Ultrastructure of degenerating cerebellothalamic terminals in the ventral medial nucleus of the cat

Summary Terminal degeneration of cerebellar afferents in the ventral medial thalamic nucleus (VM) was studied in cats at the ultrastructural level after uni- or bilateral lesions in the brachium conjunctivum (BC). To achieve discrete lesions within the BC, a new very accurate stereotaxic technique was used. Numerous large terminals belonging to a population of so-called LR boutons were observed degenerating in the VM. The boutons displayed a wide variety of degenerative changes. Some revealed the features of the classical neurofilamentous type of degeneration. Others, although containing a slightly increased number of neurofilaments, featured much more prominently large numbers of coated vesicle shells and heavy accumulations of a flocculent electrondense material. Degeneration in a third group of boutons similar to some extent to the light type of degeneration was characterized by tight clumping of enormously swollen or distorted synaptic vesicles within a light matrix. At later stages, however, all these boutons were believed to become shrunken and electron-dense since intermediate stages between the light- and dark-appearing boutons were observed. The degenerating cerebellar boutons formed asymmetrical synaptic contacts. Groups of 3 or 4 boutons terminated upon dendrites of projection neurons synapsing more frequently on spines than on dendritic stems. The synaptic contacts between cerebellar boutons and the vesicle-containing dendrites of local circuit neurons were encountered as often if not more than the contacts on projection neuron dendrites. Triads consisting of cerebellar boutons and dendrites of both types of neurons were observed very regularly. This synaptic arrangement provides the anatomical basis for the modification of cerebellar input in the VM by interneurons.     

Dual axonal terminations from the retrosplenial and visual association cortices in the laterodorsal thalamic nucleus of the rat

Light and electron microscopic tracing studies were conducted to assess the synaptic organization in the laterodorsal thalamic nucleus (LD) of the rat and the laminar origins of corticothalamic terminals from the retrosplenial and visual association cortices to LD. A survey of the general ultrastructure of LD revealed at least three types of presynaptic terminals identified on the basis of size, synaptic vesicle morphology, and synaptic membrane specializations: (1) small axon terminals with round synaptic vesicles (SR), which accounted for the majority of terminal profiles and made asymmetric synaptic contacts predominantly with small dendritic shafts and spines; (2) large axon terminals with round synaptic vesicles (LR), which formed asymmetric synaptic contacts mainly with large dendritic shafts; and (3) small to medium-size axon terminals with pleomorphic synaptic vesicles (SMP), which symmetrically synapsed with a wide range of postsynaptic structures from cell bodies to small dendrites. Synaptic glomeruli were identified, whereas no presynaptic dendrites were found. To characterize and identify corticothalamic terminals arising from the retrosplenial and visual association cortices that project to LD, wheat germ agglutinin conjugated to horseradish peroxidase (WGA–HRP) was injected into these cortices. Axons anterogradely labeled with WGA–HRP ended in both SR and LR terminals. On the other hand, dextran-tetramethylrhodamine injected into LD as a retrograde fluorescent tracer labeled large pyramidal cells of layer V as well as small round or multiform cells of layer VI in the retrosplenial and visual association cortices. These findings provide the possibility that corticothalamic terminations from cortical neurons in layer V end as LR terminals, while those from neurons in layer VI end as SR boutons.     

Light and electron microscopical studies of the normal nuclei ventralis lateralis and ventralis anterior thalami in the cat

Summary A light and electron microscopical investigation of the nucleus ventralis lateralis (VL) and nucleus ventralis anterior (VA) of the cat's thalamus was made. Light microscopical examination of Golgi impregnated material revealed the existence of two types of cells based on differences in their dendritic arborization and branching of the axon. One of the cells is considered to be the thalamocortical relay cell, whereas the other is tentatively considered to be a Golgi type II neuron. Electron microscopical investigations of the two nuclei revealed the existence of a high number of profiles containing pleomorphic vesicles, and which have been identified as dendrites. Based on correlation with the Golgi material as well as on cytological features of the parent cell bodies, the dendrites containing vesicles are believed to belong to Golgi type II neurons. In addition to the vesicle-filled dendritic profiles, five different types of boutons have been identified. Two of these boutons, type LR and type SR, contain ovoid vesicles and establish asymmetrical synaptic contacts with dendrites of both types of neurons. Type F1, F2 and F3 boutons contain pleomorphic vesicles, but can be distinguished from dendritic profiles containing pleomorphic vesicles. Type F2 and F3 boutons establish symmetrical contacts with dendrites of both thalamocortical relay cells and Golgi type II neurons. Type F1 boutons establish symmetrical synaptic contact with the proximal dendrites or soma of the thalamocortical relay neurons only.Dendrites of both thalamocortical relay cells and Golgi type II neurons, as well as type LR, SR, F2 and F3 boutons, are engaged in glomeruli. Dendro-dendritic synapses between Golgi type II dendrites and relay cell dendrites are frequently seen, whereas no evidence of axo-axonic synapses have been found.Differences and similarities in the ultrastructural organization of VL and VA are described in some detail.Some of the observations reported in this and the following two papers were presented at the 6th Symposium of the International Society for Research in Stereoencephalotomy, Tokyo, October 12th & 13th, 1973.     

Quantitative ultrastructural analysis of the periaqueductal gray in the rabbit

The fine structure of the periaqueductal gray (PAG) of the rabbit was examined using the transmission electron microscope. On the basis of synaptic polarity, vesicle size, and the nature of the pre- and postsynaptic elements, 10 essentially different synaptic types could be discerned (6 axo-dendritic, 2 axo-somatic, 1 axo-axonic, and 1 dendro-dendritic). Synaptic contact on the soma of PAG neurons were small and covered, on average, only 1.6% of the soma surface. The most striking feature of the synaptic structure of the PAG was that more than 94.1% of all synapses were axo-dendritic. Of these, 83.5% were of the symmetrical type. Most of these contacts occurred on buttons of small to medium size, and contained either round vesicles of medium size or pleomorphic vesicles of medium size. Boutons containing only flattened vesicles were quite rare. Boutons contacting larger dendrites were generally small-to-medium in size, made asymmetric-type synaptic contacts, and contained pleomorphic vesicles of medium-to-large size. Medium-sized dendrites were contacted principally by small boutons exhibiting either symmetrical or asymmetrical junctions containing medium-sized pleomorphic vesicles, and in addition a few of these boutons contained both large, and small, round vesicles. Dendritic spines were generally provided with only one synaptic contact, stretching the entire width of the spinous process. Boutons and the spines on dendrites were approximately the same size. Synapses between two vesicle-containing structures (axo-axonic or dendro-dendritic synapses) were rare (1.4%). They were generally asymmetric and contained round vesicles of medium size. Complex synapses, where a glial sheet enclosed the synapse, were occasionally observed. Also seen were multiple synapses, with up to 11 contacts on a single dendritic profile. Large dense-core vesicle were seen in approximately 40% of all synapses, whereas small dense-core vesicles were only found in about 3%. Data is provided on how different synaptic features relate to ventral, lateral, dorsal, and medial PAG. Principally this is in relation to neuron size, glia cell content, axonal characterization, and vesicular type. © 1993 Wiley-Liss, Inc.     

大鼠丘脑前核内海马下托复合体谷氨酸能传入终末与皮质投射神经元的突触联系

目的 探讨大鼠丘脑前核-海马下托复合体神经元环路的突触结构及谷氨酸分布特征.方法 应用HRP束路追踪结合包埋后胶体金免疫电镜技术.结果 在丘脑前核内,可见HRP顺行标记的海马下托复合体传入轴突终末,终末多为卵圆形,内含圆形透亮突触小泡和数个线粒体.其做为突触前成分与HRP标记的树突或非HRP标记的树突形成非对称性突触.在谷氨酸胶体金免疫反应切片上,胶体金颗粒标记胞体、树突、轴突终末等.HRP标记的轴突终末和一些非HRP标记的与突触后成分形成非对称性突触的轴突终末(Gray Ⅰ型)内,胶体金颗粒密度明显大于背景(胞体、树突、Gray Ⅱ型轴突终末等)的胶体金颗粒密度.其平均胶体金颗粒密度为突触后树突的3倍多,为对称性轴突终末(Gray Ⅱ型)的6倍多.在两张邻近的连续切片,γ-氨基丁酸(GABA)胶体金免疫反应切片上,GABA胶体金颗粒浓重标记Gray Ⅱ型轴突终末,背景标记极少;而非对称性轴突终末(Gray Ⅰ型)胶体金颗粒标记极弱.谷氨酸胶体金免疫反应切片上,Gray Ⅱ型轴突终末胶体金颗粒标记极弱.GABA阳性轴突终末与HRP标记的树突形成对称性突触,在同一树突上可见GABA能轴突终末形成的对称性突触和其他轴突终末形成的非对称性突触.结论 丘脑前核内来自海马下托复合体投射神经元的轴突终末是谷氨酸能的;来自海马下托复合体皮质投射神经元轴突终末,在丘脑前核与投射至海马下托皮质的神经元树突形成非对称性轴-树突触.     

A GABA immunocytochemical study of rat motor thalamus: light and electron microscopic observations.

A light and electron microscopic study of GABA-immunoreactive neurons and profiles in the ventroanterior-ventrolateral and ventromedial nuclei of rat dorsal thalamus was conducted using antiserum raised against GABA. Less than 1% of the neurons in these motor-related nuclei exhibited GABA immunoreactivity, confirming previous reports that these nuclei are largely devoid of interneurons. Immunoreactive neurons in the ventral anterior-ventral lateral complex and ventromedial nucleus were bipolar or multipolar in shape, and tended to be smaller than non-immunoreactive neurons. GABA immunoreactivity in the neuropil consisted of labeled axon terminals and myelinated and unmyelinated axons, and was lower in the ventral anterior-ventral lateral complex and ventromedial nucleus than in neighboring thalamic nuclei. The density of neuropil immunolabeling was slightly higher in ventral anterior-ventral lateral complex than in ventromedial nucleus. GABA-immunoreactive axon terminals, collectively termed MP boutons for their medium size and pleomorphic vesicles (and corresponding to "F" profiles of some previous studies of thalamic ultrastructure), formed symmetric synapses and puncta adhaerentia contacts predominantly with large and medium-diameter (i.e. proximal) non-immunoreactive dendrites. Approximately 12 and 18% of boutons in the ventral anterior-ventral lateral complex and ventromedial nucleus, respectively, were GABA-immunopositive. Many of these immunoreactive profiles probably arose from GABAergic neurons in the thalamic reticular nucleus, substantia nigra pars reticulata and entopeduncular nucleus. Two types of non-immunoreactive axon terminals were distinguished based on differences in morphology and synaptic termination sites. Boutons with small ovoid profiles and round vesicles that formed prominent asymmetric synapses onto small-diameter dendrites were observed. Mitochondria were rarely observed within these boutons, which arose from thin unmyelinated axons. These boutons composed approximately 82 and 74% of boutons in the ventral anterior-ventral lateral complex and ventromedial nucleus, respectively, and were considered to arise predominantly from neurons in the cerebral cortex. In contrast, boutons with large terminals that contained round or plemorphic vesicles and formed multiple asymmetric synapses predominantly with large-diameter dendrites were also observed. Puncta adhaerentia contacts were also common. Mitochondria were numerous within large boutons with round vesicles, which arose from myelinated axons. Many of the large boutons were likely to have originated from neurons in the cerebellar nuclei. Approximately 6% of the boutons in the ventral anterior-ventral lateral complex and 8% in ventromedial nucleus were of the large type.(ABSTRACT TRUNCATED AT 400 WORDS)     

A golgi and a combined Golgi/GABA immunogold study of local circuit neurons in the homing pigeon hippocampus

Three types of local circuit neurons have recently been reported in the homing pigeon hippocampus. The principal type appears to be constituted by the medium-sized angular or ovoid local circuit neurons that occur in all layers of the hippocampus. The current Golgi study has revealed that these neurons can be classified according to their axonal arborisation extension: (1) in all directions, (2) principally medio-laterally, or (3) dorso-ventrally. The local circuit neurons with dorso-ventral axon arborisation are present only in the subpyramidal layer. Serial sections of a Golgi-impregnated medium-sized, multiangular local circuit neuron in the pyramidal layer and a small, ovoid neuron in the suprapyramidal layer were investigated in the electron microscope. Some of these sections were processed for GABA immunogold cytochemistry. The soma and large dendrites of both neurons displayed GABA immunogold labelling. On the soma of medium-sized local circuit neuron there were numerous terminals; on the soma of the small one relatively fewer terminals were observed. The terminals contained round and/or flat synaptic vesicles. The long axonal branches of the neurons exhibited varicosities containing flattened or pleomorphic vesicles. Axo-dendritic, axo-somatic and a few axo-axonic synapses were observed. The large dense axon arborisation field of medium-sized local circuit neurons is properly situated to modulate intrinsic hippocampal activity and that of the small local circuit neurons is well situated to modulate the hippocampal input in the suprapyramidal layer.     

Local circuit neurons in both the dentate gyrus and Ammon's horn establish synaptic connections with principal neurons in five day old rats: a morphological basis for inhibition in early development

Summary Glutamate decarboxylase (GAD)-positive and Golgi impregnated local circuit neurons of the hippocampal formation of five day old rats were examined in light and electron microscopic preparations. The ultrastructural features of these neurons were similar in both the dentate gyrus and CA1 area of Ammon's horn. Somata displayed a perikaryal cytoplasm rich in organelles but lacked organized Nissl bodies. Most nuclei showed intranuclear infoldings of varying degrees but no intranuclear sheets or rods were found. Somata and dendrites were contacted by relatively immature axon terminals that formed mainly symmetric synapses. The axons of local circuit neurons in both the dentate gyrus and Ammon's horn formed symmetric synapses with somata and dendrites of the principal neurons in these regions. Thus, both GAD-positive and Golgi-impregnated terminals of local circuit neurons were observed to form synapses with pyramidal and granule cells. These terminals were usually small and contained relatively few pleomorphic synaptic vesicles. The results show that a circuitry for inhibition is established in the 5 day old dentate gyrus and Ammon's horn, even though the local circuit neurons lack some of the typical adult ultrastructural features at this age.     

Electron microscopic identification of reticulo-subthalamic axon terminals in the cat

Synaptic boutons emanating from axons of nucleus tegmenti pedunculopontinus origin were identified by electron microscopy in the neuropil of the subthalamic nucleus. Such boutons measure 1.5-3 microns, contain round synaptic vesicles and make asymmetrical axodendritic and axosomatic synaptic contacts with large subthalamic neurons. Very few contacts with vesicle-containing dendrites, and no contacts with the perikarya of the small neurons were observed. The present findings, in keeping with the relevant light microscopic and electrophysiologic data, furnish evidence for a substantial bilateral tegmenti pedunculopontinofugal projection that excites monosynaptically the relay subthalamic neurons.     

Ultrastructural features of non-commissural GABAergic neurons in the medial vestibular nucleus of the monkey.

The ultrastructural characteristics of non-degenerating GABAergic neurons in rostrolateral medial vestibular nucleus were identified in monkeys following midline transection of vestibular commissural fibers. In the previous papers, we reported that most degenerated cells and terminals in this tissue were located in rostrolateral medial vestibular nucleus, and that many of these neurons were GABA-immunoreactive. In the present study, we examined the ultrastructural features of the remaining neuronal elements in this medial vestibular nucleus region, in order to identify and characterize the GABAergic cells that are not directly involved in the vestibular commissural pathway related to the velocity storage mechanism. Such cells are primarily small, with centrally-placed nuclei. Axosomatic synapses are concentrated on polar regions of the somata. The proximal dendrites of GABAergic cells are surrounded by boutons, although distal dendrites receive only occasional synaptic contacts. Two types of non-degenerated GABAergic boutons are distinguished. Type A terminals are large, with very densely-packed spherical synaptic vesicles and clusters of large, irregularly-shaped mitochondria with wide matrix spaces. Such boutons form symmetric synapses, primarily with small GABAergic and non-GABAergic dendrites. Type B terminals are smaller and contain a moderate density of round/pleomorphic vesicles, numerous small round or tubular mitochondria, cisterns and vacuoles. These boutons serve both pre- and postsynaptic roles in symmetric contacts with non-GABAergic axon terminals. On the basis of ultrastructural observations of immunostained tissue, we conclude that at least two types of GABAergic neurons are present in the rostrolateral portion of the monkey medial vestibular nucleus: neurons related to the velocity storage pathway, and a class of vestibular interneurons. A multiplicity of GABAergic bouton types are also observed, and categorized on the basis of subcellular morphology. We hypothesize that "Type A" boutons correspond to Purkinje cell afferents in rostrolateral medial vestibular nucleus, "Type B" terminals represent the axons of GABAergic medial vestibular nucleus interneurons, and "Type C" boutons take origin from vestibular commissural neurons of the velocity storage pathway.     

Quantitative ultrastructure of synapses on functionally identified primary afferent neurons in the cat trigeminal mesencephalic nucleus

Though a number of studies have reported the presence of synapses on neurons in the trigeminal mesencephalic nucleus (Vmes), there have been no quantitative studies of either the density of innervation, or the ultrastructure, of the synapses on single, physiologically identified neurons in this nucleus. In this study we recorded from single neurons in the Vmes, identified them as being either muscle spindle afferents (MS) or periodontal ligament mechanoreceptor afferents (PL), and then labeled the neurons by intra-axonal injection of horseradish peroxidase (HRP). The material was first processed to reveal the HRP activity, following which ultrathin sections through the labeled somata were cut and examined under the electron microscope. Complete serial reconstructions were made through the soma of one MS neuron and one PL neuron, and the contacts on the neurons reconstructed. Boutons were found on the soma, spines, appendages and the axon hillock and the initial segment of the axon. The numbers of boutons terminating on the two neurons were 198 (PL) and 424 (MS), giving a packing density of 4.4 and 10.7 boutons respectively (i.e., number of boutons/100 micron 2 of the postsynaptic membrane). Boutons could be separated into two types on the basis of their vesicles: those containing clear, round vesicles (i.e., S-type) and those containing a mixture of round, oval and flattened vesicles (P-type). Ninety-five (PL neuron) and 99% (MS neuron) of terminals on the two neurons were P-type. All the S-type boutons and 80% of the P-type boutons formed asymmetric synaptic contacts while 10% of the P-type boutons made symmetric contacts. Quantitative measurements of the P-type boutons on the labeled neurons, in which the data of MS and PL neurons were pooled, revealed that bouton volume was highly correlated with bouton surface area, active zone number, total active zone area, vesicle number, and mitochondrial volume. However, comparing the quantitative measurements of the P-type boutons with those of previously reported vibrissa afferent terminals and their associated axon terminals revealed that all the parameters were smaller for the P-type boutons (on Vmes neurons) than those of the vibrissa afferent terminals but similar to those of axon terminals presynaptic to the vibrissa afferents. Taken together, our results emphasize the wide scope for synaptic interactions in the Vmes and suggest that it may be more fruitful to view the Vmes as an integrating center.     

Ultrastructure of cultured rat neostriatum

Four different types of neurons were identified in cultures of newborn rat neostriatum. Small and medium-sized neurons were most numerous. A few large neurons and some very small ‘microneurons’ were observed. The morphology of medium-sized neurons varied, and this group may contain more than one functional subgroup. Axosomatic synapses were associated with all types of neurons, but most of them made contacts with medium-sized neurons. All axodendritic synapses made symmetrical contacts, with or without synaptic membrane thickenings. A great majority of terminal boutons contained small, round, clear vesicles. A few terminals with large pleomorphic clear vesicles were seen. Large granular vesicles were found in the peripheral cytoplasm of some medium-sized neurons, dendrites and axon terminals. No terminals contained exclusively large granular vesicles, but in some terminals they were more numerous than the small clear vesicles. The dense core of the large granular vesicles was resistant to reserpine treatment. Kainic acid did not cause specific degenerative changes. The presence of several morphologically distinct populations of neurons renders it possible to study the nature of these cells in different experimental conditions. Intrinsic neostriatal synaptic contacts appeared to be symmetrical, although it is possible that some of them have the capacity to develop asymmetrical contacts. The lack of effect of kainic acid may be explained by the early maturational stage of the cells or by the lack of extrinsic contacts. More functional studies are necessary before the usefulness of these cultures for investigating neostriatal function can be assessed.     

Quantitative analysis of the ultrastructural distribution of GABA-like immunoreactivity in the intermediate and medial part of hyperstriatum ventrale of chick

Summary The intermediate and medial part of the hyperstriatum ventrale of the chick telencephalon plays a crucial role in the learning processes of imprinting. The distribution within the intermediate and medial part of the hyperstriatum ventrale of the neurotransmitter -amino butyric acid was studied with light and electron microscopy using an antibody against this amino acid. The antibody labelled 18.4% of neuronal somata. GABA-labelled terminals made symmetrical synapses onto somata and dendrites of labelled and unlabelled neurons. Labelled somata received about three times as many synaptic boutons as unlabelled somata. Approximately 21% of synaptic terminals on labelled somata were themselves labelled; unlabelled somata received a higher proportion (37.6%) of such terminals. Most labelled terminals synapsing with dendrites were confined to the shafts; very few labelled terminals contributed to axospinous synapses. Synaptic contacts made on dendritic shafts by labelled boutons were intermingled with symmetrical and asymmetrical contacts from non-immunoreactive terminals. The proportion of labelled terminals received by labelled dendrites (33.1%) was approximately twice that received by unlabelled dendrites (15.9%). Labelled neurons therefore received a higher proportion of labelled terminals on their dendrites and a lower proportion on their somata compared with unlabelled neurons. No immunoreactivity was seen in glial cells or ependyma.     

Immunocytochemical study of GABAergic neurons containing the calcium-binding protein parvalbumin in the rat hippocampus

Summary The structural features of PV-immunoreactive (PV-I) neurons, a particular subpopulation of GABAergic neurons, in the hippocampus were studied by immunocytochemistry. The PV-I cell bodies were concentrated within the stratum pyramidale (SP) and stratum oriens (SO) in the hippocampus. PV-I puncta were frequent in SP, while they were rarely seen in other layers. The dendritic arborization of PV-I neurons resembled that of some of the nonpyramidal cells observed after Golgi-impregnation. The most commonly observed PV-I neurons had their perikarya located in SP with dendrites extending into SO and the stratum radiatum (SR). Most of the dendrites in SR had typical beaded or varicose segments. The dendrites extending into SO had few beaded parts. There were many bipolar and multipolar neurons with smooth dendrites in SO, but only a small number of such multipolar neurons in SR. An electron microscopic analysis revealed that PV-I products were located to perikarya, dendrites, myelinated axons and synaptic boutons. The perikarya of PV-I neurons exhibited several ultrastructural features of nonpyramidal cells, e.g., abundant cisternae of endoplasmic reticulum, mitochondria and other perikaryal organelles, an infolded nuclear envelope and intranuclear inclusions. They received many asymmetric synapses with round presynaptic vesicles. There were numerous PV-I boutons, presumably axonal endings, covering the pyramidal cell bodies. The PV-I boutons also contacted the axon initial segments and proximal dendrites of the pyramidal cells. In addition PV-I terminals were found on somata and dendrites of both PV-I or PV-negative nonpyramidal cells. The results suggest that PV-containing neurons include basket and axo-axonic cells.     

Synaptic terminals in the dorsal lateral geniculate nucleus from neurons of the thalamic reticular nucleus: A light and electron microscope autoradiographic study

(³H)-proline was injected in the caudodorsal part (visual segment) of the thalamic reticular nucleus to study its projection to the dorsal lateral geniculate nucleus. This was done by autoradiographic tracing of anterograde axonal transport of tritium at the light- and electron microscopic level. The results of the light-microscope autoradiography show that connections of the thalamic reticular nucleus are distributed along lines of projections in the dorsal lateral geniculate nucleus, indicating a retinotopic arrangement of this projection. The results of the electron microscope autoradiography provide direct evidence that axons of cells in the thalamic reticular nucleus terminate in the dorsal lateral geniculate nucleus as synaptic boutons that contain flattened synaptic vesicles, dark mitochondria and establish symmetrical synapses with perikarya and with proximal, intermediate and distal dendrites. They do not take part in intraglomerular synapses (as boutons with pleomorphic synaptic vesicles do) and are not postsynaptic to other vesicle-containing boutons in the dorsal lateral geniculate nucleus.The present results, taken in conjunction with physiological studies that have shown postsynaptic inhibitory effects of the thalamic reticular nucleus on dorsal lateral geniculate nucleus relay cells in the rat, establish a correlation of an inhibitory synapse with the presence of flattened synaptic vesicles in the corresponding synaptic boutons. Also, the observation that thalamic reticular nucleus terminals in the dorsal lateral geniculate nucleus avoid forming synapses with boutons containing pleomorphic vesicles, believed to be synaptic processes of interneurons, is indicative that the inhibitory effects are exerted monosynaptically on geniculate relay cells.     

On the identification of the afferent axon terminals in the nucleus lateralis of the cerebellum an electron microscope study

Summary Axon terminals in the neuropil of the lateral nucleus can be divided into six classes, each with a specific constellation of characteristics that consistently occur together. Two of these classes have synaptic varicosities with elliptical synaptic vesicles, one in a dense, the other in a sparse matrix, and both make axosomatic and axodendritic synapses. The remaining four classes all have round synaptic vesicles and do not make axosomatic synapses. In the first of these four, the vesicles are tightly packed in a dense matrix, in another they are loosely dispersed, and in the third they are clustered. In the fourth, large granular vesicles predominate. Of these six classes, the most numerous belong to the axons of the Purkinje cell terminal arborization. These boutons resemble their counterparts in the cerebellar cortex, the recurrent collaterals of the Purkinje axon. They have elliptical and flat synaptic vesicles in a dark matrix. The varicosities terminate on somata and dendrites of large and small neurons and constitute the majority of their input. Purkinje axons constitute 86% of the total population of terminals on large neuronal perikarya and 50% of those on their dendrites, but only 78% on the somata of small neurons and 31% on their dendrites. The terminals of climbing fiber collaterals are recognized by their resemblance in electron micrographs to the terminals of the climbing fiber arborization in the cerebellar cortex. They bear round synaptic vesicles packed into a dense axoplasmic matrix and make Gray's type 1 axodendritic synapses with large and small neurons. These axons are restricted to the lateral and ventral aspects of the nucleus and constitute 5% of the terminals on large cell dendrites and 6% of those on small neurons. The axons tentatively identified as collaterals of mossy fibers are myelinated fibers with a light axoplasm containing round synaptic vesicles, dispersed throughout their varicosities. They make Gray's type 1 synapses and constitute a fair percentage of the total axodendritic contacts in the neuropil, 22% on large neurons and 28% on small neurons. The bases for these tentative identifications are discussed in detail, as are the various synaptic relationships undertaken by each class of axon. The remaining 4 classes of axons of the neuropil will be described in subsequent papers.Supported in part by U.S. Public Health Service grants NS 10536 and NS 03659, Training grant NS 05591 from the National Institute of Neurological Diseases and Stroke, and a William F. Milton Fund Award from Harvard University.     

A light and electron microscopic study of NADPH-diaphorase-, calretinin- and parvalbumin-containing neurons in the rat nucleus accumbens

The rat nucleus accumbens contains medium-sized, spiny projection neurons and intrinsic, local circuit neurons, or interneurons. Sub-classes of interneurons, revealed by calretinin (CR) or parvalbumin (PV) immunoreactivity or reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry, were compared in the nucleus accumbens core, shell and rostral pole. CR, PV and NADPH-diaphorase-containing neurons are shown to form three non-co-localising populations in these three areas. No significant differences in neuronal population densities were found between the subterritories. NADPH-diaphorase-containing neurons could be further separated morphologically into three sub-groups, but CR- and PV-immunoreactive neurons form homogeneous populations. Ultrastructurally, NADPH-diaphorase-, CR- and PV-containing neurons in the nucleus accumbens all possess nuclear indentations. These are deeper and fewer in neurons immunoreactive for PV than in CR- and NADPH-diaphorase-containing neurons. CR-immunoreactive boutons form asymmetrical and symmetrical synaptic specialisations on spines, dendrites and somata, while PV-immunoreactive boutons make only symmetrical synaptic specialisations. Both CR- and PV-immunoreactive boutons form symmetrical synaptic specialisations with medium-sized spiny neurons and contact other CR- and PV-immunoreactive somata, respectively. A novel non-carcinogenic substrate for the peroxidase reaction (Vector Slate Grey, SG) was found to be characteristically electron-dense and may be distinguishable from the diaminobenzidine reaction product. We conclude that the three markers used in this study are localised in distinct populations of nucleus accumbens interneurons. Our studies of their synaptic connections contribute to an increased understanding of the intrinsic circuitry of this area.     

Ultrastructural identification of synaptic terminals from cortical axons and from collateral axons of geniculo-cortical relay cells in the perigeniculate nucleus of the cat

Summary Electron microscopic analysis of sections of the perigeniculate nucleus (PGN) of the cat processed with horseradish peroxidase (HRP) histochemistry after massive injections of this enzyme in the visual cortex showed two types of synaptic terminals labeled with HRP reaction products. One type (RLD terminals) is characterized by round synaptic vesicles, large size, dark mitochondria and asymmetrical synaptic contacts with somata and dendrites. The second type (RSD terminals) is characterized by round synaptic vesicles, small size, dark mitochondria and asymmetrical synaptic contacts with dendrites. The HRP + RSD terminals, which were also found in the dorsal lateral geniculate nucleus (LGN), are interpreted as terminals of cortical origin both in the PGN and LGN, since previous studies have identified cortical terminals as being of RSD type in the LGN and in other thalamic nuclei. The HRP + RLD terminals are interpreted as synaptic terminals of collaterals axons of geniculo-cortical relay cells in the PGN labeled by retrograde transport of HRP from the cortex. In addition, in semithin and ultrathin sections somata in the PGN were never found labeled with HRP products indicating the absence of a PGN projection to the visual cortex.     

GABAergic cells in the dentate gyrus appear to be local circuit and projection neurons

Immunocytochemical results indicate that GAD-positive neurons are found in the molecular and granule cell layers of the dentate gyrus as well as in the hilar region. GAD-positive cells in the molecular and granule cell layers are identified as various types of local circuit neurons. Most of the GAD-positive puncta found throughout the molecular layer and within the granule cell layer are interpreted as axon terminals of these neurons, including five types of basket cells. This interpretation is based on data that indicate the axons of basket cells form synapses with the somata and proximal dendrites of granule cells. The results in the hilus show that 60% of the hilar neurons are GAD-positive. Since previous studies have indicated that 80% of hilar neurons give rise to both associational and commissural pathways, many GABAergic neurons in the hilus are probably projection neurons. This finding is consistent with recent physiological data which suggest that commissural pathway stimulation directly inhibits granule cells. Therefore, GABAergic cells in the dentate gyrus appear to be both projection and local circuit neurons.     

Aspinous and sparsely-spinous stellate neurons in the visual cortex of rats contain glutamic acid decarboxylase

Summary Glutamic acid decarboxylase (GAD), the enzyme that synthesizes the neurotransmitter -aminobutyric acid (GABA), has been localized in the rat visual cortex by immunocytochemical methods with both light and electron microscopy. In both colchicine-injected and non-injected preparations of the visual cortex, GAD-positive reaction product was observed in somata, proximal dendrites and axon terminals of non-pyramidal neurons. The GAD-positive terminals were observed to form symmetric synaptic junctions most commonly with dendritic shafts and somata of pyramidal and stellate neurons and less frequently with initial axon segments of pyramidal neurons and dendritic spines. In colchicine-injected preparations, GAD-positive somata were located in all cortical layers including the immediately subjacent white matter. In contrast, sections from non-injected rats displayed GAD-positive somata within a superficial and a deep cortical band. The GAD-positive somata observed in both types of preparations received both symmetric and asymmetric synaptic junctions, lacked apical dendrites, and had radially oriented dendrites of small diameter. These characteristics of GAD-positive neurons indicate that they are aspinous and sparsely-spinous stellate neurons. The localization of GAD within these neurons in combination with physiological and pharmacological data indicate that these local circuit neurons mediate GABA-ergic inhibition in the neocortex.