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Heterosynaptic changes accompany long-term but not short-term potentiation of the perforant path in the anaesthetized rat.

Brief high-frequency trains of electrical stimulation delivered to the perforant path result in long-term potentiation (l.t.p.) of field potentials recorded extracellularly from granule cells of the dentate gyrus. L.t.p. of the population spike is often disproportionately greater than l.t.p. of the population excitatory post-synaptic potential (e.p.s.p.). We have investigated the basis of this effect in rats anaesthetized with sodium pentobarbitone. A series of graded stimuli were given before and after tetanization of the perforant path. From data obtained in this way, we plotted stimulus-response curves, and the relation (E-S curve) between the slope of the population e.p.s.p. (E) and the amplitude of the population spike (S). Curves relating spike onset latency to the slope of the e.p.s.p. were also constructed. Tetanization of the combined medial and lateral components of the perforant path led to long-term changes in the relation between the e.p.s.p. and the population spike. For a given e.p.s.p., the corresponding population spike was of greater amplitude and earlier onset. This E-S potentiation was marked by a shift to the left of the E-S amplitude curve and a downward displacement of the E-S latency curve. Tetanization of the lateral component of the perforant path had two long-term effects on responses evoked by test stimuli to the untetanized medial component: (1) long-term depression of the medial e.p.s.p. and (2) long-term E-S potentiation. The net result of these two heterosynaptically induced effects was to leave unaltered information transfer across medial perforant path-granule cell synapses; for a given test volley the e.p.s.p. was smaller, but because of E-S potentiation the population spike remained relatively unaffected. Short-term potentiation, which has a time course of only a few minutes and is presumed to be mediated by presynaptic mechanisms, was not accompanied by E-S potentiation or by corresponding changes in spike latency. Possible mechanisms of long-term heterosynaptic depression of the e.p.s.p. and of homo- and heterosynaptic E-S potentiation, are discussed. We conclude that although these effects probably reflect a generalized post-synaptic change, this change is unlikely to be a prolonged reduction in the membrane potential of granule cells.

Action Potentials↗

Activity patterns and synaptic organization of ventrally located interneurons in the embryonic chick spinal cord.

To investigate the origin of spontaneous activity in developing spinal networks, we examined the activity patterns and synaptic organization of ventrally located lumbosacral interneurons, including those whose axons project into the ventrolateral funiculus (VLF), in embryonic day 9 (E9)-E12 chick embryos. During spontaneous episodes, rhythmic synaptic potentials were recorded from the VLF and from spinal interneurons that were synchronized, cycle by cycle, with rhythmic ventral root potentials. At the beginning of an episode, ventral root potentials started before the VLF discharge and the firing of individual interneurons. However, pharmacological blockade of recurrent motoneuron collaterals did not prevent or substantially delay interneuron recruitment during spontaneous episodes. The synaptic connections of interneurons were examined by stimulating the VLF and recording the potentials evoked in the ventral roots, in the VLF, or in individual interneurons. Low-intensity stimulation of the VLF evoked a short-latency depolarizing potential in the ventral roots, or in interneurons, that was probably mediated mono- or disynaptically. At higher intensities, long-latency responses were recruited in a highly nonlinear manner, eventually culminating in the activation of an episode. VLF-evoked potentials were reversibly blocked by extracellular Co2+, indicating that they were mediated by chemical synaptic transmission. Collectively, these findings indicate that ventral interneurons are rhythmically active, project to motoneurons, and are likely to be interconnected by recurrent excitatory synaptic connections. This pattern of organization may explain the synchronous activation of spinal neurons and the regenerative activation of spinal networks when provided with a suprathreshold stimulus.

Activity Cycles↗

Morphological and electrophysiological characterization of layer III cells of the medial entorhinal cortex of the rat.

Entorhinal cortex layer III cells send their axons into hippocampal area CA1, forming the less well studied branch of the perforant path. Using electrophysiological and morphological techniques within a slice preparation, we can classify medial entorhinal cortex layer III cells into four different types. Type 1 and 2 cells were projection cells. Type 1 cells fired regularly and possessed high input resistances and long membrane time constants. Electrical stimulation of the lateral entorhinal cortex revealed a strong excitation by both N-methyl-D-aspartate and non-N-methyl-D-aspartate receptor-mediated excitatory postsynaptic potentials. Type 2 cells accommodated strongly, had lower input resistances, faster time constants and featured prominent synaptic inhibition. Type 1 and 2 cells responded to repetitive synaptic stimulation with a prolonged hyperpolarization. We identified the two other, presumed local circuit, cell types whose axons remained within the entorhinal cortex. Type 3 cells were regular firing, had high input resistances and slow membrane time constants, while type 4 cells fired at higher frequencies and possessed a faster time constant and lower input resistance than type 3 neurons. Type 3 cells presented long-lasting excitatory synaptic potentials. Type 4 neurons were the only ones with different responses to stimulation from different sites. Upon lateral entorhinal cortex stimulation they responded with an excitatory postsynaptic potential, while a monosynaptic inhibitory postsynaptic potential was evoked from deep layer stimulation. In contrast to type 1 and 2 neurons, none of the local circuit cells could be antidromically activated from deep layers, and prolonged hyperpolarizations following synaptic repetitive stimulation were also absent in these cells. Together, the complementing morphology and the electrophysiological characteristics of all the cells can provide the controlled flexibility required during the transfer of cortical information to the hippocampus.

2-Amino-5-phosphonovalerate↗

Early nerve-muscle synapses in vitro release transmitter over postsynaptic membrane having low acetylcholine sensitivity.

Functional nerve-muscle synapses form rapidly in cultures of embryonic chicken spinal cord and muscle cells. As early as 30 min after nerve processes first contact muscle fibers they are able to release stimulus-evoked neurotransmitter. This release was detected only after wave-form averaging because of the exceedingly low amplitude of the synaptic potentials. This small size was likely due to a postsynaptic effect, because the electrophysiologically assayed acetylcholine sensitivity of the synaptic muscle membrane was low and did not differ significantly from extrasynaptic levels. Transmitter release was elicited both from along the lengths of nerve processes and from active growth cones.

Acetylcholine↗

Age-dependent long-term potentiation of inhibitory synaptic transmission in rat visual cortex.

Long-term potentiation (LTP) of inhibitory synaptic transmission in rat visual cortex was investigated using intracellular recording in slice preparations. The inhibitory postsynaptic potential (IPSP) of layer V cells evoked monosynaptically by layer IV stimulation was recorded while excitatory postsynaptic potentials (EPSPs) were blocked by adding NMDA and non-NMDA receptor antagonists to the perfusate. Separate presynaptic fiber groups were activated by two stimulating electrodes placed in layer IV. One electrode was used to test the effect of conditioning stimulation; the other served as a control. Trains of high-frequency stimulation (50 Hz, 1 sec) were applied at an interval of 10 sec as a conditioning stimulation. The LTP of IPSP induced by the conditioning stimulation had properties similar to those known for LTP of EPSP in hippocampal CA1: (1) it was specific to the conditioned pathway; (2) repetitive application of the conditioning stimulation saturated the LTP; (3) LTP was induced by conditioning stimulation at intensities higher than a threshold; (4) weak conditioning stimulation to one pathway, which alone failed to induce LTP, induced associative LTP when strong conditioning stimulation was simultaneously applied to the other pathway; (5) weak conditioning stimulation sometimes induced short-term potentiation. In contrast to these similarities, LTP of IPSP was induced by a wider range of stimulus frequencies than that capable of generating LTP of EPSPs in CA1. It was induced not only by a brief high-frequency stimulation but also by an intermediate-frequency (2 Hz) stimulation continued for 5 min. In addition, the LTP induction was not dependent on postsynaptic membrane potential. Finally, the LTP of IPSP was more easily induced in developing than in mature rats. This age dependence suggests that LTP of IPSPs may contribute to the improvement of response selectivity of visual cortical cells through visual experience during postnatal development.

Aging↗

Evidence for a serotonin-mediated slow excitatory potential in the guinea-pig coeliac ganglia.

The nature of the putative transmitter(s) mediating the non-cholinergic excitatory post-synaptic potential (e.p.s.p.) described in the preceding paper was investigated by means of electrophysiological, pharmacological and immunohistochemical methods. Serotonin (1-10 microM) when applied by superfusion caused a slow depolarization that closely mimicked the synaptic response in about 60% of the coeliac neurones that exhibited a non-cholinergic e.p.s.p. The serotonin depolarization evoked in low-Ca2+, high-Mg2+ solution or in a Krebs solution containing cholinergic antagonists was quantitatively similar to that elicited in normal Krebs solution. When compared in the same neurones the membrane resistance change during the course of the serotonin depolarization and of the non-cholinergic e.p.s.p., as well as their respective responses to conditioning polarization, were similar. The non-cholinergic e.p.s.p. was reversibly abolished during serotonin-induced depolarization; the blockade persisted when the membrane potential was restored to the resting level by hyperpolarizing current. The serotonin depolarization as well as the non-cholinergic e.p.s.p. were reversibly suppressed by cyproheptadine (20-50 microM), a serotonin antagonist, and enhanced by fluoxetine (30-50 microM), a serotonin reuptake inhibitor. On the other hand, pre-treating the ganglia with L-tryptophan (50 microM), a precursor of serotonin, preferentially augmented the synaptically induced response. A portion of the neurones (15%) were depolarized by substance P (1 microM) which also reversibly desensitized the non-cholinergic e.p.s.p. elicited in these neurones. The remaining neurones (25%) were insensitive to either serotonin or substance P, and the non-cholinergic e.p.s.p.s elicited in these cells were likewise not appreciably affected by these two agents. Furthermore, cyproheptadine, fluoxetine and L-tryptophan had no significant effect on the non-cholinergic e.p.s.p.s elicited in serotonin-insensitive neurones. Using the immunohistofluorescent techniques, dense but unevenly distributed serotonin immunoreactive nerve fibres could be observed surrounding many coeliac neurones. Immunoreactivity was not observed in the ganglia incubated with antisera pre-absorbed with excess serotonin. Collectively our results suggest that serotonin is the mediator of non-cholinergic e.p.s.p.s. elicited in about 60% of coeliac neurones sampled in this study, and that in the remaining neurones the slow depolarization may be generated by substance P and/or some unknown transmitter(s).

Action Potentials↗

Multiple mechanisms for the potentiation of AMPA receptor-mediated transmission by alpha-Ca2+/calmodulin-dependent protein kinase II.

Some forms of activity-dependent synaptic potentiation require the activation of postsynaptic Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). Activation of CaMKII has been shown to phosphorylate the glutamate receptor 1 subunit of the AMPA receptor (AMPAR), thereby affecting some of the properties of the receptor. Here, a recombinant, constitutively active form of alphaCaMKII tagged with the fluorescent marker green fluorescent protein (GFP) [alphaCaMKII(1-290)-enhanced GFP (EGFP)] was expressed in CA1 pyramidal neurons from hippocampal slices. The changes in glutamatergic transmission onto these cells were analyzed. AMPA but not NMDA receptor-mediated EPSCs were specifically potentiated in infected compared with nearby noninfected neurons. This potentiation was associated with a reduction in the proportion of synapses devoid of AMPARs. In addition, expression of alphaCaMKII(1-290)-EGFP increased the quantal size of AMPAR-mediated responses. This effect reflected, at least in part, an increased unitary conductance of the channels underlying the EPSCs. These results reveal that several key features of long-term potentiation of hippocampal glutamatergic synapses are reproduced by the sole activity of alphaCaMKII.

Animals↗

Nickel induces oscillatory behavior and enhanced synaptic and electrotonic transmission between stomatogastric neurons of Panulirus interruptus.

The pyloric pattern generator network of the stomatogastric ganglion uses a mixture of burst-inducing plateau potentials, synaptic transmission, and electrical coupling to produce its patterned output. This study examines the effects of two divalent, calcium channel blockers, nickel and cadmium, on voltage oscillations, synaptic transmission, and electrical coupling between the two pyloric dilator (PD) neurons and lateral pyloric (LP) neuron of Panulirus interruptus. The in vitro stomatogastric ganglion was bathed in saline containing tetrodotoxin (TTX) to eliminate Na-spikes and the spontaneous voltage oscillations of the pyloric rhythm, resulting in a steady resting potential. Addition of 50-100 microM Ni2+ to the TTX-saline induced voltage oscillations of similar amplitude and frequency as the endogenous rhythmic activity (before the application of TTX). 25-50 microM nickel enhanced graded synaptic transmission and electrical coupling and altered voltage waveforms, while producing little change in the input resistance measured in the soma. 10-1000 microM Cd2+ acted as a dose-dependent blocker of graded synaptic transmission, but had no other detectable effects. We propose that nickel, in contrast to cadmium, exerts a modulator-like effect deep in the pyloric neuropil.

Animals↗

Spontaneous network activity transiently depresses synaptic transmission in the embryonic chick spinal cord.

We examined the effects of spontaneous or evoked episodes of rhythmic activity on synaptic transmission in several spinal pathways of embryonic day 9-12 chick embryos. We compared the amplitude of synaptic potentials evoked by stimulation of the ventrolateral funiculus (VLF), the dorsal or ventral roots, before and after episodes of activity. With the exception of the short-latency responses evoked by dorsal root stimulation, the potentials were briefly potentiated and then reduced for several minutes after an episode of rhythmic activity. Their amplitude progressively recovered in the interval between successive episodes. The lack of post-episode depression in the short-latency component of the dorsal root evoked responses is probably attributable to the absence of firing in cut muscle afferents during an episode of activity. The post-episode depression of VLF-evoked potentials was mimicked by prolonged stimulation of the VLF, subthreshold for an episode of activity. By contrast, antidromically induced motoneuron firing and the accompanying calcium entry did not depress VLF-evoked potentials recorded from the stimulated ventral root. In addition, post-episode depression of VLF-evoked synaptic currents was observed in voltage-clamped spinal neurons. Collectively, these findings suggest that somatic postsynaptic activity and calcium entry are not required for the depression. We propose instead that the mechanism may involve a form of long-lasting activity-induced synaptic depression, possibly a combination of transmitter depletion and ligand-induced changes in the postsynaptic current accompanying transmitter release. This activity-dependent depression appears to be an important mechanism underlying the occurrence of spontaneous activity in developing spinal networks.

Animals↗

Reorganization of corticorubral synapses following cross-innervation of flexor and extensor nerves of adult cat: a quantitative electron microscopic study.

A quantitative electron microscopic study of corticorubral synapses was performed in the red nucleus (RN) of adult cats to determine the morphological correlates for the changes in time course of corticorubral excitatory post-synaptic potentials, which occur following cross-innervation of forelimb extensor and flexor nerves. Corticorubral synaptic endings were identified by anterograde degeneration after lesions of the ipsilateral sensorimotor cortex. Rubrospinal neurons innervating upper spinal segments were electrophysiologically identified and filled with horseradish peroxidase (HRP). These cells were mainly situated in the dorsomedial part of RN. Electron micrographs of the degenerating corticorubral synaptic endings were taken in the region surrounding HRP-filled neurons and the diameter of the dendrites contacted by such terminals was measured. In the cross-innervated animals many degenerating terminals were found to synapse on dendrites with large diameter and the somata of neurons in RN. This is in contrast to the previous observations in normal cats, in which very few corticorubral synapses were found to synapse on proximal dendrites and somata of RN neurons. The diameter of HRP-filled neurons in cats which were cross-innervated was slightly smaller than those observed in normal animals. These results indicate that new corticorubral synapses were formed on proximal dendrites and somata of RN neurons as a consequence of cross-innervation.

Animals↗

Heterosynaptic facilitation and post-tetanic potentiation in Aplysia nervous system.

1. Heterosynaptic facilitation was defined as an increase of amplitude of a test excitatory post-synaptic potential (EPSP) after the activation of a pathway (heterosynaptic pathway) different from that which produced the test EPSP. This phenomenon has been studied in Aplysia central nervous system under conditions which excluded the participation of post-tetanic potentiation.2. A unitary test was produced in the left and right giant cells, by indirect stimulation of an interneurone located in the peri-oesophageal ring.3. During heterosynaptic stimulation, orthodromic and antidromic activation of the test interneurone was prevented by (1) isolating the synaptic afferent region of the test interneurone from the tested synapse on the right giant cell by a sucrose block applied on the left pleurovisceral connective, and (2) using physiological stimulation of a piece of skin as a heterosynaptic stimulus. Under these conditions which prevented any firing in the test interneurone, heterosynaptic facilitation is observed as a 200% increase of amplitude of the test EPSP in the right cell which lasted more than 15 min. When instead of the physiological stimulus a supramaximal electrical stimulation of the nerves afferent to the abdominal ganglion was used, the increase of amplitude of the test EPSP could reach as much as 500% of its original amplitude. The effectiveness of such heterosynaptic stimulus was smaller when it was applied in the absence of a block of the left pleuro-visceral connective.4. It was possible to produce heterosynaptic facilitation when the preparation was cooled to 7-9 degrees C or if Li(+) replaced Na(+) in the medium. Both of these changes suppressed post-tetanic potentiation.5. It was concluded that heterosynaptic facilitation is a phenomenon different from post-tetanic potentiation. Heterosynaptic facilitation is similar to heterosynaptic inhibition seen in other cells in the same preparation, except for the polarity of action. Both phenomena seem to result from comparable mechanisms, probably acting on the quantity of transmitter released.

Action Potentials↗

Protein synthesis is required for synaptic immunity to depotentiation.

De novo protein synthesis and transcription are necessary for the expression of long-lasting synaptic potentiation [long-term potentiation (LTP)] in hippocampal area CA1 and for the consolidation of long-term memory. The stability of LTP and its longevity require macromolecular synthesis at later stages, but a specific role for early protein synthesis has not been identified. Using electrophysiological recording methods in mouse hippocampal slices, we show that multiple trains of high-frequency stimulation provide immediate synaptic immunity to depotentiation. This immunity to depotentiation is dependent on the amount of synaptic stimulation used to induce LTP, it is input specific, and it is prevented by inhibitors of protein synthesis. We propose that local translation mediates input-specific synaptic immunity against depotentiation. We also present evidence suggesting that, in addition to translation, products of transcription can provide cell-wide immunity to depotentiation via heterosynaptic transfer of synaptic immunity between distinct pathways in area CA1. Protein synthesis and transcription may importantly regulate long-term storage of information by conferring synaptic immunity to depotentiation at previously potentiated synapses.

Animals↗

The electrophysiological effects of vasoactive intestinal polypeptide in the guinea-pig inferior mesenteric ganglion.

1. The effects of vasoactive intestinal polypeptide (VIP) on the inferior mesenteric ganglion of the guinea-pig were studied in vitro. 2. In 67% of the neurones tested, application of VIP (1-7.5 X 10(-5) M) by pressure ejection caused a depolarization of the membrane potential which averaged 8.6 +/- 0.4 mV. 3. In 52% of the cells that were responsive to VIP, the membrane depolarization was accompanied by a decrease in membrane input resistance. In another 48% of the cells tested, there was an increase in membrane input resistance. 4. Membrane depolarization caused by VIP enhanced the excitability of post-ganglionic neurones and converted subthreshold electrotonic and subthreshold synaptic potentials to action potentials. 5. The effects of VIP persisted during nicotinic and muscarinic synaptic blockade. The effects of VIP also persisted in a low-Ca2+, high-Mg2+ solution. Thus, the site of action of VIP was on the postsynaptic membrane. 6. Electrical stimulation of the lumbar colonic nerves evoked a slow noncholinergic depolarization of the membrane potential. 7. VIP appeared to be one of the transmitters involved in the electrically evoked e.p.s.p. because both prior desensitization with exogenous VIP and VIP antiserum reduced the amplitude of the slow, non-cholinergic e.p.s.p. 8. Radial distension of a segment of colon attached to the inferior mesenteric ganglion (i.m.g.) evoked a non-cholinergic depolarization of the membrane potential in neurones in the i.m.g. 9. The distension-induced non-cholinergic depolarization was reduced by VIP antiserum. 10. The data support the hypothesis that a population of the mechanosensory afferent nerves running between the colon and the i.m.g. utilize VIP or a VIP-like peptide as a transmitter to modulate reflex activity between the colon and the i.m.g.

Afferent Pathways↗

Postsynaptic potential and postsynaptic current in muscle fibres with large time-constant. EPSP amplitude is independent of membrane resistance.

1. Equations have been developed for a computer programmed calculation of synaptic current from synaptic potentials (epsps). 2. The method permits separation of presynaptic and postsynaptic effects in experiments involving drugs which affect synaptic transmission. It is particularly applicable where the postsynaptic cells are large as in the case of commonly employed crustacean muscle fibres. 3. Contrary to a widely held view the theoretical approach used predicts that epsp-amplitude is relatively independent of membrane resistance. 4. Confirmation is provided by experiments involving the application of barium which increases, and of GABA which decreases membrane resistance.

Animals↗

Neural networks and synaptic transmission in immature hippocampus.

The results reviewed in this chapter indicate that local circuit synaptic interactions are surprisingly well-developed in the rat hippocampal CA3 subfield during the second postnatal week. Intracellular recordings reveal large spontaneous epsps and ipsps. Synchronized bursts of synaptic potentials are observed in most paired intracellular recordings. Antidromic and orthodromic electrical stimulation evokes synaptic responses that are reminiscent of recordings from mature hippocampus. However, following brief trains of electrical stimuli and during bath application of a GABAA receptor antagonist, large prolonged depolarizations are recorded. These are suppressed by excitatory amino acid antagonists. In comparison, slices from mature rats do not produce these events under the same conditions. Thus, a hypothesis has been presented that the degree of recurrent synaptic interaction between pyramidal cells may be enhanced during this critical period in hippocampal development. An analysis of recurrent epsps using dual intracellular recordings is consistent with this contention. The degree of excitatory synaptic interaction between CA3 neurons appears to be at least equal to and likely in excess of that reported in mature hippocampus. One possible explanation for this is that the number of recurrent excitatory synapses may increase transiently during hippocampal development, only later to regress to numbers found in the adult. Recent studies of others suggest that activation of NMDA receptors may play a key role in the maintenance of synapses during development (Cline et al., 1987; Kleinschmidt et al., 1987). In this regard it is interesting that the characteristic of the NMDA receptor-ion channel complex on immature and mature rat CA3 hippocampal pyramidal cells appear to be quite different. These differences may play a role in synapse formation and maintenance. The role of NMDA receptors in LTP and learning (for review see Cotman et al., 1989; Lynch, 1986) are widely discussed. In other circles recurrent excitatory neuronal networks are thought to be substrates for memory (Lynch, 1986; Haberly and Bower, 1989). Both NMDA receptors and recurrent excitation are well represented in the CA3 subfield of immature hippocampus. One challenging area for future study will be the clarification of the interrelations between synaptogenesis and synaptic plasticity within neural networks of the developing hippocampus.

Animals↗

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%).

Action Potentials↗

Activation of calcium-dependent chloride channels causes post-tetanic depolarization in rabbit parasympathetic neurons.

Intracellular recordings were made from neurons in rabbit and feline vesical parasympathetic ganglia in vitro. In response to cathodal current injection (0.1-1 nA for 2-20 ms) the majority of rabbit neurons (229 out of 250) exhibited a single action potential that was followed by a fast and slow after-hyperpolarization (sAHP neuron). The remainder of the cells exhibited an action potential followed by only a fast after-hyperpolarization (fAHP neuron). fAHP neurons did not exhibit anomalous rectification and a spontaneous rhythmic hyperpolarization, which were common membrane properties in sAHP neurons. In response to a train of cathodal current pulses (5-20 Hz for 0.1-10 s), fAHP neurons exhibited action potentials followed by a post-tetanic depolarization (PTD). The PTD was associated with a decrease in membrane input resistance. The amplitude and duration of the PTD were a function of the number of action potentials in the train. The amplitude of the PTD was increased by membrane hyperpolarization and its estimated reversal potential was approximately -30 mV. Low-chloride solution and intracellular injection of chloride ions augmented the amplitude and duration of the PTD, whereas low-sodium, high-potassium and low-potassium solutions did not affect them. Tetraethylammonium (5-10 mM) and barium (0.5-1 mM) increased the amplitude and duration of the PTD. Nominal calcium-free solutions and omega-conotoxin (500 nM) abolished the PTD. The data suggest that activation of chloride channels by calcium influx through omega-conotoxin-sensitive calcium channels mediates the PTD. Repetitive stimulation of the pelvic nerve evoked a train of orthodromic action potentials followed by the PTD of fAHP neurons. (+)-Tubocurarine (10 microM) and hexamethonium (200 microM), but not atropine (1 microM), abolished orthodromic action potentials and the PTD, whereas these cholinergic antagonists did not depress the PTD evoked by direct action potentials. In summary, the data suggest that the PTD may function as a slow synaptic potential in fAHP neurons. This appears likely because neither slow excitatory nor inhibitory postsynaptic potentials are present in neurons of rabbit vesical parasympathetic ganglia. In contrast, slow inhibitory and excitatory postsynaptic potentials were recorded from neurons in feline vesical parasympathetic ganglia.

Action Potentials↗

Cellular and synaptic physiology and epileptogenesis of developing rat neocortical neurons in vitro.

The cellular and synaptic physiology of developing rat neocortical neurons was studied using the in vitro slice method. Rats aged 1-28 days were used for analysis. During the first two postnatal weeks several sequential changes occur in membrane properties and evoked synaptic potentials. Immature neurons had higher input resistances, more linear I-V characteristics, longer membrane time constants, and slower rising and falling phases of action potentials. The developmental increase in rate of rise of the action potential suggests an increasing density of voltage-dependent Na+-channels are inserted in neuronal membranes during postnatal development. The higher input resistance of young cells might be due to their small size and differences in membrane properties. The long time constant indicates a higher specific membrane resistivity of immature neurons. Postsynaptic potentials (PSPs) recorded in young neurons were longer in latency, longer in duration, and more fragile during repetitive activation than their mature counterparts. In addition, PSPs evoked in neurons of animals less than 1 week old did not contain inhibitory postsynaptic components. These physiological features of immature neocortical neurons help explain the pattern of epileptogenesis in young animals. When neonatal cortical slices were exposed to the gamma-aminobutyric acid (GABA) antagonists penicillin or bicuculline, the frequency of occurrence of discharges resembling epileptiform depolarization shifts approached that found in mature slices only during the second postnatal week. Depolarization shifts at younger ages were less stereotyped and more sensitive to stimulus parameters than those in mature neurons.

Animals↗