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Afferent connections of dorsal and ventral agranular insular cortex in the hamster Mesocricetus auratus.

The agranular insular cortex is transitional in location and structure between the ventrally adjacent olfactory allocortex primutivus and dorsally adjacent sensory-motor isocortex. Its ventral anterior division receives major afferent projections from olfactory areas of the limbic system (posterior primary olfactory cortex, posterolateral cortical amygdaloid nucleus and lateral entorhinal cortex) while its dorsal anterior division does so from non-olfactory limbic areas (lateral and basolateral amygdaloid nuclei). The medial segment of the mediodorsal thalamic nucleus projects to both the ventral and dorsal divisions of the agranular insular cortex, to the former from its anterior portion and to the latter from its posterior portion. Other thalamic inputs to the two divisions arise from the gelatinosus, central medial, rhomboid and parafascicular nuclei. The dorsal division, but not the ventral division, receives input from neurons in the lateral hypothalamus and posterior hypothalamus. The medial frontal cortex projects topographically and bilaterally upon both ventral and dorsal anterior insular cortex, to the former from the ventrally located medial orbital and infralimbic areas, to the latter from the dorsally-located anterior cingulate and medial precentral areas, and to both from the intermediately located prelimbic area. Similarly, the ipsilateral posterior agranular insular cortex and perirhinal cortex project in a topographic manner upon the two divisions of the agranular insular cortex. Commissural input to both divisions originates from pyramidal neurons in the respective contralateral homotopical cortical area. In each case, pyramidal neurons in layer V contribute 90% of this projection and 10% arises from layer III pyramidals. In the brainstem, the dorsal raphe nucleus projects to the ventral and dorsal divisions of the agranular insular cortex and the parabrachial nucleus projects to the dorsal division. Based on their cytoarchitecture, pattern of afferent connections and known functional properties, we consider the ventral and dorsal divisions of the agranular insular cortex to be, respectively, periallocortical and proisocortical portions of the limbic cortex.

Afferent Pathways↗

Distribution of preproenkephalin messenger RNA in the basal ganglia and limbic-associated regions of the monkey telencephalon.

We studied the distribution of preproenkephalin messenger RNA in the monkey forebrain, using a free-floating method for in situ hybridization histochemistry. Autoradiographs reveal a high level of specific hybridization to preproenkephalin messenger RNA in the monkey striatum and forebrain regions. In the monkey striatum, the distribution of preproenkephalin messenger RNA is heterogeneous. There is variation in the general labeling pattern between regions of the striatum. For example, a particularly densely labeled area of preproenkephalin messenger RNA is observed in the ventral part of the caudal putamen. In addition, at the macroscopic level, there are patches of specific hybridization intermingled with areas containing less specific labeling. This forms a mosaic-like pattern. At the microscopic level, densely labeled individual cells are found among those with little or no specific labeling. Adjacent sections, processed for in situ hybridization and immunohistochemistry, show some correlation between the perikarya containing preproenkephalin messenger RNA and enkephalin-positive fibers in the striatum. Specific hybridization to preproenkephalin messenger RNA is evident throughout the cortical mantle, primarily concentrated in layers 2 and 3. Particularly high levels of preproenkephalin messenger RNA are found in specific limbic-associated cortices, including the piriform allocortex, the agranular area of the orbitofrontal cortex, the agranular insular cortex and the caudal field of the entorhinal cortex. Specific labeling is also present in the granular cell layer of the dentate gyrus and in the amygdaloid complex. This study reveals heterogeneous distribution of dense preproenkephalin messenger RNA in the basal ganglia and high levels of preproenkephalin messenger RNA in specific limbic-associated regions of the monkey telencephalon.

Animals↗

Region specific regulation of NR1 in rhesus monkeys following chronic antipsychotic drug administration.

BACKGROUND: Altered NMDA receptor subunit protein levels have been reported in various regions of the schizophrenic brain; however, chronic antipsychotic administration in schizophrenic subjects may confound interpretation. METHODS: The effects of chronic antipsychotic drug administration (haloperidol and clozapine) on protein levels of NR1, NR2A and NR2B proteins were evaluated in the nucleus accumbens (NAc), putamen (PUT), dorsolateral prefrontal cortex (DLPFC), superior temporal gyrus (STG), and entorhinal cortex (EC) of rhesus monkeys using Western blot analysis. RESULTS: Haloperidol administration significantly decreased NR1 expression in the DLPFC. In contrast, NR2B expression was not affected by antipsychotic administration in any brain region examined. NR2A was not reliably detected in any of the brain regions. CONCLUSIONS: Results indicate that the NR1 subunit in the DLPFC may be a substrate for antipsychotic action and that glutamatergic hypofunction in the DLPFC commonly associated with cognitive dysfunction in schizophrenia may be associated with haloperidol administration.

Animals↗

Cortical areas abundant in extracellular matrix chondroitin sulphate proteoglycans are less affected by cytoskeletal changes in Alzheimer's disease.

In the human brain, the distribution of perineuronal nets occurring as lattice-like neuronal coatings of extracellular matrix proteoglycans ensheathing several types of non-pyramidal neurons and subpopulations of pyramidal cells in the cerebral cortex is largely unknown. Since proteoglycans are presumably involved in the pathogenesis of Alzheimer's disease, we analysed the distribution pattern of extracellular chondroitin sulphate proteoglycans in cortical areas, including primary motor, primary auditory and several prefrontal and temporal association areas, in normal human brains and in those showing neuropathological criteria of Alzheimer's disease. In both groups, neurons with perineuronal nets were most numerous in the primary motor cortex (approximately 10% in Brodmann's area 4) and in the primary auditory cortex as a representative of the primary sensory areas. Their number was lower in secondary and higher order association areas. Net-associated pyramidal cells occurred predominantly in layers III and V in motor areas, as well as throughout lower parts of layer III in the primary auditory cortex and neocortical association areas. In the entorhinal cortex, net-associated pyramidal cells were extremely rare. In brains showing hallmarks of Alzheimer's disease, the characteristic patterns of hyperphosphorylated tau protein, stained with the AT8 antibody, largely excluded the zones abundant in perineuronal nets and neuropil-associated chondroitin sulphate proteoglycans. As shown in double-stained sections, pyramidal and non-pyramidal neurons ensheathed by perineuronal nets were virtually unaffected by the formation of neurofibrillary tangles even in severely damaged regions. The distribution patterns of amyloid B deposits overlapped but showed no congruence with that of the extracellular chondroitin sulphate proteoglycans. It can be concluded that low susceptibility of neurons and cortical areas to neurofibrillary changes corresponds with high proportions of aggregating chondroitin sulphate proteoglycans in the neuronal microenvironment.

Aged↗

Neural inputs into the nucleus basalis of the substantia innominata (Ch4) in the rhesus monkey.

Neurons of the nucleus basalis-substantia innominata-nucleus of the ansa peduncularis complex (Ch4) provide the major source of cholinergic innervation for the entire neocortical surface. In contrast to their widespread projections to all parts of the neocortex, these neurons receive reciprocal projections from only very few cortical areas. Most of the sensory, motor, and association areas in the frontal, parietal, occipital and temporal lobes do not seem to project back to the Ch4 complex. The Ch4 neurons receive their cortical input from prepyriform cortex, orbitofrontal cortex, the anterior insula, the temporal pole, entorhinal cortex and the medial temporal cortex. There are also subcortical inputs from septal nuclei, the nucleus accumbens-ventral pallidum complex and the hypothalamus. This organization suggests that the Ch4 complex is in a position to act as a cholinergic relay for transmitting predominantly limbic and paralimbic information to the neocortical surface. It would also appear that the cortical areas which do not project into Ch4 have no direct way of controlling the cholinergic input which they receive, whereas the limited set of cortical areas which do project into Ch4 can control not only the cholinergic innervation that they receive but also the cholinergic innervation into the entire neocortical mantle.

Alzheimer Disease↗

Exposure to behaviourally relevant odour reveals differential characteristics in rat central olfactory pathways as studied through oscillatory activities.

This study investigated how changes in nutritional motivation modulate odour-related oscillatory activities at several levels of the olfactory pathway in non-trained rats. Local field potential recordings were obtained in freely moving animals in the olfactory bulb (OB), anterior and posterior parts of the piriform cortex (APC and PPC respectively) and lateral entorhinal cortex (EC). Dynamic signal analysis detected changes in power during odour presentation for several frequency bands The results showed that in most cases odour presentation was associated with changes in a wide 15-90 Hz frequency band of activity in each olfactory structure. However, nutritional state modulated initial responses to food odour (FO) in the OB and EC selectively in the 15-30 Hz frequency band. Changes in nutritional state also modulated responses to repeated FO stimuli. Habituation was expressed differentially across structures with a clear dissociation between the two parts of the piriform cortex. Finally, systemic injections of scopolamine (0.125 mg/kg) selectively blocked expression of the nutritional modulation in the OB found in the beta band. These results suggest that internal state can differentially modulate odour processing among different olfactory areas and point to a cholinergic-sensitive beta band oscillation during presentation of a behaviourally meaningful odorant.

Animals↗

Mnemonic firing of neurons in the monkey temporal pole during a visual recognition memory task.

1. We examined single-neuronal activity in the temporal pole of monkeys, including the anterior ventromedial temporal (VMT) cortex (the temporopolar cortex, area 36, area 35, and the entorhinal cortex) and the anterior inferotemporal (IT) cortex, during a visual recognition memory task. In the task, a trial began when the monkey pressed a lever. After a waiting period, a visual sample stimulus (S) was presented one to four times on a monitor with an interstimulus delay. Thereafter, a new stimulus (R) was presented. The monkeys were trained to remember S during the delay period and to release the lever in response to R. Colored photographs of natural objects were used as visual stimuli. 2. About 70% of the recorded neurons (225 of 311) responded to at least one of the Ss tested. Thirty percent of these neurons (68 of 225) continued to fire during the subsequent delay periods. In 75% of these neurons (51 of 68), the firing during the delay period strongly correlated with the response to S. 3. The discharge rate during the delay period did not correlate with the monkey's eye movements, pressing or releasing of the lever, or the reaction time. 4. If the monkey erroneously released the lever in response to S or during the delay period, the firing disappeared after the erroneous lever release. If the monkey failed to release the lever in response to R, the firing persisted even after R was withdrawn. The discharge rate in incorrect trials was comparable with that in correct trials. The neurons were considered to fire for as long as the memory of S was necessary. 5. Firing persisted even when an achromatic version or half (even a portion) of S was presented, indicating that the color, a particular portion, or the entire shape of S was not always necessary to elicit firing. 6. An S that elicited firing during the delay period invariably elicited a visual response. Neurons that fired during the delay period showed a higher stimulus selectivity than other visually responsive neurons in the anterior VMT cortex. Thus neurons that fire during the delay period represent a subgroup of visually responsive neurons that are selectively tuned to a certain stimulus. 7. More neurons fired during the delay period in the anterior VMT cortex than in the anterior IT cortex. 8. We conclude that firing during the delay period by neurons in the temporal pole reflects the short-term storage of visual information regarding a particular S.

Animals↗

Entorhinal axons perforate hippocampal field CA3 in organotypic slice culture.

In growing towards their hippocampal targets, incoming afferent axons from the entorhinal cortex arrive at the subicular pole of the hippocampus and normally turn pialwards from the alvear path, crossing (perforating) the subiculum and field CA1, but never the more distally situated field CA3. To address the question of whether a specific repulsive characteristic of field CA3 might explain this behaviour, artificial confrontation were set up in vitro. Embryonic entorhinal explants were placed in restricted contact with 8-day-old rat hippocampal slices, orientated so that outgrowing axons could only grow into either the dentate gyrus, the subiculum/field CA1, or field CA3. Anterograde biotin-dextran labelling of projections after 2 weeks in culture showed that entorhinal axons perforated the stratum oriens, pyramidal cell layer, and stratum radiatum of CA3 just as readily as they did along their normal trajectory across CA1/subiculum. It is therefore concluded that spatiotemporal cues are more likely than specific chemorepulsive molecules to be involved in setting up this part of the entorhinal pathway.

Animals↗

Myelin does not influence the choice behaviour of entorhinal axons but strongly inhibits their outgrowth length in vitro.

Myelin is crucial for the stabilization of the entorhinohippocampal projection during late development and is a non-permissive substrate for regrowing axons after lesion in the adult brain. We used two in vitro assays to analyse the impact of myelin on rat entorhinohippocampal projection neurons. A stripe assay was used to study the impact of myelin on the choice behaviour of axons from the entorhinal cortex (EC). Given a choice between alternating hippocampal membrane lanes from developmental stages ranging from early postnatal to adult, EC axons preferred to extend on early postnatal hippocampal membranes. Neither the neutralization of myelin-associated factors by a specific antibody (IN-1) nor the separation of myelin from membranes interfered with the axons' choice behaviour. The entorhinal axons showed no preference in the membrane combination of adult and myelin-free adult hippocampal membranes. These stripe assay experiments demonstrate that support for EC axon choice in the developing hippocampus is maturation-dependent and is not influenced by myelin. The application of IN-1 in the outgrowth assay and the separation of myelin from membranes, enhanced elongation of outgrowing entorhinal axons on adult hippocampal membranes, whereas a control antibody did not. This shows that myelin-associated factors have a strong inhibitory effect on the outgrowth length of entorhinal axons. In conclusion, we suggest that axonal elongation in the entorhinohippocampal system during development is strongly influenced by myelin-associated growth inhibition factors and that specific target finding of entorhinal axons is regulated by a different mechanism.

Age Factors↗

Basic fibroblast growth factor enhances axonal sprouting after cortical injury in rats.

The trophic factors responsible for initiating and guiding the outgrowth of axons have proven to be elusive throughout most of this century. Entorhinal cortex injury, which denervates the hippocampal formation of rats, induces axonal sprouting by several surviving hippocampal afferents and results in a significant elevation of growth factors, one of which is basic fibroblast growth factor (bFGF). The possibility that bFGF may regulate lesion-induced hippocampal sprouting was examined by making i.v. bFGF infusions into rats with unilateral entorhinal lesions. Basic FGF treatment significantly increased sprouting by the cholinergic septodentate pathway. Thus, the increase in bFGF following central nervous system injury may signal its role in the regulation of injury-related axonal remodeling of a cholinergic pathway.

Animals↗

New illumination technique for IR-video guided patch-clamp recording from neurons in slice cultures on biomembrane.

Slice cultures on biomembrane are the method of choice for studying Ca2+-dependent plastic changes occurring over several days to weeks. Using IR-differential interference contrast, good visualization of neurons in biomembrane slice cultures has been achieved despite a negative optical effect of the biomembrane, but epifluorescence imaging requires removal of a Wollaston prism and the analyzer. Here, we describe a novel illumination method to overcome this problem. Using optic fiber illumination at a shallow angle from the top of the slice culture, with or without additional illumination from the bottom, we obtained good cellular resolution of neurons in biomembrane slice cultures as well as in acute slices with an infrared-video camera. With this technique, we demonstrate visually guided whole-cell patch-clamp recording of Na+- and K+-currents as well as combination of whole-cell recording with fluorescence imaging of hippocampal and entorhinal cortex neurons in biomembrane slice cultures. Our inexpensive method should prove very useful for studying in vitro effects of long-term manipulations on membrane currents and intracellular Ca2+-signaling.

Animals↗

Delayed peripheral nerve regeneration and central nervous system collateral sprouting in leucocyte common antigen-related protein tyrosine phosphatase-deficient mice.

Cell adhesion molecule-like receptor-type protein tyrosine phosphatases have been shown to be important for neurite outgrowth and neural development in several animal models. We have previously reported that in leucocyte common antigen-related (LAR) phosphatase deficient (LAR-deltaP) mice the number and size of basal forebrain cholinergic neurons, and their innervation of the hippocampal area, is reduced. In this study we compared the sprouting response of LAR-deficient and wildtype neurons in a peripheral and a central nervous system lesion model. Following sciatic nerve crush lesion, LAR-deltaP mice showed a delayed recovery of sensory, but not of motor, nerve function. In line with this, neurofilament-200 immunostaining revealed a significant reduction in the number of newly outgrowing nerve sprouts in LAR-deltaP animals. Morphometric analysis indicated decreased axonal areas in regenerating LAR-deltaP nerves when compared to wildtypes. Nonlesioned nerves in wildtype and LAR-deltaP mice did not differ regarding myelin and axon areas. Entorhinal cortex lesion resulted in collateral sprouting of septohippocampal cholinergic fibres into the dentate gyrus outer molecular layer in both genotype groups. However, LAR-deltaP mice demonstrated less increase in acetylcholinesterase density and fibre number at several time points following the lesion, indicating a delayed collateral sprouting response. Interestingly, a lesion-induced reduction in number of (septo-entorhinal) basal forebrain choline acetyltransferase-positive neurons occurred in both groups, whereas in LAR-deltaP mice the average cell body size was reduced as well. Thus, regenerative and collateral sprouting is significantly delayed in LAR-deficient mice, reflecting an important facilitative role for LAR in peripheral and central nervous system axonal outgrowth.

Acetylcholinesterase↗

Increased density of entorhinal glutamate-immunoreactive vertical fibers in schizophrenia.

Gluatamtergic fibers have been immunocytochemically localized in the entorhinal cortex of postmortem schizophrenic brains. The density of small caliber vertical fibers was higher in schizophrenics than controls, with no significant increase in the number of large caliber fibers. Increased glutamatergic fiber density has been previously reported in the cingulate cortex. It is proposed that increases in glutamatergic fibers from the amygdala may be responsible for these changes and that they may play a central role in the pathophysiology of schizophrenia.

Cadaver↗

MRI predictors of risk of incident Alzheimer disease: a longitudinal study.

OBJECTIVE: To determine if baseline entorhinal and hippocampal volumes and their rate of atrophy could predict the risk of incident Alzheimer disease (AD). METHODS: The authors used proportional odds models to assess the relationship between entorhinal and hippocampal size and risk of incident AD among 58 nondemented elderly people. All participants were followed with annual clinical evaluations and structural MRI scans for up to 5 years (baseline and 5 years of follow-up). At baseline, 23 of 58 participants received a diagnosis of amnestic mild cognitive impairment (MCI) and 35 of 58 were healthy control subjects with no cognitive impairment. Structural MRI scans were acquired with a T1-weighted three-dimensional spoiled gradient-recalled echo pulse sequence in a 1.5 T scanner. Entorhinal and hippocampal volumes were derived from 1.6-mm gapless coronal images reformatted to be perpendicular to the long axis of the hippocampus and were normalized by dividing with intracranial volume. RESULTS: Fourteen of 58 nondemented participants developed AD during the follow-up period. Initial diagnosis of MCI was a significant predictor of incident AD. In addition, both baseline entorhinal volume and its slope of decline were independent predictors of incident AD, but initial hippocampal size and its rate of decline were not, after controlling for entorhinal volume. CONCLUSION: In nondemented individuals, entorhinal cortex atrophy is associated with risk of Alzheimer disease.

Aged↗

Changes in hippocampal SNAP-25 expression following afferent lesions.

Reductions in SNAP-25 immunohistochemistry were found after removing the glutamatergic and cholinergic inputs to the rat hippocampus. SNAP-25 levels were normalised by 1 month after afferent lesions. Surprisingly, a superimposed cholinergic lesions did not affect the return to normal SNAP-25 levels after a long-term entorhinal cortex lesion. It is concluded that changes in SNAP-25 may represent early markers of synaptic loss following afferent lesions to the hippocampus.

Analysis of Variance↗

Computational experiments support a competitive function in the CA3 region of the hippocampus.

The comprehension of activities and functions of complex brain structures requires, among other things, information on simultaneous activities in several regions. Results reported in the literature using multi(micro/macro)electrode recordings or imaging techniques provide incomplete information due either to the small size and/or small number of investigated regions or to the poor spatiotemporal resolution, respectively. This is particularly true for the hippocampus and its subfields, and mathematical modeling and computer simulation have been used with the aim of obtaining information when this is lacking. Global activities in the CA3 field of the hippocampus, and in particular the genesis of theta rhythm and sharp waves, have been investigated here by a mathematical model formulated within the frame of a kinetic theory of neural systems. The model has taken into account data of experimental results both on different PSPs recorded in hippocampal neurons and on recurrent pyramidal collateral geometries. The computational 'experiments' to which the model was subjected suggest that the sharp waves arise through a selective and short block of the fast inhibitory neurons of CA3, produced by a medial septum inhibitory input, whereas the theta activity is produced by a durable, continuous inhibition of the slow inhibitory neurons. Information obtained also suggests that the recurrent pyramidal collaterals subserve a competitive, rather than a cooperative, organization. Based on these results a hypothesis on the possible functional organization of the CA3 field and of the entire hippocampus has been formulated. According to this hypothesis, the CA3 imposes a serial order on the flow of activity arriving at the hippocampus from the entorhinal cortex and from its connected polymodal cortical regions. This ordering permits cortical activities, arriving at CA3 on appropriate time intervals, to produce effects in regions of brain to which the CA3 projects. The competing cortical activities are lost.

Animals↗

Domoic acid-induced neuronal degeneration in the primate forebrain revealed by degeneration specific histochemistry.

Domoic acid is a potent excitotoxin produced by diatoms which is subsequently passed along the marine food chain. Its chemical structure and toxicological properties are similar to kainic acid. Like kainic acid, exposure results in extensive hippocampal degeneration. The effect of domoic acid on other primate brain structures, however, is less resolved. In an attempt to clarify this issue, the present study applied a degeneration specific histochemical technique (de Olmos' cupric-silver method) to reveal degeneration within the brains of domoic acid-dosed cynomolgus monkeys. Degenerating neuronal cell bodies and terminals were found not only within the hippocampus, but also within a number of other 'limbic' structures including the entorhinal cortex, the subiculum, the piriform cortex, the lateral septum, and the dorsal lateral nucleus of the thalamus. Although the hippocampus is a component of the original limbic circuit of Papez, other components such as the mammillary bodies, the anterior nucleus of the thalamus and the cingulate cortex contained no degeneration, while a number of more recently documented efferent targets of the hippocampal formation revealed extensive degeneration. The pattern of degeneration generally correlated with those regions containing high densities of kainate receptors.

Animals↗

Memory impairment in temporal lobe epilepsy: the role of entorhinal lesions.

Temporal lobe epilepsy (TLE) patients are frequently afflicted with deficits in spatial and other forms of declarative memory. This impairment is likely associated with the medial temporal lobe, which suffers widespread damage in the disease. Physiological and lesion studies, as well as examinations of the complex connectivity of the medial temporal lobe in animals and humans, have identified the entorhinal cortex (EC) as a key structure in the function and dysfunction of this brain region. Lesions in EC layer III, which normally provides monosynaptic input to area CA1 of the hippocampus, frequently occur in TLE and may be causally related to the memory impairments seen in the disease. Lesions that are initially largely restricted to EC layer III can be produced in rats by focal intra-entorhinal injections of 'indirect excitotoxins' such as aminooxyacetic acid or gamma-acetylenic GABA. These animals eventually show more extensive neurodegeneration in temporal lobe structures and, after a latent period, exhibit spontaneously recurring seizure activity. These progressive features, which may mimic events that occur in TLE, provide new opportunities to explore the role of the EC in memory deficits associated with TLE. These animals will also be useful for evaluating new treatment strategies that focus on the prevention of pathological events in the EC.

Animals↗