Intrinsic and commissural connections within the entorhinal cortex. An anterograde and retrograde tract-tracing study in the cat

Intrinsic and commissural connections within the entorhinal cortex (EC) were examined in the cat by the anterograde and retrograde tract-tracing methods with Phaseolus vulgaris leucoagglutinin and cholera toxin B subunit. Intrinsic axons to the superficial layers (layers I-III) arose mainly from layers II, III, Vd (deep part of layer V), and VI, were distributed more widely in the superficial layers than in the deep layers, and terminated progressively more densely in more superficial layers; most densely in layer I. In the medial entorhinal area (MEA) and the ventromedial and the ventrolateral divisions of the lateral entorhinal area (VMEA and VLEA), the longitudinal connections through the intrinsic fibers to the superficial layers is often more restricted in rostral direction than in caudal direction. In the dorsolateral division of the lateral EC (DLEA), the longitudinal connections through the intrinsic fibers to the superficial layers extended distantly in both rostral and caudal directions. Intrinsic fibers to the deep layers (layers IV-VI) originated mainly from layers IV and Vs (superficial part of layer V) and were distributed rather sparsely and diffusely; they were distributed more widely in the deep layers than in the superficial layers. Commissural axons to the homotopic EC regions originated from layers II and III of the MEA and DLEA and terminated in all EC layers, most densely in layer I.     

Zinc-positive afferents to the rat septum originate from distinct subpopulations of zinc-containing neurons in the hippocampal areas and layers

The purpose of the present study was to examine whether zinc-positive and zinc-negative hippocampal neurons in rats differed with respect to their projections to the septum. By combining retrograde axonal transport of the fluorescent tracer Fluoro-Gold with histochemical demonstration of zinc selenide complexes in zinc-containing neurons after intraperitoneal injection of sodium selenite, we were able to visualize the distribution of retrogradely Fluoro-Gold labeled neurons and zinc-containing neurons in the same sections. After unilateral injection of Fluoro-Gold into the rat septum a few retrogradely labeled cells were observed in layer IV of the ipsilateral medial entorhinal area, and numerous labeled cells were observed mainly in the superficial layers of the ipsilateral subicular areas and throughout the CA1 and CA3 pyramidal cell layers, as well as in the contralateral CA3 pyramidal cell layer. Zinc-containing neurons were observed in layers IV–VI of the medial entorhinal area, layers II and III of the parasubiculum, layers II, III and V of presubiculum, and in the superficial CA1 and deep CA3 pyramidal cell layers. Cells double-labeled with Fluoro-Gold and zinc selenide complexes were primarily located in distal (relative to the area dentata) parts of the superficial CA1 pyramidal cell layer and distal parts of the deep CA3 pyramidal cell layer and in layers II and III of presubiculum. Only a very few double-labeled cells were seen in the contralateral CA3. The result demonstrates that the hippocampo-septal projection of rats is a mixture of zinc-positive and zinc-negative fibers. Where-as zinc-negative fibers originate from neurons throughout the hippocampal and retrohippocampal areas, zinc-positive fibers originate from distinct subgroups of zinc-containing cells in different areas and layers.     

A description of the amygdalo-hippocampal interconnections in the macaque monkey

The interconnections between the amygdala and the hippocampal formation were investigated in the macaque monkey using anterograde tracers. The hippocampal inputs to the amygdala arose from the subicular and entorhinal cortices and passed through the angular bundle to terminate principally in the medial basal and lateral basal nuclei, with lighter termination in the lateral nucleus, the periamygdaloid cortex, and the cortical-transition area. The majority of these amygdaloid inputs arose from the rostral hippocampal formation although there was equivocal evidence of an additional projection from the caudal hippocampus to the central nucleus. Projections arose from many of the amygdaloid nuclei to terminate in the molecular layer of the amygdalo-hippocampal area and the adjacent stratum moleculare of the uncal portion of the hippocampus. The accessory basal, lateral basal, and medial basal nuclei also projected to the most rostral portions of the stratum moleculare of fields CA1-3, the heaviest termination occurring in field CA3. Additional projections from the basal nuclei terminated in the prosubiculum, presubiculum, and parasubiculum. The heaviest entorhinal inputs arose from the accessory basal and lateral nuclei and terminated in layers I, II, and III of areas 28b, 28i, and the prorhinal cortex. The major amygdaloid input to the perirhinal cortex arose from the lateral basal nucleus.     

Projections from the presubiculum and the parasubiculum to morphologically characterized entorhinal-hippocampal projection neurons in the rat

The relations between the inputs from the presubiculum and the parasubiculum and the cells in the entorhinal cortex that give rise to the perforant pathway have been studied in the rat at the light microscopical level. Projections from the presubiculum and the parasubiculum were labeled anterogradely, and, in the same animal, cells in the entorhinal cortex that project to the hippocampal formation were labeled by retrograde tracing and subsequent intracellular filling with Lucifer Yellow. The distribution and the number of appositions between the afferent fibers and hippocampal projection neurons in the various layers of the entorhinal cortex were analyzed. The results show that layers I–IV of the entorhinal cortex contain neurons that give rise to projections to the hippocampal formation. The morphology of these projection neurons is highly variable and afferents from the presubiculum and the parasubiculum do not show a preference for any specific morphological cell type. Both inputs preferentially innervate the dendrites of their target cells. However, presubicular and parasubicular projections differ with respect to the layer of entorhinal cortex they project to. The number of appositions of presubicular afferents with cells that have their cell bodies in layer III of the entorhinal cortex is 2–3 times higher than with cells in layer II. In contrast, afferents from the parasubiculum form at least 2–3 times as many synapses on the dendrites of cells located in layer II than on neurons that have their cell bodies in layer III. Cells in layers I and IV of the entorhinal cortex receive weak inputs from the presubiculum and parasubiculum. Not only is the presubiculum different from the parasubiculum with respect to the distribution of projections to the entorhinal cortex, they also differ in their afferent and efferent connections. In turn, cells in layer II of the entorhinal cortex differ in their electrophysiological characteristics from those in layer III. Moreover, layer II neurons give rise to the projections to the dentate gyrus and field CA3/CA2 of the hippocampus proper, and cells in layer III project to field CA1 and the subiculum. Therefore, we propose that the interactions of the entorhinal-hippocampal network with the presubiculum are different from those with the parasubiculum.     

Neurons and terminals in the retrohippocampal region in the rat''s brain identified by anti-gamma-aminobutyric acid and anti-glutamic acid decarboxylase immunocytochemistry

Summary The distribution of gamma-aminobutyric acid (GABA) containing nerve cells and terminals was studied at the light and electron microscopic levels in the retrohippocampal region of the rat by using anti-glutamic acid decarboxylase (GAD) and anti-GABA antibodies in immunocytochemistry. Large numbers of GAD and GABA stained cells were found in all retrohippocampal structures. At the ultrastructural level, the immunoreactivity against GABA and against the synthesizing enzyme GAD was localized to cytoplasmic structures, including loose clumps of rough endoplasmic reticulum, ribosomal arrays, outer mitochondrial surfaces and in axonal boutons.The GAD- and GABA-immunorective(-i) cells were found in all subfields of the retrohippocampal region (e.g., the subicular complex, the entorhinal area). Within the entorhinal area a slightly larger number of immunoreactive cells could be detected in layers II and III than in the other layers. In the subiculum, pre- and parasubiculum the GAD and GABA-i cells were present in relatively large numbers in all layers, except the molecular layer, which contained only a small number of GABA cells. Within the entorhinal area, GAD and GABA stained cells ranged in size from small (13 m in diameter) to large (22 m in diameter). A large number of different morphological classes of cells were found, except pyramidal and stellate cells. In the pre- and parasubiculum, on the other hand, the GABA cells were generally small to medium in size and morphologically more homogeneous than in the subiculum and entorhinal area.The entire retrohippocampal region was densely innervated by GABA preterminal processes, with little variation in the regional density of innervation. Within the entorhinal area, presubiculum and subiculum, a clear difference was found in the laminar pattern of innervation. In all three subfields the densest innervation was in layer II. In the entorhinal area both GAD- and GABA-i axons form palisades of fibers around the somata of neurons, which are tightly packed together in this layer. In the electron microscope both GAD-i and GABA-i were demonstrated in these axons. Axosomatic synaptic contacts were common between axons and the stellate neurons and other cells of this layer. Layers IV and VI appeared less dense in GAD-i terminals but appeared more densely innervated than layers III and V. The lamina dessicans was relatively poor in GAD-i. In the subiculum and presubiculum, as well as all other subfields of the hippocampal region, the innervation is dominated by axo-somatic innervation of layer II cells. The outer third of the molecular layer was more densely innervated than the inner part. Taken together, the present study has shown that the retrohippocampal region is rich in GABAergic neurons as well as axon terminals, some of which form numerous synapic contacts with cells of the region. GABAergic neurotransmission is an important mechanism in retrohippocampal circuits not only for the resident interneuronal population but in the surround as well.     

Morphological details of the projection from the presubiculum to the entorhinal area as shown with the novel PHA-L immunohistochemical tracing method in the rat

Iontophoretic injections of the lectin, phaseolus vulgaris leucoagglutinin (PHA-L) were made into the presubiculum of rats. The anterogradely transported lectin was visualized by using an anti-PHA-L antibody in combination with immunohistochemistry. The PHA-L tracing method revealed morphological details of the projection of the presubiculum to the ipsi- and contralateral medial entorhinal area usually not seen with other anterograde transport techniques. Fine varicose fibers form a dense terminal plexus in the deep parts of layer III. In layer II and deep layer I, the fibers form column-like axonal bundles, terminating in patches in the deep part of layer I. Some fibers reach the outer three layers of the entorhinal area (EA) from collaterals of axons running in the molecular layer, while a majority enter from the deep layers.     

Terminal distribution of projections from the retrosplenial area to the retrohippocampal region in the rat, as studied by anterograde transport of biotinylated dextran amine

The terminal distribution of projections from the retrosplenial area to the retrohippocampal region was examined in the rat with anterograde transport of biotinylated dextran amine. Projections from the retrosplenial granular area (RSG) to the retrohippocampal region terminate predominantly ipsilaterally in layers I, III, V and VI of the presubiculum, layers I and IV–VI of the parasubiculum, the molecular and pyramidal cell layers of the subiculum, and layers I, III, V and VI of the entorhinal area. On the other hand, projections from the retrosplenial agranular area (RSA) terminate predominantly ipsilaterally in layers I and III of the presubiculum and layers V and VI of the entorhinal and perirhinal areas, and ipsilaterally in layers IV–VI of the parasubiculum. The results show that projections from the RSG to the retrohippocampal region are as massive as those from the RSA, and that each retrosplenial area has distinct projection fields in the retrohippocampal region. This suggests that each retrosplenial area may play some distinct functional roles in memory and learning processes such as spatial behavioral learning.     

The distribution and orientation of serotonin fibers in the entorhinal and other retrohippocampal areas. An immunohistochemical study with anti-serotonin antibodies in the rats brain

Summary The serotonin (5-hydroxytryptamine; 5-HT) innervation of the retrohippocampal region (subiculum, pre-and parasubiculum, area 29e, medial and lateral entorhinal area) in the rat brain has been examined with antibodies against 5-HT used in combination with fluorescence histochemistry. Analysis of consecutive sections cut in the coronal, sagittal, and horizontal planes revealed a widespread distribution of 5-HT immunoreactive fibers throughout the retrohippocampal region. This innervation was heterogeneous with regard to the morphological characteristics of the 5-HT fibers, their density and their spatial orientation.On the basis of morphological criteria, four different types of 5-HT positive processes were distinguished: (a) fine, convoluted fibers with small (0.5–0.8 m), round and evenly spaced varicosities; (b) fine fibers with elongated, irregularly distributed varicosities; (c) thick, possibly myelinated fibers, and (d) a terminal plexus with large (5–10 m), irregularly spaced varicosities. Analysis of the laminar distribution of the 5-HT fibers showed that whereas all layers contain 5-HT positive fibers, the molecular layer was the most densely innervated. The 5-HT fibers were found to be oriented both parallel and transverse to the longitudinal axis of medial and lateral entorhinal area. This grid-like arrangement was less pronounced in the presubiculum. Although the 5-HT innervation of the retrohippocampal region was found to be dominated by a widespread and apparently diffuse pattern, several areas contained dense clusters of preterminal 5-HT processes: area 29e, dorsal presubiculum (layer II), lateral entorhinal area (layer III and ventral layer II) and the transitional zone of the ventral entorhinal area. The 5-HT fibers were found to enter the retrohippocampal region primarily by three different routes; from the ventral and dorsal aspects and from the piriform and lateral neocortex (via the perirhinal area). Most of the fibers enter the region by the ventral route and these were found to ascend in all layers but predominantly in layer I.The location of the 5-HT cells giving rise to the innervation of the entorhinal area was studied by combining retrograde transport of fluorescent tracers with immunohistochemistry on the same tissue section. Both ipsi-and contralaterally located cells in the dorsal and median raphe nuclei were found to project to the entorhinal area. Most, but not all, of these retrogradely labeled cell bodies also contain 5-HT immunoreactivity.Abbreviations Used for Text and Figures ab angular bundle - ad area dentata - cc corpus callosum - dcp decussation of bracium conjunctivum - dr dorsal raphe nucleus (B7) - flm medial longitudinal fasciculus - mr median raphe nucleus (B8) - occ occipital cortex - rtp nucleus reticularis tegmentipontis - sub subiculum - vtG ventral tegmental area of Gudden - III third nerve nucleus - 28L entorhinal area, lateral part - 28L entorhinal area, lateral part (ventral) - 28M entorhinal area, medial part - 28M entorhinal area, medial part (ventral) - 49(a,b) parasubiculum - AMYG amygdala - Prs Presub-presubiculum - TA transitional area - TZ transitional zone     

The substance P innervation of the rat hippocampal region

Summary The distribution of substance P (SP) immunoreactive nerve cell bodies and preterminal processes was studied in the rat brain by using several anti-SP-antibodies in combination with immunohistochemical techniques. In normal rats and in rats pretreated with colchicine, SP immunoreactive preterminal processes were found in the hippocampal region, but SP positive cellbodies could be detected only after colchicine pretreatment. Medium-sized to large, multipolar cells immunoreactive for SP were found in stratum oriens of the hippocampal subfield CA3 and in the hilus of the area dentata. Medium-sized to small, round or fusiform cells were detected in the pyramidal layer of the ventral subiculum and in layers III–VI of the ventral entorhinal area. The SP stained preterminal processes were of two types. Numerous fine, varicose axons were stained in different parts of Ammon's horn, while in the retrohippocampal structures, the SP immunoreactivity was present in small distinctly stained puncta. These frequently formed pericellular arrangements around unstained cells, indicative of axosomatic contacts between SP terminals and cells in the hipocampus. In Ammon's horn, the densest SP innervation was found in strata oriens, radiatum and moleculare of subfields CA3a and CA2. Scattered fibers were also present in the stratum oriens of CA3a-c and in the hilus, in particular at ventral levels. In retrohippocampal structures, the SP innervation predominated in the deep pyramidal layer of the subiculum, the second layer of the presubiculum and in layers VI and IV of the medial and lateral entorhinal area. Many of these terminals may arise from local interneurons as well as from sources outside the hippocampal region.Taken together, these studies demonstrate a far more extensive innervation by SP, or a closely related peptide, of the rat hippocampal region than was previously recognized. This suggests that SP may play an important role in neurotransmission within the hippocampal region.Stephen Davies was supported by Travel grants from the Wellcome Trust and the Gurantors of Brain.     

Contacts between medial and lateral perforant pathway fibers and parvalbumin expressing neurons in the subiculum of the rat

The entorhinal cortex (EC) projects via the perforant pathway to all subfields in the hippocampal formation. One can distinguish medial and lateral components in the pathway, originating in corresponding medial and lateral subdivisions of EC. We analyzed the innervation by medial and lateral perforant pathway fibers of parvalbumin-expressing neurons in the subiculum. A neuroanatomical tracer (biotinylated dextran amine, BDA) was stereotaxically injected in the medial or lateral entorhinal cortex, thus selectively labeling either perforant pathway component. Transport was allowed for 1 week. Transported BDA was detected with streptavidin-Alexa Fluor 488. Parvalbumin neurons were visualized via immunofluorescence histochemistry, using the fluorochrome Alexa Fluor 594. Via a random systematic sampling scheme using a two-channel, sequential-mode confocal laser scanning procedure, we obtained image series at high magnification from the molecular layer of the subiculum. Labeled entorhinal fibers and parvalbumin-expressing structures were three dimensionally (3D) reconstructed using computer software. Further computer analysis revealed that approximately 16% of the 3D objects ('boutons') of BDA-labeled fibers was engaged in contacts with parvalbumin-immunostained dendrites in the subiculum. Both medial and lateral perforant pathway fibers and their boutons formed such appositions. Contacts are suggestive for synapses. We found no significant differences between the medial and lateral components in the relative numbers of contacts. Thus, the medial and lateral subdivisions of the entorhinal cortex similarly tune the firing of principal neurons in the subiculum by way of parvalbumin positive interneurons in their respective terminal zones.     

Cortical afferents of the nucleus accumbens in the cat,studied with anterograde and retrograde transport techniques

The cortical afferentation of the nucleus accumbens in the cat was studied with the aid of retrograde tracing techniques. Retrograde experiments were carried out with horseradish peroxidase or one of the fluorescent tracers Bisbenzimid, Nuclear Yellow and Fast Blue. In the anterograde experiments [³H]leucine and [³⁵S]methionine were used as tracers.Following injections in the nucleus accumbens, retrogradely-labelled cells were found in the medial frontal cortex, the anterior olfactory nucleus, the posterior part of the insular cortex, the endopiriform nucleus, the amygdalo-hippocampal area, the entorhinal and perirhinal cortices and the subiculum of the hippocampal formation. In the medial frontal cortex most of the labelled cells were found in layers III and V of the prelimbic area (area 32 of Brodmann), but retrogradely-filled neurons were also present in the infralimbic area and in the caudoventral part of the lateral bank of the proreal gyrus. Retrogradely-labelled cells in the entorhinal and perirhinal cortices were located in the deep cellular layers. Following large injections in the nucleus accumbens, retrograde labelling in the subiculum extended from the most dorsal, septal pole to the most ventral, temporal pole.Injections of anterograde tracers were placed in the frontal cortex, the entorhinal and perirhinal cortices and the hippocampal formation. The prelimbic area was found to project via the internal capsule to mainly the rostral half of the nucleus accumbens, whereas in the caudal half of the nucleus only a lateral region receives frontal cortical fibres. Following injections in the infralimbic area only fibres passing through the nucleus accumbens were labelled. Afferents from the entorhinal and perirhinal cortices reach the nucleus accumbens by way of the external capsule and terminate mainly in a ventral zone of the nucleus accumbens.Afferents from the entorhinal area are distributed to the entire accumbens, whereas the termination field of the perirhinal afferents is largely restricted to the lateral part of the nucleus accumbens. Both the frontal cortex and the entorhinal and perirhinal cortices appear to project also to the nucleus caudatus and the tuberculum olfactorium. These cortical areas also project to the contralateral striatum.Both anterograde and retrograde tracing experiments demonstrated a topographical relationship between the subiculum and the nucleus accumbens. The ventral pole of the subiculum projects via the fornix to the medial part of the caudal half of the nucleus accumbens and to a small dorsomedial area in its rostral half. Successively more dorsal portions in the subiculum project to successively more ventrolateral parts in the rostral nucleus accumbens. The projection from the hippocampus was found to extend also to the tuberculum olfactorium. The results of the present study do not provide unambiguous criteria for the delimitation of the nucleus accumbens in the cat.     

Afferents to the seizure-sensitive neurons in layer III of the medial entorhinal area: a tracing study in the rat

Neurons in layer III of the medial entorhinal area (MEA) in the rat are extremely vulnerable to local injections of amino-oxyacetic acid and to exprimentally induced limbic seizures. A comparable specific pathology has been noted in surgical specimens from patients with temporal lobe epilepsy. Efforts to understand this preferential neuronal vulnerability led us to study the neural input to this layer in the rat. Iontophoretic injection of the retrograde tracer fast blue, aimed at layer III of the MEA, resulted in retrogradely labeled neurons in the presubiculum in all the injected hemispheres. The nucleus reuniens thalami, the anteromedial thalamic nucleus, the ventral portion of the claustrum (endopiriform nucleus), the dorsomedial parts of the anteroventral thalamic nucleus, and the septum-diagonal band complex were labeled less frequently. In only one experiment, retrogradely labeled neurons were observed in the ventrolateral hypothalamus and in the brainstem nucleus raphe dorsalis. Since projections from claustrum to the entorhinal cortex has not been studied in the rat with modern sensitive anterograde tracing techniques, iontophoretic injections of the anterograde tracer Phaseolus vulgaris-leucoagglutinin were placed into the ventral portion of the claustrum. Anterogradely labeled fibers in the entorhinal area proved not to be confined to the MEA, since a prominent projection distributed to the lateral entorhinal area as well. In both areas, the densest terminal labeling was present in layers IV–VI, whereas layer III appeared to be only sparsely labeled. The present data indicate that of all potential afferents only those from the presubiculum distribute preferentially to layer III of the MEA. This, in turn, suggests a potentially important role of the presubiculum in the seizure-related degeneration of neurons in layer III of the MEA.     

Distribution of [3H]cholecystokinin octapeptide binding sites in the hippocampal region of the rat brain as shown by in vitro receptor autoradiography

The distribution of binding sites for the neuropeptide cholecystokinin octapeptide in the rat hippocampal region was studied by using quantitative in vitro receptor autoradiography. Biochemical analysis of [3H]cholecystokinin octapeptide binding to tissue sections of the hippocampal region showed it to be of high affinity, to be saturable and approximately 50% specific at saturating concentrations. The binding of [3H]cholecystokinin octapeptide to hippocampal sections was dose-dependently blocked by cholecystokinin octapeptide, cholecystokinin and by pentagastrin. The autoradiographic analysis showed high densities of [3H]cholecystokinin octapeptide binding sites in the hilus of the area dentata, the outer three layers of the retrosplenial area and the presubiculum, layer 3 of the medial, but not the lateral, entorhinal area and the deep and superficial parts of layer 1 and 2, respectively of both the medial and the lateral entorhinal area. Medium binding densities were found in the parasubiculum and remaining layers of the entorhinal area and low densities occurred in the subiculum and in all subfields of Ammon's horn. The angular bundle and fornix-fimbria lacked specific [3H] cholecystokinin octapeptide binding sites. A very similar pattern of binding densities was found for [3H]pentagastrin. Comparisons of the cholecystokinin octapeptide receptor distribution with the cholecystokinin octapeptide innervation of the hippocampal region suggest that there exists a relatively good concordance in some hippocampal subfields such as the presubiculum and the entorhinal area between binding sites for [3H]cholecystokinin octapeptide and cholecystokinin-immunoreactive afferent input.     

Senile plaques are concentrated in the subicular region of the hippocampal formation in Alzheimer's disease

The distribution of senile plaques within the hippocampal formation was examined at autopsy in the brains of 18 patients with Alzheimer's disease, ranging in age from 62 to 89 years. Wax sections were cut at the level of the central part of the cornu ammonis and stained by a silver impregnation method. Plaque surface density was determined in the stratum pyramidale of the presubiculum, subiculum, prosubiculum and CA1-4. Senile plaques were present in all 7 regions of the hippocampal formation. However, the number of senile plaques/mm2 was significantly greater in the subicular complex than in the cornu ammonis. There was no difference in plaque density between the individual regions of the subicular complex or between the 4 CA fields.     

Effects of phencyclidines on signal transfer from the entorhinal cortex to the hippocampus in rats

The information transfer from the superficial layers of the entorhinal cortex (EC) to the hippocampus is regulated in a frequency dependent manner. Phencyclidine and related compounds such as MK-801 produce psychotic symptoms that closely resemble schizophrenia. We studied the effects of systemic administration of MK-801 on the signal transfer from the EC layer III to the hippocampal area CA1. High frequency (above 10 Hz) activation of the bi-synaptic entorhinal input in control animals results in a strong suppression of the field potentials in the stratum lacunosum-moleculare of the area CA1. In contrast, in MK-801 pretreated rats the field response was less reduced. The field potential responses evoked in these two groups of animals by high-frequency activation of the monosynaptic input were similar suggesting selective alterations in layer III of the medial EC. We suggest, that MK-801 causes disinhibition of layer III projection cells and, therefore, may cause strong, pathological activation of direct layer III-CA1 pathway.     

The localization of leucine-enkephalin immunoreactivity within the guinea pig hippocampus

Summary The distribution of enkephalin immunoreactive fibres has been studied in the hippocampus, subiculum and entorhinal cortex of the guinea pig. Two immunoreactive enkephalin fibre systems were found. One system corresponds to the mossy fibre system from the fascia dentata to CA3 and courses at the level of the mossy fibre end bulb in a longitudinal direction along the main axis of the hippocampus. Another system originates in the medial and lateral entorhinal cortex, traverses the subiculum, and then courses in the stratum molecu-lare/lacunosum to CA1 and CA3; part of these fibres crosses the hippocampal fissure and reaches the stratum moleculare of the fascia dentata. In the fascia dentata intense immunoreactivity was observed in the distal and middle one-third of the stratum moleculare at the side of the terminations of the lateral and medial perforant path fibres. Various types of immunoreactive cell bodies were found in the fascia dentata, CA3, CA1, subiculum and in the entorhinal cortex.Supported in part by the Dutch Organization of Pure Scientific Research, FUNGO/ZWO     

Topographic projections from the subiculum to the limbic regions of the medial frontal cortex in the cat.

When WGA-HRP was injected into the subicular cortex of the ventral hippocampal formation (Hv) in the cat, terminal labeling was seen ipsilaterally in the caudoventral parts of the medial frontal cortex: in the infralimbic cortex (area 25), ventral part of the prelimbic cortex (area 32), and caudoventral part of the orbitofrontal cortex (area 12). The terminal labeling was observed in all cortical layers except layer 1. The cells of origin of these projections were then confirmed to be pyramidal neurons in the subiculum of the Hv. The projections were topographically organized: the temporal-septal shift along the long axis of the subiculum of the Hv corresponded to a rostroventral-caudodorsal shift in the anterogradely labeled limbic regions of the medial frontal cortex.     

Calbindin-D28k在出生前人海马本部及下托神经元的表达及其变化

本实验采用免疫组织化学方法研究了１３～３８ 周人胎儿海马本部及下托含Ｃａｌｂｉｎｄｉｎ－Ｄ２８ｋ 神经元的分布和发育。结果表明：在１３～１４ 周时,许多含Ｃａｌｂｉｎｄｉｎ－Ｄ２８ｋ 锥体细胞可见于ＣＡ１ 区锥体细胞层中部及深部,随着胎龄增大,ＣＡ１ 区含Ｃａｌ－ｂｉｎｄｉｎ－Ｄ２８ｋ 锥体细胞的数量及密度逐渐下降,最终消失,并且这种下降及消失首先从含Ｃａｌｂｉｎｄｉｎ－Ｄ２８ｋ 锥体细胞区浅部开始,然后向深部推进;在１３～２８ 周期间,ＣＡ２ 和ＣＡ３ 区也有许多含Ｃａｌｂｉｎｄｉｎ－Ｄ２８ｋ 锥体细胞,但至３２ 周以及其后,ＣＡ３ 和ＣＡ２ 区则不见含Ｃａｌｂｉｎｄｉｎ－Ｄ２８ｋ 锥体细胞,仅在ＣＡ２ 与ＣＡ１ 交界区见到少量弱染的含Ｃａｌｂｉｎｄｉｎ－Ｄ２８ｋ 锥体细胞。此外,在２８～３８ 周期间,ＣＡ３ 和ＣＡ２ 区锥体细胞层周围可见许多含Ｃａｌｂｉｎｄｉｎ－Ｄ２８ｋ 的苔藓纤维,其密度随胎龄增大而增加。１４～３８ 周期间,许多含Ｃａｌｂｉｎｄｉｎ－Ｄ２８ｋ 的锥体细胞也出现于下托锥体细胞层全层及前下托锥体细胞层浅部（细胞岛区）及中部。这些区域含Ｃａｌ－ｂｉｎｄｉｎ－Ｄ２８ｋ 锥体细胞的数量及染色强度在１４～２４ 周期间逐渐增     

Parvalbumin-immunoreactive neurons in the hippocampal formation of Alzheimer''s diseased brain

The number and topographic distribution of immunocytochemically stained parvalbumin interneurons was determined in the hippocampal formation of control and Alzheimer's diseased brain. In control hippocampus, parvalbumin interneurons were aspiny and pleomorphic, with extensive dendritic arbors. In dentate gyrus, parvalbumin cells, as well as a dense plexus of fibers and puncta, were associated with the granule cell layer. A few cells also occupied the molecular layer. In strata oriens and pyramidale of CA1–CA3 subfields, parvalbumin neurons gave rise to dendrites that extended into adjacent strata. Densely stained puncta and beaded fibers occupied stratum pyramidale, with less dense staining in adjacent strata oriens and radiatum. Virtually no parvalbumin profiles were observed in stratum lacunosum-moleculare or the alveus. Numerous polymorphic parvalbumin neurons and a dense plexus of fibers and puncta characterized the deep layer of the subiculum and the lamina principalis externa of the presubiculum. In Alzheimer's diseased hippocampus, there was an approximate 60% decrease in the number of parvalbumin interneurons in the dentate gyrus/CA4 subfield (P<0.01) and subfields CA1–CA2 (P<0.01). In contrast, parvalbumin neurons did not statistically decline in subfields CA3, subiculum or presubiculum in Alzheimer's diseased brains relative to controls. Concurrent staining with Thioflavin-S histochemistry did not reveal degenerative changes within parvalbumin-stained profiles. These findings reveal that parvalbumin interneurons within specific hippocampal subfields are selectively vulnerable in Alzheimer's disease. This vulnerability may be related to their differential connectivity, e.g., those regions connectionally related to the cerebral cortex (dentate gyrus and CA1) are more vulnerable than those regions connectionally related to subcortical loci (subiculum and presubiculum).