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Relations between early prespike magnetic field changes, interictal discharges, and return to basal activity in the neocortex of rabbits.

To evaluate possible prespike field synchronizations, its relation to both interictal discharges and postspike return to baseline, penicillin-induced cortical interictal discharges were recorded in anaesthetized rabbits by magnetoencephalography (MEG) and electrocorticography (EcoG). Statistical parameters of spatial (global field power (GFP)) and temporal properties (Z-parameter) of field synchronization were calculated. In our previous report, three types of prespike field synchronization were found before the onset of interictal spike. We report here that the continuous and fluctuating, but not the abrupt prespike increases, were more often associated with a spike and wave pattern of interictal discharge than with a spike alone. Furthermore, the postspike return of these statistical parameters shows the same three patterns as the prespike field synchronizations, but in the inverse time sequence. More often than not pre- and postspike pattern were of the same type. The results suggest an influence of prespike field synchronization upon interictal discharge and subsequent field return dynamics.

Animals↗

Tangential synaptic distribution of NMDA and AMPA receptors in rat neocortex.

We performed an electron microscopic study in layers II-III of S-1 in rats, using postembedding immunogold histochemistry to compare the synaptic distribution of N-methyl D-aspartate (NMDA) receptors (assessed with an antibody for the NMDAR1 subunit) with that of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors (assessed with an antibody for the GluR2/3 subunit). Labeling for each receptor was concentrated at active zones of asymmetric synapses. Analysis of the tangential position of gold particles along the postsynaptic active zone revealed that NMDA receptors were at highest concentration in the middle of the synaptic apposition, whereas AMPA receptors were concentrated in an annulus away from its center. These data support the view that the two types of receptors are anchored by distinct subsynaptic assemblies, and raise the possibility of independent synaptic microdomains.

Animals↗

Labeling of N-acetylaspartate and N-acetylaspartylglutamate in rat neocortex, hippocampus and cerebellum from [1-13C]glucose.

Both N-acetylaspartate (NAA) and N-acetylaspartylglutamate (NAAG) are localized almost exclusively to neurons, and have become important markers of neuronal viability in a number of cerebral pathological conditions. Using nuclear magnetic resonance spectroscopy combined with [1-13C]glucose administration (200 min infusion) we show that the synthesis of both NAA and NAAG can be observed. Label was incorporated into NAA from labeled acetate and from labeled aspartate, while NAAG was labeled from labeled glutamate. The low fractional enrichment of NAA (ca. 3%) relative to aspartate (20%) suggests a slow turnover rate, while NAAG (20.0%) and glutamate (25.2%) labeling were nearly equal, suggesting that NAAG labeling is near steady state. The rapid turnover of NAAG suggests an important role in glutamate delivery, while the slow rate of NAA turnover implies that its major role is as substrate for the formation of NAAG.

Animals↗

Synaptic transmission in the neocortex during reversible cooling.

We studied the effects of reversible cooling on synaptic transmission in slices of rat visual cortex. Cooling had marked monotonic effects on the temporal properties of synaptic transmission. It increased the latency of excitatory postsynaptic potentials and prolonged their time-course. Effects were non-monotonic on other properties, such as amplitude of excitatory postsynaptic potentials and generation of spikes. The amplitude of excitatory postsynaptic potentials increased, decreased, or remain unchanged while cooling down to about 20 degrees C, but thereafter it declined gradually in all cells studied. The effect of moderate cooling on spike generation was increased excitability, most probably due to the ease with which a depolarized membrane potential could be brought to spike threshold by a sufficiently strong excitatory postsynaptic potential. Stimuli that were subthreshold above 30 degrees C could readily generate spikes at room temperature. Only at well below 10 degrees C could action potentials be completely suppressed. Paired-pulse facilitation was less at lower temperatures, indicating that synaptic dynamics are different at room temperature as compared with physiological temperatures. These results have important implications for extrapolating in vitro data obtained at room temperatures to higher temperatures. The data also emphasize that inactivation by cooling might be a useful tool for studying interactions between brain regions, but the data recorded within the cooled area do not allow reliable conclusions to be drawn about neural operations at normal temperatures.

Action Potentials↗

Changes of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate receptors in layer V of epileptogenic, chronically isolated rat neocortex.

In vivo chronic partial isolation of neocortical islands results in epileptogenesis that involves pyramidal neurons of layer V. To test whether an alteration in glutamate receptors might contribute to the epileptiform activity, we analysed the time-course of light microscopic changes in expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptors using subunit-specific antibodies. The isolation caused a rapid down-regulation of immunoreactivity for GluR1 and GluR2/3 subunits in deep layer V pyramidal neurons within the neocortical island which was evident 24h post-lesion, and within three days was reduced to about 40-60% of the control level. Many pyramidal cells in deep layer V completely lacked GluR2. Between one and four weeks of survival, down-regulation of GluR2/3 and GluR2 involved the majority of pyramidal layer V neurons, except for cells in the upper part of layer V, and those within narrow areas of all sub-laminae of layer V ("micro-islands"). Initial down-regulation was also observed one to three days post-lesion for subunits 1 and 2 of the N-methyl-D-aspartate receptor, but in contrast to GluR2/3 immunoreactivity, NMDAR2A/B immunoreactivity was enhanced three weeks post-lesion. The present data provide evidence for plastic changes in glutamate receptors in neurons of partially isolated neocortical island. A sub-population of layer V neurons remains relatively unaffected, and would presumably be capable of generating fast glutamatergic synaptic potentials necessary for the development of synchronous epileptiform activity.

Animals↗

Impact of antipsychotic drug administration on the expression of nuclear receptors in the neocortex and striatum of the rat brain.

We have recently shown that the expression of the nerve growth factor-inducible gene B (NGFI-B, or Nur77), a transcription factor belonging to the large ligand-activated nuclear receptor family, is modulated by antipsychotic drugs in the rat forebrain. In the present work, we have investigated the impact of antipsychotic drugs on a series of transcription factors also belonging to the nuclear receptor family. The receptors investigated include retinoid X receptor (RXR), thyroid hormone receptor (TR), retinoic acid receptor (RAR), RAR-related orphan receptor (RZR) and Rev-erb receptor isoforms in addition to the NGFI-B transcript. We have used in situ hybridization to monitor their mRNA levels after acute and chronic antipsychotic drug administration. RZRbeta and NGFI-B mRNA levels are down-regulated after chronic haloperidol or clozapine treatment in the primary somatosensory cortex. The TRbeta1 isoform mainly expressed in the cingulate cortex is modulated only after chronic clozapine treatment, whereas TRalpha isoform mRNAs are modulated by both antipsychotics in the cingulate cortex and nucleus accumbens shell; two brain areas associated with limbic functions. The RXRgamma1 isoform, mostly expressed in the dorsolateral portion of the striatum is modestly affected by antipsychotics. Modulation of the expression of transcription factors belonging to the ligand-activated nuclear receptor family by antipsychotics represents an additional molecular event in the mechanism of action of these drugs. We suggest that modification of the pattern of transcription factor expression may play a role in long-term cellular responses to these drugs.

Animals↗

Laminar properties of 4-aminopyridine-induced synchronous network activities in rat neocortex.

We examined the effects of 4-aminopyridine (4-AP) on isolated horizontal (superficial, middle and deep) rat neocortical slices in order to study laminar synchronous network behavior directly. Application of 4-AP induced spontaneous synchronized activity in all of these types of slices. In middle and deep layer slices the activities were similar to those of coronal slices, consisting of periodic short- and long-duration discharges. In superficial slices distinct spontaneous rhythmic multiphasic burst discharges were induced. Ionotropic glutamate receptor antagonists blocked the 4-AP-induced synchronous activities in middle and deep layer slices, but those in superficial slices persisted. The GABA(A) receptor antagonist picrotoxin suppressed this spontaneous synchronous activity resistant to 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (a NMDA receptor antagonist) and 6-cyano-7-nitroquinoxaline-2,3-dione (a non-NMDA receptor antagonist), in superficial slices, leaving small, slow spontaneous events. In superficial slices with intact excitatory amino acid transmission, picrotoxin attenuated the 4-AP-induced spontaneous synchronous discharges, even in this highly convulsant environment. By contrast, conventional coronal slices showed robust spontaneous epileptiform discharges under these circumstances. In intact coronal slices focal 4-AP application in superficial layers induced spontaneous inhibitory GABAergic events, while delivery into deep layers led to epileptiform discharges. From these results we conclude that: (1) 4-AP-induced population discharges are driven by glutamatergic transmission in middle and deep layer horizontal slices, and by GABAergic transmission in superficial layers; (2) only superficial layers are capable of supporting synchronized GABAergic activity independent of excitatory amino acid transmission; (3) superficial layers do not sustain epileptiform activity in the absence of deep layer neurons; and (4) synchronized superficial networks can inhibit deep layer neuronal activity.

4-Aminopyridine↗

Abnormal distributions of callosal commissural and corticothalamic neurons in the cerebral neocortex of Shaking Rat Kawasaki.

Shaking Rat Kawasaki (SRK) is an autosomal recessive mutant rat recognized by unstable gait and tremor and by early death around the time of weaning. We previously reported that corticospinal tract neurons are malpositioned in the motor cortex of the SRK rat [Ikeda and Terashima (1997) J. Comp. Neurol. 383, 370-380]. In the present study, we examined the distribution pattern of callosal commissural (CC) and corticothalamic (CT) neurons of SRK and normal rats with the injection of horseradish peroxidase (HRP) into the contralateral hemisphere or wheat germ agglutinin-conjugated HRP into the ventral lateral thalamic nucleus. The intracortical distribution pattern of retrogradely labeled CC and CT neurons in the motor cortex of SRK rat was abnormal: CC neurons were more deeply situated and CT neurons were more superficially situated in the SRK cortex than the corresponding components in the normal cortex. Most of labeled CC and CT neurons had abnormal dendritic configurations. Statistical analysis revealed that the difference of the mean intracortical position of CC and CT neurons of the SRK was significantly different from the normal counterparts (Student's t-test, P<0.01). Taken together with previous findings, our data demonstrate that the abnormal cytoarchitecture of SRK cortex resembles the reeler cortex.

Agenesis of Corpus Callosum↗

Acute neural damage in the rat neocortex in vitro induced by a combination of anoxia and mechanical stress.

To elucidate the mechanisms of neural damage after brain ischemia, rat neocortical slices were exposed to anoxia at room temperature for 1 h, and other slices were prepared from the neocortical blocks exposed to anoxia at room temperature for 1 h. Field potentials elicited by the stimulation of layer IV were recorded in supragranular layers in these slices. No clear damage was observed electrophysiologically or morphologically in these slices. In contrast, a complete loss of the trans-synaptic field potentials and a decrease in the density of the cells stained with Neutral Red were elicited by injecting an anoxic medium into the neocortical blocks at room temperature for 1 h. In the slice preparations, the injection of the anoxic medium failed to reproduce clear neural damage, while a combination of mechanical stress and anoxia elicited a complete loss of trans-synaptic potentials; this was alleviated by Gd3+ (50 microM) and D(-)-2-amino-5-phosphonovaleric acid (100 microM). These results indicate that a combination of mechanical stress and anoxia produces acute and severe neural damage even at room temperature in vitro. The mechanism of the damage and the relationship between the neural damage in vitro and in vivo are discussed.

Animals↗

Involvement of interleukin-1beta in the mechanism of human immunodeficiency virus type 1 (HIV-1) recombinant protein gp120-induced apoptosis in the neocortex of rat.

The effect of subchronic intracerebroventricular injection of the human immunodeficiency virus type 1 (HIV-1) recombinant protein gp120 (100 ng, given daily for up to seven consecutive days) on interleukin-1beta expression was studied by immunohistochemistry in the brain of adult rats. In comparison to control, bovine serum albumin (300 ng, given intracerebroventricularly for up to seven days) -treated animals (n=6), interleukin-1beta immunoreactivity increased in the brain cortex and hippocampus of rats (n=6) receiving a single injection of the viral protein 24 h before analysis with more substantial increases being observed in these regions of the brain (n=6) after seven days treatment. Double-labelling immunofluorescence experiments support a neuronal and, possibly, a microglial cell origin for gp120-enhanced interleukin-1beta expression. Transmission electron microscopy analysis of brain tissue sections revealed that combination treatments (given intracerebroventricularly daily for seven days) with gp120 (100 ng) and interleukin-1 receptor antagonist (80 ng) or with the interleukin converting enzyme inhibitor II (100 pmol), but not with leupeptin (100 pmol), prevented apoptotic death of rat (n=6/group) brain cortical cells typically elicited by the viral protein. These data demonstrate that gp120 enhances interleukin-1beta expression in the brain and this may be involved in the mechanism underlying apoptosis induced by gp120 in the brain cortex of rat. Further support to this hypothesis comes from the evidence that intracerebroventricular injection of murine recombinant interleukin-1beta (200 U, given daily for seven consecutive days) produces DNA fragmentation in the brain cortex of rat (n=6). Interestingly, the latter treatment enhanced nerve growth factor level in the hippocampus but not in the cerebral cortex and this coincides with a similar effect recently reported in identical brain areas of rats treated likewise with gp120. In conclusion, the present data demonstrate that treatment with gp120 enhances interleukin-1beta expression and this participates in the mechanism of apoptotic cell death in the brain cortex of rat. By contrast, in the hippocampus, gp120-enhanced interleukin-1beta expression elevates nerve growth factor that may prevent or delay apoptosis in this plastic region of the rat brain.

Animals↗

Relationship between locus coeruleus discharge rates and rates of norepinephrine release within neocortex as assessed by in vivo microdialysis.

The relationship between discharge rates of locus coeruleus noradrenergic neurons and rates of norepinephrine release was examined in the anesthetized rat. Neuronal discharge rates of locus coeruleus neurons were altered and quantified using a combined recording-infusion probe. Peri-locus coeruleus infusions of either the cholinergic agonist, bethanechol, or the alpha2-agonist, clonidine, were used to enhance or suppress neuronal discharge activity, respectively. Alterations in concentrations of extracellular norepinephrine within the prefrontal cortex were determined using in vivo microdialysis and high-pressure liquid chromatography with electrochemical detection. A linear relationship between locus coeruleus activity and norepinephrine dialysate concentration was observed between complete suppression of locus coeruleus discharge activity and approximately 300-400% of basal discharge levels (1.58+/-0.29 Hz). Above these levels, increases in locus coeruleus discharge rates were not accompanied by similar increases in dialysate norepinephrine concentrations. In general, neither activation nor suppression of locus coeruleus neuronal discharge rates appeared to alter the relationship between discharge activity and norepinephrine efflux during subsequent epochs. The one exception to this was observed during recovery from relatively high-magnitude locus coeruleus activation. In two out of three cases in which locus coeruleus discharge rates were increased greater than 450%, a recovery of norepinephrine concentrations to basal levels occurred more quickly than the recovery of locus coeruleus neuronal discharge rates to basal levels. Although limited, these latter observations suggest that dysregulation of norepinephrine release may occur following sustained activation of locus coeruleus at the highest rates examined, which may mimic those associated with intense arousal or stress.

Animals↗

A comparison of intermediate filament markers for presumptive astroglia in the developing rat neocortex: immunostaining against nestin reveals more detail, than GFAP or vimentin.

The present study compares the immunopositive elements in the developing rat cortex between the day of birth (P0) and the 18th postnatal day (P18), after immunostaining against nestin, vimentin and glial fibrillary acidic protein (GFAP). Nestin immunostaining revealed more structural details than either vimentin or GFAP, or they together. While vimentin immunostaining preferred radial glia and GFAP preferred astrocytes, nestin immunostaining detected both. Stellate-shaped astrocyte-like cells were already seen at P0 and cells of typical astrocytic morphology were numerous at P3, and were predominating elements from P7, whereas GFAP-immunopositive astrocytes were very scarce even at P7, and became numerous only by P11, when nestin immunopositivity started to disappear. Nestin immunostaining revealed such structures which were not seen in GFAP- or vimentin immunostained sections: cell body-like structures 'hanging' at the end the radial fibers, seeming to divide with their fibers, or having astrocyte-like processes. Nestin immunostaining is therefore highly recommended for studies of the glial architecture in the early post-natal brain development.

Age Factors↗

Separate progenitors for radial and tangential cell dispersion during development of the cerebral neocortex.

Cell lineage analyses suggest that cortical neuroblasts are capable of undertaking both radial and tangential modes of cell movement. However, it is unclear whether distinct progenitors are committed to generating neuroblasts that disperse exclusively in either radial or tangential directions. Using highly unbalanced mouse stem cell chimeras, we have identified certain progenitors that are committed to one mode of cell dispersion only. Radially dispersed neurons expressed glutamate, the neurochemical signature of excitatory pyramidal cells. In contrast, tangential progenitors gave rise to widely scattered neurons that are predominantly GABAergic. These results suggest lineage-based mechanisms for early specification of certain progenitors to distinct dispersion pathways and neuronal phenotypes.

Animals↗

Synchronous activity of inhibitory networks in neocortex requires electrical synapses containing connexin36.

Inhibitory interneurons often generate synchronous activity as an emergent property of their interconnections. To determine the role of electrical synapses in such activity, we constructed mice expressing histochemical reporters in place of the gap junction protein Cx36. Localization of the reporter with somatostatin and parvalbumin suggested that Cx36 was expressed largely by interneurons. Electrical synapses were common among cortical interneurons in controls but were nearly absent in knockouts. A metabotropic glutamate receptor agonist excited LTS interneurons, generating rhythmic inhibitory potentials in surrounding neurons of both wild-type and knockout animals. However, the synchrony of these rhythms was weaker and more spatially restricted in the knockout. We conclude that electrical synapses containing Cx36 are critical for the generation of widespread, synchronous inhibitory activity.

Animals↗

The timing and laminar profile of converging inputs to multisensory areas of the macaque neocortex.

Two fundamental requirements for multisensory integration are convergence of unisensory (e.g. visual and auditory) inputs and temporal alignment of the neural responses to convergent inputs. We investigated the anatomic mechanisms of multisensory convergence by examining three areas in which convergence occurs, posterior auditory association cortex, superior temporal polysensory area (STP) and ventral intraparietal sulcus area (VIP). The first of these was recently shown to be a site of multisensory convergence and the latter two are more well known as 'classic' multisensory regions. In each case, we focused on defining the laminar profile of response to the unisensory inputs. This information is useful because two major types of connection, feedforward and feedback, have characteristic differences in laminar termination patterns, which manifest physiologically. In the same multisensory convergence areas we also examined the timing of the unisensory inputs using the same standardized stimuli across all recordings. Our findings indicate that: (1) like somatosensory input [J. Neurophysiol., 85 (2001) 1322], visual input is available at very early stages of auditory processing, (2) convergence occurs through feedback, as well as feedforward anatomical projections and (3) input timing may be an asset, as well as a constraint in multisensory processing.

Acoustic Stimulation↗

Dynamic programming generation of boundaries of local coordinatized submanifolds in the neocortex: application to the planum temporale.

Dynamic programming is used to define boundaries of cortical submanifolds with focus on the planum temporale (PT) of the superior temporal gyrus (STG), which has been implicated in a variety of neuropsychiatric disorders. To this end, automated methods are used to generate the PT manifold from 10 high-resolution MRI subvolumes ROI masks encompassing the STG. A procedure to define the subvolume ROI masks from original MRI brain scans is developed. Bayesian segmentation is then used to segment the subvolumes into cerebrospinal fluid, gray matter (GM), and white matter (WM). 3D isocontouring using the intensity value at which there is equal probability of GM and WM is used to reconstruct the triangulated graph representing the STG cortical surface, enabling principal curvature at each point on the graph to be computed. Dynamic programming is used to delineate the PT manifold by tracking principal curves from the retro-insular end of the Heschl's gyrus (HG) to the STG, along the posterior STG up to the start of the ramus and back to the retro-insular end of the HG. A coordinate system is then defined on the PT manifold. The origin is defined by the retro-insular end of the HG and the y-axis passes through the point on the posterior STG where the ramus begins. Automated labeling of GM in the STG is robust with L(1) distances between Bayesian and manual segmentation in the range 0.001-0.12 (n = 20). PT reconstruction is also robust with 90% of the vertices of the reconstructed PT within about 1 voxel (n = 20) from semiautomated contours. Finally, the reliability index (based on interrater intraclass correlation) for the surface area derived from repeated reconstructions is 0.96 for the left PT and 0.94 for the right PT, thus demonstrating the robustness of dynamic programming in defining a coordinate system on the PT. It provides a method with potential significance in the study of neuropsychiatric disorders.

Algorithms↗

Experience-dependent changes in dendritic arbor and spine density in neocortex vary qualitatively with age and sex.

Male and female Long-Evans hooded rats were placed in the complex environments for 3 months either at weaning (22 days), in young adulthood (120 days), or in senescence (24 months). The dendritic morphology of both the apical and basilar fields of layer III pyramidal cells was analyzed in both parietal and visual cortex. There were two novel results. First, although spine density was increased significantly with complex-housing in adulthood, it was decreased significantly by the same housing during development. Second, dendritic length was increased in both parietal and occipital cortex at all ages in males and was increased in adult females as well, but juvenile females showed no change in dendritic length in the occipital cortex and only a small effect on the apical field in parietal cortex. Thus, there are qualitative differences in the changes in spine density at different ages and the dendritic changes in response to complex versus isolated housing vary with sex, and in females, the changes vary with age as well. These results may explain some of the apparent inconsistencies in reports of spine and dendrite changes in the literature.

Age Factors↗

Linked spike activity of rabbit neocortex neurons during self-stimulation of the lateral hypothalamus.

Cross-correlation and autocorrelation histograms were constructed to study the linkage between the spike activity of neurons in the visual and sensorimotor areas of both cerebral hemispheres in rabbits over periods of 10 sec after self-stimulation of the right and left lateral hypothalamus. Self-stimulation was followed by an increase in the number of pairs of cells in which neurons in the sensorimotor area fired after cells in the visual cortex (from 24 +/- 6% to 44 +/- 6%). Analysis of the spectrum of linked discharges revealed increases in the alpha and theta frequency ranges: the main increases in the right hemisphere were in the alpha range, while the main increases in the left hemisphere were in the theta range. Interhemisphere asymmetry in the interaction of cortical neurons was not associated with the side of the hypothalamus subjected to stimulation.

Animals↗