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  • 1
    ISSN: 1432-0568
    Keywords: Serotonin ; Entorhinal cortex ; Hippocampus ; Immunofluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The distribution of serotonin (5-hydroxytryptamine, 5HT) containing nerve terminals in the lateral entorhinal cortex (LEC) has been studied using antibodies against 5-HT in combination with fluorescence histochemistry. Thin, varicose, branching fibers were found to be distributed in a relatively even, diffuse pattern throughout all layers of the LEC. The largest amount of this type of 5HT innervation was in Layer I. This diffuse pattern of 5-HT terminals was supplemented by a dense network of 5HT terminals restricted to Layer III of a small (∼1 mm) strip of the LEC. The fibers in this layer were thicker and more convoluted and contained larger varicosities than fibers in any other layer. The existence of a distinct innervation by 5-HT terminals of only a small portion of the LEC demonstrates a hitherto unrecognized and important principle of heterogeneity in 5-HT innervation of cortex. It suggests that 5-5HT neurons in the raphe can selectively influence specific, narrow regions of the lateral perforant path system, which, in turn, affect the intrinsic hippocampal circuits.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0568
    Keywords: Antigen/antibody ; Immunocytochemistry ; Area dentata ; Interneurons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The retrograde axonal labeling of hippocampal GABA-ergic neurons was studied after in vivo injections of a characterized antibody against glutamic acid decarboxylase (GAD) into different parts of the hippocampal region. Small injections (50 nl) of undiluted GAD antibody into the area dentata (AD) labelled fusiform and dentate pyramidal basket cells within the AD and fusiform and multipolar cells in subfields CA2/CA3 (a and b) of Ammon's horn. The labeled cells were characterized by intense immunoreactivity of the soma and proximal parts of the dendrites, while the nucleus contained little or none. The morphological appearance and laminar positions of these cells corresponded to hippocampal GAD-positive neurons, as shown previously (Ribak et al. 1978) with immunocytochemistry. Injections of anti-GAD into the medial entorhinal area, subiculum, and CA1 region labeled cells in strata oriens, pyramidale and radiatum of CA2 and CA3a, but injections of the antibody into these latter areas failed to label cells in the medial CA1 and subiculum, thus suggesting a preferential organization of hippocampl GABA neuronal projections in a lateral to medial direction. Injections of preimmune sera or antiserum preabsorbed with the pure enzyme antigen GAD failed to label cells in a manner similar to that described for the anti-GAD injections. These observations, taken together with the finding that injections of anti-GAD into the terminal field of non-GABA-ergic pathways never resulted in retrograde axonal transport of the antibody-antigen complex, suggest that the in vivo injection of GAD antibody is a useful method to study the organization of hippocampal GABA-ergic neurons and their projections.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0568
    Keywords: Retrohippocampal structures ; Immunohistochemistry ; Vasoactive intestinal polypeptide ; Somatostatin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The retrohippocampal region of the rat brain was analyzed by using immunohistochemistry with specific antibodies against somatostatin (SOM) and vasoactive intestinal polypeptide (VIP). Specifically immunoreactive neurons and terminal processes were labeled with either the anti-SOM or anti-VIP antiserum and they were referred to as SOM-like immunoreactive (SOM-LI) or VIP-like immunoreactive (VIP-LI) neurons and processes, respectively. The retrohippocampal region was rich in neuronal cell bodies and terminal processes showing immunoreactivity for SOM and VIP. In the entorhinal area SOM-LI neurons were located mainly in layers IV through VI and the VIP-LI neurons were found mainly in layers I through III. Thick (70–120 μm) sections treated with the immunoperoxidase method to achieve a Golgi-like staining pattern showed that cytological differences existed between SOM- and VIP-positive neurons. SOM-LI neurons were usually multipolar, fusiform, or occasionally pyramidal while VIP-LI neurons were usually bipolar, stellate, or fusiform. SOM-LI and VIP-LI axons and preterminal processes were differentially distributed within the laminae of the retrohippocampal region. VIP-LI terminals were found throughout all layers except layer I. SOM-LI terminals were found primarily in the molecular layers of all areas, layer IV of the medial and lateral entorhinal areas, and in the angular bundle. Thus, SOM-LI and VIP-LI neurons are distinguished by their morphology and their different distribution within the cortical layers and areas of the retrohippocampal region.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0568
    Keywords: Hippocampus ; Septum ; GAD ; Immunocytochemistry ; GABA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Injections of the fluorescent dyes Fast Blue or Granular Blue into either the hippocampus (volume approximately 50 nl) or the entorhinal area (100–150 nl) resulted in labeling by retrograde axonal transport of cells in the diagonal band of Broca (dbB) and the medial septum (MS). A large number (approximately 30%) of these cells contained glutamic acid decarboxylase (GAD)-like immunoreactivity, as determined by combined retrograde fluorescent tracing and GAD-immunohistochemistry. Not all GAD positive cells in the dbB and MS were labeled by fluorochromes in a single experiment. The GAD-stained and fluorochrome-containing cells were present at all rostro-caudal levels of the septum and appeared not to belong to any single morphological class of cells. Double staining experiments showed that the GAD-positive cells did not contain acetylcholinesterase reaction product. These findings provide evidence that a significant portion of the septohippocampal projection may utilize gamma-aminobutyric acid as a neurotransmitter.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 173 (1985), S. 45-52 
    ISSN: 1432-0568
    Keywords: Substance P ; Hippocampus ; Entorhinal area ; Immunohistochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: 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.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0568
    Keywords: Fluorescence histochemistry ; Raphe nuclei ; Allocortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: 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 μm), 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 μm), 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.
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  • 7
    ISSN: 1432-0568
    Keywords: Somatostatin ; Neurotensin ; Immunohistochemistry ; Septum ; Basal forebrain ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Antibodies to the neuropeptides somatostatin (SOM) and neurotensin were used to study the distribution of the two peptides within the septum of the rat brain. In colchicine treated rats, numerous somatostatin-positive cell bodies were found in the dorsal and ventral subdivisions of the alteral septum, along the border of the nucleus accumbens, in the ventral tip of the horizontal limb of the diagonal band of Broca as well as in the anterior hippocampal rudiment, infralimbic area and several other structures of the basal forebrain (e.g., nucleus accumbens, olfactory tubercle and substantia innominata). Cell bodies containing immunoreactivity for neurotensin were situated in the intermediate and ventral subdivisions of the lateral septum, the medial septal nucleus, the diagonal band of Broca, the rostro-medial continuation of the substantia innominata and the olfactory tubercle. In untreated rats, somatostatin positive processes formed terminal plexuses in the medial septal nucleus and along an area close to the ventricular wall of the lateral ceptal nucleus. Other septal nuclei, such as the diagonal band of Broca contained a sparse innervation by somatostatin positive fibers. In contrast, the nucleus accumbens olfactory tubercle, and the substantia innominata contained a rich innervation by somatostatin positive axons and terminals. Within these structures the density of SOM positive processes show great variations with patches of densely packed terminals separated by areas of sparser or no innervation. The neurotensin positive terminals were situated predominantly within the intermediate part of the lateral septum and the medial septal nucleus. Both of these regions contained numerous pericellular baskets of neurotensin positive terminals around septal neurons. In addition to the septal innervation, several of the basal forebrain structures were rich in neurotensin positive processes with the densest innervation found in the nucleus accumbens and substantia innominata. Like the SOM-immunoreactivity distinct islands of dense neurotensin innervation separated by less or no innervation occur throughout the basal forebrain. Taken together, these findings suggest that somatostatin and neurotensin occur in separate neuronal populations and that each may influence important physiological functions within the individual septal nuclei.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-0568
    Keywords: Light, electron microscopy ; Entorhinal cortex ; Retrohippocampal region ; Pericellular plexus ; Axosomatic synapses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: 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 μm in diameter) to large (22 μm 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.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 167 (1983), S. 53-65 
    ISSN: 1432-0568
    Keywords: GAD immunohistochemistry ; Septum ; Islands of Calleja
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The distribution of gamma aminobutyric acid (GABA)-containing neurons and nerve terminals was examined in the rat septal area by using specific antibodies to the enzyme glutamic acid decarboxylase (GAD) in combination with the avidin-biotin immunoperoxidase method. Whereas only a few GAD positive neurons were present in the septum of normal rats, the septal area of rats treated with colchicine, an inhibitor of fast axonal transport, showed numerous GAD-immunoreactive neurons. These neurons were evenly filled with GAD-immunoreactive material throughout the cytoplasm of the soma and proximal parts of the dendrites. Although GAD-positive neurons were present in most parts of the septal area, their density differed greatly in the different septal subnuclei. Both the diagonal band of Broca (vertical and horizontal parts) and the lateral septum were rich in GAD positive cell bodies, whereas the medial septal nucleus and the intermediate parts of the lateral septum contained relatively few. Within the lateral septum itself a larger number of labeled cell bodies was present in its ventral subdivision. The anterior hippocampal rudiment (taenia tecta) contained numerous GAD-positive neurons, while the septal component of the island of Calleja (insula magna) was devoid of them. GAD-immunopositive neurons found within the septum ranged from small (15 μm) to large (30–35 μm). They were round or multipolar in the diagonal band, medium-sized multipolar in the lateral septum, and pyramidal, round or fusiform in the anterior hippocampal rudiment. GAD-immunoreactive nerve terminals are present in most subdivisions of the septal nuclei, with the exception of myelinated fiber tracts, and throughout all rostrocaudal levels of the septum. However, the density of the innervation is not the same within all individual nuclei. The lateral septum (dorsal and ventral parts) contained high density innervation but the diagonal band of Broca had a lower density of GAD-positive terminals. The lateral border of the islands of Calleja was rich in thick GAD-positive processes that appeared to be continuous with GAD-immunoreactive processes of the substantia inominata. The inner portion of the molecular layer adjacent to the granule cells of the anterior hippocampal rudiment contained a rich GAD-positive terminal field.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: In this study, we describe the cloning and characterization of a soluble form of kynurenine aminotransferase (KAT, EC 2.6.1.7) present in rat brain. Soluble KAT was purified from rat kidney and the amino acid sequences of four tryptic peptides determined. These peptides were found to belong to the amino acid sequence reported for rat kidney soluble cysteine conjugate β-lyase, indicating that rat kidney KAT and β-lyase represent the same molecular entity. Oligonucleotide probes derived from the β-lyase cDNA were then used as primers for PCR of reverse-transcribed rat brain poly(A)+ RNA. After subcloning of the resulting PCR fragment and sequencing of the isolated rat brain clone, its oligonucleotide sequence was found to be identical to that reported for the β-lyase cDNA. Further evidence that the isolated rat brain clone encoded for KAT was obtained by transfecting HEK-293 cells with a construct containing the coding sequence for the enzyme. The transfected cells exhibited KAT activity and, in the presence of 2 mM pyruvate and 2-oxoglutarate, the Km values for l-kynurenine were 1.2 mM and 86.3 µM, respectively. Northern blot analysis of rat kidney, liver, and brain RNA revealed a single species of KAT/β-lyase mRNA of ∼2.1 kb.
    Type of Medium: Electronic Resource
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