Efferent projections from the median preoptic nucleus to sleep- and arousal-regulatory nuclei in the rat brain

The median preoptic nucleus (MnPO) has been implicated in the regulation of hydromineral balance and cardiovascular regulation. The MnPO also contains neurons that are active during sleep and in response to increasing homeostatic pressure for sleep. The potential role of these neurons in the regulation of arousal prompted an analysis of the efferent projections from the MnPO. Anterograde and retrograde neuroanatomical tracers were utilized to characterize the neural connectivity from the MnPO to several functionally important sleep- and arousal-regulatory neuronal systems in the rat brain. Anterograde terminal labeling from the MnPO was confirmed within the core and extended ventrolateral preoptic nucleus. Within the lateral hypothalamus, labeled axons were observed in close apposition to proximal and distal dendrites of hypocretin/orexin immunoreactive (IR) cells. Projections from the MnPO to the locus coeruleus were observed within and surrounding the tyrosine hydroxylase-IR cell cluster. Labeled axons from the MnPO were mostly observed within the lateral division of the dorsal raphé nucleus and heavily within the ventrolateral periaqueductal gray. Few anterogradely labeled appositions were present juxtaposed to choline acetyltransferase-IR somata within the magnocellular preoptic area. The use of retrogradely transported neuroanatomical tracers placed within the prospective efferent terminal fields supported and confirmed findings from the anterograde tracer experiments. These anatomical findings support the hypothesis that MnPO neurons function to promote sleep by inhibition of orexinergic and monoaminergic arousal systems and disinhibition of sleep regulatory neurons in the ventrolateral preoptic area.     

Ultrastructure and synaptic connections of vasoactive intestinal polypeptide-like immunoreactive non-pyramidal neurons and axon terminals in the rat hippocampus

In the hippocampus, antibody raised against vasoactive intestinal polypeptide (VIP) labeled perikarya and processes of non-pyramidal neurons whereas these structures remained unlabeled in pyramidal cells and granule cells. In the present study, VIP-immunostaining was used to investigate the fine structure and synaptic connections of identified non-pyramidal neurons and of imrnunoreactive axon terminals in the CA1 region of the rat hippocampus by means of electron microscopic immunocytochemistry.From a number of cells studied, two VIP-like imrnunoreactive non-pyramidal neurons in the regio superior were selected for an electron microscopic analysis of serial thin sections. These cells were different with regard to the location of their cell bodies and the orientation of their dendrites. One cell was located in the stratum lacunosum-moleculare with dendritic processes oriented parallel to the hippocampal fissure. The second neuron was found in the inner one-third of the stratum radiatum. The dendrites of this cell ran nearly parallel to the ascending apical dendrites of the pyramidal cells. Both cells had a round or ovoid perikaryon and an infolded nucleus. The aspinous dendrites of both neurons were densely covered with synaptic boutons. These terminals were small, filled with spherical vesicles and established asymmetric synaptic contacts. No variations in the fine structure of the presynaptic boutons were found along the course of the labeled dendrites through the various hippocampal layers, although different afferents are known to terminate in these layers.Vasoactive intestinal polypeptide-like immunopositive axon terminals course through all layers of the hippocampus. In the stratum pyramidale they established symmetric synaptic contacts with the perikarya of pyramidal cells. In the stratum radiatum they made symmetric contacts with the shafts of apical dendrites of pyramidal cells but never contacted dendritic spines.The symmetric contacts with pyramidal cell perikarya suggest an involvement of the VIP-like immunoreactive axon terminals in pyramidal cell inhibition.     

Quantitative autoradiographic evidence for axonal transport of imipramine receptors in the central nervous system of the rat

Interruption of the ascending serotonin axons of the medial forebrain bundle (MFB) in the rat brain produced a progressive time-dependent accumulation of imipramine receptors (labeled for autoradiography with [3H]imipramine). The largest accumulation of receptors occurred during the first 12 h at the caudal aspect of the lesion. An electrolytic lesion of the nucleus raphe dorsalis, administered 24 h prior to interruption of the medial forebrain bundle, markedly reduced the number of imipramine receptors on the caudal side of the lesion, while a significant accumulation was still evident on the rostral aspect. These results suggest that imipramine receptors are undergoing the process of orthograde axonal transport to terminals in the forebrain from the neuronal perikarya found in the nucleus raphe dorsalis. These receptors may also be undergoing retrograde transport back to their cell bodies of origin.     

Ultrastructural identification of somata and neural processes immunoreactive to antibodies against glutamic acid decarboxylase (GAD) in the dorsal lateral geniculate nucleus of the cat

Summary The cat dorsal lateral geniculate nucleus (LGN) was examined at the light- and electron-microscopic level after immunocytochemistry for GAD (the synthesizing enzyme of the inhibitory neurotransmitter GABA), to identify cells and processes with GAD-like immunoreactivity. GAD-positive perikarya were distributed throughout the A and C laminae, constituting a moderate proportion of cells in the LGN. Labeled cells were characterized by small size, scant cytoplasm, relatively large nuclei with common indentations, small mitochondria, few organelles and few strands of rough endoplasmic reticulum. Unlabeled cells were of large, medium and small size. GAD-positive terminals were identified as F1 and F2 types (Guillery's nomenclature) on the basis of their synaptic relations and ultrastructure. Labeled F2 terminals were postsynaptic to retinal (RLP) boutons and presynaptic to unlabeled dendrites in synaptic glomeruli. Labeled F1 terminals made synapses on unlabeled somata and dendrites, and on labeled dendrites and F2 terminals. Presumably, most labeled F1 terminals originate from GABAergic perigeniculate axons. Retinal (RLP) and cortico-geniculate (RSD) boutons remained unlabeled in the reative zone. These terminals made synapses with labeled and unlabeled dendrites and with labeled F2 boutons. In conjunction with previous studies on GAD-positive cells in the perigeniculate nucleus, these results provide immunocytochemical and morphological evidence suggesting that the GABAergic intrinsic and extrinsic (perigeniculate) interneurons mediate the different inhibitory phenomena which occur in relay cells of the cat LGN. The ultrastructural features and synaptic relations of GABAergic cells and processes in the cat LGN are similar to those of equivalent neural elements in the LGN of rat and monkey, suggesting general principles of organization and morphology for GABAergic neurons in the thalamus of different mammals.Supported in part by grants EY 02877 and HD 03352 from the National Institutes of Health     

Ultrastructure and synaptic relations of neural elements containing glutamic acid decarboxylase (GAD) in the perigeniculate nucleus of the cat

Summary The perigeniculate nucleus of the cat (PGN) was examined at light and electron microscopic levels after immunocytochemical labeling for the gamma-aminobutyric acid (GABA) synthesizing enzyme, glutamic acid decarboxylase (GAD). In light microscopic sections, virtually all perikarya were found to be labeled (GAD+), as well as proximal dendrites, fibres and punctiform elements. Cells in the thalamic reticular nucleus (TRN) dorsal to PGN were also labeled. Ultrastructural analysis of PGN showed immunoreactivity in all somata, in dendrites and in the following vesicle containing profiles: 1.) F1 terminals, which are characterized by large size, dark mitochondria, and pleomorphic vesicles. These terminals form symmetrical synaptic contacts with somata, somatic spines and with dendrites of GAD+ PGN cells. 2.) F2 terminals, which are smaller than F1 terminals, contain also pleomorphic vesicles and frequently make serial synapses of the symmetric type with other F2 terminals. Presumably, F1 terminals are formed by collaterals of PGN-cell axons and F2 terminals by vesicle containing dendrites of PGN cells. Terminals devoid of immunoreactivity included: 1.) RLD terminals characterized by large size, round vesicles, dark mitochondria, and by asymmetric synaptic contacts with somata, especially with somatic spines, and with dendrites of GAD+ perigeniculate neurons; 2.) RSD terminals, characterized by small size, round vesicles and dark mitochondria, which make asymmetric synapses with GAD+ dendrites of medium and small size; 3.) Multivesicular (MV) terminals with variably shaped vesicles including dense core vesicles synapsing on GAD+ dendrites. There are reasons to believe that RSD terminals belong to corticofugal axons and RLD terminals to collateral axons of LGN relay cells. The origin of MV terminals remains to be determined. The GABAergic nature of the PGN cells conforms with the presumed function of these cells as mediators of inhibition of LGN relay cells. The complex synaptic relations observed between GAD+ elements in the PGN would allow for reciprocal inhibition between perigeniculate cells.Supported in part by NIH grants EY02877 to V.M. Montero and HD 03352 to the Waisman Center     

Morphology of cortically projecting basal forebrain neurons in the rat as revealed by intracellular iontophoresis of horseradish peroxidase

The intracellular horseradish peroxidase technique was employed to study the morphology of basal forebrain neurons that were identified as cortically projecting by antidromic invasion from the cerebral cortex. Four neurons were examined in detail; they were located at different rostrocaudal levels within the basal forebrain. Their somata were large, 30-50 microns in longest dimension, and gave rise to three to eight primary dendrites, which ramified into third- to fifth-order dendrites. The longest observed dendrite in each neuron terminated at a distance of 600-900 microns from the soma. The sizes of soma and dendritic field of the two most rostrally located cells were smaller than those of the other two cells located more caudally. Dendritic spines were seen in all four cortically projecting basal forebrain neurons. Spines had shafts of variable lengths, and usually had spherical or elongated heads. The density of spines varied among the four neurons; one neuron, a type II cortically projecting basal forebrain neurons as defined physiologically by Reiner et al., had a much greater number of dendritic spines than the other three neurons, which were type I neurons. No somatic spines were observed. Presumptive axons were identified in three of the four cortically projecting basal forebrain neurons. These axons originated from either the soma or a primary dendrite, and two of them gave off local collaterals, which displayed occasional bouton-like swellings. The above observations confirm and extend previous findings that cortically projecting neurons in the basal forebrain are large multipolar cells, and provide evidence to support the conclusion that these cells, although somewhat variable in size, generally have extensive dendrites which display frequent spines.     

Spatial relationship between neurotensinergic axons and cholinergic neurons in the rat basal forebrain: a light microscopic study with three-dimensional reconstruction

Cholinergic neurons of the basal forebrain are known to project to the hippocampus and cerebral cortex wherein they play an important role in cortical activation, attention and memory. These neurons have been shown to possess neurotensin binding sites and to respond electrophysiologically to local application of neurotensin, indicating the presence of functional receptors on their membrane. In the present light microscopic study, the spatial relationship between neurotensinergic axons and cholinergic nerve cell bodies and proximal dendrites was investigated in the basal forebrain of the rat by dual immunostaining for neurotensin and choline acetyltransferase. Rostrally, neurotensinergic fibres were concentrated in the lateral septum and anterior substantia innominata, whereas cholinergic neurons were located in the medial septum, diagonal band of Broca and magnocellular preoptic nucleus. At high magnification, a few neurotensinergic axonal varicosities were observed in the region of cholinergic neurons, and fewer still in close proximity to cholinergic perikarya and proximal dendrites. Caudally, neurotensinergic fibres formed a dense plexus of varicose axons in the same region where cholinergic neurons were located in the posterior substantia innominata and in the ventral and caudal aspects of the globus pallidus. At high magnification, many of these neurotensinergic varicosities were seen in close proximity to the cholinergic perikarya. These results suggest that cholinergic cells receive a much denser neurotensinergic innervation in the caudal than in the rostral aspect of the basal forebrain. This differential distribution is not reflected in the uniform density of neurotensin receptors and potent responses to neurotensin through the cholinergic cell population, suggesting the possibility that neurotensin's effects are mediated in part by a paracrine mechanism.     

Nitrergic neurons make synapses on dual-input dendritic spines of neurons in the cerebral cortex and the striatum of the rat: implication for a postsynaptic action of nitric oxide

Pre-embedding electron microscopic immunocytochemistry was used to examine the ultrastructure of neurons containing nitric oxide synthase and to evaluate their synaptic relationships with target neurons in the striatum and sensorimotor cerebral cortex. Intense nitric oxide synthase immunoreactivity was found by light and electron microscopy in a type of aspiny neuron scattered in these two regions. The intensity of the labeling was uniform in the soma, dendrites and axon terminals of these neurons. In both forebrain regions, nitric oxide synthase-immunoreactive neurons received synaptic contacts from unlabeled terminals, which were mostly apposed to small-caliber dendrites. The unlabeled symmetric contacts were generally about four times as abundant as the unlabeled asymmetric contacts on the nitric oxide synthase-immunoreactive neurons. Terminals labeled for nitric oxide synthase were filled with synaptic vesicles and were observed to contact unlabeled neurons. Only 54% (in the cerebral cortex) and 44.3% (in the striatum) of the nitric oxide synthase-immunoreactive terminals making apposition with the target structures were observed to form synaptic membrane specializations within the plane of the randomly sampled sections. The most common targets of nitric oxide synthase-immunoreactive terminals were thin dendritic shafts (54% of the immunoreactive terminals in the cortex and 75.7% of the immunoreactive terminals in the striatum), while dendritic spines were a common secondary target (42% of the immunoreactive terminals in the cortex and 20.6% of the immunoreactive terminals in the striatum). The spines contacted by nitric oxide synthase-immunoreactive terminals typically also received an asymmetric synaptic contact from an unlabeled axon terminal.These findings suggest that: (i) nitric oxide synthase-immunoreactive neurons in the cortex and striatum preponderantly receive inhibitory input; (ii) nitric oxide synthase-containing terminals commonly make synaptic contact with target structures in the cortex and striatum; (iii) spines targeted by nitric oxide synthase-containing terminals in the cortex and striatum commonly receive an asymmetric contact as well, which may provide a basis for a synaptic interaction of nitric oxide with excitatory input to individual spines.     

Projections from the hypothalamus and its adjacent areas to the posterior pituitary in the rat

Cholera toxin conjugated horseradish peroxidase was injected into the posterior pituitary and its afferents traced in 21 albino rats. The neuronal processes as well as the perikarya were elaborately displayed. The principal and retrochiasmatic supraoptic nuclei and the magnocellular paraventricular subnuclei were densely labelled. The accessory cell groups or nuclei labelled included: the medial preoptic and anterior hypothalamic areas, the anterior and posterior fornical nuclei, the lateral hypothalamic area, the nucleus circularis and nucleus of the forebrain bundle and hitherto unknown or not fully appreciated retrochiasmatic area, the dorsal accessory groups in an area between the stria medullaris and fornix, on the one hand, and the stria terminalis and internal capsule, on the other, and a well developed subependymalperiventricular zone. The medial preoptic nucleus, subfornical organ and organ vasculosum laminae terminalis were also weakly stained. Dendrites of the magnocellular paraventricular nucleus have been said by some to be largely confined to the subnuclei in which they lie. Immunohistochemical studies have proved that they extended beyond their nuclear confinement. The present study has found much wider extension of their dendritic fields. In fact, dendrites of the magnocellular neurosecretory cells in general were long and had a certain degree of directional bias. Several sites projecting to the posterior pituitary were closely related to the cerebrospinal fluid. Namely, the subependymal neuronal plexuses along the third ventricle and beneath the interventricular foramen, and the subpial dendritic plexuses of the supraoptic and retrochiasmatic supraoptic nuclei. Neurons were seen to squeeze in-between the ependymal cells, bringing themselves very close to the cerebrospinal fluid. No direct cerebrospinal fluid-contacting elements, either cell bodies or processes, however, could be ascertained. It is proposed that these plexuses may monitor changes in the cerebrospinal fluid. Besides the principal neurohypophysial tract the posterior pituitary was found in the present study to receive its afferents via two accessory fasciculi, one coursing in the medial forebrain bundle and the other running along the lateral wall of the infundibular recess subependymally.     

Histaminergic neurons receive substance P-ergic inputs in the posterior hypothalamus of the rat.

The synaptic connections between histaminergic neurons and substance P (SP) afferents in the caudal magnocellular nucleus (CM) of the hypothalamus were examined using an immunoelectron microscopic mirror method. SP-immunoreactive (SP-IR) terminals made synaptic contacts with the somata, somatic spines and dendrites of histidine decarboxylase immunoreactive (HDC-IR) neurons. This suggests that SP afferents exert monosynaptic influence on the central histaminergic neuronal system.     

Association between pituitary adenylate cyclase-activating polypeptide and thyrotropin-releasing hormone in the rat hypothalamus

Pituitary adenylate cyclase-activating polypeptide (PACAP) is present in many regions of the hypothalamus including the paraventricular nucleus (PVN). In this study the anatomical relationship between PACAP- and thyrotropin-releasing hormone (TRH)-immunoreactive neuronal elements was investigated in the rat hypothalamus. Using a well-characterized mouse monoclonal antibody against PACAP and a rabbit polyclonal antiserum against TRH, we found numerous nerve fibers with PACAP-immunoreactivity (ir) closely apposed to TRH neurons in the PVN suggesting synaptic contacts. Electron microscopy confirmed the presence of synapses between PACAP-ir terminals and TRH-ir perikarya and various dendritic profiles as well as between PACAP-ir terminals and unlabeled perikarya and small- to medium-sized dendrites. Coexistence of the two peptides in perikarya of the PVN was limited to only a few neurons in the periventricular subdivision, but PACAP-ir and TRH-ir extensively coexisted in perikarya of the perifornical cell group, medial preoptic area, lateral hypothalamus and dorsomedial nucleus. The interactions between PACAP-containing neuronal processes and TRH neurons in the PVN raise the possibility that PACAP modulates the secretion of TRH destined for regulation of anterior pituitary TSH. The more general association between PACAP and TRH in other regions of the hypothalamus suggests a further role for PACAP as a cofactor in the function of TRH neurons.     

Ca2+/calmodulin-dependent protein kinase II in the rat cerebellum: an immunohistochemical study with monoclonal antibodies specific to either alpha or beta subunit.

Monoclonal antibodies specific to either alpha or beta subunit of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) of the rat brain were produced and the distribution of each subunit in the rat cerebellum was examined immunohistochemically. Each antibody detected solely the corresponding subunit in immunoblot analysis of crude homogenates of the rat forebrain and cerebellum, and purified CaM kinase II from the rat forebrain. Immunoreactivity for alpha subunit was present selectively in Purkinje cells: perikarya, dendrites with their spines, axons and their terminal-like structures in the cerebellar cortex, cerebellar nuclei and lateral vestibular nucleus. Many of these alpha subunit-immunoreactive axons from the cerebellum were traced only through the inferior cerebellar peduncle. beta Subunit was detected in perikarya and dendrites of a limited number of Purkinje cells, many granule cells and neurons in the cerebellar nuclei. Thus, different distributions of alpha and beta subunits of CaM kinase II in the cerebellum were demonstrated.     

Plasmalemmal appositions between cholinergic and non-cholinergic neurons in rat caudate-putamen nuclei

We have observed that in rat caudate-putamen nuclei, neurons immunolabeled for choline acetyltransferase were sometimes in direct apposition to unlabeled perikarya and dendrites [Pickel V. M. and Chan J. (1990) J. Neurosci. Res. 25, 263-280]. Similar juxtapositions between plasmalemmas of nerve cells each receiving input from one common terminal have been associated with activation of certain central neurons [Theodosis D. T. and Poulain D. A. (1989) Brain Res. 484, 361-366]. Thus, we sought to determine the relative abundance and ultrastructure of the appositions and the frequencies of shared synapses between choline acetyltransferase-labeled and unlabeled neurons in the rat striatum. A monoclonal antibody raised against choline acetyltransferase was localized in semi-adjacent ultrathin sections through 24 neurons in the dorsolateral caudate-putamen nuclei. Five of these choline acetyltransferase-labeled perikarya showed direct somatic appositions with unlabeled neurons. The remaining 19 of the choline acetyltransferase-labeled perikarya did not show somatic appositions with unlabeled perikarya; however, when traced through multiple (20-100) semi-adjacent sections their dendrites always showed extensive plasmalemmal juxtapositions with one or more unlabeled perikarya. The apposed perikarya had round nuclei and other characteristics of medium, spiny neurons. The majority of the apposed cholinergic and non-cholinergic neurons were postsynaptic to at least one common unlabeled terminal. These terminals usually formed symmetric junctions. At sites of appositions, the plasmalemmas of choline acetyltransferase-immunoreactive soma or dendrites and unlabeled neurons were closely spaced without intervening astrocytic processes. The appositions lacked the ultrastructural features typical of gap-junctions, but did occasionally show parallel arrays of thin (1-2 nm) electron-dense bands. In both labeled and unlabeled perikarya, the nuclei were separated from the appositional zones by narrow (0.7-3.3 microns) rims of cytoplasm. This cytoplasmic rim contained subsurface cisternae and other less specialized smooth and rough endoplasmic reticulum, and vesicular structures. The findings suggest that in the caudate-putamen nuclei (1) the tonically active cholinergic neurons [Wilson C. J. et al. (1990) J. Neurosci. 10, 508-519] may modulate or be modulated by non-cholinergic spiny neurons through non-synaptic somatic or dendritic appositions, and (2) that both neurons may be simultaneously inhibited by shared afferent input. Activation of this system could facilitate coordinated movements through synchronization of cholinergic interneurons and spiny projection neurons containing GABA or other transmitters.     

The neurophysin-containing innervation of the forebrain of the mouse

The oxytocinergic and vasopressinergic innervation of the forebrain of normal mice was studied immunocytochemically by use of a set of mouse monoclonal anti-neurophysins applied to serial vibratome sections. The extensive hypothalamic and extra-hypothalamic location of these neuropeptides was revealed, with, or without colchicine pretreatment. Magnocellular perikarya immunoreactive for either oxytocin-neurophysin or vasopressin-neurophysin were concentrated mainly: in the anterior commissural nucleus; in various subdivisions of the paraventricular nucleus; in a profuse array in the periventricular region; in the supraoptic nucleus including its retrochiasmatic division; in various accessory nuclei; and as a number of cells scattered throughout the preoptic and hypothalamic regions. Extensive groups of parvocellular neurons, containing only vasopressin-neurophysin, were located in the suprachiasmatic nucleus including a ventromedial division, in the bed nucleus of the stria terminalis, and in the medial amygdaloid nucleus. Perikarya in the magnocellular nuclei were of generally similar size distribution and there was no evidence that distinct populations of magnocellular and parvicellular neurons, separable on the basis of size, had been labelled within these nuclei. Within the paraventricular nucleus, however, neurons in the posterior part were smaller than those located more anteriorly, and the cells containing oxytocin-neurophysin were slightly smaller than those containing vasopressin-neurophysin. Within the generally similar size distribution, magnocellular neurons of the anterior commissural nucleus were the largest. During processing, shrinkage of the tissue and immunolabeled cells had occurred. The immunocytochemical procedure delineated neuronal processes, in particular dendrites, very effectively. The dendrites were shown to project for far greater distances than is generally recognized, some were of a characteristic corkscrew-like morphology, and most were oriented in a well-defined pattern. Many dendrites of paraventricular neurons passed medially than caudally towards and then along the third ventricle. Most dendrites of supraoptic neurons, in particular those containing vasopressin-neurophysin, had an extensive anteroposterior course beneath the pia of the base of the brain. The axons containing oxytocin- and vasopressin-neurophysin were shown to take rather different paths from the paraventricular nucleus towards the median eminence.(ABSTRACT TRUNCATED AT 400 WORDS)     

大鼠最后区内神经降压肽电镜免疫细胞化学定位

采用电镜免疫细胞化学技术观察了大鼠最后区内神经降压肽样免疫反应神经成分的超微结构。免疫反应产物要见于核周质，树突，轴突及其终末内。神经降压肽样免疫反应轴突终末与末标记树突形成轴－树突触。未标记轴突终末与神经降压肽样免疫反应树突呆构成轴－树突触。     

Morphology of intracellularly stained spiny neurons in rat striatal grafts.

Two to six months after implantation of fetal striatal primordia into the kainic acid-lesioned neostriatum of adult rats, spiny neurons in the grafts were stained intracellularly with biocytin. To determine whether the spiny neurons in the grafts differentiate morphologically as in the host neostriatum, the intracellularly stained spiny neurons in the grafts were studied with light and electron microscopy and compared with that of spiny neurons in the host neostriatum. The spiny neurons in the grafts had ovoid or polygonal cell bodies with dendrites radiating in all directions. The somata were smooth and the dendrites, except for their most proximal portions, were rich in spines. All these features resembled the appearance of spiny neurons in the intact neostriatum. However, quantitative studies showed that the somata of spiny neurons in the grafts were larger than those in the host neostriatum (projected cross-sectional areas of 230 +/- 64.6 microns 2 in the grafts and 158 +/- 28.9 microns 2 in the host) and the spine density of graft neurons was lower than that of host neurons. Cells near the border of the grafts had dendrites extending both into the graft and into the host neostriatum. In these cells, the dendrites in the grafts had fewer spines than the dendrites in the host tissue. The axons of spiny neurons in the grafts had very large and dense intrastriatal collateral arborizations, which occupied a much larger volume than that of the dendritic domain of the parent cells. The local axonal arborizations of each of these cells filled almost the entire graft. In some cells, axonal branches were traced outside the grafts and were seen to enter the internal capsule fascicles. Unlike spiny neurons in the normal adult neostriatum, the spiny cells of the graft could have nuclear indentations. With this exception, the ultrastructural features of spiny neurons in the grafts were very similar to those in the hosts. Many unlabeled boutons made synapses on identified spiny neurons in the grafts. Terminals with small round vesicles made synaptic contacts on dendritic shafts and dendritic spines, while terminals with flattened or pleomorphic vesicles contacted somata, dendrites, and dendritic spines. Labeled axon collaterals of graft neurons made symmetrical synapses on somata, dendrites and spines in the grafts and in the host neostriatum. In the grafts, more than 60% of the axon terminals contacted dendritic shafts. The proportion of axosomatic and axospinous synapses varied substantially from cell to cell.(ABSTRACT TRUNCATED AT 250 WORDS)     

Synaptic connections of morphologically identified and physiologically characterized large basket cells in the striate cortex of cat

Neurons were studied in the striate cortex of the cat following intracellular recording and iontophoresis of horseradish peroxidase. The three selected neurons were identified as large basket cells on the basis that (i) the horizontal extent of their axonal arborization was three times or more than the extent of the dendritic arborization; (ii) some of their varicose terminal segments surrounded the perikarya of other neurons. The large elongated perikarya of the first two basket cells were located around the border of layers III and IV. The radially-elongated dendritic field, composed of beaded dendrites without spines, had a long axis of 300-350 microns, extending into layers III and IV, and a short axis of 200 microns. Only the axon, however, was recovered from the third basket cell. The lateral spread of the axons of the first two basket cells was 900 microns or more in layer III and, for the third cell, was over 1500 microns in the antero-posterior dimension, a value indicating that the latter neuron probably fulfills the first criterion above. The axon collaterals of all three cells often branched at approximately 90 degrees to the parent axon. The first two cells also had axon collaterals which descended to layers IV and V and had less extensive lateral spreads. The axons of all three cells formed clusters of boutons which could extend up a radial column of their target cells. Electron microscopic examination of the second basket cell showed a large lobulated nucleus and a high density of mitochondria in both the perikarya and dendrites. The soma and dendrites were densely covered by synaptic terminals. The axons of the second and third cells were myelinated up to the terminal segments. A total of 177 postsynaptic elements was analysed, involving 66 boutons of the second cell and 89 boutons of the third cell. The terminals contained pleomorphic vesicles and established symmetrical synapses with their postsynaptic targets. The basket cell axons formed synapses principally on pyramidal cell perikarya (approximately 33% of synapses), spines (20% of synapses) and the apical and basal dendrites of pyramidal cells (24% of synapses). Also contacted were the perikarya and dendrites of non-pyramidal cells, an axon, and an axon initial segment. A single pyramidal cell may receive input on its soma, apical and basal dendrites and spines from the same large basket cell.(ABSTRACT TRUNCATED AT 400 WORDS)     

Dopaminergic innervation of interneurons in the rat basolateral amygdala

The basolateral nuclear complex of the amygdala (BLC) receives a dense dopaminergic innervation that plays a critical role in the formation of emotional memory. Dopamine has been shown to influence the activity of BLC GABAergic interneurons, which differentially control the activity of pyramidal cells. However, little is known about how dopaminergic inputs interface with different interneuronal subpopulations in this region. To address this question, dual-labeling immunohistochemical techniques were used at the light and electron microscopic levels to examine inputs from tyrosine hydroxylase-immunoreactive (TH+) dopaminergic terminals to two different interneuronal populations in the rat basolateral nucleus labeled using antibodies to parvalbumin (PV) or calretinin (CR). The basolateral nucleus exhibited a dense innervation by TH+ axons. Partial serial section reconstruction of TH+ terminals found that at least 43-50% of these terminals formed synaptic junctions in the basolateral nucleus. All of the synapses examined were symmetrical. In both TH/PV and TH/CR preparations the main targets of TH+ terminals were spines and distal dendrites of unlabeled cells. In sections dual-labeled for TH/PV 59% of the contacts of TH+ terminals with PV+ neurons were synapses, whereas in sections dual-labeled for TH/CR only 13% of the contacts of TH+ terminals with CR+ cells were synapses. In separate preparations examined in complete serial sections for TH+ basket-like innervation of PV+ perikarya, most (76.2%) of TH+ terminal contacts with PV+ perikarya were synapses. These findings suggest that PV+ interneurons, but not CR+ interneurons, are prominent synaptic targets of dopaminergic terminals in the BLC.     

大鼠臂旁核内接受孤束核内脏传入之神经元同时接受中央杏仁核的反馈调节──溃变、HRP法结合包埋后免疫电镜研究

为研究来自孤束核的内脏传导信息在臂旁核水平是否接受中央杏仁核的反馈调节及其递质性质，以及孤束核—臂旁核—中央杏仁核传导通路中，在臂旁核水平是否接受ＧＡＢＡ的调节，本文将ＨＲＰ注入中央杏仁核进行顺、逆行标记，同时将兴奋性氨基酸毒素海人酸注入孤束核进行损毁，观实其顺行溃变终末，取外侧臂旁核超薄切片后结合抗ＧＡＢＡ的免疫电镜染色，观察发现有下列几种标记；（１）顺行溃变终末，所有的都与臂旁核神经元形成非对称性突触；（２）ＨＲＰ标记终末有两类：第一类和臂旁核神经元形成对称性突触，占ＨＲＰ标记终末总数的８０％以上，第二类与臂旁核神经元形成非对称性突触，另外有大量的ＨＲＰ标记的胞体和树突；（３）胶体金标记的ＧＡＢＡ阳性终末，皆与突触后结构形成对称性突触；（４）ＧＡＢＡ／ＨＲＰ双标记终末，具有ＧＡＢＡ免疫阳性终末和第一类ＨＲＰ标记终末的共同特征。上述几种标记在臂旁核内有以下几种关系：（１）溃变终末和ＧＡＢＡ阳性终末与同一个ＨＲＰ标记或非标记的突形成轴－树突触；（２）溃变终末和第一类ＨＲＰ标记终末共同终止于同一非标记讨突；（３）溃变终末与ＨＲＰ标记树突或胞位形成非对称性突触；（４）ＧＡＢＡ／ＨＲＰ双标记终末与非标记树突或胞体?