Changes In Postsynaptic Potentials Research Articles

Inhibitory synapse formation in a co-culture model incorporating GABAergic medium spiny neurons and HEK293 cells stably expressing GABAA receptors.

Inhibitory neurons act in the central nervous system to regulate the dynamics and spatio-temporal co-ordination of neuronal networks. GABA (γ-aminobutyric acid) is the predominant inhibitory neurotransmitter in the brain. It is released from the presynaptic terminals of inhibitory neurons within highly specialized intercellular junctions known as synapses, where it binds to GABAA receptors (GABAARs) present at the plasma membrane of the synapse-receiving, postsynaptic neurons. Activation of these GABA-gated ion channels leads to influx of chloride resulting in postsynaptic potential changes that decrease the probability that these neurons will generate action potentials. During development, diverse types of inhibitory neurons with distinct morphological, electrophysiological and neurochemical characteristics have the ability to recognize their target neurons and form synapses which incorporate specific GABAARs subtypes. This principle of selective innervation of neuronal targets raises the question as to how the appropriate synaptic partners identify each other. To elucidate the underlying molecular mechanisms, a novel in vitro co-culture model system was established, in which medium spiny GABAergic neurons, a highly homogenous population of neurons isolated from the embryonic striatum, were cultured with stably transfected HEK293 cell lines that express different GABAAR subtypes. Synapses form rapidly, efficiently and selectively in this system, and are easily accessible for quantification. Our results indicate that various GABAAR subtypes differ in their ability to promote synapse formation, suggesting that this reduced in vitro model system can be used to reproduce, at least in part, the in vivo conditions required for the recognition of the appropriate synaptic partners and formation of specific synapses. Here the protocols for culturing the medium spiny neurons and generating HEK293 cells lines expressing GABAARs are first described, followed by detailed instructions on how to combine these two cell types in co-culture and analyze the formation of synaptic contacts.

Inhibitory Synapse Formation in a Co-culture Model Incorporating GABAergic Medium Spiny Neurons and HEK293 Cells Stably Expressing GABA<sub>A</sub> Receptors

Inhibitory neurons act in the central nervous system to regulate the dynamics and spatio-temporal co-ordination of neuronal networks. GABA (γ-aminobutyric acid) is the predominant inhibitory neurotransmitter in the brain. It is released from the presynaptic terminals of inhibitory neurons within highly specialized intercellular junctions known as synapses, where it binds to GABAA receptors (GABAARs) present at the plasma membrane of the synapse-receiving, postsynaptic neurons. Activation of these GABA-gated ion channels leads to influx of chloride resulting in postsynaptic potential changes that decrease the probability that these neurons will generate action potentials. During development, diverse types of inhibitory neurons with distinct morphological, electrophysiological and neurochemical characteristics have the ability to recognize their target neurons and form synapses which incorporate specific GABAARs subtypes. This principle of selective innervation of neuronal targets raises the question as to how the appropriate synaptic partners identify each other. To elucidate the underlying molecular mechanisms, a novel in vitro co-culture model system was established, in which medium spiny GABAergic neurons, a highly homogenous population of neurons isolated from the embryonic striatum, were cultured with stably transfected HEK293 cell lines that express different GABAAR subtypes. Synapses form rapidly, efficiently and selectively in this system, and are easily accessible for quantification. Our results indicate that various GABAAR subtypes differ in their ability to promote synapse formation, suggesting that this reduced in vitro model system can be used to reproduce, at least in part, the in vivo conditions required for the recognition of the appropriate synaptic partners and formation of specific synapses. Here the protocols for culturing the medium spiny neurons and generating HEK293 cells lines expressing GABAARs are first described, followed by detailed instructions on how to combine these two cell types in co-culture and analyze the formation of synaptic contacts.

Simultaneous NMDA-Dependent Long-Term Potentiation of EPSCs and Long-Term Depression of IPSCs in Cultured Rat Hippocampal Neurons

A fundamental issue in understanding activity-dependent long-term plasticity of neuronal networks is the interplay between excitatory and inhibitory synaptic drives in the network. Using dual whole-cell recordings in cultured hippocampal neurons, we examined synaptic changes occurring as a result of a transient activation of NMDA receptors in the network. This enhanced transient activation led to a long-lasting increase in synchrony of spontaneous activity of neurons in the network. Simultaneous long-term potentiation of excitatory synaptic strength and a pronounced long-term depression of inhibitory synaptic currents (LTDi) were produced, which were independent of changes in postsynaptic potential and Ca2+ concentrations. Surprisingly, miniature inhibitory synaptic currents were not changed by the conditioning, whereas both frequency and amplitudes of miniature EPSCs were enhanced. LTDi was mediated by activation of a presynaptic GABAB receptor, because it was blocked by saclofen and CGP55845 [(2S)-3-{[(15)-1-(3, 4-dichlorophenyl)ethyl]amino-2-hydroxypropyl)(phenylmethyl)phosphinic acid]. The cAMP antagonist Rp-adenosine 3 ', 5 ' -cyclic monophosphothioate abolished all measured effects of NMDA-dependent conditioning, whereas a nitric oxide synthase inhibitor was ineffective. Finally, network-induced plasticity was not occluded by a previous spike-timing-induced plasticity, indicating that the two types of plasticity may not share the same mechanism. These results demonstrate that network plasticity involves opposite affects on inhibitory and excitatory neurotransmission.

Spatiotemporal Dynamics of Sensory Responses in Layer 2/3 of Rat Barrel Cortex MeasuredIn Vivoby Voltage-Sensitive Dye Imaging Combined with Whole-Cell Voltage Recordings and Neuron Reconstructions

The spatiotemporal dynamics of the sensory response in layer 2/3 of primary somatosensory cortex evoked by a single brief whisker deflection was investigated by simultaneous voltage-sensitive dye (VSD) imaging and whole-cell (WC) voltage recordings in the anesthetized rat combined with reconstructions of dendritic and axonal arbors of L2/3 pyramids. Single and dual WC recordings from pyramidal cells indicated a strong correlation between the local VSD population response and the simultaneously measured subthreshold postsynaptic potential changes in both amplitude and time course. The earliest VSD response was detected 10-12 msec after whisker deflection centered above the barrel isomorphic to the stimulated principal whisker. It was restricted horizontally to the size of a single barrel-column coextensive with the dendritic arbor of barrel-column-related pyramids in L2/3. The horizontal spread of excitation remained confined to a single barrel-column with weak whisker deflection. With intermediate deflections, excitation spread into adjacent barrel-columns, propagating twofold more rapidly along the rows of the barrel field than across the arcs, consistent with the preferred axonal arborizations in L2/3 of reconstructed pyramidal neurons. Finally, larger whisker deflections evoked excitation spreading over the entire barrel field within approximately 50 msec before subsiding over the next approximately 250 msec. Thus the subthreshold cortical map representing a whisker deflection is dynamic on the millisecond time scale and strongly depends on stimulus strength. The sequential spatiotemporal activation of the excitatory neuronal network in L2/3 by a simple sensory stimulus can thus be accounted for primarily by the columnar restriction of L4 to L2/3 excitatory connections and the axonal field of barrel-related pyramids.

Plasticity in an electrosensory system. II. Postsynaptic events associated with a dynamic sensory filter.

1. This report summarizes studies of the changes in postsynaptic potentials that occur as pyramidal cells within the primary electrosensory processing nucleus learn to reject repetitive patterns of afferent input. The rejection mechanism employs "negative image inputs" that oppose or cancel electroreceptor afferent inputs or patterns of pyramidal hyperpolarization or depolarization caused by intracellular current injection. Feedback pathways carrying descending electrosensory as well as other types of information provide the negative image inputs. This study focuses on the role of a directly descending projection from a second-order electrosensory nucleus the nucleus praeeminentialis (nP), which provides excitatory and inhibitory inputs to the apical dendrites of electrosensory lateral line lobe (ELL) pyramidal cells. 2. Electrical stimulation of the pathway linking the nP to the ELL was used to activate descending inputs to the pyramidal cells. Pyramidal cell activity was typically increased due to stimulation of this pathway. Tetanic stimulation of the descending pathway paired with either electrosensory stimuli that inhibited pyramidal cells, or hyperpolarizing current injection, increased the excitation provided by subsequent stimulation of this pathway. Pairing tetanic stimulation with excitatory electrosensory stimuli or depolarizing current injection had the opposite effect. Subsequent activation of the descending pathway inhibited pyramidal cells. 3. Intracellular recordings showed that the increased firing of pyramidal cells evoked by stimulation of the descending pathway following tetanic stimulation paired with postsynaptic hyperpolarization resulted from larger amplitude and longer-duration excitatory postsynaptic potentials (EPSPs). The shift in the effect of activity in this descending pathway to providing net inhibitory input to the pyramidal cells after paired presynaptic activity and postsynaptic depolarization probably results from the potentiation of inhibitory postsynaptic potentials (IPSPs). The EPSP and IPSPs evoked by activity in this descending pathway can be continuously adjusted in amplitude, thereby counterbalancing patterns of pyramidal cell excitation and inhibition received from the periphery with the result that repetitive patterns of afferent activity are strongly attenuated.

Synaptic activation of N-methyl- d-aspartate receptors induces short-term potentiation at excitatory synapses in the striatum of the rat

We examined the effect of augmenting the activation of N-methyl- d-aspartate receptors during tetanic stimulation of the corpus callosum. Excitatory postsynaptic potentials were recorded from striatal neurons in an in vitro brain slice bathed in either normal or Mg 2+-free artificial cerebrospinal fluid. Post-tetanic potentiation was followed by a long-lasting depression of the excitatory postsynaptic potential in normal artificial cerebral spinal fluid. Mg 2+-free artificial cerebrospinal fluid increased the duration of the pretetanus excitatory postsynaptic potential and the amplitude and duration of the direct response to the tetanus. Mg 2+-free artificial cerebrospinal fluid also enabled the expression of a short-term potentiation of the excitatory postsynaptic potential amplitude and duration. N-methyl- d-aspartate receptor antagonists blocked the induction of all tetanus-induced excitatory postsynaptic potential changes specific to the Mg 2+-free artificial cerebral spinal fluid. These results indicate that maintained activation of N-methyl- d-aspartate receptors by synaptically released glutamate can produce a sustained enhancement of the excitatory postsynaptic potential. This use-dependent increase in synaptic efficacy may contribute to basal ganglia-related motor performance.

Postsynaptic modulation of rectifying electrical synaptic inputs to the LG escape command neuron in crayfish

The lateral giant (LG) tail-flip escape system of crayfish is organized to provide a massive convergence of mechanosensory inputs onto the LG command neuron through electrical synapses from both mechanosensory afferents and interneurons. We used electrophysiological techniques to show that the connections between three major mechanosensory interneurons and LG rectify, and that their inputs to LG can be reduced by postsynaptic depolarization and increased by postsynaptic hyperpolarization. The mechanosensory afferents and interneurons are excited by sensory nerve shock, and the components of the resulting LG PSP can be similarly modulated by the same postsynaptic potential changes. Because these inputs are all made through electrical synapses, we conclude that they are rectifying connections, as well. To test the physical plausibility of this conclusion, we developed an electrical model of the rectifying connection between a mechanosensory interneuron and LG, and found that it can reproduce all the qualitative features of the orthodromic and antidromic experimental responses. The ability of postsynaptic membrane potential to modulate inputs through rectifying electrical synapses is used in the escape system to enhance LG's relative sensitivity to novel, phasic stimuli. Postsynaptic depolarization of LG produced by earlier inputs "reverse-biases" the rectifying input synapses and reduces their strength relative to times when LG is at rest.

Characterization of postsynaptic potentials evoked by sural nerve stimulation in hindlimg motoneurons from acute and chronic spinal cats

The purpose of this study was to characterize the changes in postsynaptic potentials recorted in ankle extensor motoneurons resulting from activation of the sural nerve spinal cord transection in the adult cat. Eight acute and nine chronic animals were spinalized at T 12. Intracellular recordings from motoneurons innervating the triceps surae were performed. Sural nerve stimulations evoked complex synaptic potentials consisting of early and late components in all motoneurons. Early excitatory and inhibitory postsynaptic potentials (PSPs), as well as long latency excitatory postsnnaptic potentials were recorded and averaged for assessment of PSP amplitude and duration. Early PSPs, both excitatory and inhibitory, were significantly larger in the motoneurons of cats spinalized 4–6 months earlier. Central latency of excitatory potentials were similar in the two samples of motoneurons, but the central latency associated with the initial inhibitory PSP was significantly shorter in the recordings from motoneurons of chronic spinal cats. In most recordings, an additional inhibitory PSP followed the initial excitatory PSP in motoneurons, and this secoddary inhibitory PSP was similar in peak amplitude and duration in both samples of motoneurons. Also, a long latency excitatory PSP was recorded in a large percentage of motoneurons from both samples. This potential was typically of greater amplitude and longer duration in the motonuurons from chronic animals, when compared to recordings from acute animals. Although changes in amplitude and duration of PSP activity could be documented, there was no marked alteration in the frequency of occurrence of each PSP pattern recorded from the two preparations. This suggests that the synaptic pathways mediating the sural nerve reflexes have not qualitatively changed in the chronic spinal animals. The changes in amplitude and durations of the PSPs in the chronic spatial cat indicate, however, the quantitative changes have occurred. The quantitative changes have probably occurred in the interneuronal networks activated by cutaneous nerve (sural) stimulation, since it was shown that only minor changes in motoneuron membrane properties could be recorded in these same chronic spinal animals.

Long-term potentiation in guinea pig hippocampal slices monitored by optical recording of neuronal activity

Pre- and postsynaptic potential changes evoked by stimulation of the Schaffer collateral-commissural input to CA 1 pyramidal neurons were optically recorded in guinea pig hippocampal slices after staining the preparation with a suitable voltage-sensitive fluorescent dye. Brief tetanic stimulation induced long-term potentiation of synaptic transmission as monitored both optically and electrically. The results demonstrate that non-invasive optical techniques can be used to study long-term changes in spatial neuronal interactions, possibly involved in learning and other higher functions of the nervous system.

γ-Aminobutyrate sensitivity does not change during long-term potentiation in rat hippocampal slices

Long-term potentiation is a long-lasting enhancement of synaptic efficacy following brief, high-frequency, repetitive stimulation of a monosynaptic input. Intracellular recordings have shown that the inhibitory postsynaptic potential changes in amplitude during long-term potentiation. Yet how this may occur is unclear. To test for a possible alteration in postsynaptic sensitivity to the recurrent inhibitory transmitter γ-aminobutyrate, we have examined the effect of γ-aminobutyrate, focally applied to the hippocampal CA1 cell-body layer, on the extracellular recorded action potential (population spike). We found that the degree, duration, dose-dependence and time-course of inhibition produced by γ-amino-butyrate are unchanged during long-term potentiation. This suggests that a change in sensitivity of CA1 pyramidal cells to the transmitter γ-aminobutyrate is not the reason for the alteration in the inhibitory postsynaptic potential during long-term potentiation.

Experimental approaches to understanding the role of protein phosphorylation in the regulation of neuronal function.

Studies by Earl Sutherland and his colleagues on hormonal regulation of the breakdown of glycogen in liver resulted in the discovery that the first step in the action of many hormones is to increase the synthesis of cAMP by activating adenylate cyclase (Raft et al 1957, Sutherland & Rall 1958, Robison et al 1968). It was later established that cAMP exerts its effects by stimulating protein kinases that catalyze the phosphorylation of specific functional proteins and thereby regulate their activity (Walsh et al 1968, Kuo & Greengard 1969, Krebs & Beavo 1979). The discovery that the brain contains a high concentration of cAMP-dependent protein kinase led to the proposal that protein phosphorylation might play an important role in regulation of neuronal properties by neurotransmitters and neurohormones (Miyamoto et al 1969). In particular, it seemed that protein phosphorylation, which usually takes place on a time scale of hundreds of milliseconds or longer, might be a mechanism underlying relatively long-lasting changes in neuronal properties such as slow changes in post-synaptic potentials (McAfee & Greengard 1972), changes in the rate of transmitter synthesis (Morgenroth et al 1975), or changes in gene expression (Klein & Berg 1970). The biochemists and neurobiologists who took up the study of brain protein phosphorylation hoped to gain insight into some of the mechanisms underlying changes in neuronal excitability and synaptic efficacy and also, perhaps, into processes that govern the development of various neuronal types during the formation of the nervous system. This line of research was bolstered by the findings that the brain contains not only high concentrations of protein kinases, but also protein phosphatases, adenylate cyclase, and phosphodiesterase (Greengard 1976), and also by the discovery that several neurotransmitters stimulate the synthesis of second messengers such as cyclic AMP and cyclic GMP by binding to specific receptors on the surfaces of neurons (for reviews see Nathanson 1977, Greengard 1981).

Transmission mediated with and without spikes at connexions between large second-order neurones of locust ocelli

Large second-order neurones of locust ocelli (L-neurones) make both excitatory and inhibitory connexions with each other. Small, graded depolarisations and hyperpolarisations are transmitted at the excitatory connexions but, at the inhibitory connexions, spikes in the presynaptic neurone are required for transmission. 1. L-neurones spike, usually once only, when hyperpolarisations end rapidly. The hyperpolarisations can be caused either by illumination of the ocellus, or by injection of current. The amplitude of a spike depends both upon the amplitude and the duration of the preceding hyperpolarisation (Fig. 1B). 2. At the excitatory connexions, the resting potential of the presynaptic neurone normally lies depolarised from the threshold for transmission, so that both small hyperpolarisations and depolarisations effect changes in postsynaptic potential (Fig. 2A, B). Over periods of several minutes, there is no sign of decrement in transmission at these connexions (Fig. 2D). Spikes in the presynaptic neurone usually ensure that the postsynaptic neurone also spikes. 3. At the inhibitory connexions, the postsynaptic potential decrements within 10–20 ms (Fig. 3A). Because of this, rapidly rising presynaptic potentials, such as spikes, are required for transmission. Also, presynaptic hyperpolarisations do not effect changes in postsynaptic potential. Following an inhibitory postsynaptic potential, transmission at an inhibitory connexion remains depressed for about a second (Figs. 3B, C). 4. All three members of one anatomical class of L-neurone (L1–3; C.S. Goodman 1976) of a lateral ocellus make reciprocal inhibitory connexions with each other (Fig. 7B; Table 1). Some of these neurones are presynaptic at excitatory connexions with another class (L4–5; Fig. 7A). Many L-neurones do not project to the whole area of the retina, and most project to the dorsal or ventral halves (Fig. 6). The excitatory connexions may sharpen responsiveness to decreases in illumination, and the inhibitory connexions may enhance the detection of rapid movements of large objects, such as the visual horizon.

Viewpoint. What the marine mollusc Aplysia can tell the neurologist about behavioral neurophysiology.

Only recently has man begun to regard himself as mundane and not divine. This conceptual liberation has allowed him to ask frank questions concerning the physical and chemical mechanisms which determine or affect his behavior. Unfortunately the answers to these questions have been slow in coming. The reasons for this are two-fold: Basic ethical considerations preclude the experiments necessary to investigate the neural substrates of human behavior in man. Further, man’s behavior and nervous system are both so enormously complex and subtle, it is therefore unlikely that much real fundamental knowledge could be gained from such experiments if performed. It is more expedient to study simple behavior in simpler organisms than man to understand how nervous systems operate in general and, it is hoped, to eventually gain a better understanding of the human in particular. This tactic is known as the “model systems” approach. By discovering the strategies adopted by less complex nervous systems to deal with simple situations one can devise a realistic model of the neural mechanisms that control more complex behavior in more advanced animals.Many animals have served as valuable sources of model systems. Among them the marine gastropod molluscAplysiahas received considerable attention. In comparison to the human nervous system with approximately 50 billion neurons, theAplysianervous system contains relatively few neurons — about 20,000. Furthermore the study of theAplysianervous system has several other advantageous characteristics. A number of forms of behavioral plasticity that are found in all higher metazoans including man are also found in theAplysia. These simple but non-trivial types of behavioral plasticity include habituation, sensitization and associative learning as well as easily defined qualities of neural function which we choose to call “behavioral states”. In addition the nervous system is composed of neurons which are large and, in many cases, easily identified by anatomical and physiological criteria so that the “same” cell can be studied in more than one animal under more than one set of experimental conditions. The cell bodies of the neurons inAplysia, from which electrical recordings can be fairly easily obtained, are electrically close to their dendrites so that changes in postsynaptic potentials occurring during modifications of behavior can be monitored.

The non-impulsive stretch-receptor complex of the crab: a study of depolarization-release coupling at a tonic sensorimotor synapse

A new preparation for the study of synaptic transmission is described from the thoracic ganglion of the crabCallinectes sapidus. The central anatomy of the nonimpulsive stretch-receptor neurons of the thoracic-coxal joint and that of the promotor motoneurons with which they form synaptic junctions was studied with intracellular cobalt staining and light and electron microscopy. Attention was centred on the interaction of the stretch-receptor T-fibre and the four large motoneurons supplying the promotor muscle which have their cell-bodies on the dorsal surface of the ganglion. The presynaptic terminal region of the T-fibre appeared to be a simple cylinder in form with a diameter of 40-60 μm and containing large stores of synaptic vesicles at its periphery, opposite the complex of motoneuron dendrites. The transmission characteristics of the junctions between receptor cell and motoneurons were studied by transmembrane current injection into the isolated T-fibre by means of a sucrose gap and simultaneous intracellular recording with microelectrodes from the presynaptic terminal and the somata of postsynaptic cells. It was shown that depolarization-release coupling in the T-fibre has similar properties to those that have been demonstrated in the squid giant synapse, with the same values for ‘threshold’, peak release and ‘suppression potential’. The crab synapses differ from that of the squid in that they normally transmit prolonged, graded depolarizations (i.e. receptor potentials) which are decrementally conducted from the periphery. Consistent with this role, the junctions were found to be capable of continuous tonic transmission over many seconds without the strong depletion seen in more phasic synapses. In a study of the relation between the synaptic properties and the stretch reflex it was shown that some of the time- and amplitude-dependent behaviour of the overall reflex can be encoded at the level of the synaptic transmission, largely through the parameter of transmitter availability. Conduction of electrical signals in the proximal and presynaptic part of the sensory fibre was also investigated. Transient responses to step depolarizing currents in the fibre indicate the existence of a mechanism for the partial compensation of capacitative distortion in the decrementally conducted receptor potential. This is the first example of intracellular recording from presynaptic terminals of nonimpulsive neurons with simultaneous monitoring of postsynaptic potential changes, allowing for a direct analysis of depolarization-release coupling characteristics. The use of the preparation for further study of synaptic physiology and sensorimotor systems is discussed.

Graded synaptic transmission between local interneurones and motor neurones in the metathoracic ganglion of the locust.

1. In the metathoracic ganglion of the locust some neurones can effect changes in the membrane potential of identified post-synaptic motor neurones without themselves spiking. 2. These 'non-spiking' neurones have processes only within the metathoracic ganglion, and therefore are local intraganglionic interneurones. 3. The absence of spikes in the interneurones reflects their normal physiological state and is not due to the experimental conditions. 4. When the interneurones are depolarized by the injection of current pulses lasting several hundred milliseconds, post-synaptic motor neurones are either depolarized, or hyperpolarized, for the duration of the pulse. 5. The magnitude of the change in post-synaptic voltage is graded according to the amount of presynaptic current. 6. A number of physiological tests indicate that the graded effects upon motor neurones are mediated by chemical synaptic transmission. For example, an evoked hyperpolarization of a motor neurone can be reversed in polarity by simultaneously hyperpolarizing the motor neurone with injected current. 7. At their resting potential some interneurones tonically release sufficient transmitter to have a measurable post-synaptic effect. The injection of depolarizing and hyperpolarizing currents into these interneurones effects opposite changes in post-synaptic potential. 8. Other interneurones must be depolarized from resting potential before a post-synaptic effect is observed, and hyperpolarizing currents have no post-synaptic effect. In these interneurones it is estimated that a depolarization of only 2 mV is sufficient to effect the release of transmitter. 9. The membrane potentials of non-spiking interneurones can fluctuate by as much as 15 mV during active movements of the hind legs and individual p.s.p.s as large as 5 mV can be recorded. Therefore, summed p.s.p.s or even single ones are expected to be the electrophysiological signals effecting transmitter release from these interneurones.

Changes in rubral multiunit activity after lesions in the interpositus nucleus of the cat

The initial effect of an acute lesion in the central nervous system is loss of neural function. With time, however, restorative processes, generally termed 'compensation', may result in some degree of recovery of neural function. The mechanisms involved in recovery may include morphological changes in synaptic connections, a process described as neuroplasticity. Direct evidence for neuroplasticity in the mature mammalian central nervous system has been presented (for recent reviews, see refs. 3-6). Electrophysiological studies of the rubrospinal tract in the cat have shown evidence of neuroplasticity in which sprouting of axonal terminals (collateral sprouting) is thought to occur in the red nucleus following partially deafferenting lesions ~4. The demonstration of synaptic reorganization in this nucleus is based on evidence that the two major inputs to this nucleus, corticorubral and interpositorubral fibers, make synaptic connection with two separate portions of the membranes of rubral neurons. Tsukahara and co-workers12, la showed in the intact cat that excitatory postsynaptic potentials (EPSPs) evoked in rubral neurons by stimulating the ipsilateral pericruciate cortex had relatively slow rise times, long decay phases, and low peak amplitudes. EPSPs evoked by stimulating the contralateral interpositus nucleus displayed more rapid rise times, shorter decay phases, and greater peak amplitudes. They attributed these differences to segregation of synaptic terminals on rubral neurons, cortical fibers on distal dendrites and interpositus fibers on proximal dendrites and somas. The reorganization of cortical synaptic contacts on rubral neurons after partial deafferenration was investigated in chronic cats with lesions of the contralateral interpositus nucleus 14. They observed a progressive shortening of the rise times of cortically evoked EPSPs for 4~10 days after the lesions and no further change after longer intervals. These changes in cortically evoked EPSPs were thought to result from collateral sprouting of cortical fibers to innervate more proximal portions of rubral neurons. If neuroplasticity participates in the recovery of motor function after a lesion, it must do so not only through changes in postsynaptic potentials, but also through changes in patterns of spike activity directed toward brain stem and spinal nuclei. It is not clear what effect changes in EPSP wave forms may have on rubral spike activity. We have investigated this issue by recording spontaneous multiunit spike activity in

Quantitative description of end plate current and its reconstruction.

An attempt was made to quantify the postsynaptic current based on the experimental data of the voltage clamp method. The conductance change in postsynaptic membrane was derived from the postsynaptic conductance of voltage clamped postsynaptic membrane, then it's temporal characteristics and dependence on the clamped voltage have been quantified. The temporal characteristics was found to be explained by the introduction of two schematic operators, active and inactive. This idea was applied to a simple electrical circuit model of the postsynaptic cells. Besides the change in postsynaptic potentials of normal synapse in excitable state was culculated as its application.

Structural and functional changes in an identified cricket neuron after separation from the soma. I. Structural changes.

The morphological effects of separation from the soma were examined in isolated arborization and isolated axon segments of an identified motor neuron in the Polynesian field cricket, Teleogryllus oceanicus. The identified neuron, the contralateral dorsal longitudinal motor neuron of the metathoracic ganglion (CDLM), possesses an arborization most of which lies contralateral to its soma within the metathoracic ganglion. Midline surgical lesions in the metathoracic ganglion separated CDLM into a distal segment composed of the axon and most of the arborization, and a proximal segment comprised of the remaining arborization, neuritie, and soma. Isolated axonal segments were produced by cutting the nerve root containing the axon of CDLM close to the ganglion. The normal anatomy of CDLM was determined by axonal dye-fills using cobaltous chloride. Morphological changes in the isolated arborization of CDLM were examined by axonal dye-fills at successive time intervals. Changes in the isolated CDLM axon were examined via dissection and histological cross-sections of the distal nerve at graded time intervals. In one example, a remnant of the isolated CDLM arborization survived to 168 days postoperative, a time comparable to the longest previously-reported physiological and morphological survival times of distal axonal segments of invertebrates. In general the isolated arborization does not survive this long. Normally-occurring branches of the arborization can be preserved about 0 to 50 days. After this period branches of the arborization seem to be lost in progressive fashion from smaller to larger, leading to complete loss of the arborization and axon in most cases at 100 or more postoperative days. There is evidence for the presence of supernumerary fibers in the isoalted CDLM arborization between 0 to 63 days postoperative. Such supernumerary fibers indicate an independent capacity for outgrowth of the isolated arborization without connection to the nucleus. The distal axonal segment of CDLM degenerates physiologically and morphologically within 4 to 15 days after peripheral nerve section. This time course is close to that of Wallerian degeneration of vertebrate peripheral nerve axons.

Structural and functional changes in an identified cricket neuron after separation from the soma. II. Functional changes

Physiological and behavioural effects of separation from the soma were examined in isolated arborization and isolated axon segments of an identified motor neuron in the Polynesian field cricket, Teleogryllus oceanicus. The identified neuron, the contralateral dorsal longitudinal motor neuron of the metathoracic ganglion (CDLM), has an arborization most of which lies contralateral to its soma within the ganglion. Midline lesions in the ganglion separated CDLM into a distal segment composed of the axon and most of the arborization, and a proximal segment made up of the remaining arborization, neurite and soma. Isolated axonal segments were produced by cutting the nerve containing the CDLM axon. The function of the neuron-muscle system composed of CDLM, its pre-synaptic inputs, and its innervated muscle bundle was examined in contrl and experimentally operated animals. Extracellular recording assessed function in the axon. Electrical or tactil stimulation was used to excite pre-synaptic inputs to the CDLM arborization. Intracellular recording determined changes in post-synaptic potentials and miniature end-plate potentials in the muscle bundle innervated by CDLM. Normal axonal conduction, competence to respond to pre-synaptic input, neuron-muscle transmission, and miniature end-plate potential appearance can remain in the isolated arborization preparation. Physiological viability is longer in the cricket isolated arborization than in other insect distal segments described. Survival times of axonal conduction and the competence of the isolated arborization to respond to pre-synaptic input are roughly correlated with disappearance of the whole distal segment at 100 or more postoperative days. A naturally-occurring breakdown of the metathoracic dorsal longitudinal muscles in Teleogryllus eventually prevents measurements of post-synaptic potentials and miniature end-plate potentials. Normal post-synaptic function mediated by the distal arborization is maintained up to this breakdown, to a maximum of 44 days postoperative. The distal axonal segment of CDLM degenerates physiologically within four days postoperative, a time course approximating that of degeneration in vertebrate peripheral nerve distal axons.

Characteristics of the output from the dentate nucleus to spinal neurons via pathways which do not involve the primary sensorimotor cortex.

Experiments were performed to determine the action of the dentate output on neurons in the spinal cord mediated by pathways which do not involve the primary sensorimotor and premotor cortices. The dentate nucleus was electrically stimulated by stereotaxically placed electrodes in Rhesus monkeys whose contralateral sensorimotor and premotor cortices were ablated. The resultant changes in excitability of lumbar alpha motorneurons activated by Ia afferents from nerves innervating femoral, hamstring, gastrocnemius-soleus and peroneal muscles were measured by intracellular recordings and by determining the percent change in the amplitude of the monosynaptic reflex recorded from ventral roots. The effect of stimulation of the dentate nucleus on proprioceptive reflexes was determined by recording the changes in postsynaptic potentials evoked by selective stimulation of Ia and Ib afferent fibers. The results demonstrated that dentate nucleus exerts a significant action on the excitability of spinal neurons via pathways which do not include the sensorimotor and premotor cortices. Whether the dentate stimulus produced an increase or decrease in the excitability of these neurons was dependent upon the site within the dentate nucleus at which the stimulus was applied, demonstrating that, in the decorticate preparation, the output from this nucleus is quite heterogeneous. In addition, stimulation of the dentate nucleus in these monkeys did not affect the Ia reflex pathway but significantly changed the amplitude of the inhibitory postsynaptic potential evoked by Ib afferents in lumbar alpha motorneurons.