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G F Tseng

Publications and source records attributed to G F Tseng.

At least 19 recordsLinked to original sources
PubMed2000

Fate of the supraspinal collaterals of cord-projection neurons following upper spinal axonal injury.

In investigating the fate of the cord-projecting CNS neurons following spinal axonal injury, we have demonstrated that surviving rat rubrospinal neurons have altered electrical membrane properties so that their input/output relationship was increased. Further, we found that the synaptic inhibition they received from nearby reticular formation was also reduced following injury. Whether or not these property changes were functional was dependent on the output connections of injured neurons. In the current communication, we examined the supraspinal efferents of the injured neurons recognizing that normal neurons innervate not only spinal but also supraspinal targets. To this end we conducted anterograde tracing on the injured red nucleus 8 weeks following spinal lesion. Results showed that injured rubrospinal neurons still innervated the same supraspinal targets, targeted by normal neurons. We subsequently evaluated the relative intensity of the sustained supraspinal connectivity by examining, in detail, the cerebellar projection of rubrospinal neurons of similarly injured animals using retrograde tracing technique. Here our data revealed that the number, distribution and labeling intensity of rubrospinal neurons projecting to the cerebellum were unchanged following cord injury. In conclusion, although spinal cord injury deprive cord-projecting CNS neurons of their spinal targets, injured neurons survived with altered electrical membrane properties and intact supraspinal projections. The sustained supraspinal connections might allow injured cord-projecting CNS neurons to exert a different weight of influence on higher centers following spinal cord injury.

Animals↗
PubMed1997

Membrane properties and inhibitory connections of normal and upper cervically axotomized rubrospinal neurons in the rat.

Membrane properties and inhibitory synaptic connections of normal and axotomized rat rubrospinal neurons were examined using a coronal slice preparation. Rubrospinal neurons were axotomized at the C2 vertebral level in vivo. Retrograde labelling in vivo and intracellular biocytin injection following recording were combined to identify recorded axotomized rubrospinal neurons. Their input resistances decreased three and four days and became higher than normal four and 10 weeks following lesioning which coincided with a sequential increase and decrease of their soma area. On the other hand, although their membrane time-constant was reduced three and four days following lesioning, it returned to normal value four and 10 weeks following axotomy. Other than these, their membrane current-voltage relationship including an inward rectification in the hyperpolarizing direction was not altered. Normal rubrospinal neurons generated very fast spikes which were not affected by axotomy. Both normal and axotomized cells generated trains of repetitive spikes with a fast spike frequency adaptation at the beginning upon suprathreshold current injection. However, the slope of the steady-state spike frequency and applied current relationship was increased four and 10 weeks following axotomy which also showed an increased steady-state spike frequency in response to high-amplitude current injection. Synaptically, the amplitude and duration of the monosynaptic inhibitory potential evoked from nearby reticular formation were reduced following axotomy. In addition, fewer rubrospinal neurons were found to receive this inhibition 10 weeks following axotomy. Thus, our results show that spinal axotomy induces a time-dependent modification of the membrane properties and spike generating behaviour of rubrospinal neurons which probably represents an initial decrease and a later increase of their excitability. This is accompanied by a persistent decrease of synaptic inhibition which is expected to affect structures that remained innervated by the undamaged axon collaterals of these spinally axotomized neurons.

Animals↗
PubMed1996

Rubral astrocytic reactions to proximal and distal axotomy of rubrospinal neurons in the rat.

Spinal tractotomy-induced perineuronal astrocytic reaction of the rat rubrospinal system was studied using an antiserum to the astrocyte-specific glial fibrillary acidic protein as a marker. The effect of the proximity of axonal cut to cell bodies was also studied by comparing astrocytic reactions elicited by upper cervical and lower thoracic tractotomy. Fast blue was used as a retrograde tracer to identify axotomized neurons, which were found to concentrate in the caudal part of the contralateral red nucleus. The length of reactive astrocytic processes in the dorsomedial and ventrolateral parts of the nucleus was quantified separately since neurons in these two parts project to cervical and lumbar spinal cord, respectively. Those of the ipsilateral nucleus were also quantified. Sham operation caused a transient increase in reactive astrocytic processes one day after surgery. An early and a late increase of reactive astrocytic processes was found 2-5 days and 2-8 weeks following both thoracic and cervical tractotomy. Cervical axotomy of lumbar-cord-projecting rubral neurons caused an increase of reactive astrocytic processes similar in magnitude to that generated by thoracic axotomy. Following thoracic axotomy, the uninjured dorsomedial area of the contralateral nucleus also displayed an increase concomitant with that which occurred within the neighboring, injured ventrolateral nuclear area suggesting the action of diffusible factor(s). Surprisingly, cervical and thoracic tractotomy also elicited a similar increase of reactive astrocytic processes in the ipsilateral nuclei, independent of the number of ipsilaterally projected neurons present in each nucleus. This may be attributed to the retrograde influence from the denervated spinal target sites which were carried by fibers of the intact rubrospinal tract known to terminate bilaterally. In the lesioned nucleus, reactive astrocytic processes were often located close to axotomized cell bodies as early as 3 days following upper cervical and also, to a lesser extent, lower thoracic tractotomy. However, reactive astrocytic processes in the ipsilateral nucleus usually remained in the neuropil. These results suggest that axotomy induces two levels of retrograde astrocytic reactions within the soma area of intrinsic central neurons. Reactive astrocytic processes located proximally to axotomized cell bodies may have a different functional role from those distributed in the neuropil.

Animals↗
PubMed1996

Perineuronal microglial reactivity following proximal and distal axotomy of rat rubrospinal neurons.

Microglial reactivity in the red nucleus of rats was studied following upper cervical and lower thoracic rubrospinal tractotomy using the lectin binding method. Following axotomy, the contralateral nucleus containing the axotomized neurons was identified using the retrograde tracer Fast blue. It was subdivided into dorsomedial (DM) and ventrolateral (VL) portions known to project to the cervical and lumbar spinal cord, respectively. Lectin-labeled microglial cells and processes on the contralateral as well as in the ipsilateral nucleus were then quantified. An early and a late increase in microglial reactivity was observed in the nucleus at 2-5 days and 2-8 weeks following thoracic and cervical tractotomy with the latter producing a more pronounced reactivity. In rats subjected to thoracic axotomy, a similar microglial increase also occurred in the intact contralateral DM nuclear area suggesting the possible action of diffusable factor(s) that might have triggered the microglial activation from the axotomized VL nuclear area. The uninjured ipsilateral nucleus also exhibited a similar pattern of microglial reactivity irrespective of the number of ipsilaterally projecting neurons following both cervical and thoracic axotomy. This could have been elicited by the retrograde influence from the denervated targets carried by the intact rubrospinal fibers of the opposite side since many of them in fact terminate bilaterally (Antal, M. et al., J. Comp. Neurol., 325 (1992) 22-37). In all the axotomized or intact nucleus, microglial processes did not appear to surround neuronal cell bodies. The characteristic responses of microglial cells in the red nucleus may be related to the failure of rubrospinal neurons to regenerate following the severance of their axons.

Amidines↗
PubMed1996

Extrinsic inhibitory innervation to rubral neurons in rat brain-stem slices.

Synaptic connections between the neurons in the red nucleus (RN) and its extrinsic neurons were studied using rat brain-stem slices. Intracellular records were obtained from the RN neurons. Ipsilateral stimuli to areas in the dorsolateral mesencephalic reticular formation (DLMRF) or substantia nigra (SN) elicited monosynaptic hyperpolarizing postsynaptic potentials (PSPs) in about 95% of RN neurons recorded. The hyperpolarizing PSPs could be reversibly blocked by bicuculline, indicating that they were GABAA receptor-mediated-Cl(-)-inhibitory PSPs. The sites of most inhibitory synapses arising from DLMRF and SN are possibly located on the proximal half of the soma-dendritic membrane of RN neurons, according to the analysis of the IPSPs with Rall's model. In addition, tracing dyes were employed to examine the morphological pathways. After rhodamine B, a retrograde tracer, was applied to the RN in brain slices, the cell bodies of a number of neurons in DLMRF and SN were labeled. These labeled neurons were also immunopositive for glutamic acid decarboxylase (GAD) as revealed from double labeling with an anti-GAD antiserum. The anterograde tracer, tetramethylrhodamine dextran, was applied to the DLMRF or SN and taken up by many neurons in the areas. A portion of these cells extended their processes toward and terminated within the RN. Moreover, electron microscopic examination confirmed that the tetramethylrhodamine dextran-decorated synaptic terminals were present in the RN. The results indicate that the rubral neurons receive direct GABAA receptor-mediated inhibitory inputs from neurons in the DLMRF and SN, which may participate in modulation of rubral outputs.

Animals↗
PubMed1996

Axotomy affects the retrograde labeling of cervical and lumbar-cord-projecting rubrospinal neurons differently.

The effect of axotomy at cervical and lumbar spinal levels upon the ability of rubrospinal neurons to retrogradely transport tracer was compared. Unilateral rubrospinal tractotomy was performed first at C5 and, after a few days, at C2 vertebral levels. Different retrograde tracers were applied at the lesioned sites right after tractotomy. Tracer applied at C5 labeled both cervical and lumbar-cord-projecting neurons. Tracer applied at C2 also labeled both groups of neurons if performed 2 days after that at C5; however, only cervical-cord-projecting neurons were labeled when it was performed 3 or 5 days after that at C5. In another set of experiments, a T10 tractotomy without tracer application was performed 2 or 5 days prior to the C5/C2, series of tract lesions. When preceded by a T10 lesion 2 days in advance, tracer applied at C5 labeled both cervical and lumbar-cord-projecting neurons. However, a T10 lesion 5 days in advance resulted in the labeling of only cervical-cord-projecting neurons by the tracer applied at C5. In either case, tracer applied at C2 consistently labeled only cervical-cord-projecting neurons, irrespective of the intervals-2, 3, or 5 days-allowed between C5 and C2 lesions. Most neurons labeled from C2 were also double-labeled by the tracer applied at C5. Thus, unlike lumbar-cord-projecting counterparts, cervical-cord-projecting rubrospinal neurons retain the ability to uptake and/or transport retrograde tracer several days following axotomy. This implies that cervical-cord-projecting rubrospinal neurons survive in a different functional state from their lumbar-cord-projecting counterparts following axonal injury.

Animals↗
PubMed1996

Compartmentalization of calbindin and parvalbumin in different parts of rat rubrospinal neurons.

The distribution of calbindin-immunoreactive neurons in the red nucleus and the subcellular distribution of the calbindin and parvalbumin in tracer-identified rubrospinal neurons of the rat were studied. Only a fraction of the retrogradely labelled rubrospinal neurons was found to contain calbindin. These neurons filled the caudal part of the red nucleus and also appeared sporadically along the ventromedial border of the middle segment of the red nucleus. In addition to the somata, calbindin was found in the dendritic arbors of tracer-identified rubrospinal neurons, revealed by injecting the fluorescent dye Lucifer Yellow into their cell bodies. The axons of rubrospinal neurons located in the caudal red nucleus were marked by performing anterograde tracing with fluorescent dextran tracer in freshly prepared brainstem slices. Parvalbumin was found to locate in swellings along these tracer-identified axons as well as at their cut ends. The results indicate that calbindin and parvalbumin are segregated to the somadendritic and axonal compartments of the rat rubrospinal neurons, respectively. This anatomical segregation suggests that they may have different functions in neurons.

Animals↗
PubMed1996

Structural and functional alterations in rat corticospinal neurons after axotomy.

1. The electrophysiological properties of rat corticospinal neurons (CSNs) were studied 3, 9, and 12 mo after axotomy in the cervical spinal cord, with the use of a combination of the in vitro neocortical slice technique, intracellular recordings, and a double-labeling method that allowed identification of CSNs studied in vitro. 2. CSNs retained the rhodamine-labeled microspheres employed as a retrograde marker and were functionally active in the longest survival group (1 yr). 3. The somatic area of axotomized CSNs became progressively smaller, a reduction that amounted to 37% for all cells at 1 yr. There were no obvious differences between normal and axotomized cells in terms of apical dendritic widths, numbers of apical dendritic branches, or basal dendritic arbors. Intracortical axonal arborizations of axotomized neurons were in general similar to those of normal CSNs in that most axons ended in layers V and VI with only occasional collaterals reaching supragranular layers. 4. Axotomized CSNs were grouped according to their spike firing patterns during depolarizing current pulses so that their electrophysiological behavior could be compared with that of regular spiking and adapting groups of normal CSNs. No significant differences were found in resting membrane potential, or spike parameters between axotomized neurons in any survival group and normal controls. Neurons surviving 1 yr after axotomy had a higher input resistance (RN) than normal CSNs. There was a reduction in the percentage of CSNs that generated prominent spike depolarizing afterpotentials in the axotomized group. 5. The steady-state relationship between spike frequency and applied current (f-I slope) became steeper over time and was significantly greater 9 mo after axotomy in regular spiking (RS) and adapting neurons than in normal CSNs in the same groups. The increase in steady-state f-I slope was in part related to increases in the RN of axotomized neurons. 6. There was a significant decrease in the generation of slow afterhyperpolarizations following trains of spikes in axotomized versus normal RS neurons, first detected at 3 mo and also present in 9 mo and 1 yr survival groups. 7. Biphasic inhibitory postsynaptic potentials (IPSPs) were evoked in only 1 of 11 axotomized neurons in the 3-mo group, 2 of 12 cells examined at 9 mo, and 3 of 15 neurons 1 yr after axotomy. The proportions of neurons generating IPSPs were significantly smaller than in comparable groups of control CSNs. As a consequence, longer duration evoked excitatory postsynaptic potentials were generated by axotomized CSNs. 8. Results show that axotomized CSNs undergo alterations in intrinsic membrane properties and inhibitory synaptic electrogenesis that would tend to make them more responsive to excitatory inputs.

Animals↗
PubMed1996

Axotomy induces retraction of the dendritic arbor of adult rat rubrospinal neurons.

The effect of distal axonal injury on the soma-dendritic morphology of intrinsic central neurons was examined using adult rat lumbar spinal cord-projecting rubrospinal neurons as a model. The soma-dendritic morphology was revealed using an improved Golgi-aldehyde method. Impregnated neurons were reconstructed in the two-dimensional plane for analysis. Four weeks after axotomy, neurons had reduced soma sizes and remained multipolar in shape. Some dendrites were found to end not far from their cell bodies. In addition, no long dendrite was identified following axotomy. Sholl's analysis [The Organization of the Cerebral Cortex. London, Methuen, [1956] revealed that axotomized neurons had fewer dendritic branches than control neurons. Total dendritic length was also reduced. Subsequent analyses showed that the average number of dendritic trunks was not altered however the mean number of terminal branches per dendritic trunk was reduced. The dendritic membrane of the normal neurons was usually smooth with occasional short protuberances on the proximal dendrites and spines on the distal dendrites, which did not change after axotomy. In control neurons, we identified an elaborate type of dendritic structure named dendritic appendage aggregates. These aggregates were located preferentially on terminal dendrites and were classified into three categories according to their complexity. The incidence of occurrence for these aggregates decreased following distal axotomy. These phenomena indicate that rat lumbar spinal cord-projecting rubrospinal neurons retract their distal dendrites in response to distal axotomy. The observed anatomic restructuring following axonal injury is likely to be accompanied by an alteration of afferents which normally synapse on distal dendrites.

Animals↗
PubMed1995

A time-dependent loss of retrograde transport ability in distally axotomized rubrospinal neurons.

Studies on the effect of axotomy on adult intrinsic central projection neurons have generally assumed that the severed proximal axonal stumps were still capable of retrogradely transporting tracer at varying times after injury. Failure of transport was interpreted as neuronal death, which is at odds with current understanding that central projection neurons survived distal axotomy. We used lumbar spinal cord-projecting rubrospinal neurons of the rat as a model to evaluate the ability of injured neurons to transport tracer retrogradely at different times after distal axotomy. We examined only the caudal part of the red nucleus, since rubrospinal neurons are concentrated here. In control animals, tracer applied to the rubrospinal tract at the T10 vertebral level labeled ventrolateral rubral neurons, while C3 application marked all rubral neurons. From 3 days after a T10 axotomy and tracer application, most ventrolateral neurons were no longer labeled by another tracer application at the C3 vertebral level via an axonal cut. The phenomenon was not caused by tracer toxicity, since a T10 tractotomy without tracer application also prevented these axotomized neurons from being labeled when treated similarly. Thus, neuronal retrograde transport capability was seriously retarded 3 days after a distal axotomy. Loss of retrograde transport may merely suggest that a mechanism no longer in service has been switched off, or perhaps it may insulate injured neurons from the effect of lesion site-derived factors. Using this property, we were able to localize cervical spinal cord-projecting rubrospinal neurons in the caudal red nucleus.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Axonal sprouting in layer V pyramidal neurons of chronically injured cerebral cortex.

We performed experiments to determine whether axonal sprouting occurs in neurons of chronic neocortical epileptogenic lesions. Partially isolated somatosensory cortical islands with intact pial blood supply were prepared in mature rats. Neocortical slices from these lesions, studied 6-39 d later, generated spontaneous and/or evoked epileptiform field potentials (Prince and Tseng, 1993) during which neurons displayed prolonged polyphasic excitatory and inhibitory synaptic potentials/currents. Single electrophysiologically characterized layer V pyramidal neurons in control and epileptogenic slices were filled with biocytin using sharp and patch-electrode techniques, their axonal arbors reconstructed and compared quantitatively. Neurons in injured cortex had a 56% increase in total axonal length, a 64% increase in the number of axonal collaterals and more than a doubling (115% increase) of the number of axonal swellings. The presumed boutons were smaller and more closely spaced than those of control cells. In some neurons the main descending axon had hypertrophic segments from which branches arose. These highly significant changes were most marked in the perisomatic region of layer V. The axonal sprouting was associated with a decrease in somatic area but no significant change in dendritic arbors. Results suggest that a significant degree of axonal reorganization takes place in the chronically injured cortex where it might be an adaptive mechanism for recovery of function after injury, or might be maladaptive and play an important role in the generation of epileptiform events by increasing the numbers and density of synaptic contacts between neurons.

Animals↗
PubMed1994

Efferent connections from the external cuneate nucleus to the medulla oblongata in the gerbil.

The present study revealed the efferent projections from the external cuneate nucleus (ECN) to various medullary nuclei in the gerbil as demonstrated in fresh living brainstem slices by using in vitro anterogradely tracing with the dextran-tetramethyl-rhodamine-biotin. The tracer-labelled ECN axon terminals were observed (1) in most of the vital autonomic-related nuclei: the nucleus solitary tractus, nucleus ambiguus, rostroventrolateral reticular nucleus and C2 adrenergic area, (2) in the reticular formation: the medullary, parvocellular, intermediate, gigantocellular, dorsal paragigantocellular and lateral paragigantocellular reticular nuclei and medullary linear nucleus, and (3) in sensory nuclei: the cuneate nucleus, spinal trigeminal nuclei caudalis and interpolaris, paratrigeminal nucleus, medial and spinal vestibular nuclei, inferior olive and prepositus hypoglossal nucleus. These new findings are discussed in relation to possible roles of the ECN in cardiovascular, respiratory and sensorimotor controls.

Animals↗
PubMed1994

The postnatal development of the GABAA/benzodiazepine receptor in the rat red nucleus.

The development of the GABAA/Benzodiazepine receptor (GABAAR) in the red nucleus was studied using 3H-flunitrazepam (FNZ) as the probe. Saturation binding assay showed that the Bmax of the ligand to the membranes of the nucleus increased from 0.50 +/- 0.04 nmol/mg protein at postnatal day 4, to 0.71 +/- 0.1 and 0.78 +/- 0.08 at day 7 and day 10. At day 20 the Bmax decreased to a level near day 4 and persisted until day 40. However, the affinity of 3H-FNZ to the receptor remained quite constant. At least 4 proteins of 51kD, 53kD, 59kD and 62kD in the nucleus were labeled by 3H-FNZ, as revealed from photoaffinity binding and SDS-PAGE. The labeling of 53kD, 59kD and 62kD was high at earlier ages than day 10, whereas the 51kD was predominent from day 10 to day 40. Receptor binding autoradiography of the nucleus also showed that the most dense labeling was seen around day 10. The early transient increase in the GABAAR of the red nucleus may indicate the plasticity of the nucleus in response to environmental changes after birth.

Age Factors↗
PubMed1993

Heterogeneity of rat corticospinal neurons.

In order to examine the degree of diversity within a population of cortical projection neurons, rat corticospinal cells were retrogradely labeled in vivo by injecting rhodamine-tagged microspheres into the cervical spinal cord, and subsequently studied electrophysiologically and anatomically in neocortical slices maintained in vitro, by use of standard current clamp techniques and a double-labeling protocol (Tseng et al., J. Neurosci. Meth. 37:121-131, 1991). Three different subgroups were distinguished on the basis of their spiking behavior: (1) Adapting cells had a marked fast (50 ms) and slow phase (200 ms) of spike frequency adaptation; (2) regular spiking (RS) cells had only a period of fast adaptation; (3) some regular spiking neurons had prominent depolarizing afterpotentials (DAPs) and could generate bursts of spikes, often in repetitive fashion (RSDAP cells). Subgroups of RSDAP cells had different patterns of burst responses to depolarizing current pulses, suggesting differences in the types and/or sites of underlying ionic conductances. Adapting cells had a slightly higher membrane input resistance and more prominent slow hyperpolarizing afterpotentials than RS and RSDAP neurons; however, the activation of presumed anomalous rectifier current by intracellular hyperpolarizations was less prominent in adapting neurons. Orthodromic stimulation in layer I evoked presumed excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs)in all three types of cells, but prominent short-latency IPSPs were found in a higher percentage of adapting neurons. The morphology of electrophysiologically characterized corticospinal neurons was studied following intracellular injection of biocytin. All three spiking types were typical layer V pyramids with apical dendrites reaching layer 1, basal dendrites in infragranular layers, and deep-directed axons that had a moderate density of local collaterals in lower cortical layers. The profuseness of dendrites, examined by Sholl's analysis of two-dimensional, camera lucida-reconstructed neurons was comparable in the three neuronal subgroups, although a smaller somatic area and more slender apical dendritic trunk were found in adapting neurons. Our results suggest that corticospinal cells in rats are a heterogeneous population of projection neurons with respect to their spiking behavior, membrane properties, synaptic connections, and, to a lesser extent, their morphology. This diversity revealed in vitro adds new complexity to the classification of corticospinal neurons.

Action Potentials↗
PubMed1993

Epileptogenesis in chronically injured cortex: in vitro studies.

1. Field potentials and intracellular activities were examined in neocortical slices obtained through areas of chronic cortical injury produced by cortical undercutting and transcortical lesions made in vivo 7-122 days before the terminal in vitro slice experiment. 2. Abnormal field potentials characterized by long- and variable-latency multiphasic events could be evoked by layer VI-white matter or subpial stimulation in 9 of 15 animals that had adequate partial cortical isolations. These "epileptiform" field potentials were recorded in layers II-V and propagated across the cortex. They appeared at threshold in an all-or-none fashion and, in most slices, could be blocked by increasing stimulus intensity. In one slice, spontaneous epileptiform events occurred that were similar to those evoked by extracellular stimulation. 3. Intracellular activities during the epileptiform field potentials consisted of polyphasic synaptic events that were predominantly depolarizing and that could last < or = 400-500 ms, synchronous with the field potential activities. A variety of observations suggested that the neuronal activities underlying epileptiform field potentials were relatively asynchronous and much less intense than those previously found in chemically induced epileptogenesis within the neocortex. 4. Inhibitory postsynaptic potentials (IPSPs) were not prominent in neurons when threshold stimuli evoked epileptiform events; however, suprathreshold stimuli could elicit biphasic IPSPs and block the long-latency polysynaptic activity and abnormal field potential in most slices. Depolarizing components of the polysynaptic activity had the appearance of excitatory postsynaptic potentials in terms of their responses to alterations in membrane potential. 5. Comparison of spike parameters in layer V neurons of epileptogenic slices with those in control layer V neurons showed no significant differences in spike height, threshold, duration, or rise time. Resting membrane potentials were also not significantly different. 6. There was a highly significant difference in input resistance (RN) between layer V neurons in control and injured slices; the mean value for neurons in lesioned cortex was 68.1 M omega, whereas that in control cells was 30.5 M omega. There was also a significant prolongation of the slow membrane time constant in neurons of injured cortex (19.4 ms) as opposed to that in control cells (12.2 ms), suggesting that a change in specific resistivity or capacitance contributed to the higher RNS. 7. The relationship between adapted spike frequency and applied current (f-I slope) was steeper in layer V neurons from injured cortical slices (44.3 Hz/nA) than in normal layer V cells (28.2 Hz/nA).(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1991

Double-labelling with rhodamine beads and biocytin: a technique for studying corticospinal and other projection neurons in vitro.

Corticospinal neurons retrogradely labelled with rhodamine-labelled latex microspheres (RLMs) in vivo were studied intracellularly in a slice preparation up to 13 months later with electrodes containing biocytin. The physiological properties of these double-labelled corticospinal neurons were indistinguishable from those of comparable neurons which were impaled with biocytin-containing electrodes without prior RLM-labelling, and neurons studied with potassium acetate-filled electrodes in similar areas. Thus, neither labelling with RLMs nor injection of biocytin affected neuronal properties. This important advantage of RLMs makes them suitable for prelabelling projection neurons in vivo for subsequent studies that take advantage of the versatility of a brain slice preparation. In addition to its lack of effects on neuronal properties, intracellular labelling with biocytin also provides high-quality morphological details ideal for anatomical analysis. The compatibility of retrograde labelling with RLMs and intracellular staining with biocytin make this a useful combined technique for tracking electrophysiological and anatomical changes in identified projection neurons over time.

Animals↗
PubMed1989

Deep neurons in piriform cortex. I. Morphology and synaptically evoked responses including a unique high-amplitude paired shock facilitation.

1. Synaptic responses of cells in layer III of the piriform cortex and the subjacent endopiriform nucleus (layer IV) were analyzed with intracellular recording techniques in a slice preparation from the rat, cut perpendicular to the pial surface. 2. Micropipettes containing Lucifer yellow (LY) were used to correlate response properties with morphology. An antiserum to LY was used to intensify staining and to prevent fading during detailed morphological study. Response properties were also examined with potassium acetate-containing electrodes. 3. Morphologically, two cell types were identified: pyramidal cells that were confined to layer III of the piriform cortex and multipolar cells that were in layer III and the endopiriform nucleus. 4. In morphology, deep pyramidal cells in layer III closely resembled superficial pyramidal cells in layer II, with the exception that primary apical dendritic trunks were longer and basal dendritic arborizations were more extensive than apical. Like superficial pyramidal cells, apical dendrites of all deep pyramidal cells stained extended through the afferent fiber termination zone in layer Ia and gave rise to local axonal arbors that were concentrated in layer III and the endopiriform nucleus. 5. Multipolar cells were morphologically indistinguishable in layer III and the endopiriform nucleus. All gave rise to nonvaricose spiny dendrites that never extended into layer II and local axonal arbors. 6. Response properties of deep pyramidal and multipolar cells were similar; responses of both of these populations were very different from those of superficial pyramidal cells. The primary difference between responses of deep pyramidal and multipolar cells was a shorter latency of postsynaptic potentials evoked in deep pyramidal cells by stimulation of afferent fibers, consistent with the extension of their dendrites into layer Ia. 7. Responses of most deep cells to stimulation of afferent and association fibers at sufficiently high strength consisted of an initial excitatory postsynaptic potential (EPSP), followed by a fast Cl- -mediated and a slow K+-mediated inhibitory postsynaptic potential (IPSP). 8. A characteristic feature of deep cells, which was only rarely observed in superficial pyramidal cells, was the presence of variable EPSPs evoked at long latencies (greater than 100 ms) by stimulation of afferent or association fibers. 9. A striking finding for deep pyramidal and multipolar cells, when studied with LY-containing pipettes, was a variable slowly rising depolarizing potential triggered at depolarized membrane potentials by stimulation of afferent or association fibers.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗