stimulation and 2.10 +/- 0.53 ms to R stimulation respectively. None of these mean values were significantly different from one another. Conduction time in the unbranched portion of the branching axon was estimated according to the equation reported by Anderson and Yoshida. The mean conduction time for 20 B-NA neurons was 0.45 +/- 0.35 ms. The branching point in B-NA neurons was estimated on the basis of the conduction time in the unbranched stem portion and those times in two branches of a branching axon measured electrophysiologically. The results suggest that the majority of B-NA neurons bifurcate within a half axonal length.(ABSTRACT TRUNCATED AT 250 WORDS)     

A new aspect of the collision test principle: cell body distance from recording site

A theoretical method is described for estimating the distance between a spike recording-site, possibly axonal, and the corresponding cell body of unknown location. The method requires that an orthodromic spike be recorded following an antidromic spike, with estimation of a collision interval analogous to that used for establishing antidromicity. To calculate the distance between recording-site and cell body, values are needed for the collision interval between antidromic and succeeding orthodromic spikes, the refractory period of the spike, and the antidromic conduction speed. Problems may arise in determining the last value. The method is illustrated with antidromic spikes recorded in the medial thalamus of the cat upon stimulating the caudate nucleus.     

Large identified pyramidal cells in macaque motor and premotor cortex exhibit "thin spikes": implications for cell type classification

Recent studies have suggested that extracellular recordings of putative cortical interneurons have briefer spikes than those of pyramidal neurons, providing a means of identifying cortical cell types in recordings from awake monkeys. To test this, we investigated the spike duration of antidromically identified pyramidal tract neurons (PTNs) recorded from primary motor (M1) or ventral premotor cortex (area F5) in 4 awake macaque monkeys. M1 antidromic latencies (ADLs) were skewed toward short ADLs (151 PTNs; 0.5-5.5 ms, median 1.1 ms) and significantly different from that of F5 ADLs (54 PTNs; 1.0-6.9 ms, median 2.6 ms). The duration of PTN spikes, recorded with a high-pass filter of 300 Hz and measured from the negative trough to the positive peak of the spike waveform, ranged from 0.15 to 0.71 ms. Importantly, we found a positive linear correlation between ADL and spike duration in both M1 (R(2) = 0.40, p < 0.001) and F5 (R(2) = 0.57, p < 0.001). Thus PTNs with the shortest ADL (fastest axons) had the briefest spikes, and since PTN soma size is correlated with axon size and conduction velocity, it is likely that the largest pyramidal neurons (Betz cells in M1) have spikes with short durations (0.15-0.45 ms), which overlap heavily with those reported for putative interneurons in previous studies in non-primates. In summary, one class of physiologically identified cortical pyramidal neuron exhibits a wide variety of spike durations and the results suggest that spike duration alone may not be a reliable indicator of cell type.     

In vivo Intracellular Recording of Neurons in the Supraoptic Nucleus of the Rat Hypothalamus

Intracellular recordings were made from cells in the hypothalamic supraoptic nucleus in the urethane-anaesthetized male rat using the ventral surgical approach. Impalements lasted from 5 min to 1 h and recorded cells had an input resistance of 55 to 170 megohms. Spikes of over 50 mV were recorded from 14 cells which could be antidromically activated by stimulation of the neural stalk. The spikes showed a hyperpolarizing afterpotential and the broadening characteristic of rapidly firing magnocellular neurons, which recovered rapidly (<200 ms). When depolarized, the cells showed evidence of a transient potassium current. Recurrent synaptic coupling between the recorded cell and adjacent cells would be expected to alter the hyperpolarizing afterpotential of an antidromic spike as compared with a spontaneous spike; no perceptible difference in the waveforms of the different types of spike could be detected in 11 spontaneously active cells. Application of just subthreshold stimuli to the neural stalk did not evoke depolarizing or hyperpolarizing potentials. Suprathreshold shocks to the neural stalk, when the antidromic spike was prevented by collision, also had no discernible effect on membrane potential. Thus intracellular recordings from magnocellular neurons in vivo revealed electrophysiological properties similar to those seen in vitro. No evidence for synaptic interconnection between magnocellular neurons was found in male rats.     

Rubrospinal tract of the cat: superposition of antidromic responses and changes in axonal excitability following orthodromic activity

Activity was recorded from rubrospinal neurons (RSNs) in anesthetized, paralyzed, artificially ventilated cats. Multiple-unit microelectrodes were used to simultaneously record the activity of neighboring RSNs. When antidromically activated, the RSNs responded forming 'stacks' of superimposed spikes. By using appropriate collision tests, it was found that the spikes forming a stack arose from different neurons. In addition, single extracellular and intracellular recordings were obtained from RSNs. The changes in the axonal excitability of rubrospinal axons were tested following synaptically evoked (by contralateral interpositus (IP) stimulation) and/or directly evoked (by injection of current through the intracellular electrode) action potentials at different postspike delays. Subthreshold stimuli for antidromic activation in absence of orthodromic activity were well suprathreshold for most fibers in a wide range of postspike delays. The supernormal axonal periods were longer-lasting when tested after synaptic spikes (up to an average delay of 100.4 ms; range, 10-500 ms) than after directly evoked spikes (mean delay, 78.8 ms; range, 10-296 ms). If synaptic stimulation fires more RSNs than direct stimulation, then the longer-lasting supernormal periods might be due to the activity of adjacent fibers. An additional increase in external potassium concentration in the vicinity of the axon would explain these results.     

Central vagal organization in rats: An electrophysiological study

The organization of the vagal nuclei was studied electrophysiologically in chloralose-anesthetized rats by analyzing the field potentials and unitary responses evoked in the nuclei by stimulation of the cervical vagus nerve. The rostral part of the nucleus commissuralis yielded only a long-latency response to stimulation of this nerve, suggesting that this region receives projections solely of nonmyelinated afferent fibers. In the nucleus tractus solitarius the stimulation elicited both short-latency and long-latency responses, indicating converging projections of myelinated and nonmyelinated afferents. A long-latency response was recorded diffusely within n. commissuralis and n. tractus solitarius of the contralateral side, whereas a short-latency response was restricted to a midline area, the caudal n. commissuralis, and the most medial part of n. tractus solitarius adjacent to it. These observations also suggest a difference in projections of myelinated and nonmyelinated afferents. Two types of motor neurons were identified in the dorsal vagal nucleus by antidromic activation: one with B-fiber axons and the other with C-fiber axons. C-Fiber motor neurons were characterized by the large positivity of the spike and the presence of an inflection in the rising phase of the spike, presumably between the initial segment and somatodendritic components. The latter component was readily blocked by repetitive stimulation. In the nucleus ambiguus, stimulation of the vagus nerve produced the earliest antidromic response of A-fiber motor neurons accompanied by multiple orthodromic responses of short and long latencies. Electrolytic lesions of the dorsomedial medulla oblongata abolished all potentials in n. ambiguus except the antidromic one, indicating that all the orthodromic responses were generated via the vagal sensory nuclei sinuated dorsomedially.     

Properties of antidromically activated callosal neurons and neurons responsive to callosal input in rabbit binocular cortex

Extracellular spikes were recorded from the cell bodies of antidromically activated callosal axons in the binocular visual cortex of unanesthetized, unparalyzed rabbits. Callosal axons were stimulated near their terminals in the contralateral cortex. Recordings were also obtained from neurons which responded synaptically to contralateral cortical stimulation. The primary method for differentiating antidromic from synaptic activation was the test for collision of impulses. Additional tests provided further confirmation of antidromic activation. Units which sent an axon across the corpus callosum (callosal neurons) were thereby distinguished from units which responded synaptically to callosal input. Eighteen percent of units sampled sent an axon across the corpus callosum. The median conduction velocity of callosal axons was less than 2 m/sec. An additional 18% of units encountered were synaptically activated by contralateral cortical stimulation. Callosal neurons were found to differ from synaptically activated units in three distinct ways. Callosal neurons had very low spontaneous firing rates (median =< 1.0 spike/sec), responded with a single spike to contralateral cortical stimulation and never responded to diffuse flash illumination. In contrast, most synaptically activated units demonstrated high spontaneous firing rates (median = 10.2 spikes/sec), responded with a burst of spikes to contralateral cortical stimulation and were also driven by diffuse flash illumination.     

Nucleus basalis neurons exhibit axonal branching with decreased impulse conduction velocity in rat cerebrocortex

Single neurons in the basal forebrain (nucleus basalis area) were antidromically activated from the frontal or parietal cortex in anesthetized rats. Wide ranges of antidromic latencies were observed overall, with frontal and parietal stimulation yielding values ranging from 1.0 to 26.0 ms and 1.6-24.0 ms, respectively. Individual neurons often exhibited multiple antidromic latencies, such that deeper sites of stimulation or greater stimulation amplitudes generally yielded discretely different, shorter latencies than more superficial sites or lower amplitudes of stimulation. Single neurons were also often driven from neighboring sites (1-2 mm apart) within the frontal cortex, but no cell was coactivated from both frontal and parietal cortices. Finally, patterns and rates of spontaneous activity varied markedly among these cortically projecting neurons, with some cells being non-spontaneous and others exhibiting tonic rates of 30-40 Hz. Impulse waveforms also differed among driven cells, from relatively low-amplitude, negative spikes to large-amplitude, entirely positive spikes in unfiltered signals. These results indicate that cortically projecting, putatively cholinergic neurons in the basal fore-brain form a physiologically heterogeneous population in terms of impulse conduction velocity, spontaneous discharge, and spike waveforms. Our finding of multiple antidromic latencies and driving from neighboring sites indicate that these fibers may be highly branched in local terminal fields, but that individual cells may project exclusively to a single cortical area. Faster conduction velocities for deep compared to superficial cortical stimulation sites imply that these fibers may become non-myelinated upon entering cortical terminal fields, or that they may become markedly thinner as they travel within the cortex. This system of cholinergic cortical afferents differs in many physiologic aspects from the other non-thalamic cortical input systems of catecholamine or indoleamine neurons.     

Antidromic activation of dorsal raphe neurons from neostriatum: Physiological characterization and effects of terminal autoreceptor activation

Three types of neurons, distinguished on the basis of their spontaneous firing rates and patterns, extracellularly recorded waveforms and responses to neostriatal stimulation, were observed in the dorsal raphe nucleus in urethane-anesthetized rats. Type 1 neurons (presumed to be serotonergic) fired spontaneously from 0.1 to 3 spikes/s in a regular pattern, with initial positive-going bi- or triphasic action potentials. Type 1 cells exhibited long-latency antidromic responses to neostriatal stimulation (mean +/- S.E.M. 24.9 +/- 0.3 ms) that sometimes occurred at discrete multiple latencies, and supernormal periods persisting up to 100 ms following spontaneous spikes. Type 2 cells fired spontaneously in an irregular, somewhat bursty pattern from 0 to 2 spikes/s with initial negative-going biphasic spikes, and were antidromically activated from neostriatal stimulation at shorter latencies than Type 1 cells (21.8 +/- 0.9 ms). Type 3 cells were characterized by initial positive-going biphasic waveforms and displayed a higher discharge rate (5-30 spikes/s) than Type 1 or Type 2 cells. Type 3 cells could not be antidromically activated from neostriatal stimulation. The relatively long conduction time to neostriatum of the Type 1 presumed serotonergic neuron is discussed with respect to previous interpretations of the synaptic action of serotonin in the neostriatum. In conjunction with these antidromic activation studies, the neurophysiological consequences of serotonergic terminal autoreceptor activation were examined by measuring changes in the excitability of serotonergic terminal fields in the neostriatum following administration of the serotonin autoreceptor agonist, 5-methoxy-N,N-dimethyltryptamine (5-MeODMT). The excitability of serotonergic terminal fields was decreased by intravenous injection of 40 micrograms/kg 5-MeODMT, and by infusion of 10-50 microM 5-MeODMT directly into the neostriatum. These results are interpreted from the perspective of mechanisms underlying autoreceptor-mediated regulation of serotonin release.     

Intracellular study of rat globus pallidus neurons: membrane properties and responses to neostriatal, subthalamic and nigral stimulation.

Physiological properties of globus pallidus (GP) neurons were studied intracellularly in anesthetized rats. More than 70% of the neurons exhibited continuous repetitive firing of 2-40 Hz, while others exhibited periodic burst firing or no firing. The repetitively firing neurons exhibited the following properties: spike accommodation; spike frequency adaptation; continuous firing with a frequency of about 100 Hz generated by intracellular current injections; fast anomalous rectification; ramp-shaped depolarization upon injection of depolarizing current; and post-active hyperpolarization. The burst firing neurons evoked a large depolarization with multiple spikes in response to depolarizing current, and a similar response was observed after the termination of hyperpolarizing current. The few neurons which did not fire spontaneous spikes exhibited strong spike accommodation when they were stimulated by current injections. The continuously firing neurons were antidromically activated by stimulation of the neostriatum (Str) (23 of 68), the subthalamic nucleus (STh) (55 of 75), and the substantia nigra (SN) (25 of 46). The antidromic latencies of the 3 stimulus sites were very similar (about 1 ms). None of the burst firing neurons were antidromically activated. Three non-firing neurons evoked antidromic responses only after Str stimulation. Only repetitively firing neurons evoked postsynaptic responses following stimulation of the Str and the STh. Stimulation of the Str evoked initial small EPSPs with latencies of 2-4 ms and strong, short duration IPSPs with latencies of 2-12 ms. Stimulation of the STh evoked short latency EPSPs overlapped with IPSPs. Frequently, these responses induced by Str and STh stimulation were followed by other EPSPs lasting 50-100 ms. These results indicated: (1) that the GP contains at least 3 electrophysiologically different types of neurons; (2) that GP projections to the Str, the STh, and the SN are of short latency pathways; (3) that Str stimulation evokes short latency EPSPs followed by IPSPs and late EPSPs in GP neurons; and (4) that STh stimulation evokes short latency EPSPs overlapped with short latency IPSPs and late EPSPs in GP neurons.     

Caudate evoked synaptic activities in nigral neurons

Intracellular recordings from neurons in the substantia nigra have revealed three varieties of monosynaptic PSPs in response to stimulation of the ipsilateral head of the caudate nucleus: short (3–5 msec) latency EPSPs; short (3–5 msec) latency IPSPs; and long (15–20 msec) latency IPSPs. The data indicate that at least three efferent fiber systems link the caudate to the nigra: two fast conducting axonal systems of comparable diameters mediate the short-latency PSPs whereas a slow conducting axonal system mediates the long-latency PSPs. The caudate evoked long-latency IPSPs in nigral neurons were preceded by antidromic and orthodromic potentials in the motor cortex; these cortical potentials are regarded as epiphenomena. The dual, facilitatory-inhibitory control of caudate on nigral neurons is consonant with the proposal that the caudate self-regulates its input from the nigra. The caudate-evoked EPSPs in nigral neurons are sine qua non for the operation of the caudato-nigrothalamic projection system.     

Reactions of neurons of the reticular and ventral anterior thalamic nuclei to electrical stimulation of the centrum medianum of the thalamus

Responses of 145 reticular (R) and 158 ventral anterior (VA) thalamic neurons to electrical stimulation of centrum medianum (CM) were studied in cats anaesthetized with thiopental sodium (30-40 mg/kg intraperitoneally) and immobilized with d-tubocurarine (1 mg/kg). 4.1% of R and 4.4% of VA neurons under study responded to CM stimulation by antidromic spike (latency 0.3-2.0 ms). The conduction velocity of antidromic excitation in axons of those neurons was found to be 1.7-7.6 m/s. There were neurons which responded by antidromic spike to the other thalamic nuclei stimulation as well as to CM. This fact is the electrophysiological proof of the axonal branching in these neurons. 53.8% of R and 46.9% of VA neurons responded to CM stimulation with orthodromic excitation. Two groups of cells were separated among neurons excited orthodromically. The first group neurons responded to CM stimulation by discharges composed of 6-12 spikes with frequency of 130-640 per second. The neurons of the second group generated a single spike. Inhibitory reactions were noticed only in 0.7% of R and in 4.4% of VA neurons. It is shown that afferent impulses from relay nuclei, lateral posterior nucleus and motor cortex converged to some R and VA neurons responding to CM.     

Direct projection of pause neurons to nystagmus-related excitatory burst neurons in the cat pontine reticular formation

Brain-stem pause neurons (PNs) are inhibitory neurons which cease their tonic firing about 20 ms prior to the quick phase of horizontal vestibular nystagmus in either direction. One group of nystagmus-related burst neurons just rostral to the abducens nucleus exhibits a burst of spikes before and during the quick phase to the ipsilateral side--excitatory burst neurons (EBNs). The present study supported the conclusion that PNs project to, and tonically inhibit EBNs during the slow phase and that the burst of activity of EBNs at the quick phase is partly caused by the abrupt release from pauser inhibition. The evidence leading to this conclusion is: simultaneous recording of PNs and EBNs showed close alternation of firing; PNs were antidromically activated from the EBN region; systematic microstimulation tracks within the EBN region showed an antidromic activation pattern of low threshold sites separated by high threshold sites consistent with PN axonal branching in the EBN region; during the nystagmus slow phase there were positive field potentials in the EBN region, followed by an abrupt negative deflection whose onset was synchronous with the last pauser spike; when single PN spikes were used to trigger averages of extracellular field potentials in the EBN region (postspike averaging), a consistent short-latency positivity was observed. This study shows an additional connection in the premotor neural network responsible for the generation of the quick phase of horizontal vestibular nystagmus.     

Mode of activation of thalamic neurons by stimulation of the cerebellar fastigal nuclei in the cat

In thalamic neurons of Nembutal-anesthetized cats, stimulation of the fast igial nucleus facilitated the antidromic invasion of cortically-evoked spikes over a considerable period and induced a spike train with a wide variability of latencies. Intracellular recordings suggested that excitatory inputs through polysynaptic routes should underlie these effects of fastigial stimulation on thalamic neurons.     

Differences in Penicillin-Induced Synchronous Bursts in the CA1 and CA3 Regions of the Hippocampus

Epileptiform field potentials were compared in the CA3 and CA1 regions of penicillin-treated hippocampal slices. The CA3 field bursts usually began with decrementing spike patterns similar to reported single-unit bursts, whereas spike amplitude gradually increased in spontaneous and long-latency CA1 bursts. Stimuli close to CA1 recording sites also evoked short-latency, decrementing CA1 responses. We postulate that these patterns reflect a more rapid recruitment of CA3 neurons into synchronous bursts and a gradual sequential activation of the CA1 neurons by Schaffer collateral input from CA3. Stimulation of stratum radiatum close to CA1 also produced long-latency "all-or-none" bursts in CA3 and then CA1, identical to spontaneous bursts and those produced by stimulation remote from CA1. At threshold, 76% of the latency to the CA1 burst occurred between the stimulus and the onset of the CA3 burst. The latency to the CA3 burst decreased with increasing stimulus intensity but the intervals from CA3 to CA1 bursts remained constant. Thus, long-latency CA1 bursts appear to be due to antidromic activation of CA3 followed by reexcitation of CA1.     

Reactions of neurons in the primary and secondary somatosensory zones of the cortex to stimulation of the ventral posterior nucleus of the thalamus]

Neuronal responses in the first and second somatosensory cortex (SI and SII) to stimulation of the ventroposterior nucleus of the thalamus (VP) were studied in experiments on cats immobilized with d-tubocurarine. 12.0% responding neurons in SI and 9.5% in SII were activated antidromically by VP stimulation. In the majority of antidromic responses the latencies did not exceed 1.0 ms. The minimal latency of orthodromic spikes was 1.5 ms in SI and 1.7 ms in SII. In SI the number of neurons whose orthodromic spike latencies did not exceed 3.0 ms was larger than neurons activated with latencies of 3.1-4.5 ms. In SII an inverse quantitative relationship between those two neuronal groups was observed. In SII a significantly larger number of neurons was excited with latencies of EPSPs ranged between 1.1-9.0 ms in SI and between 1.4-6.6 ms in SII and the latencies of IPSPs between 1.5-6.8 ms in SI and 2.2-9.4 ms in SII. The importance of different pathways for excitatory and inhibitory VP influences to neurons of SI and SII is discussed.     

Multiple axonal spike initiation zones in a motor neuron: serotonin activation.

The lateral gastric (LG) motor neuron of the stomatogastric nervous system of the crab Cancer borealis has a large soma in the stomatogastric ganglion (STG). The LG motor neuron makes inhibitory synaptic connections within the neuropil of the STG, and also projects to the periphery, where it innervates a series of muscles that control the movements of the lateral teeth of the gastric mill. The LG motor neuron has a spike initiation zone close to its neuropilar integrative regions, from which spikes propagate orthodromically to the muscles. Additionally, under certain conditions, the LG neuron can initiate spikes at peripheral axonal sites that can be 0.5-2.0 cm from the STG. Peripherally initiated spikes propagate antidromically into the STG and also propagate to the muscle. The peripheral spike initiation zones are often active in combined preparations in which the muscles are left attached. When the muscles are removed, depolarization of the LG soma together with 5-HT applied to the motor nerve also evokes peripheral spike initiation. At a given 5-HT concentration, the duration of the trains of antidromic spikes can be controlled by current injection into the soma, suggesting the presence of a slow voltage-dependent conductance in the LG axon. The antidromic spikes contribute to lengthening of the duration of contraction in some of the muscles innervated by the LG, but do not evoke IPSPs onto LG follower neurons. Thus, the LG neuron can send different signals to its peripheral and central targets.     

Responses of inferior salivatory neurons to stimulation of trigeminal sensory branches

A total of 84 single inferior salivatory neurons was identified by antidromic stimulation of the tympanic nerve. Their responsiveness was tested to stimulation of the ipsilateral infraorbital, lingual, and inferior alveolar nerves in urethane-chloralose-anesthetized cats. The conduction velocities of preganglionic fibers of inferior salivatory neurons ranged from 2.2 to 9.1 m/s, and 54% of those neurons responded with spikes to stimulation of at least one of the infraorbital, lingual, or inferior alveolar nerves (responsive type neurons). The latencies of spike responses to stimulation of the trigeminal sensory branches ranged from 4.0 to 21.0 ms, which were shorter than those of superior salivatory neurons. Impulses of both A-beta and A-delta afferent fibers of the trigeminal nerve were found to be effective for activation of inferior salivatory neurons. The convergence of excitatory inputs from more than one sensory nerve was found in most of the responsive type neurons (73%).     

The characteristics of the antidromic discharge of cat dorsal raphe neurons during repetitive activation

Slowly discharging neurons in the cat dorsal raphe could be classified into 3 types according to the behavior of antidromic spike discharges during repetitive stimulation of the medial forebrain bundle at 10 Hz. In the types 1 and 2, the latency of antidromic discharge was gradually prolonged to reach an asymptote, whereas no marked change occurred in the type 3. The type 2 neurons, which had a slower conduction velocity, showed a greater prolongation than the type 1 neurons. The maximum length of this prolongation was not significantly correlated with the initial latency. During 10 Hz stimulation some neurons showed repeatedly a conduction block after a sequence of initial decrease and later increase in latency. The spontaneous discharge was strongly suppressed during 10 Hz stimulation. During 1 Hz stimulation just after the cessation of 10 Hz stimulation, the prolonged antidromic latency was gradually restored in parallel with the recovery of the spontaneous discharge. Circumstantial evidences seem to be in favor of the idea that hyperpolarization of the axonal and somatic membranes is mainly responsible for the observed behavior of antidromic spikes of type 1 and 2 neurons.     

Monosynaptic and disynaptic activation of pyriform cortex neurons by synchronous lateral olfactory tract volleys in the rabbit

To elucidate the organization of synaptic inputs to pyriform cortex neurons, intracellular and extracellular responses of single units were analyzed in urethane-anesthetized rabbits. The lateral olfactory tract (LOT) or the olfactory bulb (OB) was electrically stimulated. Intracellular recordings revealed two types of cells (type I and type II cells), according to the types of EPSP evoked by the LOT or OB shock. The EPSP in the type I cells had shorter latencies (0.0 to 0.9 ms) from the onset of the component 2 (C2) wave of the field potential (which signals the onset of the synaptic depolarization of the apical dendrites of the pyramidal cells in the PC), and that in the type II cells had longer latencies (1.0 to 6.0 ms). A conditioning LOT or OB shock did not suppress the testing EPSP in the type I cells, whereas the conditioning stimulation greatly suppressed the testing EPSP in most of the type II cells. Extracellular recordings from units responding synaptically to the LOT or OB shock revealed a group of units which had short latencies (0.7 to 1.9 ms) of spike discharges. Those units, which were likely to be the same cells as the type I cells, are believed to mediate excitatory synaptic inputs to the type II cells. On the basis of these results, we concluded that type I cells are monosynaptically activated by LOT volleys, whereas type II cells are activated di- or polysynaptically by way of a relay from type I cells. The type I cells were recorded in both the superficial and the deep parts of the pyriform cortex, although they were recorded more frequently in the superficial part. On the other hand, most of the type II cells were recorded in the deep part of the PC. These results support and extend the previous model, in which the monosynaptically activated superficial pyramidal cells give rise to excitatory inputs to other pyramidal cells and neurons in deep layers.