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Non-hippocampal cortical projections from the entorhinal cortex in the rat and rhesus monkey.

The entorhinal cortices are known to give rise to powerful projections that terminate in the hippocampus and dentate gyrus. Collectively, these link the hippocampal formation to many parts of the cortex and to subcortical structures like the amygdala. Non-hippocampal projections from the entorhinal cortices are understood poorly. Such projections to neighboring temporal areas in the rat and rhesus monkey have been investigated using the autoradiographic and horseradish peroxidase (HRP) tracing procedures. In the rat, HRP-labeled neurons were observed in the intermediate and lateral fields of the entorhinal cortices after injections of temporal cortical areas 20, 35, 36 and 41. They were located predominantly in layers II, III and IV. In the monkey, HRP-labeled neurons were observed in the entorhinal cortices after injections of the rostral superior temporal gyrus (area TA or 22); the temporal polar cortex (area TG or 38); the inferior temporal cortex (area TE or 20); the perirhinal cortex (area 35) and the posterior parahippocampal cortices (areas TF and TH). Unlike the rat, labeled entorhinal neurons in the monkey were located in layer IV. Autoradiographic experiments in the monkey yielded complimentary results. In view of the fact that layer IV of the entorhinal cortex in both the rat and monkey receives a powerful projection from the subicular-CAl fields of the hippocampal formation, the results imply that this layer mediates an indirect non-fornical connection between the hippocampal formation and the temporal cortex.

Amygdala↗

Increases in transforming growth factor-beta mRNA in hippocampus during response to entorhinal cortex lesions in intact and adrenalectomized rats.

Transforming growth factor-beta mRNA was detected with a rat TGF-beta 1 coding sequence probe as a 2.5 kb band by RNA blot hybridization of total RNA from the adult male rat hippocampus. Following electrolytic lesions of the entorhinal cortex that cause hippocampal deafferentation and synaptic remodeling, TGF-beta mRNA increases 5-fold in ipsilateral hippocampus when compared with intact controls. This increase was independent of prior adrenalectomy or corticosterone-replacement. These data demonstrate that TGF-beta gene expression increases in response to hippocampal deafferentation.

Adrenal Cortex↗

Combined damage to entorhinal cortex and cholinergic basal forebrain neurons, two early neurodegenerative features accompanying Alzheimer's disease: effects on locomotor activity and memory functions in rats.

In Alzheimer's disease (AD), cognitive decline is linked to cholinergic dysfunctions in the basal forebrain (BF), although the earliest neuronal damage is described in the entorhinal cortex (EC). In rats, selective cholinergic BF lesions or fiber-sparing EC lesions may induce memory deficits, but most often of weak magnitude. This study investigated, in adult rats, the effects on activity and memory of both lesions, alone or in combination, using 192 IgG-saporin (OX7-saporin as a control) and L-N-methyl-D-aspartate to destroy BF and EC neurons, respectively. Rats were tested for locomotor activity in their home cage and for working- and/or reference-memory in various tasks (water maze, Hebb-Williams maze, radial maze). Only rats with combined lesions showed diurnal and nocturnal hyperactivity. EC lesions impaired working memory and induced anterograde memory deficits in almost all tasks. Lesions of BF cholinergic neurons induced more limited deficits: reference memory was impaired in the probe trial of the water-maze task and in the radial maze. When both lesions were combined, performance never improved in the water maze and the number of errors in the Hebb-Williams and the radial mazes was always larger than in any other group. These results (i) indicate synergistic implications of BF and EC in memory function, (ii) suggest that combined BF cholinergic and fiber-sparing EC lesions may model aspects of anterograde memory deficits and restlessness as seen in AD, (iii) challenge the cholinergic hypothesis of cognitive dysfunctions in AD, and (iv) contribute to open theoretical views on AD-related memory dysfunctions going beyond the latter hypothesis.

Acetylcholine↗

Degree of hyperinnervation of area dentata by locus coeruleus in the presence of septum or entorhinal cortex as studied by sequential intraocular triple transplantation.

In situ area dentata receives a sparse noradrenergic innervation from locus coeruleus. Embryonic area dentata co-transplanted with locus coeruleus to the anterior eye chamber receives an abundant ingrowth of nerves from the noradrenergic neurons of the locus graft. We sought to identify restrictive forces acting on coeruleo-dentate axons by arranging for the innervation of area dentata transplants by either entorhinal cortex or septal nuclei transplants prior to locus coeruleus transplantation. The noradrenergic hyperinnervation was not inhibited when locus coeruleus transplants were placed on the opposite side of area dentata from the entorhinal or septal transplant. Noradrenergic innervation of area dentata was restricted when the locus coeruleus transplant was placed in contact with the septal transplant. This inhibitory interaction seemed to take place between the septal and locus coeruleus transplants rather than in the area dentata neuropil. This type of interaction points towards one means by which axonal growth may be inhibited during development or in the adult.

Animals↗

The effect of unilateral and bilateral removal of the entorhinal cortex on the glucose utilization in various hippocampal regions in the rat.

In the present study lesion-induced changes in function of various hippocampal regions, as reflected by the metabolic rate of glucose, were measured by means of quantitative autoradiography, 4 days after unilateral or bilateral surgical removal of the entorhinal cortex. The greatest decrease (45%) was seen in the stratum lacunosum moleculare of the CA1, whereas a lesser decline (34%) was seen in the molecular layer of the dentate gyrus, stratum lucidum of the CA3 (31%) and the stratum radiatum of the CA1 (36%). These findings support the view that in addition to the indirect trisynaptic temporo-ammonic pathway, there is a functionally active direct pathway.

Animals↗

Sulfated glycoprotein-2 is increased in rat hippocampus following entorhinal cortex lesioning.

Thios study showed responses of sulfated glycoprotein-2 (SGP-2) in the rat hippocampus after deafferenting lesion. SGP-2 is a plasma protein that also occurs in many peripheral tissues. In some circumstances, elevations of SGP-2 mRNA are associated with cell degeneration and responses to injury. This study used entorhinal cortex lesions (ECL) to partially deafferent the hippocampus by damaging the perforant path and to induce synaptic remodeling. SGP-2 mRNA is increased in hippocampal astrocytes after ECL. Western blot analysis of soluble hippocampal proteins identified 3 major forms of rat SGP-2 protein: a precursor (61 kDa) and 2 reduced subunits at 39.5 and 35 kDa. These forms increased at 4 days post ECL ipsilaterally to the lesion. By immunocytochemistry (ICC), SGP-2 showed an increased immunoreactivity on the lesioned side by 2 days post ECL that continued through 14 days post ECL. Besides immunopositive astrocytes, punctate immunochemical reaction products occurred among the degenerating fibers of the perforant path. We conclude that changes of SGP-2 protein in the hippocampus after ECL occur roughly in parallel with increases of SGP-2 mRNA. The punctate immuno-deposits could represent secreted SGP-2 and may be useful as a marker for degenerating pathways.

Animals↗

Spontaneous EEG spikes in the normal hippocampus. IV. Effects of medial septum and entorhinal cortex lesions.

Spontaneous EEG spikes (SPKs) were recorded from the CA1 region of the dorsal hippocampus in normal rats during behavioral states not accompanied by rhythmical slow activity (RSA) such as awake immobility and slow wave sleep. The present experiment was designed to examine the effects of large electrolytic lesions of the 2 major hippocampo-petal systems, medial septum (MS) and entorhinal cortex (EC), on spontaneous SPK activity. MS lesions, while completely abolishing RSA, did not eliminate SPKs or change their behavioral correlates. SPKs were still suppressed during behaviors (walking, head movement, etc.) normally associated with RSA. However, SPK frequency was approximately halved after MS lesions. Total bilateral EC lesions did not, in general, change SPKs or RSA unless abnormal hippocampal activities persisted over the first post-lesion week during which most of the recordings were made. Laminar profiles of SPKs in both MS- and EC-lesioned rats showed a normal pattern; small positivity in stratum oriens, large negativity in stratum radiatum and polarity reversal around stratum pyramidale. These results suggest that (1) neither medial septal input nor entorhinal cortical input to the hippocampus is necessary for the generation of SPKs, and (2) SPK suppression still occurs in the absence of these inputs during behaviors normally associated with RSA. The possibility remains, however, that in intact animals these inputs do influence SPK activity in a behavior-dependent manner.

Animals↗

The supramammillary region of the cat sends substance P-like immunoreactive axons to the hippocampal formation and the entorhinal cortex.

A combined method of the tracing of WGA-HRP (wheat germ agglutinin-conjugated horseradish peroxidase) and the immunohistochemistry of substance P (SP) showed that many SP-like immunoreactive neurons in the supramammillary nucleus of cat hypothalamus sent their axons to the hippocampal formation. SP-like immunoreactive axons in the hippocampal formation and entorhinal cortex were markedly reduced in number ipsilaterally after placing an electrothermic lesion in the supramammillary region of the hypothalamus.

Animals↗

A projection from the entorhinal cortex to the nucleus accumbens in the rat.

A study was performed in which both anterograde ([3H]leucine radioautography) and retrograde (horseradish peroxidase (HRP) histochemistry) tracing methods were employed to identify the origin of the fimbrial projection to the nucleus accumbens in the rat. The data reveal that this pathway arises predominantly from layers II--III of the anterior two-thirds of entorhinal cortex rather than from any part of hippocampal formation.

Animals↗

Epileptiform activity in combined slices of the hippocampus, subiculum and entorhinal cortex during perfusion with low magnesium medium.

Reduction of [Mg2+]o induced spontaneous epileptiform activity consisting of 40-100-ms bursts of population spikes in hippocampal slices. This activity disappeared from area CA1 when the connections to area CA3 were cut, but persisted in isolated minislices of area CA3. Spontaneous activity was also observed in the dentate gyrus, provided that the connections to the subiculum and entorhinal cortex (EC) were intact. In the parasubiculum and EC longer lasting epileptiform events were observed which resembled seizure-like behaviour. The epileptiform activity was completely suppressed by 2-aminophosphonovalerate (30 microM) suggesting that N-methyl-D-aspartate receptors for excitatory amino acid transmitters participate in the generation of this activity. These findings show that the EC possesses properties which permit the generation of seizure-like activity in contrast to the hippocampus where the activity resembled recurrent interictal events.

Action Potentials↗

The effect of Ginkgo biloba extract (EGb 761) on gliotic reactions in the hippocampal formation after unilateral entorhinal cortex lesions.

PURPOSE: Ginkgo biloba extract (EGb 761) has been shown to facilitate behavioral and neuro-morphological recovery from brain injury, but less is known about its effects on glia. Since gliosis may be an important component of the recovery process, we tested the hypothesis that EGb 761 alters the time course and development of microglial activation and astrocytosis after brain injury. METHODS: Rats were treated with either saline or EGb 761 and killed at 2 hrs, 1, 3, 7, and 14 days following unilateral entorhinal cortex (EC) lesions. Microglia and their precursors were visualized with a silver impregnation method, and astrocytes with GFAP. RESULTS: Blood-borne monocytes/macrophages were seen as early as 2 hrs after injury in all animals. The side contralateral to the injury showed minimal microglial activation and there were no significant effects of drug treatment. On the side ipsilateral to the lesion EGb 761 enhanced microglial activation at 3, 7, and 14 days in the molecular layer and the hilus of the dentate gyrus; the areas of most profound deaf-ferentation after EC injury. Regions of the corpus callosum also showed enhanced microglial activation over the same time course. Reactive astrocytes were stained with GFAP and were found to be more numerous than activated microglia, particularly in the ipsilateral corpus callo-sum. EGb 761 treatment enhanced astrocytosis at 3 days in the molecular layer, the hilus, and the corpus callosum on the ipsilateral side. CONCLUSIONS: Taken together our results show that EGb 761 enhances, accelerates and prolongs the activation of microglia and astrocytosis at the site of injury.

Journal Article↗

Amygdaloid complex modulates neurotransmission from the entorhinal cortex to the dentate gyrus of the rat.

In urethane-anesthetized rats, single-pulse stimulation of the lateral nucleus of the amygdaloid complex evoked field responses in the dentate gyrus. These responses were similar in many respects to those evoked by stimulation of the monosynaptic perforant path input to the dentate, except for their longer latency. Paired-pulse tests revealed a potent facilitation of perforant path-evoked dentate responses by preceding stimulation of the lateral amygdaloid nucleus. These observations indicate a strong amygdaloid influence on cortico-hippocampal neurotransmission and provide some support for recent anatomical descriptions suggesting that the lateral nucleus of the amygdaloid complex projects to the dentate via a disynaptic pathway relaying in the entorhinal cortex.

Amygdala↗

The second layer neurones of the entorhinal cortex and the perforant path in physiological ageing and Alzheimer's disease.

The hippocampal formation was studied in 5 brains of younger (29 to 52 years of age) and 6 brains of elderly (61 to 89 years of age) subjects without signs of dementia, as well as in 11 brains of patients with Alzheimer's disease (65 to 91 years of age). The 8-microns-thick sections were stained either with cresyl violet, Weil method or with immunocytochemical methods for amyloid (4G8) and neurofibrillary tangles (Tau-1). Cell bodies, senile plaques and tangles were counted in all brains. In brains of patients with Alzheimer's disease a significant neuronal loss (about 56%) was observed in the second layer of the entorhinal cortex. The tangles/neurones ratio was very high (62.79.1%) in this layer. A great number of senile plaques were present in the whole hippocampal formation, especially in the molecular layer of the dentate gyrus (22.91.5 plaques/mm2) which is the termination zone of the perforant path. It seems therefore, that pathological alterations in Alzheimer's disease disrupt the main input to the hippocampal formation. In "physiological" ageing we did not observe changes in the density of neurones, although single tangles and plaques were found in all hippocampal areas. In elderly individuals 3.81.3% of neurones of the second layer revealed neurofibrillary pathology; a few plaques were found in various areas of the hippocampal formation. These observations may suggest only a slight decrease in number of neurones in the hippocampal formation. However, these changes cause a slight impairment of memory and learning often found in elderly individuals without dementia.

Adult↗

The influence of electrocoagulation of the septum and section of the entorhinal cortex on general behaviour and memory in cats.

In chronic experiments on cats it was shown that the lesion of the medial part of the septum does not result in the "septal syndrome"; the ratio of the different sleep-wakefulness cycle phases remains within the baseline values; the acquisition, retrieval and extinction of instrumental alimentary conditioned reflexes proceed normally; the delayed conditioned reflexes are impaired. Massive septal lesion, including its lateral part, leads to the development of the "septal syndrome"; there are changes in the structure and percentage of the different sleep-wakefulness cycle phases; the acquisition and extinction of instrumental alimentary reflexes with the sound discrimination are markedly retarded; the performance of delayed conditioned reflexes is completely destroyed. The section of the entorhinal cortex produces an increase in the number of repeated errors and perseverative movements during performance of instrumental alimentary reflexes, the deceleration of the acquisition and extinction of instrumental alimentary reflexes, complete disturbance of the delayed conditioned reflexes and does not affect the sleep-wakefulness cycle. The problems of the role of the hippocampus and its main inputs in the regulation of the short-term operative memory in the "pure form" as well as the significance of the descending influence of the hippocampus on the regulation of general animal behavior were also discussed.

Animals↗

Evidence that transmitter-containing dystrophic neurites precede those containing paired helical filaments within senile plaques in the entorhinal cortex of nondemented elderly and Alzheimer's disease patients.

Within the amygdala of elderly subjects and patients with Alzheimer's disease (AD), we recently found evidence suggesting amyloid beta-protein (A beta P) deposition occurs before the appearance of dystrophic neurites. Moreover, these data suggested dystrophic neurites initially lack evidence of cytoskeletal pathology although with time and further maturation, the dystrophic neurites display an altered cytoskeleton as evidenced by their immunoreactivity to Alz-50 and paired-helical filaments (PHF). These findings are of particular relevance to our understanding of the sequence of pathologic events in AD and thus it has become important to determine whether these events are unique to the amygdala or are representative of a more general pattern which can be found throughout the brain. Using a battery of antibodies to markers that are characteristic of AD pathology (i.e., A beta P, PHF, and Alz-50), three peptidergic neurotransmitters (neurotensin, somatostatin, and substance P), and one neurotransmitter biosynthetic enzyme (choline acetyltransferase), we examined the entorhinal cortex (EC) of three groups of subjects (AD, normal elderly, and a group of nondemented elderly with numerous senile plaques). The EC was studied, in part, because it is well recognized as a brain region displaying severe and, most importantly, early pathologic changes. Like the amygdala, we found evidence that amyloid beta-protein immunoreactive (A beta P-IR) and thioflavine-S-positive senile plaques occur within the EC prior to the appearance of transmitter-, Alz-50-, or PHF-immunoreactive dystrophic neurites. We also observed transmitter-immunoreactive dystrophic neurites in the absence of Alz-50 or PHF-immunolabeled dystrophic neurites and transmitter- and Alz-50-IR dystrophic neurites in the absence of those containing PHF. Collectively, these findings were similar to those seen within the amygdala and thus reinforced the concept that A beta P deposition is the primary event in plaque pathology, and this deposition is subsequently followed by the appearance of dystrophic neurites which retain their transmitter phenotype yet lack an altered cytoskeleton. With time, these dystrophic neurites develop cytoskeletal alterations and become immunoreactive to Alz-50 and PHF.

Adult↗

Cholinergic innervation in the human hippocampal formation including the entorhinal cortex.

The cholinergic innervation of the hippocampal formation is thought to play an important role in memory processes, but its organization in humans has not been described in detail. We studied the cholinergic innervation of the human hippocampal formation by means of immunohistochemistry with polyclonal antisera directed against acetylcholinesterase (AChE), choline acetyltransferase (ChAT), and the low-affinity (p75) nerve growth factor receptor (NGFR). The density of ChAT-like immunoreactive (ChAT-li) fibers differed substantially among the various regions, in general paralleling the pattern of AChE-li staining. One notable exception was the presence of AChE-li cell bodies. In contrast, ChAT immunoreactivity was associated only with fibers and terminals. NGFR-li staining corresponded closely to the ChAT-li fiber pattern. ChAT-li fibers in the CA fields diffusely filled the stratum pyramidale and extended into the stratum oriens and radiatum as well. The highest density was consistently observed in CA4 and CA3 subfields. Staining decreased from CA4 to CA1 and was substantially less dense in the subicular complex. In the entorhinal cortex, the ChAT- and NGFR-li fiber innervation displayed a laminar pattern, most intense over the nests of cells in layer II. There was a trend towards an age-related reduction in the density of ChAT- and AChE-li fibers and terminals. Nonetheless, we also found a surprisingly conserved NGFR-li innervation and the presence of occasional NGFR-li pyramidal cells, providing evidence of a plastic response in the brains of the elderly patients.

Acetylcholinesterase↗

Identification of a subpopulation of neuropeptide Y-containing locus coeruleus neurons that project to the entorhinal cortex.

A fundamental question important to the understanding of the neurochemical organization of the central nervous system focuses on the relationships between the differential phenotypic expression of multiple neurotransmitter markers in individual neuronal populations and the factors that regulate their expression. The first approach in studying this phenomenon is the determination of specific relationships between neurochemically distinct neuronal subpopulations and their efferent targets. The pontine nucleus locus coeruleus (LC) provides a useful model for addressing this question since the projections of LC neurons are topographically organized and several neuropeptides are expressed along with noradrenergic markers in subsets of these neurons. In these studies, we have focused on defining the efferent targets of LC neurons that contain neuropeptide tyrosine (NPY)-like immunoreactivity. This has been accomplished by injecting the retrograde fluorescent tracer fluorogold into specific cortical and hippocampal targets in adult rats and identifying the proportion of retrogradely labeled LC neurons that are positive for NPY-like immunoreactivity. In agreement with other investigators, no preferential cortical projections of NPY-positive LC neurons were observed. However, when fluorogold injections included or were limited to the entorhinal cortex, a discrete cluster of round or ovoid neurons in the dorsomedial portion of the LC approximately 9.8 mm posterior to bregma were found to contain NPY-like immunoreactivity. This observation demonstrates that some topographic organization of NPY-containing LC neurons does exist. In fact, these data indicate that morphologic and topographic organization exists even within neurochemically distinct subsets of neuronal populations.

Animals↗

Organization of connectivity of the rat presubiculum: I. Efferent projections to the medial entorhinal cortex.

The organization of the laminar and topographical projections from the presubiculum to the entorhinal area was studied in the rat by anterograde labeling with Phaseolus vulgaris leucoagglutinin and retrograde labeling with horseradish peroxidase conjugated to wheat germ agglutinin. We found that the pattern of presubiculo-entorhinal projections differs between the superficial and deep layers of the presubiculum. The superficial layers (layers II and III) of the presubiculum gave rise to bilateral projections to layers I-VI of the medial entorhinal area (MEA). Many terminals were distributed in layer III, fewer in layer II and the deep portion of layer I, and many fewer terminals in the deep layers (layers V and VI) of MEA. In contrast, the deep layers (layers V and VI) of the presubiculum gave rise to ipsilateral projections to the entorhinal area. Many axon terminals were distributed in layers V and VI of MEA and the most superficial portion of layer I of MEA, but very few in layers II and III. In addition, the ramifications in layer I extended to the lateral entorhinal area (LEA). Using two-dimensional unfolded maps of parahippocampal cortices, we elucidated the distinct topographical relationship in the presubiculo-entorhinal projection: 1) The septotemporal or longitudinal axis of the presubiculum corresponded to the axis on the MEA/LEA boundary, where the septal presubiculum projected toward the rhinal fissure and the temporal presubiculum projected away from the fissure. 2) The proximodistal axis of the presubiculum corresponded to the axis from the MEA/LEA boundary to the MEA/parasubiculum boundary that was virtually perpendicular to the MEA/LEA boundary, where the proximal portion of the presubiculum (close to the subiculum) projected to the region near the MEA/LEA boundary.

Animals↗