Immunocytochemical localization of gamma-aminobutyric acid (GABA) in the cat superior colliculus

This paper reports the pattern of labeling in the cat superior colliculus produced by an antiserum raised against BSA-conjugated gamma aminobutyric acid (GABA) and visualized by light and electron microscope immunocytochemistry. Neuropil labeling was densest within the zonal and superficial gray layers but was also found in the deep layers. Neurons labeled by the GABA antibody were also most dense within the zonal and superficial gray layers, although many labeled neurons were also found in the deeper layers. The ratio of labeled to unlabeled cells varied from an average of 45% in the superficial subdivision and the intermediate gray layer to less than 30% in the deeper laminae. Almost all intensely labeled cells were small (mean area = 127 micron 2) and had varied morphologies. Several types of labeled cell were observed with the electron microscope. One type had a horizontal, fusiform cell body and a deeply invaginated nucleus. Another type had a small round or ovoid cell body with cytoplasm clumped at one end. Labeled cells with other morphologies were also occasionally seen. No labeled glial cells were found. Two types of vesicle-containing dendrite were stained by the GABA antibody. One type had loose accumulations of small synaptic vesicles and often received input from retinal terminals. Another type had spines also containing small synaptic vesicles. Labeled dendrites without synaptic vesicles were also seen frequently. Putative axon terminals labeled by the GABA antibody had densely packed synaptic vesicles and formed symmetric synaptic contacts. Labeled myelinated axons were also commonly found. These results confirm those using uptake of tritiated GABA (Mize et al.: J. Comp. Neurol. 202:385-396, '81, J. Comp. Neurol, 206:180-192, '82) in that two of the same classes of GABA neuron, horizontal I and granule I cells, were identified in the superficial laminae. However, the GABA antiserum used in this study also revealed a third class of GABA neuron with vesicle-containing spines. The antiserum also labeled a significant number of putative GABAergic neurons located in the deep subdivision of the cat superior colliculus which were not previously recognized by using transmitter autoradiography.     

Superior colliculus neurons which project to the cat lateral posterior nucleus have varying morphologies

We have studied the laminar position, morphology, and synaptic relationships of neurons in the cat superior colliculus which project to the interadjacent division of the lateral posterior nucleus (LPi), using the retrograde transport of horseradish peroxidase. The neurons which project to LPi are remarkably varied in depth, size, morphology, and synaptic density and appear to consist of at least four cell types. Labeled cells were found laminae. Forty-seven percent were found in the superficial gray layer (50-550 micrometer), all but a few within its deep subdivision. Forty-seven percent were located in the optic layer (550-1,200 micrometer), the majority of these being within the upper one-half of the layer Seven percent were found in the intermediate and deep gray layers (below 1,200 micrometer). Cell body area varied widely, ranging from 37 to 768 micrometer 2 (mean of 243 micrometer2). Based on cell size, shape, and dendritic field orientation, we identified four distinct cell morphologies which were labeled. Thirty-five percent were stellate, 32% were vertical fusiform 19% were granule, and 12% were horizontal cells. Electron microscope analysis confirmed that neurons projecting to the lateral posterior nucleus are a morphologically diverse group. A sample of 71 labeled cells varied significantly in density of synaptic input as well as in size, shape, depth, dendritic distribution, and cytology.     

Enkephalin-like immunoreactivity in the cat superior colliculus: distribution, ultrastructure, and colocalization with GABA

The distribution of enkephalin (ENK) immunoreactivity has been examined in the cat superior colliculus (SC) by means of light and electron microscope immunocytochemistry. The antisera were directed against leucine enkephalin but also recognized methionine enkephalin. Colocalization of ENK with gamma aminobutyric acid (GABA) was studied with a two-chromagen double-labeling technique. Enkephalin antiserum labeling was highly specific. Dense neuropil labeling was found only in a thin band 75-100 microns wide within the upper superficial gray layer of SC. Negligible neuropil labeling was seen deeper, except for patches of label within the intermediate gray layer. Intensely labeled neurons also had a specific distribution. Forty-seven percent were located within the upper 200 microns of SC, 40% within the deep superficial gray layer, 11% in the optic layer, and only 2% below that layer. Almost all ENK-labeled cells were small (mean area of 117 microns2). Some of these had horizontal fusiform cell bodies and horizontally oriented dendrites. Others had small round somata and thin, obliquely oriented dendrites. In double-labeling experiments, 18% of anti-ENK-labeled cells were also immunoreactive for GABA. Four distinct types of ENK-labeled profile were identified with the electron microscope. Presynaptic dendrites (PSD) with loose accumulations of synaptic vesicles were densely labeled with the antiserum. Conventional dendrites were also labeled. Both types of labeled profile received input from unlabeled synaptic terminals, including those from the retina that contained pale mitochondria and round synaptic vesicles and formed asymmetric synaptic contacts. Retinal terminals were never labeled with the antisera. However, some axon terminals with round synaptic vesicles, dark mitochondria, and symmetric synaptic densities were labeled by the antisera, as were some thinly myelinated axons. These results show that there is a small population of enkephalinergic neurons in the cat SC, some of which also contain GABA. Because not all cells with identical morphologies were double labeled, it appears that neurons of like morphology are chemically heterogeneous.     

A Golgi-electron microscopic study of fusiform neurons in the hilar region of the dentate gyrus

The fusiform cells of the dentate gyrus are located in a portion of the hilus within 100 micron of the granule cell layer. They have ovoid somata and bipolar dendrites that generally run parallel to the granule cell layer. The dendrites of these cells are either spiny or sparsely spiny. The spiny fusiform cell has numerous spines along its dendrites, which are contacted by terminals with the features of granule cell axon collaterals. This cell type also displays somal spines that are contacted by similar terminals. In contrast, the sparsely spiny fusiform cell displays only a few spines, which are contacted by multiple small axon terminals that synapse with both the stalk and end bulb of the spine. Most synaptic input for this cell type is made with the smooth surfaces of the soma and dendrites. A variety of terminals form synapses with the sparsely spiny fusiform cell, including terminals that resemble the fine axon collaterals of mossy fibers. The somata of these two cell types also display differences in the amount of Nissl bodies and the degree of nuclear infolding. The results indicate that spiny fusiform cells are similar to mossy cells, another hilar cell type that receives its major synaptic input from axon collaterals of mossy fibers from granule cells. The distribution of the dendrites of spiny fusiform cells and the pattern of granule cell axon collaterals suggest a high degree of convergence from granule cells. In contrast, the variety of axodendritic synapses for sparsely spiny fusiform cells suggests that more diverse inputs affect this cell's activity. Therefore, the structure and circuitry of these two hilar cell types are probably different. This study adds further evidence to indicate that the hilus contains a large variety of cell types with different neuronal connections.     

Ultrastructural study of large efferent neurons in the superior colliculus of the cat after retrograde labeling with horseradish peroxidase

The ultrastructure of large neurons in the stratum griseum intermedium of the cat superior colliculus was examined following injections of horseradish peroxidase (HRP) into the dorsal tegmental decussation. Four HRP-labeled cells were selected, and the synaptology of their cell bodies and selected regions of proximal and distal dendrites was examined. The four neurons represent four morphologically distinct cell types: multipolar radiating, tufted, large vertical, and medium-sized trapezoid radiating. These four neurons correspond with cell types X1, X2, X3, and T1 respectively, according to the recent classification of neurons in the superior colliculus of the cat by Moschovakis and Karabelas (J. Comp Neurol. 239:276-308, '85). The three X type neurons are similar in having 83% of their somata and over 74% of their proximal dendrites contacted by synaptic profiles. Distal dendrites of the X type neurons, however, receive fewer synaptic contacts. In contrast, in the T1 cell, only 69% of the soma membrane is contacted by synaptic profiles, and the synaptic coverage on proximal and distal dendrites does not vary much from this. Of the eight types of synaptic terminals described in the stratum griseum intermedium of the cat superior colliculus by Norita (J. Comp. Neurol. 190:29-48, '80), only five are found in contact with the X and T type efferent neurons described here. There are some regional differences in terminal distribution, although each terminal is represented on each cell. Type III terminals (small, contain mostly pleomorphic vesicles, and make symmetrical contacts) are the most abundant on cell bodies and dendrites of all four cell types. Terminal types II (medium-sized, containing round and flattened vesicles, and making asymmetrical contacts), and IV (medium to large in size, containing flattened vesicles, and making symmetrical contacts) are well represented. In general, terminal types I (small, containing densely packed round vesicles, and making asymmetrical contacts) and VI (small and irregular in shape, containing flattened vesicles and making symmetrical contacts) are found infrequently. The identity of different types of synaptic terminal is discussed.     

Fine structure of identified neurons in the primate hippocampus: a combined Golgi/EM study in the baboon

The hippocampi of two 1-year-old female baboons (Papio anubis) were used for a combined Golgi/electron microscope (EM) study of characteristic cell types in the hippocampus proper and fascia dentata. Results were compared with previous Golgi/EM studies of hippocampal neurons in small laboratory animals. Cell bodies of pyramidal neurons in CA1 were more loosely distributed than known from studies on the rat or guinea pig. Numerous basal and horizontal dendrites originating from the perikaryon filled in the space between neighboring cell bodies. Apical stem dendrites were varying in length, depending on the position of the parent cell body in outer or inner portions of the pyramidal layer. Dendrites were densely covered with spines which in the EM showed very complex synaptic contacts. In contrast to our observations in rats and guinea pigs, CA3 pyramidal cells in the monkey hippocampus exhibited numerous large spines or excrescences not only on apical dendrites but also on basal dendrites running through stratum oriens. These excrescences appeared to be more complex than in small rodents. They often branched, protruding deeply into presynaptic mossy fiber boutons, and formed multiple asymmetric synaptic contacts. Granule cells of the monkey fascia dentata, in contrast to those of the rodent, occasionally had basal dendrites extending into the hilar region. In the EM, granule cells either with or without basal dendrites exhibited fine structural characteristics that were very similar to those described in Golgi/EM studies of granule cells in the rat fascia dentata. Of the various types of nonpyramidal neurons the horizontal cells in stratum oriens with dendrites parallel to the alveus were analyzed. As seen in rats, these cells exhibited large amounts of rough endoplasmic reticulum, indentations of the nuclear membrane, and nuclear inclusions. Numerous terminals formed synaptic contacts on dendritic shafts. In contrast to rodents, numerous spines arose from dendrites and cell bodies of these neurons. In the EM, often single spines were found to establish synaptic contacts with several presynaptic boutons. In summary, our correlated light and EM study of four characteristic cell types, which are present in both nonprimates and primates, demonstrates a much more complex dendritic pattern and synaptic organization of these neurons in primates than in commonly studied small laboratory animals.     

Organization of the septal region in the rat brain: a Golgi/EM study of lateral septal neurons

The combined Golgi/electron microscope (EM) technique was used to analyze the fine structure and synaptic organization of the various types of neurons in the rat lateral septum (LS), i.e., in the dorsolateral (LSd), intermediolateral (LSi), and ventrolateral (LSv) nuclei of the septal complex. Two characteristic cell types were observed in the LSd: type I with thick, short dendrites densely covered with short spines, and type II with longer and thinner dendrites exhibiting fewer but longer spines. This latter type was by far the most frequently impregnated cell type in the LSd and was also present in the LSi. Synaptic contacts on spines of either cell type were asymmetric; the majority of the presynaptic boutons contained clear round synaptic vesicles. Occasionally terminals were found that contained both clear and dense-core vesicles. Typical fusiform neurons with a low number of spines and rather long dendrites, sometimes invading other LS nuclei, were found in the LSi. The LSv contained numerous small neurons with small dendritic fields. A relatively large number of terminals with dense-core vesicles were found to establish synaptic contacts with identified LSv neurons. The morphological heterogeneity of LS neurons is discussed with regard to other studies on afferent and efferent fiber systems as well as immunohistochemical studies of this particular region of the septal complex.     

Variations in the retinal synapses of the cat superior colliculus revealed using quantitative electron microscope autoradiography

This study has examined the retinal synapses of the cat superior colliculus using electron microscope autoradiography and morphometric techniques. The depth of each retinal synapse was measured using a computer-based EM plotter. The area, perimeter, and synapse contact density of selected synapses were calculated using a computer-based digitizer. Pale mitochondria were found to be an accurate cytological marker of retinal input to the colliculus. Fifty-eight percent of pale mitochondria terminals were labeled in the colliculus contralateral to eye injections. Ten percent of pale mitochondria terminals were labeled in the ipsilateral colliculus. A few labeled terminals contained dark mitochondria. The labeled retinal terminals in the contralateral colliculus were concentrated in a 60 μm wide dense band at the top of the superficial gray layer. They were also found within the deep superficial gray and upper optic layers. This distribution corresponded exactly to a larger population of pale mitochondria terminals. The cross-sectional area and synaptic contact density of selected pale mitochondria terminals varied with depth. Within the upper superficial gray, the terminals were small (mean area= 1.26 μ²) and high contact densities (mean= 0.25 per μm). These small terminals were also found deeper within the colliculus. Below the upper subdivision of the superficial gray, some labeled terminals were much larger and had lower contact densities. These results suggest there may be two subpopulations of retinal terminal in the cat superior colliculus: (1) small terminals with scalloped contours and complex synaptic relationships which may correspond to W-type input; and (2) larger terminals with simpler synaptic relationships which are distributed deeper and may correspond to Y-type input.     

Synaptic connections of neuropeptide Y (NPY) immunoreactive neurons in the hilar area of the rat hippocampus

Synaptic connections and fine structural characteristics of neuropeptide Y-immunoreactive (NPY-i) neurons in the fascia dentata were studied using an antiserum against NPY. Normal and colchicine pretreated rats were examined to study the synaptic connections of NPY-i neurons in the normal fascia dentata. The perforant pathway and fimbria fornix were transected to label afferent fibers to NPY-positive cells. Horseradish peroxidase conjugated with wheat germ agglutinin (HRP-WGA) was injected into the contralateral hippocampus to study commissural projections of hippocampal NPY-i neurons, and to search for NPY-i synaptic contacts on immunonegative commissural cells. Since earlier reports have shown that at least half of the NPY-i neurons also contain somatostatin (SS), the distribution of NPY-i neurons in the hilar area was determined and compared with that of SS-i neurons. Four types of dentate NPY-i neurons were distinguished: Type 1: large multipolar cells in the deep hilus (9%). Type 2: medium-sized multipolar and fusiform hilar neurons with dendrites occasionally reaching the outer molecular layer (64%). Type 3: pyramidal shaped cells in the granule cell layer with long apical dendrites reaching the outer molecular layer (20%). Type 4: small multipolar NPY-i cells located in the molecular layer (7%). Our results indicate two overlapping but not identical cell populations of NPY-i and SS-i neurons. Light and electron microscopic analysis of the normal fascia dentata demonstrated that the majority of NPY-i terminals are located in the outer molecular layer of the dentate gyrus, where they establish symmetric synaptic contacts on dendritic shafts and occasionally on spines of granule cells. A moderate number of NPY-i synapses were also found on dendrites in the inner molecular layer and on the cell body of granule cells. Numerous symmetric NPY-i synapses were found on dendrites and somata of neurons in the hilar area. Some NPY-i dendrites in the hilar area received mossy axon collateral input. After transection of the perforant pathway degenerated axon terminals could be found in synaptic contact with NPY-i dendrites in the outer molecular layer. Commissurotomy revealed direct commissural input to NPY-i dendrites in the inner molecular layer and in the hilus. After injection of HRP-WGA into the contralateral hippocampus 2% of hilar NPY-i neurons were retrogradely labeled and symmetric NPY-i synapses were found on the cell bodies and dendrites of unstained HRP-WGA labeled neurons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Organization and synaptic connections of cholinergic fibers in the cat superior colliculus

The cat superior colliculus (SC) receives a dense cholinergic input from three brainstem nuclei, the pedunculopontine tegmental nucleus, the lateral dorsal tegmental nucleus, and the parabigeminal nucleus (PBG). The tegmental inputs project densely to the intermediate gray layer (IGL) and sparsely to the superficial layers. The PBG input probably projects only to the superficial layers. In the present study, the morphology of choline acetyltransferase (ChAT)-immunoreactive axons and synaptic endings in the superficial and deep layers of the SC was examined by light and electron microscopy to determine whether these cholinergic afferents form different types of synapses in the superifical and deep layers. Two types of fibers were found within the zonal (ZL) and upper superficial gray layers (SGL): small diameter fibers with few varicosities and larger diameter fibers with numerous varicosities. Quantitative analysis demonstrated a bimodal distribution of axon diameters, with one peak at approximately 0.3–0.5 μm and the other at 0.9–1.0 μm. On the other hand, ChAT-immunoreactive fibers in the IGL were almost all small and formed discrete patches within the IGL. Two types of ChAT-immunoreactive synaptic profiles were observed within the ZL and upper SGL using the electron microscope. The first type consisted of small terminals containing predominantly round synaptic vesicles and forming asymmetric synaptic contacts, mostly on dendrites. The second type was comprised of varicose profiles that also contained round synaptic vesicles. Their synaptic contacts were always symmetric in profile. ChAT-immunoreactive terminals in the IGL patches contained round or pleomorphic synaptic vescles, and the postsynaptic densities varied from symmetric to asymmetric, including intermediate forms. However, no large varicose profiles were observed. This study suggests that cholinergic fibers include at least two differnt synaptic morphologies: small terminals with asymmetric thickenings and large varicose profiles with symmetric terminals. The large varicose profile in the superficial layers is absent in the IGL. This result suggests that the cholinergic inputs that innervate the superficial layers and the patches in the IGL of the cat SC differ in their synaptic organization and possibly also in their physiological actions. © 1993 Wiley-Liss, Inc.     

Synaptic termination of afferents from the ventrolateral nucleus of the thalamus in the cat motor cortex. A light and electron microscopy study

The synaptic termination in the cat motor cortex of afferents from the ventrolateral nucleus of the thalamus (VL) has been studied with experimental light and electron microscopic methods. The distribution of normal synapses on motor cortex pyramidal, stellate, and Betz cells was also examined. Synapses in the motor cortex can be classified into two general types. The first and most prominent type contains flat vesicles, lacks a compact postsynaptic density, and corresponds to Colonnier's ('68) symmetrical synapse. Stellate neurons receive synapses of both types on their cell bodies and proximal dendritic shafts, while pyramidal cells have only symmetrical synapses at these sites. The dendritic spines of both stellate and pyramidal cells are contacted by predominantly asymmetrical synapses. Betz cells, like smaller pyramidal neurons, receive only symmetrical synapses on their cell bodies. The proximal portions of the Betz cells apical dendrites, however, receive both asymmetrical and symmetrical synapses. Following VL lesions, degenerating synapses were mainly found in three cortical layers: the upper third of layer I (18%), layer III (66%), and layer VI (13%). Degenerating synapses were not seen in the lower two-thirds of layer I or in layer II, and were only rarely seen in layer V (3%). Ninety-one percent of the VL synapses were found on spines and 8% on stellate-type dendritic shafts. Stellate cell bodies rarely received VL synapses (1%) and none occurred on pyramidal or Betz cell bodies and their proximal dendrites. A VL synapse within layer III was found on two dendritic spines of a Betz cell apical dendrite. Thus, part of the VL input to layer III synapses on the processes of both motor cortex output neurons (Betz cells in layer V) and cortical interneurons (stellate cells in layer III).     

An EM-autoradiographic analysis of the projection from cortical areas 17, 18, and 19 to the superior colliculus in the cat

The electron microscopic autoradiographic method was used to identify terminals of axons from cortical areas 17, 18, and 19 in the superficial layers of the superior colliculus. The results show that terminals of area 17 neurons contain round vesicles and made asymmetrical synaptic contacts predominantly onto one or more dendrites or dendritic appendages. Some profiles postsynaptic to labeled terminals contain vesicles and presumably are involved in serial synaptic arrangements. Terminals of area 18 and 19 neurons in the superficial collicular layers appear to comprise two populations, one similar in most respects to area 17 terminals, containing round vesicles and making asymmetrical contacts. The other contains pleomorphic vesicles and makes symmetrical contacts upon dendrites and dendritic appendages. These terminals rarely contact more than one postsynaptic profile, and rarely do the postsynaptic profiles contain vesicles. The two populations of area 18 and 19 terminals containing round and pleomorphic vesicles, respectively, are present in the ratio of approximately 3:1, although this ratio varies throughout the sublaminae of the superficial collicular layers. The presence of two distinct types of cortical terminals in the colliculus suggests that cortical modulation of collicular processing is more complex than was previously conceived.     

Structure of the piriform cortex of the opossum. I. Description of neuron types with golgi methods

The piriform cortex was studied in the adult opossum with rapid Golgi and Golgi-Cox techniques. Most pyramidal cells in the deep part of layer II and layer III resemble those in other parts of the cerebral cortex by virtue of a single apical dendritic trunk, multiple basal dendrites, a large number of small to medium dendritic spines, and a deeply directed axon. Pyramidal cells in the superficial part of layer II are similar with the exception that “secondary” apical dendrites often emerge directly from the cell body rather than from a single primary trunk. With conservative criteria for categorization, nine different types of nonpyramidal cells were distinguished, four of which have not been previously described. Layer I contains a small number of neurons with both smooth and spiny dendrites including distinctive fusiform cells with large somatic appendages. As in other species, the most common type of nonpyramidal neuron in layer II is the semilunar cell which has only apically directed dendrites. These cells have distinctive large spines confined to their distal dendritic segments. The mid to deep portion of layer III contains multipolar neurons with smooth dendrites that resemble the well-known large stellate cells in neocortex. In addition, layer III contains three non-pyramidal neuron types with spiny dendrites: (1) fusiform and multipolar cells with complex, branched dendritic appendages and somatic spines, (2) very large multipolar cells (up to 35 μm mean diameter) with large-diameter dendrites that give rise to abruptly tapering side branches and filiform spines, and (3) multipolar cells with profusely spiny dendrites. In all three layers, small neurons have been found with spherical cell bodies and “axoniform” dendrites that resemble the so called neurogliaform neurons described in a variety of brain areas. A striking feature of the organization of the piriform cortex is that, with the exception of the neurogliaform neurons, the different types of nonpyramidal cells tend to be segregated in individual layers or sublayers. Physiological implications of the results are discussed. Remarks are also made concerning the potential of the piriform cortex as a model cortical system.     

The cytoarchitecture of the dorsal cochlear nucleus in the 3-month- and 26-month-old C57BL/6 mouse: A golgi impregnation study

The cytoarchitecture of the dorsal cochlear nucleus (DCN) was compared in 3- and 26-month-old C57BL/6 mice. The effects of genetically controlled progressive hearing loss present in the CNS in this mouse strain were analyzed with Nissl-stained and Golgi-impregnated material. The DCN was divided into the superficial molecular, an intermediate fusiform-granule, and the deep polymorphic layers. The molecular layer (ML) consisted of many fibers and a few small ovoid to spherical, fusiform, and granule cells. The fusiform-granule layer (FL) contained large fusiform and many granule cells. Most FL fusiform cells were oriented with their long axes perpendicular to the DCN surface and were present as small aggregations or individually. Cartwheel cells were adjacent to the FL fusiform cells. The deep polymorphic layer (PL) contained spherical, fusiform, granule, and multipolar neurons. The granule cells formed a dorsal cap of the DCN. From this cap, sheets of granule cells separated the DCN from the posterior ventral cochlear nucleus (PVCN) and from the brainstem. The internal organization, neuronal location, orientation, and morphology were similar in both age groups. The granule cells had four to five primary dendrites, varicosities, and few to no dendritic appendages. The FL fusiform cells displayed different dendritic morphology in the two ages. One or two elaborate primary ML apical dendrites in the 3-month-old mice were covered with spikelike dendritic spines. The basal one or two PL dendrites were less elaborate and had few dendrite spines. In contrast, FL fusiform neurons in 26-month-old mice had regular dendritic varicosities and fewer spines which were short and stumpy. Basal dendrites had varicosities and interruptions. Cartwheel neurons in 3-month-old mice had elaborate ML dendritic trees covered with dendritic spines. In 26-month-old mice the dendrites had many varicosities and fewer short blunted dendritic spines. Large multipolar neurons in older mice had thinner dendrities with more varicosities than were in the 3-mcnth group. In both age groups multipolar cells had few dendritic spines limited distally. Small and large spherical cells had two to five primary dendrites with varicosities, little higher-order branching, and spines. Fusiform cells had one or two primary dendrites, little secondary branching, and few to no spines. Minor degenerative changes were noted in spherical and fusiform cells in the two age groups. These included dendritic varicosities, interruptions, and some irregularities of somata surface. Degenerative changes present in the cochlea had significant effects on a limited population of DCN neurons. Finally, the neusronal morphology and architecture of the DCN in C57BL/6 mouse is similar to other mammalian species.     

Organization of cerebellar cortex secondary to deficit of granule cells in weaver mutant mice

This study deals with some consequences of the early postnatal abnormalities of cerebellar Bergmann glial fibers and granule cell neurons. (1) Cerebellar size is mildly reduced in heterozygous weaver (+/wv) mice and markedly reduced in homozygotes (wv/wv), but the pattern of fissures is essentially normal. Comparison with other mutants displaying small cerebella suggests that cell proliferation rate in the external granular layer is a key deteminant of cerebellar cortical folding. (2) Mossy fiber terminals differentiate on schedule despite the reduced number and abnormal positions of granule cells. However, many of them enter the modified molecular layer, and as noted especially in noninbred wv/wv mice one to two years old, form synapses with dendrites of aberrant granule cells. Where granule cells are absent, mossy fibers form more than the normal number of synapses with dendrites of Golgi type II neurons. (3) Purkinje cells are only mildly affected by the disorder of neighbouring cells. Their dendrites grow abnormally into the territory occupied by external granule cells, reach the external surface, and may turn inward. They form few tertiary branches. Dendritic spines are present in profusion and show membrane thickenings akin to normal postsynaptic elements. Although they receive no axonal contacts, the spines persist, enveloped by glial processes, for at least two years. Apart from the absence of parallel fiber contacts, afferent and intrinsic axons form the normal classes of synaptic connections with Purkinje cells. (4) Interneurons of the molecular layer are generated on schedule. At the time of their earliest recognition, they reside in the external granular layer, where they receive synaptic contacts from climbing fibers and other interneurons. In the absence of parallel fibers, interneurons differentiate in situ but their dendrites are abortive and randomly oriented. Growth of their dendrites, in contrast to that of Purkinje cell dendrites, appears to be markedly influenced by the organization of the local cellular milieu.     

Somatostatin-containing neurons in rat organotypic hippocampal slice cultures: Light and electron microscopic immunocytochemistry

Light and electron microscopic immunocytochemical techniques were used to study the interneuron population staining for somatostain (SRIF) in cultured slices of rat hippocampus. The SRIF immunoreactive somata were most dense in stratum oriens of areas CA1 and CA3, and in the dentate hilus. Somatostain immunoreactive cells in areas CA1 and CA3 were characteristically fusiform in shape, with dendrites that extended both parallel to and into the alveus. The axonal plexus in areas CA1 and CA3 was most dense in stratum lacunosum-moleculare and in stratum pyramidale. Electron microscopic analysis of this area revealed that the largest number of symmetric synaptic contacts from SRIF immunoreactive axons were onto pyramidal cell somata and onto dendrites in stratum lacunosum-moleculare. In the dentate gyrus, SRIF somata and dendrites were localized in the hilus. Hilar SRIF immunoreactive neurons were fusiform in shape and similar in size to those seen in CA1 and CA3. Axon collaterals coursed throughout the hilus, projected between the granule cells and into the outer molecular layer. The highest number of SRIF synaptic contacts in the dentate gyrus were seen on granule cell dendrites in the outer molecular layer. Synaptic contacts were also observed on hilar neurons and granule cell somata. SRIF synaptic profiles were seen on somata and dendrites of interneurons in all regions. The morphology and synaptic connectivity of SRIF neurons in hippocampal slice cultures appeared generally similar to intact hippocampus. © 1994 Wiley-Liss, Inc.     

The cerebellum of the frog Rana ridibunda. An electron microscopic study

An electron microscopic study of neuronal types and different synaptic contacts has been made in the cerebellum of the frog Rana ridibunda. The Purkinje cells have a pear-shaped cell body and in their cytoplasm the organelles show a special arrangement because of the great amount of microtubules they contain. The granule cells are small, rounded neurons with a large nucleus surrounded by a thin rim of cytoplasm. The stellate cells are interneurons of the molecular layer whose large nuclei show a single finger-like invagination of its nuclear envelope. The afferent tracts to the cerebellum end either as climbing fibers or mossy fibers. The axon terminals of climbing fibers are large and the synaptic complexes exhibit all the features of a type-I Gray synapse. The mossy fibers reach the granular layer and synapses between them and granule cell dendrites are by far the most abundant. The parallel fibers establish synaptic contacts on the spines arising from the spiny branchlet units of the Purkinje cells and with the perikaryon and dendrites of stellate cells. The stellate cell axons cross the molecular layer and establish type-II Gray synapses on the Purkinje cells.     

Targets and Quantitative Distribution of GABAergic Synapses in the Visual Cortex of the Cat

The morphology and postsynaptic targets of GABA-containing boutons were determined in the striate cortex of cat, using a postembedding immunocytochemical technique at the electron microscopic level. Two types of terminals, both making symmetrical synaptic contacts, were GABA-positive. The first type (95% of all GABA-positive boutons) contained small pleomorphic vesicles, the second type (5%) contained larger ovoid vesicles. Furthermore, 99% of all cortical boutons containing pleomorphic vesicles were GABA positive, and all boutons with pleomorphic vesicles made symmetrical synaptic contacts. These results together with previously published stereological data (Beaulieu and Colonnier, 1985, 1987) were used to estimate the density of GABA-containing synapses, which is about 48 million/mm3 in the striate cortex. The postsynaptic targets of GABA positive boutons were also identified and the distribution was calculated to be as follows: 58% dendritic shafts, 26.4% dendritic spines, 13.1% somata and 2.5% axon initial segments. A total of 11% of the postsynaptic targets were GABA immunoreactive and therefore originated from GABAergic neurons. The results demonstrate that the majority of GABAergic synapses exert their action on the membrane of dendrites and spines rather than on the somata and axons of neurons.     

Synaptic patterns in the visual cortex of turtle: An electron microscopic study

The part of turtle general cortex that receives afferent fibers from the dorsal lateral geniculate nucleus and that shows evoked potentials to light stimuli has been studied with the electron microscope. This cortex consists of an outer molecular layer, a perikaryal layer, and a subcellular layer lying on a row of ependymal cell bodies. Neurons in the perikaryal lamina are characterized by long spine-bearing apical dendrites ascending through the outer molecular layer and short finer basal dendrites in the subcellular zone. Scattered neurons without apical dendrites occur in both the molecular and subcellular zones. Two types of dendritic spines can be distinguished. Some are large, have a complex irregular shape, contain a variety of membranous sacs and mitochondria, and occasionally, a single bundle of microtubules embedded in an electron-dense background opacity. These large spines are the most common postsynaptic element in the outer third of the molecular layer, where they are located on the distal tips of the apical dendrites. Other spines are small, with a simple spherical distal enlargement that contains only electron-dense fuzz. They are the most common post-synaptic element in the lower two-thirds of the molecular layer where they arise from the proximal portion of apical dendrites. Most synaptic contacts are found on the dendritic spines and are of the “round-asymmetrical” type. Not infrequently “flat-symmetrical” synapses are seen coupled to “round-asymmetrical” contacts on individual large spines. The few contacts present on spine-bearing dendritic shafts are of both types. Axo-somatic contacts are mainly of the “flat-symmetrical” variety. Thus the synaptic patterns on the principal cells of turtle visual cortex are remarkably similar to those found on pyramidal cells of mammalian neocortex. In addition, however, axon terminals, dendrites and glial (ependymal) processes were often seen to give rise to membranous pouches containing large vacuoles and invaginating into dendritic shafts or spines. Rarely, axon terminals were seen to form contacts, identical in appearance to synaptic contacts, on cell bodies in the ependymal lining. More frequently, unusual types of membrane differentiations were present at the site of apposition of the membranes of axon terminals and ependymal processes. They are interpreted as functional neuroependymal contacts.     

Neurons and glia in cat superior colliculus accumulate [3H]gamma-aminobutyric acid (GABA)

We have examined by autoradiography the labeling pattern in the cat superior colliculus following injection of tritiated gamma-aminobutyric acid (GABA). Silver grains were heavily distributed within the zonal layer and the upper 200 micrometer of the superficial gray. Fewer grains were observed deeper within the superficial gray, and still fewer were found within the optic and intermediate gray layers. The accumulation of label was restricted to certain classes of neuron and glia. Densely labeled neurons were small (8-12 micrometer in diameter) and located primarily within the upper 200 micrometer. Dark oligodendrocytes and astrocytes showed a moderate accumulation of label while pale oligodendrocytes and microglia were unlabeled. Label was also selectively accumulated over several other types of profile within the neuropil, including presynaptic dendrites, axons, and axon terminals.