Glutamate-like immunoreactivity in identified neuronal populations of insect nervous systems

Glutamate is considered to be the most likely transmitter candidate at excitatory synapses onto skeletal muscles of insects. We investigated the distribution of glutamate-like immunoreactivity (Glu-LI) in identified motor neurons of glutaraldehyde-fixed metathoracic ganglia of the locust in paraffin serial sections. The presumably glutamatergic fast and slow extensor tibiae motor neurons show Glu-LI, whereas other cells, including the GABAergic common inhibitory motor neurons and the cluster of octopaminergic dorsal unpaired median cells, show rather low levels of staining. Immunoreactivity of the fast extensor tibiae motor neuron is located in soma, neurites, axon, and the terminal arborizations. A double-labeling experiment on sections of the locust metathoracic ganglion showed that antisera against glutamate and GABA discriminate between the presumably glutamatergic and GABAergic motor neurons and that GABA-LI-positive neurons are low in Glu-LI. The results suggest that Glu-LI can be used as a marker for detecting potential glutamatergic neurons in insects under the present conditions. Application of the glutamate antiserum to sections of the honeybee brain revealed Glu-LI in motor neurons but also in certain interneurons. The most prominent populations of Glu-LI-positive cells were the monopolar cells and large ocellar interneurons, which are first-order interneurons of the visual and ocellar system. Several groups of descending interneurons also showed Glu-LI. The distributions of Glu-LI and GABA-LI are complementary in locust and bee ganglia. The high level of Glu-LI in certain interneuronal populations, as well as in identified glutamatergic motor neurons, suggests that insect central nervous systems may contain glutamatergic neuronal pathways.     

Localization of kynurenine aminotransferase immunoreactivity in the rat hippocampus.

The localization and distribution of kynurenine aminotransferase (KAT), the biosynthetic enzyme of the excitatory amino acid receptor antagonist, kynurenic acid, was studied in the rat hippocampal formation with immunohistochemical methods. The enzyme was found mainly in glial cells that could be distinguished as 3 types on the basis of their shapes and locations. Typically, these cells shared the morphological features of astrocytes and exhibited glial fibrillary acidic protein immunoreactivity as demonstrated by a double-labeling technique. The distribution of KAT-containing glial cells was heterogeneous throughout the hippocampal formation. In the hippocampus, the stratum lacunosum-moleculare of Ammon's horn and the hilus contained a higher density of KAT-positive glial cells than other regions, whereas the lowest density of KAT glial cells was observed in the granule cell layer of the dentate gyrus and in the stratum radiatum of CA subfields. In the subicular complex, the density of KAT-containing glial cells was generally higher in the superficial than in the deep layer. Hippocampal neurons exhibiting KAT immunoreactivity, distinguished as nonpyramidal cells, were very few in number and mainly distributed in strata oriens and pyramidale of Ammon's horn. Substantially more KAT-positive neurons were observed in layers II and III of the subicular complex. The organization of cellular elements containing KAT may be of relevance for the function and possible dysfunction of kynurenic acid in the rat hippocampal formation.     

Localization of mGluR1a-like immunoreactivity and mGluR5-like immunoreactivity in identified populations of striatal neurons

Metabotropic glutamate receptors are important mediators of excitatory amino acid neurotransmission in the striatum. Two-color immunofluorescence histochemistry and immunohistochemistry in combination with retrograde tract-tracing techniques were used to examine the distribution of metabotropic glutamate receptor subtypes 1a and 5 (mGluR1a and mGluR5) among identified subpopulations of striatal projection neurons and interneurons. The majority of striatopallidal and striatonigral neurons were double-labeled for both mGluR1a or mGluR5. Approximately 60% to 70% of either striatonigral or striatopallidal neurons expressed mGluR1a- or mGluR5-like immunoreactivity. The percentage of double-labeled striatopallidal or striatonigral projection neurons did not differ among striatal quadrants. Striatal interneurons expressing parvalbumin or somatostatin or choline acetyltransferase exhibited varying degrees of expression of mGluR1a or mGluR5. Virtually all (94%) parvalbumin-immunoreactive striatal neurons expressed mGluR1a-like immunoreactivity with a majority (79%) of these neurons expressing mGluR5-like immunoreactivity. A high percentage (89%) of striatal choline acetyltransferase-immunoreactive neurons were double-labeled for mGluR1a-like immunoreactivity. Approximately 65% of striatal choline acetyltransferase-immunoreactive neurons expressed mGluR5-like immunoreactivity. A majority (65%) of somatostatin-immunoreactive striatal interneurons expressed mGluR1a-like immunoreactivity with a slightly lower percentage (55%) expressing mGluR5-like immunoreactivity. These findings indicate considerable heterogeneity among striatal projection and interneurons with respect to mGluR1a and mGluR5 expression. There may be subpopulations of striatonigral and striatopallidal projection neurons. These results are consistent as well with prior data indicating subpopulations of the different classes of striatal interneurons.     

Effects of chemical and surgical lesions on levels of chromatographically identified enkephalin-like peptides in rat hippocampus

The present study quantitates the content of Met- and Leu-enkephalin in the rat hippocampus, and provides information on the localization of the enkephalins within the hippocampal neuronal circuitry. Several enkephalins were identified in rat hippocampus, two of which are shown to be Met- and Leu-enkephalin. The levels of these enkephalins, and of other unidentified enkephalin-related peptides, were not depleted by intrahippocampal colchicine, which destroyed the great majority of the hippocampal granule cells and the associated mossy fiber pathway. Entorhinal cortical lesions ablating the perforant pathway input to the hippocampus also did not significantly lower enkephalin levels in the hippocampus. Unilateral fimbrial transection caused a significant bilateral increase in both Met- and Leu-enkephalin levels. This may result from loss of a stimulatory input to putative enkephalin containing interneurons within the hippocampus. The extents of all lesions were verified histologically in hippocampi used for biochemical analysis. No evidence was seen for the presence of enkephalins in the perforant pathway, nor in nerve fibers in the fimbria/fornix, which provide the other main source of hippocampal efferents. The enkephalins are likely to be intrinsic to the hippocampus, in which neuronal cell bodies containing enkephalin-like immunoreactivity have been extensively reported.     

Acute anoxia-induced alterations in MAP2 immunoreactivity and neuronal morphology in rat hippocampus

Cerebral ischemia induces major neuronal morphological alterations. It is not clear, however, whether this directly caused by O₂ deprivation. To determine the effect of hypoxia on cytoskeletal structures and neuronal morphology, we performed experiments and examined anoxia-induced changes in microtubule-associated protein 2 (MAP2) and cell morphology in hippocampal slices in vitro. Anoxia (measured PO₂ = 0Torr) induced a marked loss in dendritic MAP2 immunoreactivity and cell swelling of hippoocampal neurons by 2 h after O₂ reinstitution. These changes were severe in CA1 and CA3 neurons and comparatively mild in dentate gyrus neurons. Quantitative analysis showed that 10 min of anoxia induced a 30% loss of MAP2-positive dendrites but this increased to 70% after 30 min of anoxia. A concurrent major increase in somata area of about 100% and 200% was observed in CA1 and CA3 neurons respectively. Somata area in the lower dentate gyrus, however, increased either insignificantly or by only 30% for the respective periods of anoxia. These results suggest that deprivation of O₂ can by itself induce a major loss in dendritic MAP2 immunoreactivity and changes in cell morphology in hippocampal neurons. These alterations occur rapidly after hypoxia, and the severity of these changes is directly related to the duration of anoxia and brain region in the hippocampus.     

Localization of delta opioid receptor immunoreactivity in interneurons and pyramidal cells in the rat hippocampus

To study possible cellular targets and subcellular sites of action of opioid ligands in the rat hippocampus, we examined the distribution of the delta opioid receptor (DOR) by immunocytochemistry. By light microscopy, numerous interneurons, or non-principal cells, were intensely labeled for DOR, whereas the CA1 and CA3 pyramidal cells were lightly labeled. DOR-immunoreactive interneurons were found throughout the hippocampus but were particularly concentrated in stratum oriens of the CA1 region. Double labeling immunofluorescence revealed that DOR-immunoreactivity was found in a subpopulation of γ-aminobutyric acid (GABA)-containing interneurons, which included most somatostatin-immunoreactive cells. Electron microscopic analysis of sections singly labeled for DOR revealed that DOR-immunoreactive profiles were abundant and widespread throughout all hippocampal lamina and had a similar distribution in CA1 and CA3. DOR-immunoreactivity was sometimes found in dendrites, which corresponded in morphology to those of interneurons. In addition, DOR-labeling was found in the shafts and spines of many dendrites, which exhibited the morphology of pyramidal cell dendrites. Within dendrites, dense DOR-immunoreactivity was associated with the plasmalemmal surface at or near the postsynaptic density, usually of asymmetric synapses. In addition, DOR labeling was present in a heterogeneous population of axon terminals, as well as in astrocytic profiles. At mossy fiber synapses, DOR labeling was occasionally found at both pre-and post-synaptic sites. These studies demonstrate that DOR is present at multiple sites on diverse cell types where it may function to regulate neuronal activity in the hippocampus. J. Comp. Neurol. 373-387, 1997. © 1997 Wiley-Liss, Inc.     

Neurohypophysial peptides and the hippocampus. I. Vasopressin immunoreactivity in the rat hippocampus

Immunocytochemical studies have demonstrated that nerve fibres containing immunoreactive vasopressin project to many areas of the central nervous system. In the present investigation, the presence of immunoreactive arginine vasopressin (IR-AVP) in the hippocampus of Wistar rats was confirmed by radioimmunoassay. The vasopressin content of the dorsal hippocampus was 30.3 ± 7.3 pg IR-AVP/mg soluble protein (mean ± SEM, n=9) and that of the ventral hippocampus was 81.4 ± 8.3 pg IR-AVP/mg soluble protein (n=9), while tissue from the cerebral cortex contained no detectable vasopressin. That the immunoreactivity was due to vasopressin was confirmed by its absence in hippocampal or cortical tissue from homozygous Brattleboro rats, which are genetically unable to synthesize vasopressin.     

Distribution of substance p and enkephalin-like immunoreactivity in the substantia nigra of rat, cat and monkey

A comparative study of the distribution of substance P (SP) and enkephalin (ENK) immunoreactivity in the substantia nigra (SN) of the rat, cat and squirrel monkey (Saimiri sciureus) was undertaken by means of the indirect immunofluorescence technique. In the rat a dense neuronal network composed of fine fibers displaying SP immunoreactivity is uniformely distributed throughout the rostrocaudal extent of the substantia nigra pars reticulata (SNr) and in the ventral part of substantia nigra pars compacta (SNc). Some coarse SP-positive fibers also occur in SNc. In addition, ENK-immunoreactive fibers are scattered amongst SNc neurons but abound particularly in the caudolateral part of SNr. In cat innumerable fine SP-positive fibers are distributed in SNr according to a pattern similar to that found in rat. ENK-immunoreactive fine fibers are densely packed in the ventromedial part of SNr whereas coarse ENK fibers are scattered in both SNc and SNr but abound particularly in the caudolateral portion of SNr. In monkey fine SP and ENK-immunoreactive fibers occur in very large number in SNr. These two types of fibers are distributed according to a similar but strikingly complex and heterogeneous pattern. In addition, coarse fibers displaying either SP or ENK immunoreactivity are scattered amongst the SNc neurons in monkey. These findings reveal that SP immunoreactive fibers are present in large number and are distributed according to a somewhat similar pattern in rat, cat and monkey. In contrast, the number of ENK-positive fibers and the complexity of their organizational feature in SN increase strikingly from rodent to primate.     

Localization of alpha-, beta-, and gamma-synuclein during neuronal development and alterations associated with the neuronal response to axonal trauma

Genetic and protein studies have indicated abnormalities in alpha-synuclein in neurodegenerative diseases. However, the developmental localization and cellular role of synuclein isoforms is contentious. We investigated the cellular localization of alpha-, beta-, and gamma-synuclein in developing cultured rat neurons and following axonal transection of relatively mature neurons, a model that disrupts the axonal cytoskeleton and results in regenerative sprouting. Cortical neurons were grown up to 21 days in vitro (DIV). Axon bundles at 21 DIV were transected and cellular changes examined at 4 and 24 h post-injury. Immunohistochemistry demonstrated that alpha- and beta-synuclein were localized to cellular cytosol and growth cones at 3DIV, with accumulating puncta-like labeling within axons and growth cones by 10-21DIV. In contrast, gamma-synuclein immunoreactivity was limited at all time points. By 21DIV, alpha- and beta-synuclein were present in the same neurons but largely in separate subregions, only 26% of puncta contained both alpha- and beta-synuclein immunoreactivity. Less than 20% of alpha-, beta-, and pan-synuclein immunoreactive puncta directly colocalized to synaptophysin profiles at 10DIV, decreasing to 10% at 21DIV. Both alpha- and beta-synuclein accumulated substantially within damaged axons at 21DIV and were localized to cytoskeletal abnormalities. At latter time points post-injury, alpha- and beta-synuclein immunoreactive puncta were localized to growth cone-like structures in regenerating neurites. This study shows that alpha- and beta-synuclein have a precise localization within cortical neurons and are generally nonoverlapping in their distribution within individual neurons. In addition, synuclein proteins accumulate rapidly in damaged axons and may have a role in regenerative sprouting.     

Changes from enkephalin-like to gastrin/cholecystokinin-like immunoreactivity in snail neurons

Coexistence of regulatory peptides is reported with increasing frequency in the mammalian neuroendocrine system. We have investigated the possible presence of such coexistence in the invertebrate Lymnaea stagnalis and have found evidence for concurrence of enkephalin- and gastrin/cholecystokinin (CCK)-like peptides in identified neurosecretory neurons. Our immunocytochemical studies reveal, however, that some of these neurons show variations in their content of immunoreactive peptides depending upon season or age. Thus, the light green cells of the Lymnaea cerebral ganglion, known to produce a body growth-stimulating hormone, contain enkephalin-like immunoreactivity during spring and gastrin/CCK-like immunoreactivity during fall. During winter, these cells contain both enkephalin-like and gastrin/CCK-like immunoreactivities. In another group of neurosecretory neurons, the caudodorsal cells, known to produce an ovulation hormone, age-dependent changes were noted. Thus, in young animals, these cells contain enkephalin-like immunoreactivity; in animals of intermediate age, they contain both enkephalin- and gastrin/CCK-like immunoreactivity and, in older animals, only gastrin/CCK-like immunoreactivity can be detected in these cells. Interestingly, the mediodorsal bodies, which are endocrine organs also implicated in regulating sexual function of Lymnaea, show age-dependent variations in immunocytochemically detectable enkephalin- and gastrin/CCK-like immunoreactivity which parallels that found in the caudodorsal cells. The changes noted in the light green cells and the caudodorsal cells are detected both in the cell bodies and in the terminals of these cells, suggesting that the immunoreactivity represents secretory material. The relation, if any, between the immunoreactivities detected and the growth and reproduction of Lymnaea remains to be established, as do the factors responsible for the observed age- and season-dependent variations.     

Design-based estimation of neuronal number and individual neuronal volume in the rat hippocampus

Tools recently developed in stereology were employed for unbiased estimation of the neuronal number and volume in three major subdivisions of rat hippocampus (dentate granular, CA1 and CA3 pyramidal layers). The optical fractionator is used extensively in quantitative studies of the hippocampus; however, the classical optical fractionator design may be affected by tissue deformation in the z-axis of the section. In this study, we applied an improved optical fractionator design to estimate total number of neurons on 100 microm thick vibratome sections that had been deformed, in the z-dimension, during histological processing. For estimating cell number, it is necessary to randomize only the location of section planes. But, in the local stereological methods, like cell volume estimation, the orientation of sections must also be randomized. We present a detailed application of a method for making vertical sections from the hippocampus. The volume of hippocampal neurons was estimated using the rotator principle on 40 microm thick plastic vertical uniform random sections and corrected for tissue shrinkage. Application of the proposed new design should result in more accurate estimates of neuron number and neuronal volumes in tissue sections affected by homogenous non-uniform shrinkage.     

Localization and quantitation of dynorphin B in the rat hippocampus

The present study measures the content of dynorphin B in the rat hippocampus, and localizes the dynorphins within the intrinsic hippocampal neuronal circuitry. The level of dynorphin B, which is representative of the prodynorphin-derived peptides, was markedly depleted by intrahippocampal injection of colchice, which destroyed the great majority of the hippocampal granule cells and the associated mossy fiber pathway. The hippocampus contralateral to the injection demonstrated a slight, non-significant rise in dynorphin B levels after colchicine. Entorhinal cortical lesions ablating the perforant pathway input to the hippocampus did not significantly alter dynorphin B levels in the hippocampus. Unilateral fimbrial transection caused a small but significant increase in dynorphin B on the side of the lesion relative to the unlesioned side, but neither side was significantly different from control.     

Localization and quantitation of proenkephalin-derived neuropeptides in the rat hippocampus

We have measured the content of met- and leu-enkephalin and dynorphin B in the rat hippocampus, and localized these opioid peptides within the intrinsic hippocampal neuronal circuitry with specific lesions. Several enkephalins, two of which were shown to be met- and leu-enkephalin, were identified in rat hippocampus. The levels of the enkephalin-related peptides were unaffected by intrahippocampal injections of colchicine, which destroyed the great majority of the hippocampal granule cells, while the level of dynorphin B, which serves as a marker for the proenkephalin B-derived peptides, was markedly depleted. Entorhinal cortical lesions ablating the perforant pathway input to the hippocampus did not significantly alter dynorphin B nor enkephalin levels in the hippocampus. Unilateral fimbrial transection caused a small but significant increase in dynorphin B on the side of the lesion relative to the non-lesioned side, although neither side was significantly different from control, while at the same time causing a significant bilateral increase in both met- and leu-enkephalin levels. This may result from loss of a direct or indirect stimulatory input to peptide-containing neurons within the hippocampus. The enkephalins appear to be located in neuronal cell bodies intrinsic to the body of the hippocampus, while the dynorphins are likely to be intrinsic only to the granule cell-mossy fiber system originating in the dentate gyrus.     

Estimated neuronal populations and volumes of the hippocampus and its subfields in schizophrenia

OBJECTIVE: It has been suggested that the primary focus of the pathological process in schizophrenia is on the limbic system, and there have been several postmortem reports of changes in the histological structure or volume of the hippocampus, as well as a larger number of MRI reports of volume reductions. There are conflicting findings, however, with both techniques. METHOD: The authors conducted a study of the gross and subfield structure and cellular composition of the hippocampus in postmortem brains from 30 patients with DSM-IV-diagnosed schizophrenia (13 women, 17 men) and 29 comparison subjects with no psychopathology (14 women, 15 men). Stereological sampling procedures were applied to 25-microm-thick coronal paraffin sections taken at 5-mm intervals throughout the formalin-fixed hippocampus. Subfields were defined as the dentate fascia, the hilus (CA4), an amalgamation of the CA2 and CA3 subfields, the CA1 subfield, and the subiculum. Volumes, cell densities, and cell numbers of the subfields were assessed microscopically, and the volume of the hippocampus was estimated from both photographs and histological slides of the coronal slices. RESULTS: As assessed from histologically stained slides, the volumes of the hippocampus and its subfields did not differ between patients and comparison subjects. Left-sided reduction in hippocampal volumes estimated from photographs, which may have included parahippocampal tissue, was not confirmed on histological examination. No significant differences were observed between patients and comparison subjects in the cellular composition of the hippocampus. CONCLUSIONS: These findings do not support a primary alteration of the hippocampus in schizophrenia.     

Postnatal development of corticosteroid receptor immunoreactivity in the rat hippocampus.

Postnatal changes in corticosteroid receptor immunoreactivity in the rat hippocampus were examined using an antiserum against a fusion protein containing an N-terminal peptide of the Type I receptor, and a monoclonal antibody against the rat liver Type II-receptor. Age-related regional differences were observed. In the pyramidal cell layer of Ammon's horn, and granule cell layer of the dentate gyrus (DG), the percentage of Type I receptor immunoreactive (Type I-ir) and Type II receptor immunoreactive (Type II-ir) cells was high perinatally, declined sharply by postnatal day 10 (P10), and showed a variable increase to adult levels subsequently. The pyramidal cells of CA1-CA2, subiculum and DG showed a selective increase in Type II-ir in late postnatal life into adulthood, while most other regions showed higher Type I-ir in both early and late postnatal life, suggesting different roles for these receptors during development. Type II-ir was predominantly nuclear in most neurons, except for a transient appearance of cytoplasmic Type II-ir in neurons of the stratum oriens and molecular layers of Ammon's horn and dentate gyrus of P20-P30. Type I-ir was diffusely nuclear and cytoplasmic at all developmental ages. This is suggestive of differential genomic and extragenomic roles for these receptors during postnatal development of the hippocampus.     

Reward modulates neuronal activity in the hippocampus of the rat

In the hippocampus of freely moving rats, neurons have been recorded that fire predominantly when the animal travels through a particular area while exploring the environment (so-called 'place cells'). This study investigates if the neuronal firing characteristics of such cells are modulated by attention, expectation of reward or memory load. A total of 16 electrodes were implanted in the CA1 region of the hippocampus of 3-month-old Long-Evans rats. Using a tetrode recording system, single neurons were recorded while a rat explored an 8-arm maze and retrieved pellets at the end of each arm. It was found that 31 out of 67 neurons showed place cell characteristics, while the other cells either fired in more than one place or fired along whole arms of the maze. Interestingly, 11 of the 31 neurons showed enhanced firing activity when the animal entered a baited arm but did not fire when the arm was visited again after the bait had been retrieved. In a second experiment, only four out of eight arms were baited. Firing rates of 46 neurons were analysed, and all cells (spatial or non-spatial) fired more in baited arms than in non-baited ones (P<0.001). In a reversal task in which the previously unbaited four arms were subsequently baited, neuronal activity was increased in the newly baited arms (42 cells analysed, P<0.001). Since no alterations to the maze or cues have been made, we interpret the increased firing probability of neurons in baited arms compared to unbaited arms as a correlate for 'attention' or 'expectation'.     

Vinconate prevents ischemic neuronal damage in the rat hippocampus

The neuroprotective effect of vinconate, a novel vinca alkaloid derivative, was examined in a rat model of forebrain ischemia induced by 4-vessel occlusion. Hippocampal cell loss was quantified histologically 3 days after 10 or 15 min of ischemia. Intraperitoneal application of vinconate (25 and 50 mg/kg) 10 min before and immediately after 10 min of ischemia significantly reduced the neuronal cell loss in the CA1 sector of the hippocampus. Protective effect of vinconate against 15 min of ischemia was reduced, but there was still significant protection at the higher dose. Autoradiography using 14C-vinconate showed that the drug easily penetrates the blood-brain barrier and distributes in the hippocampus. The result suggests that vinconate prevents ischemic neuronal damage by direct action on the hippocampal CA1 neurons.     

Dynorphin- and enkephalin-like immunoreactivity is altered in limbic-basal ganglia regions of rat brain after repeated electroconvulsive shock

In an attempt to determine whether the opioid peptides derived from prodynorphin participate in the effects of electroconvulsive shock (ECS), we used radioimmunoassay and immunocytochemistry to measure dynorphin-like immunoreactivity (DN-LI) in various rat brain regions after repeated ECS treatments. Ten daily ECSs caused a significant increase in dynorphin A (1-8)-LI in most limbic-basal ganglia structures, including hypothalamus (50%), striatum (30%), and septum (30%). No significant change was found in the frontal cortex or the neurointermediate lobe of the pituitary. In contrast, 10 ECS treatments depleted DN-LI in hippocampal mossy fibers by 64%. A detailed time-course study revealed that a single shock caused a small but significant increase in hippocampal DN-LI, whereas three consecutive shocks depleted DN-LI by 30%. The maximal decrease in DN-LI was reached after six daily ECSs. The level of DN-LI in the hippocampus partly recovered, but remained lower than the control value 4, 7, and 14 d after the cessation of six daily ECSs (50, 77, and 83% of control value, respectively). In contrast with the ECS-induced depletion of hippocampal dynorphin, 10 daily ECSs caused a significant increase (40%) in (Met5)-enkephalin-LI in the hippocampus, as well as in other limbic-basal ganglia structures. Immunocytochemistry revealed that enkephalin-LI was increased in the perforant pathway, which is presynaptic to the dynorphin-containing mossy fiber pathway in the hippocampus. These observations suggest that different mechanisms may regulate these two opioid peptide systems in the hippocampus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Three-dimensional pattern of enkephalin-like immunoreactivity in the caudate nucleus of the cat

Recent anatomical investigations of the mammalian neostriatum have uncovered discrete neurochemical zones characterized by low levels of AChE and high levels of enkephalin-like immunoreactivity (ELI) compared with the surrounding neostriatal tissue. These regions, termed striosomes, which appear as patches in individual sections, have been associated with specific afferent and efferent pathways that differ from those of the surrounding neostriatal tissue. In the present study, the 3-dimensional distribution of these enkephalin-rich compartments in the caudate nucleus of the adult cat was investigated using computer-assisted 3-dimensional reconstruction techniques. Series consisting of coronal, sagittal, and horizontal tissue sections were obtained. In individual sections, discrete patches of intense enkephalin-like immunoreactivity appeared against a lighter-staining background. Three-dimensional reconstructions revealed that these patches overlapped across sections to form a highly inter-connected structure within the head and body of the caudate nucleus. Several structural features were evident in these reconstructions that appeared to be similar across animals. One consistent pattern was the formation of enkephalin-labeled finger-like structures radiating from the ventricular edge diagonally across the width of the nucleus. Smaller crossbridges were seen connecting the fingers, which often resulted in the formation of claws or rings of enkephalin-like immunoreactivity. These elements appear to align across planes to form a lattice-like structure outlining continuous regions of matrix. This structure may provide a basis for the orderly interaction between the patch and matrix compartments.     

Distribution of alpha and beta integrin subunits in the adult rat hippocampus after pilocarpine-induced neuronal cell loss,axonal reorganization and reactive astrogliosis

Integrins are –heterodimers that act as cell-extracellular matrix (ECM) and cell-cell adhesion molecules. During development, they are involved in axonal guidance, synaptogenesis, and in astrocytic maturation and migration. Here, we have examined the potential role of the integrin subunits 1–5 and 1–5 in axonal sprouting, synaptogenesis and reactive astrogliosis in the adult rat brain caused by pilocarpine-induced status epilepticus (SE). Strong hippocampal immunoreactivity of 1–5, 1, 3, 4, and 5 was observed in the pia mater, in vascular endothelia, and in astrocytes at the pial surface. 2 immunoreactivity was found exclusively in vascular endothelia. Pyramidal cells and interneurons of CA3–CA1, as well as hilar neurons revealed moderate 5 labeling in their cell bodies. Mossy fibers were immunoreactive for 2, 4, and 5. After pilocarpine-induced SE, strong immunoreactivity for 1, 2, 4, 5, 1, 3, and 4 was observed in reactive astrocytes. Our results show that members of the integrin family are differently distributed in cellular and subcellular compartments of the hippocampus and undergo specific patterns of regulation, which may be important for lesion-induced reactive changes in the adult brain.