Biomedical subjects

R J Racine

Publications and source records attributed to R J Racine.

At least 19 recordsLinked to original sources

PubMed2003

A ligand of the p65/p95 receptor suppresses perforant path kindling, kindling-induced mossy fiber sprouting, and hilar area changes in adult rats.

Kindling, an animal model of epilepsy, results in an increased volume of the hilus of the dentate gyrus and sprouting of the mossy fiber pathway in the hippocampus. Our previous studies have revealed that chronic infusion of neurotrophins can regulate not only seizure development, but also these kindling-induced structural changes. Kindling, in turn, can alter the expression of neurotrophins and their receptors. We previously showed that intraventricular administration of a synthetic peptide that interferes with nerve growth factor stability and thus its binding to TrkA and p75(NTR) receptors suppressed kindling and sprouting. However, the precise involvement of TrkA, p75(NTR), and downstream signaling effectors of neurotrophins on kindling, sprouting and hilar changes are unknown. One of these downstream effectors is Ras. In the present study, we find that intraventricular infusion of the synthetic peptide Reo3Y, which binds to p65/p95 receptors and causes a rapid inactivation of Ras protein, impairs development of perforant path kindling, reduces the growth in afterdischarge duration, blocks kindling-induced mossy fiber sprouting in area CA3 of hippocampus and in inner molecular layer of the dentate gyrus, and prevents kindling-induced increases in hilar area. These results are consistent with a mediation of neurotrophin effects on kindling, hilar area, and axonal sprouting via Trk receptors, and suggest important roles for Ras in kindling and in kindling-induced structural changes.

Animals↗

PubMed2002

Continuous infusion of neurotrophin-3 triggers sprouting, decreases the levels of TrkA and TrkC, and inhibits epileptogenesis and activity-dependent axonal growth in adult rats.

Neurotrophin-3 (NT-3), a member of the neurotrophin family of neurotrophic factors, is important for cell survival, axonal growth and neuronal plasticity. Epileptiform activation can regulate the expression of neurotrophins, and increases or decreases in neurotrophins can affect both epileptogenesis and seizure-related axonal growth. Interestingly, the expression of nerve growth factor and brain-derived neurotrophic factor is rapidly up-regulated following seizures, while NT-3 mRNA remains unchanged or undergoes a delayed down-regulation, suggesting that NT-3 might have a different function in epileptogenesis. In the present study, we demonstrate that continuous intraventricular infusion of NT-3 in the absence of kindling triggers mossy fiber sprouting in the inner molecular layer of the dentate gyrus and the stratum oriens of the CA3 region. Furthermore, despite this NT-3-related sprouting effect, continuous infusion of NT-3 retards the development of behavioral seizures and inhibits kindling-induced mossy fiber sprouting in the inner molecular layer of the dentate gyrus. We also show that prolonged infusion of NT-3 leads to a decrease in kindling-induced Trk phosphorylation and a down-regulation of the high-affinity Trk receptors, TrkA and TrkC, suggesting an involvement of both cholinergic nerve growth factor receptors and hippocampal NT-3 receptors in these effects. Our results demonstrate an important inhibitory role for NT-3 in seizure development and seizure-related synaptic reorganization.

Animals↗

PubMed2002

Activity-dependent changes in synaptophysin immunoreactivity in hippocampus, piriform cortex, and entorhinal cortex of the rat.

Synaptophysin, an integral membrane glycoprotein of synaptic vesicles, has been widely used to investigate synaptogenesis in both animal models and human patients. Kindling is an experimental model of complex partial seizures with secondary generalization, and a useful model for studying activation-induced neural growth in adult systems. Many studies using Timm staining have shown that kindling promotes sprouting in the mossy fiber pathway of the dentate gyrus. In the present study, we used synaptophysin immunohistochemistry to demonstrate activation-induced neural sprouting in non-mossy fiber cortical pathways in the adult rat. We found a significant kindling-induced increase in synaptophysin immunoreactivity in the stratum radiatum of CA1 and stratum lucidum/radiatum of CA3, the hilus, the inner molecular layer of the dentate gyrus, and layer II/III of the piriform cortex, but no significant change in layer II/III of the entorhinal cortex, 4 weeks after the last kindling stimulation. We also found that synaptophysin immunoreactivity was lowest in CA3 near the hilus and increased with increasing distance from the hilus, a reverse pattern to that seen with Timm stains in stratum oriens following kindling. Furthermore, synaptophysin immunoreactivity was lowest in dorsal and greatest in ventral sections of both CA3 and dentate gyrus in both kindled and non-kindled animals. This demonstrates that different populations of sprouting axons are labeled by these two techniques, and suggests that activation-induced sprouting extends well beyond the hippocampal mossy fiber system.

Animals↗

PubMed2001

Sequential changes in the synaptic structural profile following long-term potentiation in the rat dentate gyrus: III. Long-term maintenance phase.

LTP has been associated with changes in synaptic morphology but the nature of these changes over the time course of the enhanced electrophysiological response has not been fully determined. The current research involved an examination of synaptic structure in the rat hippocampus during the long-term maintenance phase of LTP. Synapses were examined in the middle third of the molecular layer (MML) of the rat dentate gyrus following repeated high frequency tetanization of the perforant path. Synapses from both the ipsilateral inner third of the dentate molecular layer (IML), which was not directly stimulated during the induction of LTP, as well as implanted, nonstimulated animals, served as controls. LTP was induced over a 4-h period, and the animals were sacrificed 5 days after the final stimulation of the LTP group. Ultrastructural quantification included the total number of synapses per neuron, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synapses. No overall changes in the number of synapses per neuron, shape, or synaptic perforations were observed. There was, however, a significant increase in the length of synapses in the directly stimulated LTP tissue. This increase in synaptic length was particularly evident in the concave-shaped synapses which were also more perforated. These results, together with previous findings, describe a sequence of changes in synaptic morphology that accompany LTP in a structure that is associated with learning and memory.

Animals↗

PubMed2000

Cholinergic modulation of neocortical long-term potentiation in the awake, freely moving rat.

The neocortex has proven resistant to LTP induction using standard in vitro and acute, in vivo preparations. Because the neocortex is widely thought to be involved in long-term information storage, this resistance raises questions about the validity of LTP as a memory model. Recently, we have shown that the neocortex of freely moving rats reliably supports LTP, provided that the stimulation is spaced and repeated over days. The following experiments were designed to evaluate the neuromodulatory role played by cholinergic systems in the induction of LTP in this preparation. Chronically implanted rats received either low- or high-intensity LTP-inducing tetani in combination with the administration of either a cholinergic agonist or antagonist injected systemically. Potentiation was evidenced as amplitude changes in both early and late components of the evoked field potential, the former including population spikes. The cholinergic agonist facilitated LTP induction in the late component of both high- and low-intensity groups. The cholinergic antagonist blocked LTP induction in the early component of the high-intensity group. The possibility that there are component-specific modulatory effects of cholinergic agents on the induction of neocortical LTP is discussed.

Action Potentials↗

PubMed2000

Sequential changes in the synaptic structural profile following long-term potentiation in the rat dentate gyrus. II. Induction/early maintenance phase.

Long-term potentiation (LTP), one of the most compelling models of learning and memory, has been associated with changes in synaptic morphology. In this study, LTP was induced and animals were sacrificed 1 h after the stimulation of the LTP group (induction / early maintenance phase). Synapses in the directly stimulated middle third of the dentate gyrus molecular layer (MML) were examined while synapses from the inner third of the dentate molecular layer (IML) of the LTP animals and both the MML and the IML of implanted animals served as controls. The total number of synapses per neuron, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synaptic contact and active zone were examined. No overall change in the number of synapses per neuron was observed in the LTP tissue. LTP was associated with a significant increase in the proportion of perforated and irregular-shaped synapses compared to controls. The increase in perforated synapses was particularly apparent in the proportion of concave perforated synapses. Nonperforated concave synapses were found to be significantly larger in potentiated tissue. The total synaptic length per neuron of synapses in a concave configuration was also significantly higher following potentiation. These results suggest that the specific structural profile associated with 1-h post-LTP induction, which differed from the profile observed at 24 h post-induction, may represent a unique early phase of synaptic remodeling in a series of changes observed during LTP induction, maintenance, and decay.

Animals↗

PubMed2000

Long-term depression and depotentiation in the sensorimotor cortex of the freely moving rat.

Activity-dependent reductions in synaptic efficacy are central components of recent models of cortical learning and memory. Here, we have examined long-term synaptic depression (LTD) and the reversal of long-term potentiation (depotentiation) of field potentials evoked in sensorimotor cortex by stimulation of the white matter in the adult, freely moving rat. Prolonged, low-frequency stimulation (1 Hz for 15 min) was used to induce either depotentiation or LTD. LTD was expressed as a reduction in the amplitude of both monosynaptic and polysynaptic field potential components. Both LTD and depotentiation were reliably induced by stimulation of the ipsilateral white matter. Stimulation of the contralateral neocortex induced only a depotentiation effect, which decayed more rapidly than that induced by ipsilateral stimulation (hours vs days). Although ipsilateral LTD was effectively induced by a single session of low-frequency stimulation, multiple sessions of stimulation, either massed or spaced, induced LTD effects that were larger in magnitude and longer lasting. Previously, we showed that the induction of long-term potentiation in the neocortex of chronic preparations required multiple, spaced stimulation sessions to reach asymptotic levels. Here, we report that LTD also required multiple stimulation sessions to reach asymptotic levels, but massed and spaced patterns of low-frequency stimulation were equally effective.

Action Potentials↗

PubMed2000

Long-term potentiation in the reciprocal corticohippocampal and corticocortical pathways in the chronically implanted, freely moving rat.

The hippocampus and adjacent cortical structures, including the entorhinal, perirhinal, and parahippocampal cortices, appear to serve as an integrated memory system. This extended hippocampal system is believed to influence memory and consolidation through an extensive set of reciprocal connections with widespread areas of the neocortex. Long-term potentiation (LTP) has been well-examined in the intrinsic connections of the hippocampus and neocortex. However, LTP in the pathways and structures thought to convey information between the hippocampus and neocortex has received little attention. If these pathways and structures are involved in information storage, and if LTP reflects a general synaptic encoding mechanism, then these systems are also likely to support LTP. In this paper we discuss a series of experiments aimed at investigating LTP in the efferents between the hippocampus and neocortex in chronically implanted animals. In the first experiment, the efferents of the perirhinal cortex were stimulated. LTP in the dentate gyrus (DG) reached asymptote more slowly than is typically seen following perforant path stimulation, whereas the frontal area (M1) reached asymptote more quickly than reported following corticocortical stimulation. The DG and M1 LTP was long-lasting, but entorhinal cortex LTP had decayed to baseline levels after a week. In the second experiment, the hippocampal efferents were stimulated. The perirhinal, entorhinal, and frontal cortex showed a similar slow potentiation, with only the perirhinal cortex levels returning to baseline after a week. In the third experiment, the projections from M1 were tested. The perirhinal cortex and hippocampus showed a long-lasting LTP. Although LTP was found in all pathways examined, there were differences in the induction and decay rate, and these properties may correspond to differences in learning rate and longevity of information storage.

Animals↗

PubMed2000

GABAergic modulation of neocortical long-term potentiation in the freely moving rat.

Although the neocortex has generally been considered resistant to the induction of long-term potentiation (LTP), we have recently shown that LTP can be reliably induced in the freely moving rat provided that the stimulation sessions are spaced and repeated. Here, we report that the induction of LTP in this preparation can be modulated by both GABAergic agonism and antagonism. The delivery of stimulation trains in the presence of the GABA(A) agonist diazepam blocked the induction of neocortical LTP, while the GABA(A) antagonist picrotoxin slowed the development of potentiation. When animals that had previously received high-frequency stimulation combined with diazepam were repotentiated, they showed greater resistance to LTP induction than animals that had received diazepam alone. These data suggest that the inhibitory circuits themselves may have potentiated. The demonstration that diazepam blocks neocortical LTP provides further support for the notion that LTP plays a role in memory formation.

Animals↗

PubMed2000

Morphology of layer III pyramidal neurons is altered following induction of LTP in sensorimotor cortex of the freely moving rat.

The organization of specific cortical connections can be altered by sensory and motor experience. These changes are believed to result from activity-dependent changes in synaptic connectivity, similar to those induced in the hippocampus by high-frequency stimulation in long-term potentiation (LTP) experiments. If similar mechanisms are involved, then neocortical LTP induction may induce some of the same morphological changes that are seen following learning. We induced LTP in the contralateral sensorimotor cortex by repeated, daily tetanization of the corpus callosum in chronically implanted, freely moving rats. Anatomical results showed that the LTP induction was associated with alterations in dendrite morphology and increased spine density. These changes are qualitatively and quantitatively similar to those commonly observed in studies in which rats are housed in complex environments. The similarity of results following exposure to complex environments and after LTP induction in the neocortex may indicate a reliance on the same cellular mechanisms in both situations.

Animals↗

PubMed1999

Mossy fiber sprouting induced by repeated electroconvulsive shock seizures.

The elicitation of repeated focal seizures (kindling) induces mossy fiber sprouting in the hippocampus of the rat. The present study investigated whether repeated generalized seizures also induce mossy fiber sprouting. Human psychiatric patients receive repeated generalized seizures during electroconvulsive therapy (ECT). Male Long-Evans rats received a course of eight electroconvulsive shock (ECS) seizures administered on a 48-h schedule over a course of 2 1/2 weeks. Control subjects received matched handling, but no stimulation. Fourteen days after the last ECS trial, all subjects were sacrificed and their brains subjected to Timm staining. Cell counts and area measures were also taken in the hilus. Significant sprouting, but not significant cell loss, was seen in the fascia dentata of the subjects that had received ECS.

Animals↗

PubMed1999

Sequential changes in the synaptic structural profile following long-term potentiation in the rat dentate gyrus: I. The intermediate maintenance phase.

Changes in synaptic structure have been reported following the induction of long-term potentiation (LTP). The structure of synapses during the intermediate maintenance of LTP has yet to be fully characterized in chronically implanted freely moving animals. The present study examined synapses in the middle third of the molecular layer (MML) of the rat dentate gyrus following repeated high frequency tetanization of the perforant path. Synapses from both 1) the ipsilateral inner third of the dentate molecular layer (IML), which was not directly stimulated during the induction of LTP, as well as 2) implanted, nonstimulated animals, served as controls. LTP was induced over a 4-h period, and the animals were sacrificed 24 h after the final stimulation of the LTP group. Ultrastructural quantification included the total number of synapses, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synaptic contact. Although LTP was not associated with an overall increase in synaptic number, there was a significant increase in the proportion of presynaptically concave-shaped synapses. Further, the concave synapses in the LTP tissue were found to be significantly smaller than control concave synapses. There was also a significant increase in the number of perforated concave synapses which exceeded the overall increase in concave synapses, and occurred despite the lack of a general increase in perforated synapses. It was concluded that this specific structural profile, observed at 24 h postinduction, may help support the potentiated response observed at this stage of LTP maintenance.

Animals↗

PubMed1999

Brain-derived neurotrophic factor infusion delays amygdala and perforant path kindling without affecting paired-pulse measures of neuronal inhibition in adult rats.

Kindling is an animal model of human temporal lobe epilepsy in which excitability in limbic structures is permanently enhanced by repeated stimulations. Kindling also increases the expression of nerve growth factor, brain-derived neurotrophic factor, and brain-derived neurotrophic factor receptor messenger RNAs in both the hippocampus and cerebral cortex and causes structural changes in the hippocampus including hilar hypertrophy. We have recently shown that intraventricular nerve growth factor infusion enhances the development of kindling, whereas blocking nerve growth factor activity retards amygdaloid kindling. Furthermore, we have shown that nerve growth factor protects against kindling-induced hilar hypertrophy. The physiological role of brain-derived neurotrophic factor in kindling is not as clear. Acute injection of brain-derived neurotrophic factor increases neuronal excitability and causes seizures, whereas chronic brain-derived neurotrophic factor infusion in rats slows hippocampal kindling. In agreement with the latter, we show here that intrahilar brain-derived neurotrophic factor infusion delays amygdala and perforant path kindling. In addition, we show that brain-derived neurotrophic factor, unlike nerve growth factor, does not protect against kindling-induced increases in hilar area. To test the hypothesis that brain-derived neurotrophic factor suppresses kindling by increasing inhibition above normal levels, we performed paired-pulse measures in the perforant path-dentate gyrus pathway. Brain-derived neurotrophic factor infused into the hippocampus had no effect on the stimulus intensity function (input/output curves); there was also no significant effect on paired-pulse inhibition. We then kindled the perforant path 10 days after the end of brain-derived neurotrophic factor treatment. Once again, kindling was retarded, showing that the brain-derived neurotrophic factor effect is long-lasting. These results indicate that prolonged in vivo infusion of brain-derived neurotrophic factor reduces, rather than increases, excitability without increasing inhibitory neuron function, at least as assessed by paired-pulse protocols. This effect may be mediated by long-lasting effects on brain-derived neurotrophic factor receptor regulation.

Amygdala↗

PubMed1999

Development of kindling-prone and kindling-resistant rats: selective breeding and electrophysiological studies.

Because of the growing need for an animal model of complex partial seizures based on a genetic predisposition, we combined the kindling model of epilepsy with selective-breeding procedures to develop two new lines (or strains) of rats that are kindling-prone or kindling-resistant. The selection of these strains was based on their rates of amygdala kindling. From a parent population of Long Evans hooded and Wistar rats, the males and females that showed the fastest and slowest amygdala kindling rates were selected and bred. Similar selection procedures continued through F11, although there was little or no overlap in the distribution of kindling rates for the two new strains (FAST and SLOW) by F6. Examination of both local and propagating seizure profiles of the new strains from F6 to F10 revealed that the FAST and SLOW rats had similar amygdala afterdischarge (AD) thresholds and associated AD durations. Also, the convulsion profiles of the stage-5 responses were similar, although the severity was greater in the FAST rats. Clearly the selection was not based on local mechanisms controlling the threshold for amygdala AD evocation, but rather for the spread of AD from the focus and the recruitment of other structures, ultimately triggering convulsive seizures. Although evoked potentials and potentiation effects were similar between the strains, the SLOW rats showed a greater paired-pulse depression, raising the possibility that they differ in inhibitory mechanisms. The specificity of strain differences for the amygdala and its associated networks is described in our accompanying paper (McIntyre et al., 1999. FAST and SLOW amygdala kindling rat strains: Comparison of amygdala, hippocampal, piriform and perirhinal cortex kindling. Epilepsy Res. 35, 197-209). These strains should provide many clues to the dispositional differences between individuals for the development of epilepsy originating in temporal lobe structures.

Amygdala↗

PubMed1999

Multiple window time-frequency distribution and coherence of EEG using Slepian sequences and hermite functions.

Multiple window (MW) time-frequency analysis (TFA) is a newly developed technique to estimate a time-varying spectrum for random nonstationary signals with low bias and variance. In this paper, we describe the application of MW-TFA techniques to electroencephalogram (EEG) and compare the results with those of the conventional spectrogram. We find that the MW-TFA provide us with not only low bias and variance time-frequency (TF) distribution for EEG but also TF coherence estimation between a single realization of EEG recorded from two sites. We also compare the performance of the MW-TFA using two sets of windows, Slepian sequences, and Hermite functions. If care is taken in matching the two windows, we find no noticeable difference in the resulting TF representations.

Animals↗

PubMed1998

Time course for kindling-induced changes in the hilar area of the dentate gyrus: reactive gliosis as a potential mechanism.

Recurrent seizure activity induced during kindling has been reported to cause an increase in the hilar area of the dentate gyrus of the hippocampus. To date, very little is known about the mechanism of this increase. This study investigated the time course for kindling-induced changes in the hilar area of the dentate gyrus at seven days, one month, and two months post-kindling. Hilar area of the dentate gyrus was significantly increased by approximately 46% at seven days and remained elevated at one month, but declined back to control levels by two months. Glial fibrillary acidic protein (GFAP) immunostaining was also evaluated at the same time points to determine whether kindling-induced changes in the hilar area of the dentate gyrus are related to kindling-induced glial cell changes. Increases in hilar GFAP immunostaining by approximately 57% were observed at seven days and at one month post-kindling, but not at two months post-kindling. These findings indicate that kindling-induced changes in the hilar area of the dentate gyrus and kindling-induced glial cell changes follow a similar time course, and that kindling-induced glial cell changes may mediate the observed changes in the hilar area of the dentate gyrus.

Animals↗

PubMed1998

The degree of potentiation is associated with synaptic number during the maintenance of long-term potentiation in the rat dentate gyrus.

There is a considerable degree of variation in the amount of potentiation induced in different animals following the induction of long-term potentiation (LTP). This variation provided us with the opportunity to determine what types of synaptic changes were dependent upon the degree of induced potentiation. To examine possible 'degree of potentiation' effects on synapses, we conducted a multiple regression analysis examining the relationship between the degree of potentiation in LTP animals and a series of synaptic structural measures. We examined synapses in the middle third of the molecular layer (MML) of the rat dentate gyrus following repeated high frequency tetanization of the perforant path. LTP was induced over a 4 h period, and the animals were sacrificed 24 h after the final stimulation. Synapses from the ipsilateral inner third of the dentate molecular layer (IML) and from implanted only animals were also examined for comparison. Ultrastructural quantification included the total number of synapses per neuron, synaptic curvature, the presence of synaptic perforations, and the maximum length of the synaptic apposition. The only structural change that was significantly associated with the degree of potentiation was a positive correlation between the degree of LTP and the number of synapses per neuron. Therefore, synaptic number, while not appearing to be significantly associated with the induction of LTP, appears to be important for the degree of LTP expressed.

Animals↗

PubMed1998

Development of spontaneous seizures over extended electrical kindling. I. Electrographic, behavioral, and transfer kindling correlates.

The present study was aimed at evaluating an extended kindling model of spontaneous epilepsy. Behavioral and electrographic responses to repeated kindling of either the perforant path or amygdala were monitored for up to 300 trials. Kindling initially led to generalized convulsions equivalent to the level 5 seizure on the rating scale developed by Racine. The evoked seizures became progressively more complex with additional kindling, which was described by a 10-stage classification system. The highest stage (stage 10) was achieved when the kindling stimulation evoked two or more bouts of level 5 seizures combined with running and jumping fits. These more complex seizures developed over the course of amygdala, but not perforant path kindling. Electrographic seizures from both the amygdala and dentate gyrus increased in duration and amplitude during the early phase of kindling, but did not correlate with motor seizure development beyond level 5. During the late phase of kindling, the dentate gyrus afterdischarge amplitude decreased and became dissociated from the behavioral seizures. Manifestations of spontaneously recurring seizures were seen in the majority of animals, but spontaneous seizures of level 4 or greater were observed in only five rats. The second part of this study examined kindling transfer effects, the efficacy of kindling a new site after the completion of the initial (in this case extended) kindling protocol. The effect depended on both primary and secondary site location. When the amygdala served as primary site, perforant path transfer was complete in some animals but absent in others. No transfer occurred in the opposite direction, from the perforant path to the amygdala. Finally, transfer effects in the dentate gyrus, which was tested as tertiary site, were complete. Previous studies have found weaker transfer effects in the dentate when kindling to the standard stage 5 level.

Animals↗