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V Chan-Palay

Publications and source records attributed to V Chan-Palay.

At least 37 records · Page 2Linked to original sources

Distribution of neurotensin receptors in the primate hippocampal region: a quantitative autoradiographic study in the monkey and the postmortem human brain.

The distribution of [3H]neurotensin ([3H]NT) binding sites in the monkey and the postmortem human brain was studied by using quantitative in vitro receptor autoradiography. Biochemical experiments carried out on tissue sections of the monkey hippocampus showed that the binding of [3H]NT was saturable, reversible and of high specificity. The hippocampal [3H]NT binding was displaced by fragment NT 8-13 but not fragment NT 1-8 of the peptide. The anatomical analysis showed a highly heterogeneous distribution of [3H]NT binding sites within both the monkey and the human hippocampal region. In both species the highest density of [3H]NT binding sites was found in the presubiculum (rank order of binding density: layer 2 greater than 6 greater than 1 greater than 3, 4, 5 in both monkey and man) and the entorhinal area (monkey: layer 4 greater than 6 greater than 5 greater than 1 greater than 2 greater than 3; human: layer 1 = 2 greater than 5 greater than 3). The subiculum and Ammon's horn were relatively poor in [3H]NT binding sites in both species. In the area dentata the highest density of [3H]NT binding sites was found in the hilar region.

Animals↗

Action and location of neuropeptide tyrosine (Y) on hippocampal neurons of the rat in slice preparations.

The action of bath applied NPY (1-1,000 nM) was investigated on hippocampal slices of the rat with extra- and intracellular recording. Neuropeptide Y (NPY) at 10-1,000 nM caused a concentration-dependent, long-lasting reduction of excitatory postsynaptic potentials (EPSPs) in the hippocampal subfield CA1 and the area dentata, and an even stronger reduction of population spikes. Paired pulse experiments with low intensity, stimulation-evoked PSPs showed a marked increase in facilitation in the presence of NPY, indicating a presynaptic action. Spontaneous burst firing of CA1 pyramidal cells in low calcium, high magnesium medium was reduced, indicating a partially postsynaptic inhibitory action of NPY on their dendrites. Intracellular recording from CA1 somata during NPY administration revealed a reduction of the amplitudes of excitatory-inhibitory postsynaptic potential (EPSP-IPSP) sequences in the absence of changes in membrane potential and conductance. Accommodation of firing during long depolarizing pulses and afterhyperpolarizations were unchanged. The innervation pattern of NPY immunoreactive fibers in the same regions was studied in slices adjacent to the ones used for electrophysiology by using antisera against NPY and light and electron microscopy. There is a dense innervation of CA1 by NPY-immunoreactive axons and terminals, particularly in the stratum moleculare. NPY-immunoreactive neurons are present in the stratum oriens and pyramidale. The NPY labeled axons of the stratum moleculare participate in numerous synaptic contacts with the smaller dendritic elements in this layer, many of which belong to pyramidal neurons. These observations provide evidence for a dendritic NPY-immunoreactive innervation of CA1 neurons, which is in keeping with the electrophysiological effects of NPY on pyramidal neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Distribution of serotonin-1A receptors in the monkey and the postmortem human hippocampal region. A quantitative autoradiographic study using the selective agonist [3H]8-OH-DPAT.

Serotonin-1A receptors were visualized and their anatomical distribution mapped within the monkey and the human hippocampus by using in vitro receptor autoradiography of the selective agonist [3H]8-OH-N,N-dipropyl-2-aminotetralin ([3H]8-OH-DPAT). The results show high densities of serotonin-1A receptors heterogeneously distributed in different subfields and layers of the monkey and the human hippocampal region. High densities are found in the molecular layer of area dentata, all layers of regio superior and the subiculum, parasubiculum, and layers 2, and 4 through 6 of the entorhinal area. In the human hippocampus, a distinct band of [3H]8-OH-DPAT binding sites is present in the subgranular zone of the area dentata. The similar anatomical distribution of [3H]8-OH-DPAT binding sites in the monkey and the human hippocampal region suggests that the serotonin-1A receptor is phylogenetically well preserved and indicates that this receptor may mediate action(s) of serotonin in the primate, including the human hippocampal region.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Galanin immunoreactivity in hypothalamic neurons: further evidence for multiple chemical messengers in the tuberomammillary nucleus.

By using a specific antibody against the 29 amino-acid peptide galanin (Gal) with light and electron microscopic immunocytochemistry, we have studied the distribution of Gal immunoreactivity in the posterior hypothalamic magnocellular neurons of the rat. In colchicine-treated rats, a large number of Gal-immunoreactive cells were identified within all subdivisions of the tuberomammillary nucleus. The majority of these cells are large multipolar or fusiform neurons, with long, sparsely branching dendrites. A small number project to the ventral hippocampus, as shown by experiments with the retrograde tracing of Fast Blue. Ultrastructural examination of the Gal-immunoreactive cells confirms their indentity as magnocellular neurons, with dense deposits of immunoreaction product, particularly in small ribosomal arrays and in large, dense-cored vesicles. Axosomatic synapses occur on these neurons. The axonal boutons synapse with asymmetric and symmetric junctions and contain small synaptic vesicles as well as numerous large, dense-cored vesicles, which display Gal immunoreactivity. Sequential staining of thin, alternate sections with antibodies against Gal and L-histidine decarboxylase (HDCase; EC 4.1.1.22) showed colocalization of Galand HDCase-immunoreactivities in a majority of tuberomammilary neurons. The finding of Gal immunoreactivity within histamine-producing neurons of the tuberomammillary nucleus adds to the multiplicity of potential neuronal messengers utilized by these cells.

Animals↗

Distribution of neurons and axons immunoreactive with antisera against neuropeptide Y in the normal human hippocampus.

The detailed distribution of neuropeptide tyrosine (neuropeptide Y; NPY) immunoreactive neurons and fibers is given for the normal human hippocampus. These neuronal elements are detected by a polyclonal antibody raised against the unconjugated peptide and controls were obtained by using liquid phase absorption immunocytochemistry. The description covers the distribution in the area dentata, the hippocampal subfields CA3 and CA1, the subicular complex, and the entorhinal area. Each region is distinct in its NPY content. In general, the hippocampal NPY immunoreactive neurons fall into distinct classes--large hilar neurons; cortical small bipolar or bitufted neurons; medium-sized multipolar neurons in the deep cortical layers; and finally the distinct, small bipolar NPY neurons of the white matter bundles. None of the NPY neurons are pyramidal; many are likely to be local circuit neurons, but some appear to have extrinsic connections. The NPY immunoreactive axonal innervation is dense throughout the hippocampus but shows distinct regional differences in the hippocampal subdivisions. The area dentata has hilar NPY immunoreactive neurons and radial varicose fibers scattered throughout without a clear laminar preference. Subfield CA3 is comparatively the weakest NPY-containing region and contrasts with CA1, which is well endowed with reactive neurons and a rich and unusual axonal innervation, with distinct laminar axonal specializations. The subicular complex is well endowed with cells and fibers and the parasubiculum consistently displays unusually heavy NPY innervation. The entorhinal area exhibits a rich cortical distribution pattern, like that previously described for the human cerebral cortex (Chan-Palay et al; J. Comp. Neurol. 238:382-390, '85a,b). The fimbria, alveus, and angular bundle have NPY neurons embedded within the white matter. Like the NPY immunoreactive innervation of the hippocampal regions of laboratory animals, the human NPY innervation seems to follow a common fundamental pattern with respect to cell locations, cell morphology, and axonal innervation. The difference, however, is the greater complexity and profusion of the NPY-immunoreactive axonal plexuses in the human hippocampus. This rich peptide network within the hippocampus with likely extrahippocampal interconnections raises questions concerning coexistence with other neuroactive substances, the functions of such substantial networks, and how they are altered in human neurological disease.

Aged↗

Distribution of altered hippocampal neurons and axons immunoreactive with antisera against neuropeptide Y in Alzheimer's-type dementia.

This paper provides detailed information on the distribution of neuropeptide tyrosine (neuropeptide Y; NPY) immunoreactive neurons and fibers in the hippocampal region of eight neuropathologically confirmed cases of Alzheimer's-type dementia (ATD) at postmortem. These neuronal networks are detected by a polyclonal antibody raised against the unconjugated peptide and controls were obtained by using liquid phase absorption immunocytochemistry. The description covers the subfields area dentata, CA3 and CA1, the subicular complex, and the entorhinal area. The hippocampal regions in which the NPY-i neuron networks are most severely affected are the hilus, CA1, the parasubiculum, and the entorhinal cortex. Less obvious reductions occurred in CA3, subiculum, and the presubiculum. Parallel semiquantitative estimates were made of the numbers of neuritic plaques and neurofibrillary tangles in the other hippocampus of the brains in every ATD case. The areas of heaviest pathological changes by these indices are CA1 and the entorhinal cortex. The subicular complex CA3 and the area dentata are less affected. These findings show that the areas with the most severe loss of NPY-i neurons and axons, CA1 and the entorhinal cortex, are the same as those areas most severely affected by the other indices of ATD. Thus NPY-i networks are involved in the ATD disease process. However, other NPY-i networks survive, in some subfields better than in others. The cumulative evidence suggests a population of hippocampal peptide neurons that are remarkably resistant in terminal neurological disease. These neurons have the capability to participate in the maintenance of minimal functioning circuits in target areas of the disease and as such hold significant links for our understanding of synaptic plasticity in disease.

Aged↗

Neuropeptide Y innervation of the hippocampal region in the rat and monkey brain.

Using antibodies to neuropeptide Y (NPY) in combination with immunohistochemical techniques we have studied the distribution of cell bodies and nerve terminals containing NPY immunoreactivity (-i) in the hippocampal region of rats and monkeys (cynomolgus). In colchicine-pretreated rats a large number of NPY-positive cells are present in all areas of the hippocampal region. The NPY-i cells range in size from small (diameter across soma: 10-15 micron) to large (approximately 20 micron). Most of the NPY-i cells are situated in the hilus, in the subgranular zone of the area dentata, and in the stratum oriens of Ammon's horn. A majority of these are polymorphic cells but cells of different morphology are present in these layers as well. These include small spheroid cells and dentate pyramidal basket cells that are distinct from the polymorphic cells in the subgranular zone. The subicular complex (e.g., the subiculum, pre-, and parasubiculum) and the entorhinal area contain fewer NPY-i cells than the rest of the hippocampal region. In the dorsal parts of the pre- and parasubiculum numerous small cells are scattered throughout all layers, while in the entorhinal area the NPY-stained cells are situated primarily in the deep layers (V and VI). In the ventral part of the lateral entorhinal area large multipolar and bitufted cells are found in layers II-VI. In the untreated monkey brain NPY-positive cells are found in the hilus of the area dentata and in the deep (IV through VI) layers of both the medial and lateral entorhinal area. Fewer NPY-stained cells are present in the subicular complex and in the entorhinal area. In the monkey as well as in the rat, NPY-stained cells are present in the angular bundle and in the alveus. A dense network of NPY-i fibers innervates the entire hippocampal region in both the rat and the monkey. The hippocampal NPY-i preterminal processes are present primarily in stratum moleculare of Ammon's horn and in the outer one-third of this layer in the area dentata. The NPY-positive innervation of the dentate molecular layer is far more prominent in the monkey than in the rat brain. Numerous NPY-stained fibers are scattered in other areas as well. In all retrohippocampal structures, and in particular the entorhinal area, the NPY-i fibers form a massive network that innervates all layers to about the same extent, with the exception of the molecular layer, which is more densely innervated than the other layers.

Animals↗

Immunocytochemistry of human brain tissue with a polyclonal antiserum against neuropeptide Y.

NPY-containing neuronal structures in the cerebral cortex of surgical tissue samples were compared to those in postmortem material by immunocytochemical methods. However, the quality of preservation of individual neurons and axonal and dendritic plexuses in the neuropil is unusually fine in the surgical specimens. This result is most likely attributable to the excellent fixation that can be regularly achieved by rapid and careful handling of tissue during and after surgical removal. The tissue is suitable for both light and electron microscopy, and the superior preservation also leads to intense, reliable antibody reactions. Postmortem tissue samples can provide good specimens for immunocytochemistry when properly handled as previously described. However the postmortem delays prior to fixation disrupt neuronal integrity in the immunostained structures. Nevertheless, postmortem material from carefully studied subjects of neurological diseases compared with age matched controls can provide valuable information.

Adolescent↗

Cytology and distribution in normal human cerebral cortex of neurons immunoreactive with antisera against neuropeptide Y.

The frontal, parietal, and temporal cortices in normal human brains (Brodmann areas 10, 7a, 7b, and 21) are well endowed with numerous neurons, identifiable by immunoreactivity with antisera against the 36-amino acid brain peptide neuropeptide Y (NPY). These neurons with rare exception are small, intracortical, nonspiny neurons, 12-20 microns in somatic size, with long slender dendrites and exuberant axon plexuses exhibiting finely beaded varicosities. The cells are rarest in layers I and II, are found with frequency in the lower cortical layers (IV-VI) and in significant numbers in the subcortical white matter. Within the cortex the axonal plexuses of these peptide neurons rise straight up into the upper cortical layers or descend deep into the white matter. In layers I and II, numerous other lengthy axons, some possibly of extracortical afferent origin, run along the pial surface at right angles to the axial ones running perpendicular to the cortex. This endowment of peptide neurons and their processes forms a rich network in the cerebral cortex, relating with one another in complex fashion within palisades of terminals as well as with the other cortical neurons not labeled by these methods. It remains to be shown what functions these NPY neurons have individually and in their remarkable networks, and how they are altered in neurological disease.

Aged↗

Cortical neurons immunoreactive with antisera against neuropeptide Y are altered in Alzheimer's-type dementia.

Neurons identified by their immunoreactivity with antisera against neuropeptide Y (NPY) were studied in three selected areas of the cerebral cortex in brains from controls and in senile dementia of the Alzheimer type (ATD). Changes were more profound in temporal cortex than in parietal cortex, and more severe in parietal cortex than in frontal cortex, paralleling the severity of neuritic plaque formation and incidence of neurofibrillary tangles in these regions. NPY-i neurons became distorted, with enlarged misshapen cell somata and reduced, thickened, and gnarled dendrites. There was a sharp reduction in the extensiveness and delicacy of the axonal plexus; the reorganized axons were haphazard compared to the normal symmetry of these fibers. Besides the alteration in form and sizes, there were also appreciably fewer cells. Nevertheless, the NPY population is not eliminated. Double-label studies of NPY-i and thioflavin indicate that NPY-i fibers can participate in neuritic plaque formation although not all neuritic plaques contained NPY-i axons and not all NPY-i axons were associated with plaques. The surviving NPY cells were evident in all cortices examined, thus giving rise to the speculation that these peptide neurons may have unusual survival and reorganization potential even in terminal neurological disease.

Aged↗

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

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-immunoreactive(-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 micron in diameter) to large (22 micron 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.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Serotonin neurons on the ventral brain surface.

Serotonin neurons and fibers on the subpial surface of the ventral medulla oblongata in the rat are described by immunohistochemistry and autoradiography. The neurons are concentrated in the area encompassed by the origins of the abducens, hypoglossal, glossopharyngeal, and vagus nerves. The highest number of serotonin surface neurons appears along the median medullary fissure or basilar sulcus, where they may represent the most ventral extensions of the raphe pallidus group. As these cells lie on the surface of the brain, they could be directly affected by alterations in the chemical composition of the cerebrospinal fluid and, depending on their connections, could influence important medullary functions.

Animals↗

Distribution of tyrosine-hydroxylase-immunoreactive neurons in the hypothalamus of rats.

The distribution and morphology of cells containing tyrosine hydroxylase (TH) immunoreactivity in the hypothalamus of rats were studied by using a modified immunoperoxidase technique. The TH cell system is more complexly organized than was previously thought. On the basis of their clustering patterns, hypothalamic TH neurons could be subdivided into two groups: dorsal and ventral. The ventral group consists of a prominent aggregate of cells located in the caudal part of the arcuate nucleus. From here, cells extend around the caudal part of the ventromedial and dorsomedial nuclei and the base of the diencephalon. Tyrosine hydroxylase-positive cells are present throughout the arcuate nucleus, except in its ventromedial part. Anteriorly, immunoreactive cells appear in the suprachiasmatic and supraoptic nuclei, in the retrochiasmatic area, and in the ventral part of the anterior hypothalamic nucleus. The dorsal group has its main concentration of cells in the medial part of the zona incerta, from which two clusters of cells, one medial and one lateral, extend rostralward. The medial group comprises cells in the medial part of the dorsomedial, paraventricular, and anterior hypothalamic nuclei. These cells adjoin the periventricular cells. The lateral group of cells emanating from the zona incerta occupies the lateral part of the dorsomedial and anterior hypothalamic nuclei and the dorsal hypothalamic area. The dorsal and ventral TH cell groups are in continuity medially in the periventricular layer, and laterally through the cells that surround the ventromedial nucleus. Although the cells vary widely in size, shape, and dendritic arborization pattern, there are two main cell types. Small (21 X 11 microns), round to fusiform cells, with two or three dendrites arborizing simply, were frequently seen in the arcuate, suprachiasmatic, periventricular, supramammillary nuclei and at the borders of the ventromedial nucleus. The other cell type is larger (40 X 15 microns) and multipolar, with three to five frequently branching dendrites. The dendritic field is large and the cells are intensely TH-immunoreactive. Although the larger cells occur occasionally in every hypothalamic nucleus, their principal locations are in the dorsal parts of the dorsomedial, posterior hypothalamic nuclei and the dorsal and lateral parts of the zona incerta, and in the areas dorsal and medial to the mammillothalamic tract at caudal hypothalamic levels. In this paper we give a detailed description of TH-immunoreactive fibers and terminals in the hypothalamus and a comparison with previous studies of catecholamine cells in the hypothalamus.

Animals↗

Septal neurons containing glutamic acid decarboxylase immunoreactivity project to the hippocampal region in the rat brain.

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 septo-hippocampal projection may utilize gamma-aminobutyric acid as a neurotransmitter.

Animals↗

Somatostatin and vasoactive intestinal polypeptide-like immunoreactive cells and terminals in the retrohippocampal region of the rat brain.

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 microns) 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 medical 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.

Animals↗

Distribution of gamma aminobutyric acid containing neurons and terminals in the septal area. An immunohistochemical study using antibodies to glutamic acid decarboxylase in the rat brain.

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-immuno-positive neurons found within the septum ranged from small (15 microns) to large (30-35 micron). 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.

Animals↗

Gamma-aminobutyric acid interneurons in the rat hippocampal region studied by retrograde transport of glutamic acid decarboxylase antibody after in vivo injections.

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 hippocampal 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.

Animals↗

Immunocytochemical and autoradiographic methods to demonstrate the coexistence of neuroactive substance: cerebellar Purkinje cells have glutamic acid decarboxylase, cysteine sulfinic acid decarboxylase, and motilin immunoreactivity.

Evidence is presented from immunocytochemical experiments that Purkinje cells in the mammalian cerebellum of a number of species contain gamma aminobutyric acid (GABA), taurine and the peptide motilin. Purkinje neurons show immunoreactivity to antibodies against glutamic acid decarboxylase (GAD), the synthetic enzyme for GABA, cysteine sulfinic acid decarboxylase (CSADC ase), the synthetic enzyme for taurine and motilin. Single neurons have either unique localization for GAD or motilin or CSADC ase or two substances in combination providing evidence for coexistence of these neuroactive agents. Functional studies with iontophoresis of GABA, taurine, and motilin into rabbit lateral vestibular nucleus elicited strong depressant effect on neuronal firing. The interactions of motilin and GABA were additive and inhibitory. Thus, Purkinje neurons of the cerebellum use multiple chemical messengers in their interactions. These chemical messengers coexist in single neurons. Their effects as far as our studies indicate are inhibitory and consistent with previous demonstrations of the physiological actions of Purkinje cells.

Animals↗