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Postnatal shifts of interneuron position in the neocortex of normal and reeler mice: evidence for inward radial migration.

During development, interneurons migrate to precise positions in the cortex by tangential and radial migration. The objectives of this study were to characterize the net radial migrations of interneurons during the first postnatal week, and to investigate the role of reelin signaling in regulating those migrations. To observe radial migrations, we compared the laminar positions of interneurons (immunoreactive for GABA or Dlx) in mouse neocortex on postnatal days (P) 0.5 and P7.5. In addition, we used bromodeoxyuridine birthdating to reveal the migrations of different interneuron cohorts. To study the effects of reelin deficiency, experiments were performed in reeler mutant mice. In normal P0.5 cortex, interneurons were most abundant in the marginal zone and layer 5. By P7.5, interneurons were least abundant in the marginal zone, and were distributed more evenly in the cortical plate. This change was attributed mainly to inward migration of middle- to late-born interneurons (produced on embryonic days (E) 13.5 to E16.5) from the marginal zone to layers 2-5. During the same interval, late-born projection neurons (non-immunoreactive for GABA or Dlx) migrated mainly outward, from the intermediate zone to upper cortical layers. In reeler cortex, middle- and late-born interneurons migrated from the superplate on P0.5, to the deep cortical plate on P7.5. Late-born projection neurons in reeler migrated in the opposite direction, from the intermediate zone to the deep cortical plate. We conclude that many middle- and late-born interneurons migrate radially inward, from the marginal zone (or superplate) to the cortical plate, during the first postnatal week in normal and reeler mice. We propose that within the cortical plate, interneuron laminar positions may be determined in part by interactions with projection neurons born on the same day in neurogenesis.

Animals↗

Electron microscopical localization of guanylate cyclase activity in the neocortex of the guinea pig.

The localization of the guanylate cyclase (GC) activity has been established in the neocortex of adult guinea pigs by means of electron microscopical histochemistry [the DMSO-method of Fujimoto et al. (1981)]. Reaction product was deposited within a population of large- and medium-sized cortical neurons as well as in the cytoplasm of a part of the dendrites of variable size and in the cytoplasm and the nuclear membrane of a number of protoplasmic astrocytes. In the perikarya of the positive neurons, the reaction precipitate was mainly located within the cisterns of the rough endoplasmic reticulum and on the nuclear membrane. In the dendrites, the reaction product was usually distributed in close contact with microtubules, microfilaments, and beneath the postsynaptic membranes of a number of axodendritic synaptic contacts. The axons and all presynaptic boutons were negative. Thus, the localization of the GC could be determined as exclusively postsynaptic. The results obtained support the view for the probable participation of cyclic GMP in the cholinergic, glutaminergic or GABAergic, or peptidergic transmitter mechanisms in the central nervous system.

Animals↗

Neuronal nitric oxide has a role as a perfusion regulator and a synaptic modulator in cerebellum but not in neocortex during somatosensory stimulation--an animal PET study.

To clarify a role of neuronal nitric oxide in neurovascular coupling, we performed cerebral blood flow (CBF) and cerebral metabolic rate of glucose (CMR(glc)) measurements with positron emission tomography in somatosensory-stimulated cats using a specific neuronal nitric oxide synthase inhibitor, 7-nitroindazole (7-NI). The effect on flow-metabolism coupling were tested by global and regional-specific changes on CBF and CMR(glc), and the regional-specific effect was estimated both by regions of interest (ROI) and voxel-based (VB) analysis using globally-normalized CBF and CMR(glc) changes. The electrical somatosensory stimulation in the unilateral forepaw elicited coupled increase in CBF and CMR(glc) in the contralateral somatosensory cortex (7%) and the ipsilateral cerebellum (8%). 7-NI induced 20% decrease in global CBF both during rest and activation, but not in global CMR(glc) at simulation. Both ROI and VB analysis showed that 7-NI induced an increase in CMR(glc) (13%) in the ipsilateral cerebellum compared to control under vehicle alone, but it was accompanied by only 8% increase in CBF, suggesting uncoupling of flow-metabolism while it induced any perturbations in the contralateral somatosensory cortex. These observations suggest that neuronal nitric oxide has an important role for a mediator of regional neurovascular coupling as well as synaptic modulator in the cerebellum, but less so in the neocortex.

Animals↗

Expression of NMDAR2D glutamate receptor subunit mRNA in neurochemically identified interneurons in the rat neostriatum, neocortex and hippocampus.

NMDA receptors are composed of proteins from two families: NMDAR1, which are required for channel activity, and NMDAR2, which modulate properties of the channels. The mRNA encoding the NMDAR2D subunit has a highly restricted pattern of expression: in the forebrain, it is found in only a small subset of cortical, neostriatal and hippocampal neurons. We have used a quantitative double-label in situ hybridization method to examine the expression of NMDAR2D mRNA in neurochemically defined populations of neurons. In the neostriatum, NMDAR2D was expressed by the interneuron populations marked by preprosomatostatin (SOM), the 67-kDa form of glutamic acid decarboxylase (GAD67), parvalbumin (PARV), and choline acetyltransferase (ChAT) mRNAs but not by the projection neurons expressing beta-preprotachykinin (SP) or preproenkephalin (ENK) mRNAs. In the neocortex, NMDAR2D expression was observed in only a small number of neurons, but these included almost all of the SOM-, GAD67-, and PARV-expressing interneurons. In the hippocampus, NMDAR2D was not present in pyramidal or granule cells, but was abundant in SOM-, GAD67-, and PARV-positive interneurons. NMDAR2D expression appears to be a property shared by interneurons in several regions of the brain. The unique electrophysiological characteristics conveyed by this subunit, which include resistance to blockade by magnesium ion and long channel offset latencies, may be important for the integrative functions of these neurons. NMDAR2D-containing receptor complexes may prove to be important therapeutic targets in human disorders of movement. In addition, the presence of NMDAR2D subunits may contribute to the differential vulnerability of interneurons to excitotoxic injury.

Animals↗

Morphologic alterations of cholinergic processes in the neocortex of aged rats.

In the present study we observed enlarged cholinergic processes in the neocortex of aged Fischer 344 rats. These swollen ChAT-positive profiles appeared either as a single axon enlargement or, in many instances, the bulbous processes coalesced to form grape-like clusters of immunoreactivity. The latter structures looked similar to the immunoreactive profiles observed in the cortex of patients with Alzheimer's disease and in the rat septum following fimbria-fornix transection. Together, these data provide evidence that morphologic changes occur within processes of cholinergic neurons in the aged rat. Moreover, the similarity in appearance between the axonal alterations in the aged rat and in patients with Alzheimer's disease suggests a common pathologic process.

Aging↗

Widely integrative properties of layer 5 pyramidal cells support a role for processing of extralaminar synaptic inputs in rat neocortex.

We have compared the length, strength, conduction velocity and divergence of horizontal connections onto layers 2/3 and 5 neurons in slices of rat primary somatosensory neocortex. Slices were cut along laminar borders to eliminate most vertical connections, and excitatory postsynaptic potentials (EPSPs) were recorded from pyramidal cells in adjacent uncut tissue. When electric stimuli were delivered within the same layer as the recorded cell, EPSPs could be evoked up to 2000 microm away for both layers 2/3 and 5 pyramids. Estimates of horizontal axonal conduction velocities ( approximately 0.4 m/s) and the thresholds for activation also did not differ between layers. However, layers 2/3 cells rarely responded to stimuli delivered to isolated deeper layers, while layer 5 neurons were often excited by horizontal inputs from isolated layers 2/3 and 4.

Animals↗

Localization of KCNQ5 in the normal and epileptic human temporal neocortex and hippocampal formation.

The KCNQ family of voltage-dependent non-inactivating K+ channels is composed of five members, four of which (KCNQ2-5) are expressed in the CNS and are responsible for the M-current. Mutations in either KCNQ2 or KCNQ3 lead to a hereditary form of dominant generalized epilepsy. Using specific antisera to the KCNQ2, KCNQ3 and KCNQ5 subunits, we found that KCNQ3 co-immunoprecipitated with KCNQ2 and KCNQ5 subunits, but no association was detected between KCNQ2 and KCNQ5. Intense KCNQ5 immunoreactivity was found to be widely distributed throughout the temporal neocortex and the hippocampal formation. In these structures, both pyramidal and non-pyramidal neurons and a population of glial cells in the white matter expressed the KCNQ5 subunit. In the sclerotic areas of the CA fields of epileptic patients, a marked loss of KCNQ5 immunoreactive pyramidal neurons was found in relation with the loss of neurons in these regions. However, in the regions adjacent to the sclerotic areas, the distribution and intensity of KCNQ5 immunostaining was apparently normal. The widespread distribution of KCNQ5 subunits, its persistence in pharmacoresistant epilepsy, along with the significant role of the M-current in the control of neuronal excitability, makes this protein a possible target for the development of anticonvulsant drugs.

Adolescent↗

Neurons are generated in confluent astroglial cultures of rat neonatal neocortex.

Cells of the telencephalon are generated in specific proliferative zones from which neuronal and glial precursors migrate to their destinations. Recent evidence indicates that some precursors do not turn into differentiated cells but keep their ability to proliferate. Here, we report that neurons can originate in primary cultures of astroglial cells prepared from neocortex of newborn rats. The first neuronal cells appeared shortly before confluence, when a glial monolayer was being formed. After confluence, these still undifferentiated cells increased in number. Later, they became immunohistochemically positive for the neuron-specific marker microtubule-associated protein 2a,b. They also contained neurofilament-L protein as well as the specific messenger RNA coding for neurofilament-H. The observation that they took up bromo-deoxyuridine indicated that they synthesized DNA, i.e. they proliferated. When Dulbecco's modified essential medium was substituted with fetal calf serum, the appearance of neurons depended on the seeding density of the dispersed cells. This was no longer the case, when the cultures were maintained in Dulbecco's modified essential medium/F12 medium to which transferrin, insulin and selenium chloride had been added. It is concluded that neuronal precursors can survive in primary astroglial cultures. After confluence of the astroglial cells the precursors proliferate if appropriate conditions are present. Our observation provides a new model for the investigation of cultured neurons and neuronal-glial interactions.

Animals↗

[Functional maturation of the somatosensory neocortex S1 in the rat. Electrophysiological and histochemical study (author's transl)].

The functional maturation of the somatosensory neocortex has been studied using the projection areas of the whiskers. Natural stimulation of the whiskers evokes global potentials and unitary responses from the 4th postnatal day on. From the 2nd postnatal week the characteristics of the unit responses approximate in latency, topographic organisation of the projections, coding of the directions and temporal characteristics of the stimulation those of the adult. The histochemical activity of succinyl dehydrogenase is demonstrated in layer IV a little before the first electrophysiological response but the typical massed organisation is only seen from the 5th postnatal day. During maturation optical densitometry shows a rapid rise in enzyme activity untill the 7th postnatal day then a slower rise up to the 21st day after which it stabilises.

Aging↗

A technique based on the use of activated charcoal for easier subsequent retrieval of neocortical grafts placed in the neocortex of newborn rats.

The mechanisms underlying the differentiation of neocortical areas are still largely unknown. The development of neural connectivity constitutes one important step in neocortical differentiation. One way to study the mechanisms guiding this developmental stage is to examine the connections established by transplants of neocortical tissue of varying embryonic age placed in varying areas of the neocortex of newborn hosts. Neurotracer injection into the transplant at different intervals following transplantation is then used to identify the development of host-transplant connectivity. In most cases, however, it is rather difficult to retrieve the transplant within the host cortex even shortly after grafting. Hence, it is very difficult to perform tracer injections limited to the transplant without any involvement of the host cortex. In some instances, the transplant position can be predicted by some weaker vascularization within or at the surface of the graft. This is not, however, a reliable criterion to establish the rostrocaudal and mediolateral coordinates of the tracer injection. In this report, we describe the use of activated charcoal to mark the transplant at the time of transplantation. The transplant containing black dots can subsequently be easily distinguished from the host pale pink cortex.

Animals↗

Biochemical markers of excitability in human neocortex.

We measured biochemical markers of excitability in brain excised for neurosurgical therapy of epilepsy. Intraoperative electrocorticography was used to identify and compare samples from regions of persistent interictal spike discharges and areas of the cerebral convexity which were free of interictal spiking. We found that interictal spiking was associated with elevated tissue levels of the excitatory amino acids glutamic acid (26%, p less than 0.001) and aspartic acid (25%, p less than 0.05). There was also a significant increase in the activity of the enzymes glutamic acid dehydrogenase (20%, p less than 0.01) and aspartate acid aminotransferase (18%, p less than 0.01) which are involved in their formation. There was no change in the levels of the inhibitory neurotransmitters GABA or taurine. We also found a significant increase in the activity of tyrosine hydroxylase (52%, p less than 0.001), the rate controlling enzyme in catecholamine biosynthesis. There was a reduction in the density (Bmax) of cortical alpha-1 adrenoceptors (26%, p less than 0.01) and a concomitant diminution of receptor coupled phosphatidylinositide metabolism (21%, p less than 0.01). This blunting of inhibitory noradrenergic transmembrane signaling may contribute to a relative imbalance between excitatory and inhibitory mechanisms in epileptogenic neocortex.

Amino Acids↗

Intracortical synchronization of epileptic discharges at different stages of ultrastructural rearrangements in a completely neuronally isolated area of rat neocortex.

Cross-correlation functions were used to study the time delay (a measure of synchronization) in the appearance of epileptic discharges in distant areas of the cortex in the intact cortex and in neuronally isolated cortical strips in Wistar rats. Experiments were performed at different stages of axon sprouting 30 and 90 days after isolation of cortical areas and in intact cortex. Significant increases in the number of synapses in layer V of isolated cortical strips at 30 days correlated with significant decreases in the time delay, while decreases in the number of synapses at 90 days correlated with a significant increase in the time delay. This is evidence that newly formed synapses increase the extent of synchronization and thus affect epileptogenesis. The data obtained here suggest that large pyramidal cells in layer V of the rat neocortex form a neural network in pathological conditions, this supporting intracortical synchronization of epileptic discharges.

Animals↗

Retention of maze performance inversely correlates with N-methyl-D-aspartate receptor number in hippocampus and frontal neocortex in the rat.

The N-methyl-D-aspartate (NMDA) receptor may play a critical role in learning and memory. In the present study, a significant correlation was found between the number of NMDA-displaceable, Na+-independent L-[3H]glutamate binding sites in the hippocampus and neocortex of young rats and the mean number of errors during retention, but not acquisition, in a 14-unit T-maze.

Animals↗

Performance on two fixed-interval schedules in the absence of neocortex in rats.

Rats with complete surgical removal of the neocortex (neodecorticates) were trained, with milk as the reinforcer, on a 40-s fixed-interval (FI-40) schedule, then on an FI-120, and finally reexposed to the FI-40. Like controls, neodecorticates developed and appropriately adjusted their postreinforcement pauses and response distributions and showed scallop-type responding on the FI-120 and a positive correlation between running rate and the preceding postreinforcement pause on both FI-40 schedules but not on the FI-120. On some variables neodecorticates reached asymptotic levels of performance more slowly than controls. Extended training on the first FI-40 did not differentially affect neodecorticates' performance. The results suggest that neodecorticates retain a functional timing mechanism.

Animals↗

Odor cue mediation of alcohol aversion learning in rats lacking gustatory neocortex.

Normal rats presented with a 5% alcohol solution followed by lithium chloride-induced illness quickly learned to avoid drinking alcohol. After training, the rats also avoided drinking water in the presence of the alcohol odor alone, whether tested immediately or 1 month later. In Experiment 1, rats with gustatory neocortex (GN) ablations also developed strong alcohol aversions when the alcohol solution was paired with illness. They also showed normal avoidance of drinking in the presence of the alcohol odor alone when tested soon after training. In Experiment 2, when normal rats were trained to avoid alcohol, given GN ablations, and then tested for retention 1 month later, avoidance of drinking water in the presence of the odor alone was significant but attenuated somewhat in relation to trained control rats. These data support the hypothesis that rats lacking GN partially acquire alcohol aversions by using odor cues and confirm that associative learning is intact in these rats despite the fact that GN rats display significant deficits in aversion learning when only tastes are paired with illness.

Alcohol Drinking↗

Taste agnosia following gustatory neocortex ablation: dissociation from odor and generality across taste qualities.

In Experiment 1, rats trained to avoid drinking in the presence of a compound odor (benzyl acetate) and taste (sucrose) conditional stimulus (CS) lost the taste habit but retained the odor habit following gustatory neocortex (GN) ablation. Conversely, olfactory bulb ablation resulted in loss of the odor habit but retention of the taste habit. In Experiment 2, rats lacking GN did not retain preoperatively instated learned aversions to a suprathreshold quinine hydrochloride (bitter) taste solution which had been employed as a CS. However, rats with GN lesions that were virtually identical to those of the bitter-trained group retained a preoperatively learned aversion to a hydrochloric acid (sour) CS. Experiment 3 demonstrated that reliable agnosia for an acid CS could be produced by lesions that extended more deeply into perirhinal areas near the claustrum at the level of the GN. It is concluded that the agnosia following GN ablation is relatively specific to gustation and that agnosia for preoperatively acquired taste aversion habits occurs for all four basic gustatory stimuli following anterolateral cortex ablations centered on the GN.

Agnosia↗

Neonatal ablations of the gustatory neocortex in the rat: taste aversion learning and taste reactivity.

Rats sustaining ablations of gustatory neocortex (GN) at 2, 10, 20, or 60 days of age were compared with control rats in the acquisition and extinction of a learned taste aversion; in addition, these rats were tested for taste preference across five concentrations of sodium chloride solution. Results indicated that GN ablation disrupted aversion acquisition and extinction regardless of age at surgery. Taste response functions for the sodium chloride solutions shown by all GN groups of rats mirrored those of control rats: preference (relative to water baseline) for middle concentrations and rejection of the strongest salt concentration. There was a suggestion that the 20- and 60-day-old GN rats were hyperresponsive to the suprathreshold concentrations of NaCl (except the strongest concentration). The increased response to salt solutions in the 20- and 60-day GN rats may have been related to the significant decreases in water consumption relative to that of normal rats. Water consumption of control rats and GN rats in the 2-day and 10-day groups was essentially equal. It is concluded that infant ablation of the GN does not spare normal taste aversion learning and that rats with GN ablations, regardless of age at surgery, respond in a normal manner to the hedonic aspects of sodium chloride solutions.

Age Factors↗