Focal Synaptic Potentials Research Articles

Postnatal Development of a Segmental Switch Enables Corticospinal Tract Transmission to Spinal Forelimb Motor Circuits

Development of skilled movements and the corticospinal tract (CST) begin prenatally and continue postnatally. Because the CST is required for skilled movements in maturity, it is accepted that motor skills cannot occur until the CST develops a mature organization. We recently showed that the CST plays an essential role in postnatal development of interneurons comprising the spinal circuits it engages. We proposed that CST signals are more effectively transmitted to ventral motor circuits after interneuron maturation, thereby enabling expression of CST motor functions, suggesting development of a segmental switch promoting transmission. We tested this by recording CST-evoked focal synaptic potentials, extracellularly, in the cervical enlargement of cats before and after interneuron maturation [postnatal week 5 (PW5) to PW7]. We compared monosynaptic CST amplitude input to segmental circuits with oligosynaptic ventral horn responses, as a measure of CST-evoked segmental response transmission from input to output. The M1 primary motor cortex was unilaterally inactivated between PW5 and PW7 to determine activity dependence. CST interneuron contacts were identified using confocal microscopy. CST terminals contact diverse interneuron classes. CST stimulation strongly activated ventral motor circuits at the ages when both interneurons and CST spinal terminations have developed a mature phenotype, supporting development of segmental transmission of CST signals. CST activity blockade impeded development of effective segmental transmission by the inactivated CST and created a novel path for transmission from the ipsilateral, unaffected, CST. Our findings show that development of segmental CST signal transmission regulates nascent CST motor control functions and provide insight into systems-level mechanisms for protracted motor skill development.

Presynaptic control of transmission through group II muscle afferents in the midlumbar and sacral segments of the spinal cord is independent of corticospinal control

Transmission of information from the terminals group II muscle afferents is subject to potent presynaptic modulation by both segmental group II and cutaneous afferents and by descending monoaminergic systems. Currently it is unknown whether descending corticospinal fibres affect this transmission. Here we have examined whether corticospinal tract activation modulates the size of monosynaptic focal synaptic potentials (FSPs) evoked by group II muscle afferents, and the excitability of intraspinal terminals of group II afferents, both of which are indices used to show presynaptic control. Conditioning stimulation of corticospinal pathways had no effects on the sizes of group II evoked FSPs in the midlumbar or sacral segments at either dorsal horn or intermediate zone locations. These stimuli also had no effect on the excitability of single group II afferent terminals in the dorsal horn of the midlumbar segments. As positive controls, we verified that the corticospinal conditioning stimuli used did effectively depress FSPs evoked from cutaneous afferents recorded at the same spinal locations as the group II field potentials in all experiments. Corticospinal tract conditioning stimuli did not consistently enhance or reduce the depression of group II FSPs that was evoked by stimulation of ipsilateral segmental group II or cutaneous afferents; in the large majority of cases there was no effect. The results reveal that the control of transmission of information from group II afferents in these regions of the spinal cord is independent of direct corticospinal control.

Spinal projection of spindle afferents of the longissimus lumborum muscles of the cat

The connections and monosynaptic projections of muscle spindle afferents of individual heads of the longissimus lumborum have been studied in cats by natural stimulation, by electrical stimulation and by spike-triggered averaging from single identified afferents. The spindle afferents were classified by sensitivity to vibration and by the effect of succinylcholine on their response to ramp-and-hold muscle stretches. Axonal conduction and synaptic effects were recorded as field potentials and focal synaptic potentials during systematic exploration of the spinal cord in segments L1 to L4 with extracellular metal microelectrodes, singly and in linear arrays. Ascending branches of afferent axons within the cord had a significantly higher mean conduction velocity (CV: 56.5 m s(-1)) than descending branches (40.8 m s(-1)). The CV of ascending branches was significantly positively correlated with a measure of the strength of intrafusal bag(2) muscle fibre contacts, but not to a measure of bag(1) contacts. Two sites of monosynaptic excitatory projection in the cord were identified, namely to the intermediate region (laminae V, VI and VII) and to ventral horn region (laminae VIII and IX). In tests of 154 single afferents, signs of central projection were detected for 60, providing 122 regions of maximum negative focal synaptic potentials (FSPs) of mean amplitude 7.51 microV. Their longitudinal spacing indicated that axons gave off descending collaterals at intervals of 1.5-3.5 mm. Based on the amplitude of FSPs, the projection of secondary afferents is stronger than that of primaries in the intermediate region and possibly also in the ventral horn region. Evidence is also presented that spindle afferent input from different heads of the longissimus converges into any given spinal segment and that input in one spinal root projects to adjacent segments. It is concluded that the organization of the longissimus monosynaptic spindle input favours relatively tonic and diffuse stretch reflexes.

The influence of single VB thalamocortical impulses on barrel columns of rabbit somatosensory cortex.

Extracellular recordings were obtained from single neurons in ventrobasal (VB) thalamus of awake rabbits while field potentials were recorded at various depths within topographically aligned and nonaligned barrel columns of somatosensory cortex (S1). Spike-triggered averages of cortical field potentials were obtained following action potentials in thalamic neurons. Action potentials in a VB neuron elicited a cortical response within layer 4 with three distinct components. 1) A biphasic, initially positive response (latency <1 ms) was interpreted to reflect activation of the VB axon terminals (the AxTP). This response was not affected by infusion of an alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate receptor antagonist within the barrel. In contrast, later components of the response were completely eliminated and were interpreted to reflect focal synaptic potentials. 2) A negative potential [focal synaptic negativity (FSN)] occurred at a mean latency of 1.65 ms and lasted approximately 4 ms. This response had a rapid rise time ( approximately 0.7 ms) and was interpreted to reflect monosynaptic excitation. 3) The third component was a positive potential (the FSP), with a slow rise time and a half-amplitude duration of approximately 30 ms. The FSP showed a weak reversal in superficial cortical layers and was interpreted to reflect di/polysynaptic inhibition. The amplitudes of the AxTP, the FSN, and the FSP reached a peak near layer 4 and were highly attenuated in both superficial and deep cortical layers. All components were attenuated or absent when the cortical electrode was missaligned from the thalamic electrode by a single cortical barrel. Deconvolution procedures revealed that the autocorrelogram of the presynaptic VB neuron had very little influence on either the amplitude or duration of the AxTP or the FSN, and only a minor influence (mean, 11%) on the amplitude of the FSP. We conclude that individual VB thalamic impulses entering a cortical barrel engage both monosynaptic excitatory and di/polysynaptic inhibitory mechanisms. Putative inhibitory interneurons of an S1 barrel receive a highly divergent/convergent monosynaptic input from the topographically aligned VB barreloid, and this results in sharp synchrony among these interneurons. We suggest that single-fiber access to disynaptic inhibition is facilitated by this sharp synchrony, and that the FSP reflects a consequent synchronous wave of feed-forward inhibition within the S1 barrel.

Changes in the distribution of synaptic potentials from bulbospinal neurones following axotomy in cat thoracic spinal cord.

1. Plasticity in functional connections of expiratory bulbospinal neurones was investigated by measurement of terminal potentials (TPs) and focal synaptic potentials (FSPs), recorded with spike-triggered averaging in the thoracic spinal cord of anaesthetized, paralysed cats. These measurements were made in normal cats and in those which had previously been subjected to spinal cord lesions that transected the axons of the bulbospinal neurones in the segment below that under investigation, either about 2 weeks or about 16 weeks previously. 2. In both groups of operated animals bulbospinal neurones with firing properties and conduction velocities similar to normal were present. The extracellular recordings that were averaged to reveal TPs and FSPs were made on two standard grids, each consisting of eight sites spaced 0.25 mm apart on two electrode tracks. One grid was positioned at a rostral and one at a caudal location within one segment (T7-T9). 3. Tn the normal animals TPs and FSPs were larger and/or more common at rostral sites than at caudal sites, by a factor of about 1.7. In both 2 week and 16 week animals, TPs and FSPs were observed, both showing normal tine courses and latencies. At rostral sites in 2 week and 16 week animals the amplitudes and/or the frequency of occurrence of TPs and FSPs were similar to normal, as was the case fir caudal sites in the 2 week animals. However, at caudal sites in the 16 week animals the FSPs were mole common and/or significantly larger than normal, with the increase particularly marked on the lateral track, being equivalent to a factor of about 2. A corresponding increase in the amplitude and/or frequency of occurrence of TPs at caudal lateral sites was also seen, but was not significant. 4. The results are interpreted as evidence for short-range sprouting of the bulbospinal axons and the formation by them of new connections in the caudal parts of the segments concerned.

Field potentials generated by group II muscle afferents in the lower-lumbar segments of the feline spinal cord.

The actions of group II muscle afferents projecting to the lower-lumbar (L6 and L7) segments of the cat spinal cord were investigated by recording the cord dorsum and focal synaptic field potentials evoked by electrical stimulation of hindlimb muscle nerves. Cord dorsum potentials recorded over the lower-lumbar segments were generally much smaller than those produced by group II afferents terminating within the midlumbar and sacral segments. Only group II afferents of tibialis posterior produced potentials with an amplitude (mean maximal amplitude 39 microV, n = 7) approaching that of potentials over other segments. Focal synaptic potentials (mean maximal amplitudes 135-200 microV) were evoked by group II afferents of the following muscle nerves, listed in order of effectiveness: quadriceps, tibialis posterior (throughout L6 and L7), gastrocnemius soleus, flexor digitorum longus, posterior biceps-semitendinosus and popliteus (mainly within L7). Field potentials were recorded in the dorsal horn (laminae IV-V) and also more ventrally in a region which included the lateral part of the intermediate zone (lateral to the large group I intermediate field potentials) and often extended into the ventral horn (laminae V-VII). The latencies of the group II potentials are considered compatible with the monosynaptic actions of the fastest conducting group II muscle afferents. The results are compared with morphological evidence on the pattern of termination of group II muscle afferents in the lower-lumbar segments and with previous descriptions of the actions of group II muscle afferents in midlumbar and sacral segments.

Synaptic excitation in the thoracic spinal cord from expiratory bulbospinal neurones in the cat.

1. Synaptic actions in the thoracic spinal cord of individual expiratory bulbospinal neurones were studied in anaesthetized cats by the use of two techniques: (i) the monosynaptic connections to motoneurones were assessed by cross-correlations between the discharges of the neurones and efferent discharges in the internal intercostal nerves of several segments bilaterally; and (ii) distributions of terminal and focal synaptic potentials were measured by extracellular spike-triggered averaging in the thoracic ventral horn. 2. Monosynaptic connections were identified by both the durations and timings of observed cross-correlation peaks, taking into account accurate conduction velocity measurements derived from collision tests and from spike-triggered averaging. Discrimination was made against peaks resulting from presynaptic synchronization. 3. Monosynaptic connections to motoneurones were identified for twenty-three out of twenty-seven neurones. The connections to nerves on the side ipsilateral to the cell somata were, on average, about 36% of the strength of those on the contralateral side. The overall strength of the connections was about twice as strong as previous estimates for similar connections from inspiratory bulbospinal neurones to phrenic motoneurones. The monosynaptic pathway was calculated to be able to provide most of the depolarization for the motoneurones concerned and therefore was likely to be the main determinant of their firing patterns under the conditions of these experiments. 4. However, taking into account previous measurements it is considered possible that these connections may only involve a minority of motoneurones, perhaps only 10% of the expiratory population. Thus, in general, the control of the whole pool of expiratory motoneurones, despite the strong monosynaptic connections measured here, is suggested to be mainly dependent on spinal interneurones, as has been concluded previously for inspiratory motoneurones. 5. Spike-triggered averaging revealed that nearly all neurones gave signs of collaterals in each of the segments investigated (T7, T8 or T9), as shown by the presence of terminal potentials or focal synaptic potentials, but the projection within a given thoracic segment was non-uniform, in that large-amplitude potentials were more common in the rostral than the caudal part of the segment. This non-uniformity could be a factor involved in the apparently non-heterogeneous connections to the motoneurones.

Functional synaptic connections made by regenerated retinal ganglion cell axons in the superior colliculus of adult hamsters

Regenerated synaptic connections in the damaged mammalian visual system were studied in adult hamsters in which retinal ganglion cells (RGCs) regrew their axons through autologous peripheral nerve grafts directed from the stump of the transected optic nerve to the superior colliculus (SC). Unitary responses to illumination of small areas of the visual field were recorded within the superficial laminae of the reinnervated SC 23 to 60 weeks after grafting. Each element of a typical bursting response to light consisted of a terminal potential (TP) (half width 164 +/- 25 microseconds, amplitude up to 171 microV) arising from a regenerated RGC axon terminal arborization, followed at a latency of 268 +/- 63 microseconds by a longer duration negative focal synaptic potential (FSP) (half width 938 +/- 396 msec, amplitude up to 188 microV) reflecting EPSPs in neurons within the terminal field of the regenerated RGC axon. The FSP but not the TP was attenuated in a dose-dependent manner by iontophoretic application of GABA. In some cases spikes arose from FSPs after the first two or three impulses of a train, presumably reflecting summation of EPSPs to threshold for excitation in SC neurons contacted by the regenerated RGC axon terminals. Up to one-third of the area of the SC can be infiltrated by arborizations of the regenerated RGC axons that enter the SC through a nerve graft inserted in the lateral aspect of the SC. These experiments indicate that terminal arborizations of individual regenerated RGC axons can synapse with multiple neurons in the SC and that convergence of inputs from regenerated RGC axons is not required for activation of SC neurons in response to light.

Projection of cat jaw muscle spindle afferents related to intrafusal fibre influence.

1. A method of classification of muscle spindle afferents using succinylcholine (SCh) and ramp stretches has recently been described, which appears to estimate separately the strength of influence of bag1 (b1) and of bag2 (b2) intrafusal fibres. Increase in dynamic difference (delta DD) indicates b1 influence whilst increase in initial frequency (delta IF) indicates b2 influence. The significance of this classification has now been examined by correlation with the strength of synaptic projection of jaw muscle spindle afferents to the fifth motor nucleus (MotV) and the supratrigeminal region (STR) in anaesthetized cats. 2. Projection strength was estimated by computing the extracellular focal synaptic potential (FSP) from spike-triggered averages of 1024 sweeps at 100 microns intervals along tracks through STR and MotV. Trigger pulses were derived from spindle afferent cell bodies of the jaw-closer muscles recorded in the mesencephalic trigeminal nucleus, and characterized by the effect of SCh on their responses to ramp-and-hold stretches. 3. The maximum size of FSPs in tracks traversing STR and MotV ranged from 2.08 to 36.99 microV with a mean of 7.55 microV. The amplitudes were bimodally distributed into roughly equal-sized groups with high and low amplitude FSPs. 4. Mean values of delta IF were significantly greater for the group with large FSPs than for those with small FSPs. There were no significant differences in delta DD. FSP amplitude was significantly positively correlated with delta IF, but not with delta DD. 5. Spindle afferents with high values of FSP amplitude in MotV had a wide range of values of delta DD (b1b2c and b2c groups), while units with large FSPs in STR were all in the b2c category. Some evidence is presented to indicate that this reflects a preferential projection of secondary afferents to the STR. 6. For those units with projection to both STR and to MotV, there was a significant positive correlation between FSP amplitude in the two nuclei. 7. These results indicate that the extent of the b2 influence on spindle afferents predicts the central projection strength better than does the b1 influence. This finding is discussed from the viewpoint of possible developmental and functional issues.

Functional identities of thoracic respiratory interneurones in the cat.

1. Spike-triggered averaging was used to reveal focal synaptic potentials (FSPs) in the thoracic ventral horn resulting from impulses in individual respiratory interneurones situated in the contralateral ventral horn of the same segment (T6 or T7) in anaesthetized paralysed cats. 2. FSPs were of two types, negative-going or positive-going, and were of the same sign at each of several sites for each unit. These two types were interpreted as indicating excitatory or inhibitory actions respectively. 3. The positive-going FSPs were almost all the result of activity in the most strongly modulated, phasic respiratory interneurones, whereas most of the negative-going ones were derived from neurones with a continuous tonic component in their firing patterns. 4. Positive-going FSPs (particularly those from inspiratory neurones) were generally found at more ventral sites in the ventral horn than negative-going ones. 5. It is suggested that an important role for the phasically active interneurones is to provide phasic inhibition, which is combined with largely tonic excitation to produce strong respiratory modulation in the activity of other neurones in the thoracic spinal cord, particularly in motoneurones. However, some groups of respiratory interneurones, such as a group of tonic inspiratory ones which are located more dorsally than the phasic inspiratory ones, may not receive such phasic inhibition.

Contralateral projections of thoracic respiratory interneurones in the cat.

1. Retrograde neuronal transport following small iontophoretic injections of horseradish peroxidase was used to investigate the location of neurones projecting to the thoracic ventral horn of the cat. 2. A concentration of labelled neurones was seen in the contralateral medial ventral horn at the immediately opposite rostrocaudal position. 3. The participation of respiratory interneurones in the projection was investigated by means of spike-triggered averaging, as follows. 4. Spike trains of single interneurones whose firing patterns were related to the central respiratory cycle were recorded extracellularly in the thoracic ventral horn of anaesthetized, paralysed cats. Firing patterns were defined by the construction of cycle-triggered histograms. 5. Spike-triggered averaging of the signal from an extracellular tungsten microelectrode in the opposite ventral horn was performed to test for the presence of axonal, terminal or focal synaptic potentials. 6. At least one of these types of potential was found for 34/55 units. Terminal potentials were found for thirty-one units, accompanied by focal synaptic potentials for twenty-seven units. Potentials were found for units with all types of firing patterns. Units whose activity elicited these potentials were generally located in the medial half of the ventral horn. 7. We conclude that at least 60% of the respiratory interneurones project to the immediately opposite ventral horn.

Mechanisms of spinal reflex depressant effects of CS-722, a newly synthesized centrally acting muscle relaxant, in spinal rats

The mechanisms of the depressant action of ( R)-4-chloro-2-(2-hydroxy-3-morpholinopropyl)-5-phenyl-4-isoxazolin-3-one hydrochloride (CS-722), a newly synthesized centrally acting muscle relaxant, on spinal reflexes were investigated in spinal rats. The drug CS-722 (50 solmg/kg, i.v.) depressed the polysynaptic reflex but was less effective on the monosynaptic reflex. Eperisone-HCl (10 solmg/kg, i.v.) and baclofen (2 solmg/kg, i.v.) markedly decreased the monosynaptic and polysynaptic reflexes, with longer durations than CS-722; CS-722, eperisone and baclofen depressed the dorsal root reflex. The excitability of the motoneurone was reduced by CS-722 and eperisone. Excitability of the primary afferent fibres was reduced by CS-722, while eperisone and baclofen had no effect. Both CS-722 and eperisone did not have a depressant influence on the focal synaptic potential. These results suggest that CS-722 and eperisone but not baclofen, have a common motoneurone-membrane-stabilizing action and that this action may contribute, in part, to the spinal reflex depressant effects of CS-722 and eperisone.

Spinal distribution of extracellular field potentials generated by electrical stimulation of pudendal and perineal afferents in the cat.

Electrical stimulation of sensory pudendal and superficial perineal nerves evokes focal synaptic potentials produced by activation of spinal neurons in the lumbosacral gray matter in chloralose anesthetized or decerebrate cats. The field potentials evoked by sensory pudendal nerve stimulation were located in medial parts of laminae V and VI, and lamina X in the S1 to S3 spinal segments. The superficial perineal cutaneous field potentials partially overlapped with those produced by the pudendal nerve, but in general were localized more laterally in laminae V and VI. The central latencies of the earliest portion of the field potentials evoked by either sensory pudendal or superficial perineal nerves were less than 0.9 ms suggesting that monosynaptic activation of neurons contributed to the potentials.

Focal blockade of single unit synaptic transmission by iontophoresis of antagonists.

We have assessed the use of iontophoresis for investigating the pharmacology of synaptic transmission from individual presynaptic neurones in-vivo, by modifying extracellular focal synaptic potentials (FSPs) recorded by spike-triggered averaging. FSPs from two types of excitatory neurone (muscle spindle primary afferents and expiratory bulbospinal neurones) and from one inhibitory interneurone were studied in the thoracic ventral horn of anaesthetized cats. The antagonist of excitatory amino acids at non-NMDA receptors, DNQX, blocked the FSPs from the first two, as did bicuculline for the third. Thus the FSPs were generated by excitatory amino acids acting via non-NMDA receptors and GABA, respectively. The postsynaptic neurones were probably motoneurones. Merits and limitations of the method are discussed.

Monosynaptic excitatory connexions of reticulospinal neurones in the nucleus reticularis pontis caudalis with dorsal neck motoneurones in the cat

1. Projections of reticulospinal neurones (RSNs) in the nucleus reticularis pontis caudalis (N.r.p.c.) to dorsal neck motoneurones supplying splenius (SPL, lateral head flexor) and biventer cervicis and complexus (BCC, head elevator) muscles were studied in the cat anaesthetized with pentobarbiturate or alpha-chloralose. 2. Threshold mapping for evoking antidromic spikes revealed that most of RSNs tested projecting down to brachial segments but not to lumbar segments (C-RSNs) gave off collaterals to the gray matter of the upper spinal cord in C2-C3 segments. 3. Spike triggered averaging showed that negative field potentials were evoked after firing of a single C-RSN (single fibre focal synaptic potentials, FSPs) in the region of C2-C3 where large antidromic field potentials from nerves supplying SPL or BCC muscles were evoked. The single fibre FSPs ranged between 1 and 10 microV in amplitude and had latencies between 0.7 and 1.2 ms from the onset of the triggering spike. In most cases, a presynaptic spike preceded the negative potential by 0.3 ms. These results indicated that C-RSNs project to the SPL or BCC motor nucleus. 4. Spike triggered averaging of postsynaptic potentials revealed EPSPs (single fibre EPSPs) in 36 dorsal neck motoneurones, predominantly in SPL (25) and less in BCC (11) motoneurones, evoked from 15 C-RSNs. The amplitude of the single fibre EPSPs ranged from 5 to 310 microV, and had latencies of 0.8-2.0 ms from the onset of the triggering spikes of C-RSNs, or 0.3-0.5 ms from the presynaptic spike when recorded. The results indicated monosynaptic excitatory connexions of C-RSNs to dorsal neck motoneurones. 5. Single fibre EPSPs from a C-RSN were usually recorded from either BCC or SPL motoneurones but not from both types of motoneurones, when tested in many motoneurones. This showed that connexions of C-RSNs with dorsal neck motoneurones were muscle specific. 6. RSNs projecting down to the lumbar segment (L-RSN) also showed branching in C2-C3 segments. Excitatory monosynaptic connexion of L-RSNs with neck motoneurones were demonstrated by recording single fibre postsynaptic population potentials (p.s.p.p.s.) from the C2 ventral root perfused with sucrose. The probability of evoking monosynaptic single fibre p.s.p.p.s. was less (19%) than for C-RSNs (59%).

Ammonium decreases excitatory synaptic transmission in cat spinal cord in vivo.

1. Glutamine is thought to be a precursor of the pool of glutamate that is used as synaptic transmitter. NH4+ inhibits glutaminase, the enzyme presumed to cleave glutamine into glutamate in synaptic terminals. Therefore a decrease by NH4+ of excitatory synaptic transmission in hippocampus was suggested to be due to the inability to utilize glutamine as a precursor for glutamate and subsequent transmitter depletion. This study reexamines the effects of NH4+ on excitatory synaptic transmission. 2. The effects of NH4+ on excitatory synaptic transmission from low-threshold afferent fibers, presumably Ia-afferent fibers, to motoneurons was investigated in the spinal cord of anesthetized cats in vivo. 3. Action potentials of low-threshold afferent fibers were recorded at the entry of the dorsal roots into the spinal cord. An extracellular electrode within a motoneuron nucleus recorded the action potential of low-threshold afferent fibers and the extracellular monosynaptic excitatory postsynaptic potential, i.e., the focal synaptic potential (FSP). This extracellular electrode also recorded the antidromic field potential (AFP) in response to ventral root stimulation. Electrodes on the ventral roots recorded the monosynaptic reflex (MSR) and the monosynaptic excitatory postsynaptic potential in motoneurons electrotonically conducted into the ventral roots (VR-EPSP). 4. Intravenous infusion of ammonium acetate (AA) reversibly decreased MSR, VR-EPSP, and FSP, i.e., decreased excitatory synaptic transmission. 5. The decrease of VR-EPSP and FSP was accompanied initially by a decrease of conduction and, eventually, a conduction block in presynaptic terminals of low-threshold afferent fibers. 6. The decreases of VR-EPSP and FSP were also accompanied by the transient appearance of a reflex discharge, triggered by VR-EPSPs of decreased amplitude, and changes of the AFP indicating increased invasion of motoneuron somata by antidromic action potentials. 7. It is suggested that NH4+ depolarizes intraspinal Ia-afferent fibers and motoneurons. This depolarization initially decreases and then blocks conduction of action potentials into the presynaptic terminals of Ia-afferent fibers. The conduction block prevents the release of excitatory transmitter and decreases excitatory synaptic transmission. 8. The suggested depolarizing action of NH4+ may be due to K+-like ionic properties of NH4+ and/or an inhibition of K+-uptake into astrocytes. 9. The conduction block in presynaptic terminals of low-threshold afferent fibers can fully explain the decrease of excitatory synaptic transmission by NH4+. Because of the conduction block in presynaptic terminals, this study does not permit a conclusion as to an inhibition by NH4+ fo the utilization of glutamine as a precursor for glutamate used as synaptic transmitter.

Sites of termination and relay of pulmonary rapidly adapting receptors as studied by spike-triggered averaging

The sites of termination and relay of pulmonary rapidly-adapting receptors (RARs) were determined by averaging the extracellular field potentials produced in the nucleus tractus solitarii of the cat by individual RAR neurons. Action potentials of individual RARs were recorded extracellularly in the nodose ganglion during mechanical stimulation of the receptive field in the lung and used as triggers for the averaging. The averaged records (2000–4000 sweeps) revealed the presence of terminal and focal synaptic potentials. These potentials, indicating the presence of pre- and postsynaptic elements at the recording site, were found ipsilaterally in the caudal medial and commissural subnuclei and contralaterally in the commissural subnucleus.

Concentration of carbon dioxide, interstitial pH and synaptic transmission in hippocampal formation of the rat.

1. Interstitial pH (pHo) was measured with ion-selective microelectrodes in the fascia dentata of rats anaesthetized with urethane, while CO2 levels were controlled by varying pulmonary ventilation and CO2 content of inspired air. In the CA1 sector of hippocampal tissue slices in vitro pHo was similarly measured and altered by varying CO2 in the gas phase, or by adding HCl or NaOH to the artificial cerebrospinal fluid (ACSF) of the bath, or by changing the concentration of HCO3-. 2. Orthodromically evoked compound action potentials ('population spikes') were depressed in hypercapnia and increased in hypocapnia. In the fascia dentata of intact brains the population spike of the granule cells varied on average by more than 40% of control amplitude for each 0.1 change of pHo. In the CA1 zone of tissue slices in vitro, the change of population spike amplitude was approximately 30% per pH change of 0.1 caused by altered CO2 or HCO3- concentration, but only about 15% per pH change of 0.1 when HCl or NaOH were administered. 3. In anaesthetized rats the focal synaptic potential (FEPSP) evoked by a given stimulus intensity was weakly influenced by varying [CO2]; in tissue slices weak effects on FEPSP were inconsistent. In hippocampus both in situ and in vitro the population spike triggered by a given magnitude of FEPSP increased in hypocapnia and decreased in hypercapnia. This suggests that the main effect of CO2 is on the electric excitability of postsynaptic cells, with minor or no effect on transmitter release and on the interaction of the transmitter with its receptors. 4. Hypercapnia of anaesthetized rats was usually associated with a slight increase of [K+]o in the fascia dentata. Tissue [Ca2+]o changed little and not consistently. Neither of these two ions, nor concomitant changes of blood pressure or tissue partial pressure of oxygen, (Pt, O2), could account for the effects of pH on neuronal excitability. 5. The results show that increasing the extracellular concentration of H+ ions has a moderately depressant effect on the firing threshold of hippocampal neurones. The more powerful effects of elevated [CO2] and of lowered [HCO3-] may probably be explained by a direct effect on the neuronal membrane. The brain, by regulating breathing, controls its own excitability.

Focal synaptic potentials due to discrete mossy-fibre arrival volleys in the cerebellar cortex.

The previously described direct and relayed projections of periodontal afferents to the cerebellar cortex have been examined in detail by extracellular field-potential analysis. Advantage is taken of the very small temporal dispersion of the afferent volleys to permit identification of the presynaptic spike potential of mossy fibres, the subsequent synaptic potential and the firing of granule cells. Changes in form of the presynaptic potential with depth are compared with published descriptions of presynaptic potentials elsewhere. The negative synaptic potential in the granular layer is shown to have a positive aspect in the molecular layer. Granule-cell firing can, under some conditions, yield a population spike interrupting the synaptic potential wave. Records are presented showing all-or-none complex waves, which appear to be single glomerular potentials, not previously described in the mammalian cerebellum. Their distinction from cellular spike potentials is emphasized.

Distribution of activity in the spinal terminations of single hair follicle afferent fibers to somatotopically identified regions of the cat spinal cord

Single-unit spike-triggered averaging has been used to study the distribution of activity in the central processes of 11 single hair follicle afferent (HFA) fibers in relation to the somatotopic organization of dorsal horn neurons (DHNs). Central responses could be recorded from all but one HFA, and the waveforms of these responses were similar to the biphasic, monophasic positive and compound terminal potentials (TPs), the triphasic, positive-negative-positive axonal (preterminal) potentials (APs), and the focal synaptic potentials (FSPs) described by other workers in different preparations. No central responses could be recorded from one HFA even though this axon was shown to be intact throughout the experiment and noise levels in the averaged records were below 3.1 microV. The spatial organization of TPs and APs mirrored the anatomical organization determined using intraaxonal staining only in that the region containing these potentials was a longitudinally orientated narrow (mean width = 405 microns) strip of dorsal horn. Within this strip large TPs, APs, and FSPs were usually found only in those regions in which the receptive field (RF) of the HFA was relatively central to the RFs of DHNs. The region in which the RFs of DHNs encompassed the RF of the HFA, the somatotopically appropriate region, was also organized into a longitudinally orientated strip of dorsal horn of approximately the same width as the strip containing TPs and APs. In any single experiment the strips formed by the somatotopically appropriate region and the TP-AP region occupied the same mediolateral position, but in contrast, the rostral and/or caudal boundaries of these strips often occurred at different levels along the dorsal horn. In some cases the TP-AP strip extended rostrally and/or caudally beyond the somatotopically appropriate region or ended at the same rostrocaudal level. In other cases the somatotopically appropriate region extended rostrally and/or caudally beyond the TP-AP strip. These results are discussed in relation to the rostrocaudal spread of the dendritic trees and the RF organization of DHNs. The results show clearly that in intact cats, anesthetized with alpha-chloralose, some HFAs give rise to collaterals in somatotopically inappropriate regions of the dorsal horn and that at least some parts of these collaterals are invaded by incoming action potentials. The question of whether some HFAs give rise to collaterals that are either infrequently invaded or not invaded at all is discussed.