Biomedical subjects

C Leranth

Publications and source records attributed to C Leranth.

At least 37 records · Page 2Linked to original sources

PubMed1998

Dual (excitatory and inhibitory) calretinin innervation of AMPA receptor-containing neurons in the rat lateral septum.

A recent study demonstrated both an extrinsic and an intrinsic calretinin (CR) innervation of the rat septal complex and that a population of the extrinsic calretinin fibers is aspartate/glutamate-containing. The aim of this study was to determine which types (GluR1, GluR2/3, or both) of AMPA receptor-containing lateral septal area neurons are innervated by extrinsic and intrinsic CR neurons and whether the intrinsic CR cells are GABAergic. Light- and electron-microscopic single immunostaining for CR, GluR1, and GluR2/3, as well as light- and electron-microscopic-double immunostaining experiments for CR plus GluR1 and CR plus GluR2/3 were performed in the lateral septal area. Furthermore, the "mirror" colocalization technique was employed on consecutive vibratome sections of the septal complex to investigate whether the intrinsic septal CR neurons are GABAergic. The results are summarized as follows: (1) both GluR1- and GluR2/3-immunoreactive neurons are innervated by CR-containing fibers; (2) the majority of these synapses, observed mainly on the soma and, to a lesser extent, on proximal dendrites of AMPA receptor-containing neurons, represent asymmetric synaptic membrane specializations; (3) a minority of CR-containing axon terminals associated with both GluR1- and GluR2/3-immunoreactive neurons form symmetric contacts, predominantly on their soma; and (4) 93% of the lateral septal area CR cells are GABAergic. These observations indicate that both GluR1- and GluR2/3-containing lateral septal area neurons receive a dual intrinsic and extrinsic CR innervation. The former (intrinsic) CR boutons are GABAergic, while the latter form asymmetric synaptic contacts, are excitatory, and probably originate in the supramammillary area, since previous work has demonstrated that a population of supramammillo-septal fibers contain aspartate and/or glutamate.

Animals↗

PubMed1998

Efferent synaptic connections of dopaminergic neurons grafted into the caudate nucleus of experimentally induced parkinsonian monkeys are different from those of control animals.

This study investigated the question of whether grafted dopamine cells in the striatum of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated monkeys form synapses and, if they do, whether their postsynaptic targets were the same as those in control monkeys or in previous studies in rats. Electron-microscopic single immunostaining was performed for tyrosine hydroxylase on vibratome sections prepared from the head of the caudate nucleus of controls and MPTP-treated African green monkeys (Cercopithecus aethiops sabaeus) that received a graft. Furthermore, correlated light- and electron-microscopic double immunostaining was carried out for tyrosine hydroxylase and calbindin in the same brain area of MPTP-treated plus grafted animals. In control monkeys, the majority (97%) of dopamine boutons terminate on spines that were also synaptic targets of immunonegative boutons forming asymmetric synaptic contacts: synaptic triads. In MPTP-treated, grafted animals, the majority of transplanted dopamine cells terminate on dendritic shafts (67%) and somata (32%), and only a few (1.33%) form axospine synapses. The results of the double immunostaining experiments indicated that these newly formed axosomatic and axodendritic synapses are associated with calbindin-immunoreactive, medium-sized, spiny striatonigral projection neurons. These observations indicate that: (1) dopamine from transplanted embryonic tissue acts via synaptic contacts on host neurons; (2) the primary synaptic targets of transplanted dopamine cells are not spines but dendrites and somata of host neurons; (3) these target neurons are the same as in control animals; and (4) comparing these observations with results of control and grafted rats, there are major species differences between rats and monkeys in the dopamine innervation of both control and transplanted animals.

Animals↗

PubMed1998

A novel method for concurrent visualization of immunostain under light and electron microscopy in pancreatic islets.

We developed a simple method employing the use of flat-embedding techniques on thick frozen sections which allows correlation of light and electron microscopic immunohistochemistry. This method has been particularly useful in visualization of pancreas sections, an adaptation especially important because this tissue is not amenable to conventional vibratome sectioning. In this study we demonstrate the use of this technique to examine the same tissue section at the light and the electron microscopic level while maintaining morphology.

Animals↗

PubMed1997

Substance P innervation of the rat hippocampal formation.

Light and electron microscopic substance P (SP) immunostaining was performed on hippocampal sections of colchicine-pretreated, control, untreated fimbria-fornix-transected (5 days), as well as perforant path-stimulated Sprague-Dawley rats fixed in 5% acrolein. Numerous SP-immunoreactive neurons could be observed in the stratum oriens of the Ammon's horn and subiculum, fewer were seen in the dentate hilar area and stratum radiatum of CA2 and CA3, and even fewer were seen at the border between the CA1 strata radiatum and the lacunosum moleculare of CA1 subfield. A higher dose of colchicine resulted in SP immunoreactivity in a large population of granule cells and mossy axon terminals. The entire CA2 region, the stratum oriens of CA1, CA3, and the subiculum were densely innervated by SP-containing axon terminals. A homogeneous SP innervation was found in the stratum radiatum of CA1. Only a few SP fibers were seen adjacent to the granule cell layer. Symmetric axosomatic contacts were seen between SP-containing boutons and somata in the stratum oriens of the Ammon's horn. However, throughout the hippocampal formation, the majority of SP-containing axons formed axodendritic symmetric synapses. A dense population of SP-immunoreactive boutons that formed axodendritic asymmetric synapses was observed in the strata oriens and radiatum of the CA3a and CA2 regions, and a few were found in the supragranular and subgranular layers of the dentate gyrus. Fimbria-fornix transection resulted in a marked loss of SP fibers in the strata oriens, pyramidale, and radiatum of the CA3a and CA2 subfields. In perforant pathway-stimulated animals, a population of granule cells and a large number of mossy axon terminals were immunoreactive for SP. These observations suggest two sources of SP innervation to the hippocampal formation: one arising from intrinsic sources (interneurons and granule cells) and one arising from extrinsic sources, most likely the supramammillary region.

Animals↗

PubMed1997

Heterogeneity in the neuropeptide Y-containing neurons of the rat arcuate nucleus: GABAergic and non-GABAergic subpopulations.

Neuropeptide Y, produced in the arcuate nucleus of the hypothalamus, plays a key role in the central regulation of anterior pituitary and appetitive functions. The pleiotropic nature of neuropeptide Y in these mechanisms indicates the existence of heterogeneity in the hypothalamic neuronal population producing neuropeptide Y. In this study, we report the coexistence of neuropeptide Y and the amino acid transmitter, gamma-aminobutyric acid (GABA), in neuronal perikarya of the arcuate nucleus. Fluorescent double immunolabeling for neuropeptide Y and glutamic acid decarboxylase was carried out on vibratome sections collected through the hypothalamic arcuate nuclei of animals that were pretreated with colchicine. It was found that about one third of the neuropeptide Y-producing arcuate nucleus perikarya co-expressed glutamic acid decarboxylase. This population of neuropeptide Y-containing GABAergic neurons were distributed longitudinally within the arcuate nucleus located predominantly in its dorsomedial aspects. These results show that there are at least two distinct populations of neuropeptide Y-producing neurons in the arcuate nucleus: a subset of neuropeptide Y and GABA-co-producing neurons located in the dorsomedial arcuate nucleus and a subset of non-GABAergic neuropeptide Y cells located in the ventral arcuate nucleus. This heterogeneity in the neuropeptide Y-producing perikarya of the hypothalamus may help explain adverse neuroendocrine and behavioral effects of arcuate nucleus neuropeptide Y.

Animals↗

PubMed1997

Hypothalamo-septal enkephalinergic fibers terminate on AMPA receptor-containing neurons in the rat lateral septal area.

A large number of septal neurons express alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA)-type excitatory glutamate receptors. It has been demonstrated that in the mediolateral part of the rat lateral septum, calbindin-containing neurons are heavily innervated by hypothalamic, enkephalinergic fibers forming exclusively asymmetric synaptic contacts. This connection was suggested to be excitatory. In order to further elucidate this hypothesis, the aim of the present study was to determine whether these enkephalinoceptive neurons express GluR1 and GluR2/3 AMPA receptor subunits. Correlated light and electron microscopic analysis, using single immunostaining for GluR1 and GluR2/3, and double immunostaining for Leu-enkephalin and GluR1 or GluR2/3, was performed on vibratome sections of the rat lateral septal area. The studies revealed that while GluR1 is mainly associated with dendritic and somatic spines, GluR2/3 is mostly present in the perisomatic area. Leu-enkephalin boutons establish asymmetric synaptic contacts at the level of the soma and initial dendrites of both of these cells. A semiquantitative analysis showed that these enkephalin-targeted cells represent 50% of the total number of both GluR1 and GluR2/3-containing lateral septal neurons. These results suggest that: (1) AMPA receptor-expressing neurons appear to be the exclusive recipient of hypothalamic Leu-enkephalin boutons; (2) these enkephalinoceptive neurons contain both GluR1 and GluR2/3 AMPA receptor subunits; however, (3) only the GluR2/3 subtype, located in the perisomatic area, may be associated with Leu-enkephalin-containing inputs.

Animals↗

PubMed1997

Parvalbumin-containing cells of the angular portion of the vertical limb terminate on calbindin-immunoreactive neurons located at the border between the lateral and medial septum of the rat.

In the septal complex, both parvalbumin and calbindin neurons cocontain GABA. In the same area, a large number of GABA-GABA synaptic connections can be observed. In order to further characterize their neurochemical nature, as well as the extrinsic and/or intrinsic origin of these GABA terminals, the following experiments were performed: (1) correlated light- and electron-microscopic double immunostaining for calbindin and parvalbumin on septal sections of control rats: (2) light microscopic parvalbumin immunostaining of septal sections after surgical isolation (5 days) of the septum from its telencephalic or (3) hypothalamic afferents; and (4) parvalbumin immunostaining of sections prepared from the entire brain 2 days following horseradish peroxidase injection into the border between the lateral and medial septum. The results demonstrated that: (1) in a well-circumscribed, vertically longitudinal area located between the lateral and medial septum, 0.1-0.6 mm anterior to the bregma, a group of calbindin-containing, nonsomatospiny neurons are surrounded by parvalbumin-immunoreactive baskets; (2) these basket-forming axon terminals establish symmetric synaptic contacts with their targets; and (3) their cells of origin are not in the medial septum, but in the angular portion of the vertical limb. These observations indicate that a portion of the septal complex GABA-GABA synaptic connections represent functional interaction between two different types of GABAergic neurons. The presynaptic GABAergic neurons contain parvalbumin, and the postsynaptic GABAergic cells are immunoreactive for calbindin. Furthermore, a population of the medial septum/diagonal band parvalbumin neurons project only to the hippocampus, while others, which may also send axons to the hippocampus, terminate on lateral septum calbindin cells as well.

Animals↗

PubMed1997

Neurochemical characterization of AMPA receptor-containing neurons in the mediolateral septal area of the rat.

The lateral septum receives a massive innervation by excitatory amino acid-containing limbic cortical and hypothalamic afferents, and previous studies have described a wide distribution of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor-containing neurons in this area. The aim of this study was to determine whether different subtypes of AMPA receptors are expressed in the same neurons. Furthermore, considering the fact that a population of lateral septal cells, the "somatospiny neurons," are GABAergic calbindin-containing cells, the coexistence of each subtype of AMPA receptor with calbindin was also investigated. Colocalization experiments were performed on adjacent vibratome sections of the lateral septal area for GluR1 and GluR2/3 AMPA-receptor subunits, GluR1 and calbindin, GluR2/3 and calbindin, as well as GluR1 plus calbindin and GluR2/3 plus calbindin, using the "mirror" colocalization technique. The results are summarized as follows: (1) GluR1 is present in the soma and most intensively expressed in dendrites and somatic and dendritic spines; while GluR2/3 is associated with the soma and proximal dendrites of the neurons. (2) Forty-one percent of the AMPA receptor-containing neurons cocontain GluR1 and GluR2/3. (3) Thirty-eight percent of GluR1- and 28% of GluR2/3-labeled cells express calbindin. (4) Sixty-two percent of the calbindin-immunoreactive neurons contain GluR1 and 51% of them express GluR2/3. (5) Half of the neurons expressing both GluR1 and GluR2/3 also contain calbindin. (6) The distribution of GluR1 plus GluR2/3-containing, GluR1 plus calbindin-containing, and GluR2/3 plus calbindin-containing neurons in the lateral septum are homogeneous. This study indicates the existence of multiple populations of AMPA receptor- and calbindin-containing neurons in the lateral septal area.

Animals↗

PubMed1997

Distinct substance P- and calretinin-containing projections from the supramammillary area to the hippocampus in rats; a species difference between rats and monkeys.

Our recent studies showed the co-existence of substance P and calretinin in the supramammillo-hippocampal pathway of monkeys, as well as species differences in the synaptic targets of extrinsic substance P fibers in the hippocampi of monkeys and rats. Experiments used: (1) single and multiple stereotaxic injection of wheat germ agglutinin-conjugated HRP into the hippocampus and immunostaining for substance P in the supramammillary area; (2) colocalization of substance P and calretinin in supramammillary area cells; and (3) colocalization of these two neurochemicals in retrogradely labeled supramammillary projective cells of both male and female rats. These demonstrated: (a) many calretinin- and fewer substance P-immunoreactive neurons retrogradely labeled in the ipsilateral supramammillary area; (b) approximately 74% of all substance P cells contain calretinin and 9% of the calretinin neurons co-contain substance P; and, most importantly (c) none of the retrogradely labeled supramammillary cells colocalize calretinin and substance P. These results indicate the presence of two distinct supramammillo-hippocampal projections in the rat, one that contains substance P and the other calretinin. The latter innervates the same areas as those in the monkey, and the former terminates only in the CA2 hippocampal subfield.

Animals↗

PubMed1997

Dual role of substance P/GABA axons in cortical neurotransmission: synaptic triads on pyramidal cell spines and basket-like innervation of layer II-III calbindin interneurons in primate prefrontal cortex.

In spite of accumulating evidence on the potent neuromodulatory, neuroprotective, trophic and memory-enhancing effects of the neuropeptide substance P (SP) in the cerebral cortex, the excitatory or inhibitory nature of the cortical SP innervation remains unclear and the postsynaptic targets of SP fibers are not defined. To obtain further insight into these issues, we have examined SP-containing axons and their postsynaptic targets in the prefrontal cortex of adult monkeys with single- and double label immunocytochemistry combined with light and correlated electron microscopy. SP fibers in the primate prefrontal cortex, unlike those in the rat cortex, preferentially innervate cortical layers I, II and upper layer III. Our results demonstrate for the first time that all SP-immunoreactive boutons in all cortical layers contain GABA. Of the entire sample of SP boutons, 53% synapse on dendritic shafts, 39% on dendritic spines and 8% on cell bodies. Another new finding is that synapse-forming SP boutons, in addition to their known innervation of pyramidal cells, form pericellular baskets around interneurons in layers II and upper III, a subpopulation of which contains calbindin D28k. Finally, the study also revealed that SP boutons frequently participate in 'synaptic triads' with spines which receive another (asymmetric, putatively excitatory amino acid-utilizing) synapse. Our findings indicate that SP/GABA axons in the primate prefrontal cortex modulate excitatory amino acid-mediated neurotransmission and control feed-forward disinhibitory GABAergic circuits in supragranular cortical layers.

Animals↗

PubMed1996

A population of supramammillary area calretinin neurons terminating on medial septal area cholinergic and lateral septal area calbindin-containing cells are aspartate/glutamatergic.

The excitatory amino acid, aspartate/glutamate content of septal complex calretinin (CR)-, choline acetyltransferase plus substance P-, and Leu-enkephalin (Leu-enk)-containing extrinsic afferents was examined. Experiments were carried out using the transmitter-specific [3H]-D-aspartate retrograde tracer technique in combination with immunostaining for CR, choline acetyltransferase, and Leu-enk. The extrinsic and intrinsic CR innervation of the same brain areas were elucidated on control rats and on animals in which the septum was surgically separated from its ventral afferents. Correlated light and electron microscopic double-immunostaining experiments were used to determine the synaptic connections between CR axon terminals and lateral septal area calbindin (CB)- and medial septal area choline acetyltransferase-immunoreactive neurons. Furthermore, to determine the synaptic power of supramammilloseptal aspartate/glutamatergic neurons on the septal complex, semiquantitative analyses were performed in the supramammillary area on retrogradely (1) [3H]-D-aspartate-radiolabeled and (2) HRP-labeled material. The results demonstrated that a population of the extrinsic CR axons originating in the supramammillary area are aspartate/glutamatergic. These fibers forming asymmetric synaptic contacts terminate on both CB and cholinergic neurons. Intraseptal CR neurons, which establish symmetric synapses, innervate only lateral septal area neurons, including the CB-containing cells. These observations, together with other published data, raise the possibility of a hippocampus-lateral septal (GABAergic CB-containing neurons)-supramammillary area (aspartate/glutamatergic cells)-medial septal (cholinergic neurons)-hippocampus signal loop, which might be involved in the generation and regulation of hippocampal theta rhythm activity.

Animals↗

PubMed1996

GABAergic neurons in the rat dentate gyrus are innervated by subcortical calretinin-containing afferents.

Fibers of supramammillary origin establish putatively excitatory asymmetric synaptic connections with dentate granule cells. The present study was designed to determine whether hippocampal gamma-aminobutyric acid (GABA)-ergic nonprincipal cells are also targets of these calretinin (CR)-containing subcortical afferents. Light and electron microscopic double immunostaining for CR and parvalbumin (PA) or calbindin (CB) were performed in the rat dentate gyrus ipsilateral and contralateral to a unilateral fimbria-fornix transection. GABA-postembedding immunostaining was performed on ultrathin sections of this double-labeled material. Contralateral to the transection, CR-immunoreactive fibers formed multiple large boutons in the inner molecular layer. These fibers also impinged on PA-containing basket cells located adjacent to the granular layer and on CB-immunoreactive hilar neurons. Ipsilateral to the transection, CR-containing fibers in the inner molecular layer and boutons impinging on PA-containing or CB-immunoreactive neurons were absent. Parent cell bodies of extrinsic CR-containing afferents were traced using wheat germ agglutinin-conjugated horseradish peroxidase. Additional CR immunostaining of the subcortical region unveiled retrogradely labeled neurons that were also immunostained for CR only in the supramammillary area and the nucleus reuniens. The latter projection, however, terminates in CA1 and not in the dentate gyrus. Subcortical afferents impinging on dentate nonprincipal cells formed exclusively asymmetric synapses. Postembedding immunostaining demonstrated that CB-containing cells contain GABA, whereas CR-positive axon terminals forming asymmetric synapses are devoid of this labeling. These data indicate that dentate inhibitory neurons receive a putative excitatory input originating from the supramammillary nucleus. Thus, the supramamillo-hippocampal pathway may exert a powerful feed-forward inhibitory control of the signal flow in the rat dentate gyrus.

Animals↗

PubMed1996

Entorhinal cortical innervation of parvalbumin-containing neurons (Basket and Chandelier cells) in the rat Ammon's horn.

Physiological data suggest that in the CA1-CA3 hippocampal areas of rats, entorhinal cortical efferents directly influence the activity of interneurons, in addition to pyramidal cells. To verify this hypothesis, the following experiments were performed: 1) light microscopic double-immunostaining for parvalbumin and the anterograde tracer Phaseolus vulgaris-leucoagglutinin injected into the entorhinal cortex; 2) light and electron microscopic analysis of cleaved spectrin-immunostained (i.e., degenerating axons and boutons) hippocampal sections following entorhinal cortex lesion; and 3) an electron microscopic study of parvalbumin-immunostained hippocampal sections after entorhinal cortex lesion. The results demonstrate that in the stratum lacunosum-moleculare of the CA1 and CA3 regions, entorhinal cortical axons form asymmetric synaptic contacts on parvalbumin-containing dendritic shafts. In the stratum lacunosum-moleculare, parvalbumin-immunoreactive dendrites represent processes of GABAergic, inhibitory basket and chandelier cells; these interneurons innervate the perisomatic area and axon initial segments of pyramidal cells, respectively. A feed-forward activation of these neurons by the entorhinal input may explain the strong, short-latency inhibition of pyramidal cells.

Animals↗

PubMed1996

Potential neuronal mechanisms of estrogen actions in synaptogenesis and synaptic plasticity.

1. Studies conducted on the rat arcuate nucleus, an area involved in the development and control of LH and FSH secretion, have shown the existence of hormonally regulated developmental sex differences in synaptic patterns and estrogen-induced synaptic plasticity during adult life. Several questions raised by these findings are examined in this review: 2. The mechanisms of estrogen-regulated developmental synaptogenesis. These include the role of glycocalyx glycoproteins in neuronal membranes, neural cell adhesion molecules, and insulin-like growth factor I. 3. The relationship among circulating estrogen, gonadotropin levels, and hypothalamic synaptic plasticity. Recent evidence for the role of GABAergic and dopaminergic synaptic inputs and POMC projections from the arcuate nucleus to the GnRH cells is discussed. 4. The synaptologic basis of age-related failure of positive feedback. The role of the cumulative effect of repeated preovulatory synaptic retraction and reapplication cycles on sensescent constant estrus is analyzed.

Animals↗

PubMed1996

Distribution of substance P-immunoreactive neurons and fibers in the monkey hippocampal formation.

Substance P containing neurons was visualized by immunocytochemistry in the monkey hippocampus, subicular complex, and entorhinal cortex. Immunoreactive neurons were found solely in the hilar region of the dentate gyrus, and in strata oriens and pyramidale of Ammon's horn. In the subicular complex, immunoreactive neurons were located in those layers which were close to the alveus, whereas in the entorhinal cortex most of the substance P-positive neurons appeared in the second and third layers above the lamina dissecans. The majority of substance P-containing neurons were large multipolar cells, but small bipolar and multipolar cells also occurred in Ammon's horn, subiculum and entorhinal cortex. Dendrites of immunoreactive cells were smooth and displayed a few small, faintly stained spines which were hard to identify in the light microscopic preparations, but were visible with electron microscopy. Substance P-positive dendrites were exclusively found in the hilar region and never observed in the upper two-thirds of the molecular layer of the dentate gyrus. Moreover, immunoreactive dendrites rarely penetrated the stratum lacunosum-moleculare of Ammon's horn. In the electron microscopic preparations, somal and dendritic features of substance P-positive neurons were similar to those observed for GABAergic local circuit neurons. Axons of the substance P-immunoreactive local circuit neurons were thin and richly arborized in the upper two-thirds of the molecular layer of the dentate gyrus, in the stratum lacunosum-moleculare of Ammon's horn as well as in the subpial layers of the subicular complex and entorhinal cortex. Their terminals formed exclusively symmetric synapses with dendrites and spines. However, substance P-immunoreactive boutons were not found to make symmetric, axosomatic synapses on the granule cells of the dentate gyrus and very few were present on the pyramidal neurons of Ammon's horn, subicular complex, and entorhinal cortex. Hippocampal neurons, which were immunoreactive for substance P, also contained the neuropeptide somatostatin. However, not all of the somatostatin-containing neurons were substance P-immunoreactive. Thus, substance P-positive neurons are a subpopulation of somatostatin immunoreactive, GABAergic neurons. In conclusion, substance P-immunoreactive neurons are ideally suited for feed-back dendritic inhibition which may control the effectiveness of the main excitatory cortical input to the granule cells of the dentate gyrus and pyramidal neurons of the Ammon's horn.

Animals↗

PubMed1996

AMPA receptors in the rat and primate hippocampus: a possible absence of GluR2/3 subunits in most interneurons.

Amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors are assembled from the four subunits GluR1, 2, 3, 4 (or GluRA, B, C, D). AMPA channels that do not contain the GluR2 subunit are permeable to calcium. Recent studies indicate that excitotoxic as well as epileptic and ischemic cell damage may be mediated not only by N-methyl-Daspartate receptors, but also by AMPA receptors. The majority of interneurons in the hippocampus are resistant, but subsets of interneurons are consistently damaged in different disease states. Single immunolabeling using antibodies against AMPA receptor subunits, together with double immunolabeling for calcium-binding proteins (parvalbumin, calbindin and calretinin) and the neuropeptide somatostatin, were performed to study GluR1-4 immunoreactivity in interneuronal populations and principal cells. The ultrastructure of GluR1-4 labeled neurons was also examined using electron microscopy. With the exception of calbindin-positive interneurons, GluR2/3 was absent from hippocampal interneurons in both rat and monkey. In the rat, interneurons were more strongly immunoreactive against GluR1 than principal cells. In the monkey, immunoreactivity for GluR4 in interneurons was stronger than for GluR1. All GluR subunits were confined to spines, dendritic membrane and cytoplasm surrounding the nucleus but absent from axons and presynaptic terminals. Our findings suggest that hippocampal principal cells and interneurons express different complements of AMPA receptor subunits. Furthermore, the absence of GluR2 and/or GluR3 in both vulnerable and resistant interneurons subtypes indicates that knowledge of receptor subunit composition is not sufficient to predict neuronal vulnerability.

Animals↗

PubMed1996

Aromatase immunoreactivity in axon terminals of the vertebrate brain. An immunocytochemical study on quail, rat, monkey and human tissues.

Intraneuronal production of estradiol from testosterone has been shown to play a pivotal role in gender-specific brain development of most vertebrates, and to participate in numerous functions of the adult central nervous system. Previous biochemical and morphological approaches demonstrated that estrogen synthetase (aromatase) is present in specific limbic and hypothalamic structures. On the other hand, less attention has been paid to revealing its subcellular distribution. The possibility of aromatase presence in axonal processes has been indicated by recent biochemical and morphological observations suggesting new insights for the role of aromatase in neural functions. The objective of the present study was to provide morphological evidence for the subcellular location of aromatase in neurons of different vertebrate species including Japanese quail, rat, monkey, and human. Immunocytochemistry using a purified polyclonal antiserum against human placental aromatase localized immunoreactivity to hypothalamic and limbic cell groups in all of these species. Light and electron microscopic examination of vibratome sections revealed the presence of aromatase immunoreactivity throughout the neuronal perikarya, including dendrites and axonal processes. In each species there were numerous boutons which contained labeled small clear synaptic vesicles. Many of these axon terminals formed synapses with immuno-negative and immuno-positive dendrites and perikarya. This study furnishes the first immunolocalization of aromatase in the brains of two primate species, humans and monkeys. The provision of further evidence for estrogen synthesis in axons and axon terminals may help resolve apparent differences between the measurement of aromatase activity and the lack of aromatase-immunopositive cell bodies in previous studies. The present findings may be coupled with recent evidence regarding the molecular biology and the diversity of functional properties of P450 aromatase to indicate previously unexpected effects of brain aromatase at the synaptic level.

Adult↗

PubMed1996

Morphological and pharmacological evidence for neuropeptide Y-galanin interaction in the rat hypothalamus.

Galanin (GAL) and neuropeptide Y (NPY) have been shown to play important roles in the regulation of pituitary hormone secretion, as well as ingestive and sexual behaviors, by acting within the hypothalamus. While the mechanism of action of these regulatory peptides is under intensive investigation, less attention has been paid to the possible interaction between them in influencing these central regulatory processes. Because NPY and GAL augment pituitary gonadotropin release, the present study was undertaken to evaluate the nature of morphological and functional relationships between these excitatory hypothalamic peptidergic systems. Double immunolabeling for NPY and GAL was carried out on vibratome sections taken from the hypothalamus of colchicine-pretreated female rats. Avidinbiotin peroxidase technique and a dark blue diaminobenzidine reaction was used to visualize NPY profiles, while the GAL neurons were labeled with a light brown diaminobenzidine reaction using either the avidin-biotin peroxidase or the peroxidase antiperoxidase technique. Light microscopic examination of the immunostained material showed that in the arcuate nucleus, paraventricular nucleus, supraoptic nucleus, anterior hypothalamus, and medial preoptic area, an abundant network of NPY-immunoreactive axons surrounded GAL-immunostained cells. Numerous dark blue NPY-containing putative boutons were observed in close proximity to GAL-immunolabeled cell bodies and dendrites. Correlated light and electron microscopic examination revealed that most of the immunoreactive NPY axon terminals established synaptic connections with GAL-expressing cells. Synaptic connections were most frequently found in the medial preoptic area and in the magnocellular region of the paraventricular nucleus and arcuate nucleus. Fewer connections were observed in the supraoptic nucleus. These morphological observations demonstrate the existence of a strong NPY input to hypothalamic GAL neurons, thereby suggesting a modulatory role for NPY in monitoring GAL release. To evaluate the functional relevance of this anatomical relationship, the effects of intraventricular injection of a GAL receptor antagonist, galantide, were examined on NPY-induced LH release in ovarian steroid-primed ovariectomized rats. As expected, intraventricular injection of NPY readily stimulated LH release. Although, while on its own, galantide was ineffective in altering basal LH release, it markedly attenuated the NPY-induced LH response, thereby suggesting that GAL released in response to NPY administration may, in part, mediate the excitatory effects of NPY. These experimental results, taken together with the morphological observations, document the involvement of an NPY --> GAL signaling modality in the release of gonadotropins and, likewise, raise the possibility of a similar signaling process in the release of other pituitary hormones and elicitation of behavioral effects attributed to NPY and GAL.

Analysis of Variance↗