Axon terminals of GABAergic chandelier cells are lost at epileptic foci

Axon terminals of chandelier cells were analyzed in monkeys with cortical focal epilepsy produced by alumina gel to determine if this type of GABAergic terminal is lost at epileptic foci. These terminals form a dense plexus with the axon initial segments of pyramidal neurons, especially those in layers II and III. Axon initial segments of pyramidal neurons were traced for at least 40 μm in serial thin sections and beyond this point were observed to become myelinated. In single sections, 10–15 axon terminals were found to form symmetric synapses throughout the entire lenght of the axon initial segments from nonepileptic preparations and were observed to synapse with only these structures and not adjacent dendrites or spines. In epileptic cortex, the axon initial segments of pyramidal neurons were apposed by glial profiles that contained clusters of filaments typical of reactive astrocytes. Only a few, small axon terminals were observed to form symmetric synapses with these axon initial segments. Thus, the chandelier cell axons appeared to degenerate in epileptic cortex. The highly strategic site of GABAergic inhibitory synapses on axon initial segments suggests that they exert a strong influence on the output of pyramidal cells. The near absence of these chandelier cell axons in epileptic foci most likely contributes to the hyperexcitability of neurons.     

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 organization of GABAergic neurons in the mouse SmI cortex

Immunocytochemical methods were used to examine GABAergic neurons in the barrel region of the mouse primary somatosensory cortex. GABAergic neurons occur in all layers of the barrel cortex but are more concentrated in the upper portion of layers II/III and in layers IV and VI. Nine cells in layer IV were examined with the electron microscope, and portions of their dendrites were reconstructed from serial thin sections. These cells are of the nonspiny, multipolar or bitufted varieties, and some of them have beaded dendrites. The labeled cell bodies and their reconstructed dendrites were postsynaptic at asymmetrical synapses with thalamocortical axon terminals labeled by lesion-induced degeneration and with unlabeled axon terminals. Each cell also received symmetrical synapses from GABAergic axon terminals and from unlabeled axon terminals. Our results indicate that GABAergic cell bodies and processes receive synapses from thalamocortical axon terminals but that different cells display marked differences in the proportion of thalamocortical and other synapses they receive. These results indicate that GABAergic cells form a heterogeneous population with respect to their morphologies and patterns of synaptic inputs. The synaptic sequences revealed here for GABAergic neurons represent an anatomical substrate for various inhibitory processes known to occur within the cerebral cortex.     

The upper layers of the superior colliculus of the rat: a Golgi study

A number of questions deriving from a previous electron microscopic study of the superficial layers of the rat superior colliculus pointed out the need for a comprehensive study of the cell types of the superior colliculus and their detailed morphology. In the present Golgi study the cells of the superficial layers of the superior colliculus were found to be of five major types; marginal cells, horizontal cells, narrow field vertical cells, wide field vertical cells, and stellate cells. Some of these categories were further subdivided. Each cell type has a distinctive set of dendritic field characteristics, a regional distribution, and consistent axon characteristics. The horizontal cells were found to have a preference in the orientation of their dendrites along the vertical and horizontal axes of the visual field projection upon the superior colliculus. It is argued that the horizontal cells are the presynaptic elements in the majority of the dendrodendritic synapses.     

Retinal ganglion cell terminals in the hamster superior colliculus: an ultrastructural study.

The distribution, cross-sectional area, and presynaptic and postsynaptic characteristics of retinal ganglion cell axon terminals in the superior colliculus of normal adult female Syrian hamsters were investigated by quantitative ultrastructural techniques. After an intravitreal injection of horseradish peroxidase, most labelled axon terminals were found in the stratum griseum superficiale and stratum opticum of the contralateral superior colliculus. However, a small proportion (approximately 2%) of retinal ganglion cell axon terminals were located in deeper layers of the superior colliculus between the stratum opticum and the periaqueductal grey matter. Terminals were smaller in the upper two-thirds of the stratum griseum superficiale than in the lower one-third of this layer, the stratum opticum, and the stratum griseum intermedium. Presynaptic characteristics such as the length and number of contacts and the postsynaptic neuronal domains (somata, dendritic spines, or shafts) contacted by retinal ganglion cell axons in the superior colliculus were similar in all layers.     

A qualitative electron microscopic study of the corticopontine projections after neonatal cerebellar hemispherectomy

The present study shows that 3–5 days following leions of the dentate and interposed nuclei in normal adult rats degenerating axons and axon terminals can be detected in the contralateral pontine gray. The degenerating axon terminals form Gray's type I axo-dendritic contacts with fine intermediate dendrites measuring between 0.8–2.4 μm. The present study also investigates, by electron microscopy, the synaptic rearrangement of the sensorimotor corticopontine projections following neonatal left cerebellar hemispherectomy¹⁹. Following neonatal left cerebellar hemispherectomy, the right sensorimotor and adjacent cortex (SMC) presents a very dense ipsilateral and a modest amount of contralateral corticopontine projections in contrast with a predominantly ipsilateral corticopontine projection seen in the normal adult rat. As with the ipsilateral corticopontine projection seen in the normal adult animal, the bilateral corticopontine projections seen in the experimental animals form contacts with dendrites suggestive of Gray's type I synapses. While the corticopontine projections in normal control animals form synapses with fine dendrites measuring 0.2–1.2 μm the corticopontine projections in the experimental animals form synaptic relations with fine dendrites and with intermediate dendrites measuring 0.2–2.4 μm. As the normal cerebellopontine fibers from the dentate and interposed nuclei also form axo-dendritic synapses on fine and intermediate dendrites and the contacts formed are also of Gray's type I synapses, it is possible that some of the newly formed corticopontine fibers in the experimental animals might have replaced the cerebellopontine fibers synapsing on intermediate dendrites. Synaptic rearrangement appears to take place as suggested by the presence of synaptic complexes in which one axon terminal contacts two of more dendrites or two or more axon terminals contact one dendrite. Such complexes are frequently seen to undergo degeneration following the right SMC lesion in the experimental animals. Other complex synaptic structures are also present in both the right and left pontine gray in the experimental animals. They are not seen to undergo degeneration following the right SMC lesions. Occasional features of neuronal reaction could still be seen both sides of the pontine gray for as long as 3–6 months after the neonatal cerebellar lesions.     

Interlaminar connections of rat visual cortex: An ultrastructural study

Thermal lesions were made in layers I, II, and upper part of layer III of rat visual cortex. The distribution of degenerating axons and axon terminals in layers IV, V, and VI was studied using electron microscopic techniques. Following supragranular thermal lesions, the majority of degenerating axon terminals were found in layer V, with extension into the adjacent part of layer VI. Neural profiles postsynaptic to degenerating axon terminals were found in these layers in the following distribution: 81.7% on spines of small to medium size dendrites; 18.2% on dendrite shafts; and <1% on neuronal perikarya. Few degenerating terminals were found on or near apical dendrites. Degenerating terminals were identified on shafts of stellate-type dendrites found in the upper part of layer V. Degenerating axons oriented parallel to the cortical surface were found most often in deep layer IV and upper layer V. Degenerating axons were also seen in axon bundles coursing vertically through layer IV. Approximately 10% of the terminals within a grid square have undergone degeneration; no clustering of degenerating terminals was found in vertical or transverse sections through layers V and VI. We suggest that most axon terminals arising from pyramidal neurons in layers II and upper III synapse with spines and shafts of dendrite branches originating from pyramidal neurons in layer V and perhaps VI.     

Ultrastructure and retinal innervation of deafferentation-induced enkephalin-immunoreactive elements in the superficial layers of the rat superior colliculus

Leu-enkephalin-like immunoreactive (ENK-I) elements appearing in the superficial layers of the rat superior colliculus (SC) after eye-enucleation were examined by means of immunoelectronmicroscopy. ENK-I somata were of a single type and formed symmetric and asymmetric synapses with non-immunoreactive axon terminals. Some degenerating retinal terminals made synaptic contacts only with small ENK-I dendrites, suggesting that deafferentation-induced ENK-I neurons in the rat SC receive retinal input onto the distal portions of their dendrites.     

The synaptology of parvalbumin-immunoreactive neurons in the primate prefrontal cortex.

Electron microscopy and immunocytochemistry with a monoclonal antibody against parvalbumin (PV) were combined to analyze the distribution and morphology of PV-immunoreactive (PV-IR) neurons and the synaptology of PV-IR processes in the principal sulcus of the macaque prefrontal cortex. Parvalbumin-IR neurons are present in layers II-VI of the macaque principal sulcus (Walker's area 46) and are concentrated in a band centered around layer IV. PV-IR cells are exclusively non-pyramidal in shape and are morphologically heterogeneous with soma sizes ranging from less than 10 microns to greater than 20 microns. Well-labeled neurons that could be classified on the basis of soma size and dendritic configuration resembled large basket and chandelier cells. A novel finding is that supragranular PV-IR neurons exhibit dendritic patterns with predominantly vertical orientations, whereas infragranular cells exhibit mostly horizontal or oblique dendritic orientations. PV-IR cells within layer IV exhibit a mixture of dendritic arrangements. Vertical rows of PV-IR puncta, 15-30 microns in length, resembling the "cartridges" of chandelier cell axons were most dense in layers II, superficial III, and the granular layer IV but were not observed in the infragranular layers. Cartridges were often present beneath unlabeled, presumed pyramidal cells. PV-IR puncta also formed pericellular nests around pyramidal cell somata and proximal dendrites, suggestive of basket cell innervation. PV-IR axons were occasionally observed in the white matter underlying the principal sulcus. Electron microscopic analysis revealed that PV-IR somata and dendrites are densely innervated by nonimmunoreactive terminals forming asymmetric (Gray type I) synapses as well as by fewer terminals forming symmetric (Gray type II) synapses. The majority of terminals forming symmetric synapses with PV-IR post-synaptic structures were not immunolabeled; however, some of these boutons did contain PV-immunoreactivity. PV-IR boutons exclusively form symmetric synapses and heavily innervate layer II/III pyramidal cells. PV-IR axon cartridges formed numerous axo-axonic synapses with the axon initial segments of pyramidal cells 15-20 microns beneath the axon hillock and also terminated on large axonal spines of the initial segment. Furthermore, we failed to observe a mixture of PV-immunoreactive and non-immunoreactive boutons composing a single axon cartridge. Pyramidal cell somata and proximal dendrites were also heavily innervated by PV-IR boutons forming symmetric synapses, again, consistent with basket cell innervation. In addition, PV-IR axon terminals frequently formed symmetric synapses with dendritic shafts and spines of unidentified neurons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Variability in the terminations of GABAergic chandelier cell axons on initial segments of pyramidal cell axons in the monkey sensory-motor cortex

Chandelier cell axons were studied in the sensory-motor cortex of adult monkeys. The axonal fields of Golgi-impregnated chandelier cells in layer II in motor cortex are flattened sagittally. The vertical terminal portions of the axons varied both in length and in the numbers converging to form terminations of greater or lesser complexity. Golgi-impregnated plexuses were embedded in plastic and resectioned serially at 2.5-3.0 micrograms. A single axonal field could have as many as 400 terminal rows. All lie 3-13 micrograms beneath pyramidal cell somata. These terminations are not randomly distributed but instead, form clusters. Further resectioning the plastic sections for electron microscopy revealed that all the terminations are on the initial axon segments of pyramidal cells and all form symmetric synaptic contacts. In immunocytochemical material stained for glutamic acid decarboxylase (GAD), the enzyme involved in the synthesis of GABA, GAD-positive boutons were found to form symmetric synaptic contacts with a variety of postsynaptic elements including the axon hillocks and axon initial segments of pyramidal cells. Serial reconstructions from electron micrographs revealed GAD-positive terminals synapsing with the axon initial segment of pyramidal cells joined by cytoplasmic bridges and forming vertically oriented rows identical to those of chandelier cell terminals identified positively in the resectioned Golgi material. The GAD-positive terminals forming initial segment synapses were never continuous with GAD-positive terminals forming axo hillock synapses. The latter probably arise from basket cell axons. Initial segments of pyramidal cell axons in layers II and III were contacted by more GAD-positive terminals than the initial segments of pyramidal cell axons in layer V. The largest pyramidal cells in layer III received the most synapses. Many larger pyramidal cells, identified as callosally projecting cells by the retrograde transport of horseradish peroxidase (HRP), were shown in serial electron micrographs to possess large numbers of initial segment synapses, comparable to those seen in the immunocytochemical material. Serial reconstructions of pyramidal cell axons from axon hillock to the first myelin internode in resectioned Golgi, immunocytochemical and HRP material showed that the number of synapses varied from 2 to 52 for layers II and III and from 2 to 26 for layer V. The number of synapses on the axon hillocks varied from zero to 12. The variability in these terminations may be an important factor in the shaping of the functional properties of the pyramidal cells.     

Regenerated retinal ganglion cell axons can form well-differentiated synapses in the superior colliculus of adult hamsters

To investigate in adult animals the distribution and differentiation of the synapses made by axotomized CNS neurons whose regenerating axons are guided back to their natural targets in the brain, we attached an autologous peripheral nerve (PN) graft 2-3 cm in length to the ocular stump of a transected optic nerve (ON) in adult hamsters, inserted the distal end of the graft into the superior colliculus (SC), and, 6-8 weeks later, labeled the retinal ganglion cell (RGC) axons that entered the SC with HRP orthogradely transported from the eye. By light microscopy, regenerated RGC axons extended from the graft into the retinorecipient layers of the SC for up to 500 microns, distances that approximate the lengths of normal RGC arbors. We compared 698 control and 758 regenerated HRP-labeled axon terminals from 4 intact and 4 experimental animals by electron microscopy. The structure of the regenerated RGC terminals, the type of synaptic contacts formed, the ratios of contacts to terminal perimeter, and the domains of the postsynaptic neurons contacted were similar to those of controls. These results indicate that regenerated RGC axons can form well-differentiated synapses in the SC. Morphological differences between the regenerated and control synapses were the larger size of some regenerated terminals, the greater mean length of the regenerated synapses, and the higher proportion of contacts with dendrites that contained vesicles. The synaptic differentiation attained by these reformed retinocollicular projections suggests that regenerating CNS axons and their target neurons in the adult mammalian brain may retain or reexpress certain molecular determinants of normal connectivity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Electron microscopic immunocytochemical study of the distribution of parvalbumin-containing neurons and axon terminals in the primate dentate Gyrus and Ammon's horn

Five green monkeys were examined with light and electron microscopic preparations to explore the regional differences in the distribution of parvalbumin (PV)-positive neurons and axon terminals in the primate hippocampus. PV-positive neurons were mainly found in the hilus of the dentate gyrus and the strata oriens and pyramidale of Ammon's horn. In electron microscopic preparations, the PV-positive cells displayed nuclear infoldings, intranuclear rods, a large rim of perikaryal cytoplasm with numerous organelles and both asymmetric and symmetric axosomatic synapses. One prominent PV-positive cell type in CA1 was a large multipolar neuron that resembled the large basket cells of the neocortex. Although most PV-positive dendrites were aspiny and postsynaptic to numerous axon terminals, some PV-positive dendrites in the molecular layer of the dentate gyrus displayed filipodia-like appendages with no synapses or spines that were postsynaptic to multiple axon terminals. The PV-positive dendrites in the hilus and stratum oriens were apposed at specialized junctions that resembled gap junctions. PV-positive axons were concentrated in the principal cell layers, and formed axosomatic, axodendritic, and axon initial segment synapses. In cases where these axons were observed to appose the surface of granule cells for a long length, only one axosomatic symmetric synapse per cell was found. In the hilus, PV-positive axon terminals formed synapses onto thorny excrescences of spiny cells. Both semithin sections and electron microscopic preparations indicated that more PV-positive axon terminals formed symmetric axosomatic synapses with pyramidal cells in CA2 than in CA1 and CA3. Also, CA2 displayed a unique plexus of PV-positive axon terminals in stratum lacunosum moleculare. These results indicate that the PV-positive hippocampal cells form a subset of GABAergic local circuit neurons, including the basket and chandelier cells. The ubiquitous finding of PV-positive dendrites linked by gap junctions throughout the dentate gyrus and Ammon's horn adds further data to indicate that this subset of GABAergic neurons is linked electrotonically. The synaptic organization of PV-positive neurons in the hippocampus suggests their participation in both feedback and feedforward inhibition. The PV-positive neurons in the hippocampus are only a proportion of the basket and chandelier cells, whereas virtually all of these cells in neocortex are PV-positive. © 1993 Wiley-Liss, Inc.     

The organization of the pulvinar in the grey squirrel (Sciurus carolinensis). II. Synaptic organization and comparisons with the dorsal lateral geniculate nucleus

The purpose of these experiments was to compare the synaptic organization of the subdivisions of the pulvinar defined in the preceding paper (Robson and Hall, '77) with each other and with the organization present in the dorsal lateral geniculate nucleus. The electron microscope was used to analyze normal synaptic arrangements and degenerating axonal terminals resulting from lesions. The dorsal lateral geniculate nucleus in the grey squirrel contains synaptic clusters similar to those described previously for other species. These clusters are characterized by large optic tract terminals which form multiple contacts onto large dendritic processes and other processes containing flat or pleomorphic vesicles. The geniculate lamina adjacent to the optic tract receives projections from the superior colliculus as well are from the retina. The terminals of the superior colliculus axons are small and medium sized and lie outside of the synaptic clusters. The retinal terminals are in the clusters. In the pulvinar, the rostro-medial subdivision contains synaptic clusters which resemble those in the lateral geniculate nucleus. These clusters contain large axon terminals which make multiple contacts onto large dendrites. However, these terminals are not contributed by an ascending sensory pathway but by axons from strait cortex. The rostro-lateral and caudal subdivisions of the pulvinar also contain synaptic clusters, but these clusters consist of a segment of a large dendrite which is ensheathed by medium-sized terminals. Since only a few of these medium sized terminals in any one cluster degenerate after tectal lesions, and none degenerate after cortical lesions, it is suggested that the morphological arrangement of these clusters may permit the convergence of axons from several sources, some of which are unidentified, onto the same dendritic segment.     

Dual ultrastructural immunocytochemical labeling of μ and δ opioid receptors in the superficial layers of the rat cervical spinal cord

The δ opioid receptor (DOR) and μ opioid receptor (MOR) are abundantly distributed in the dorsal horn of the spinal cord. Simultaneous activation of each receptor by selective opiate agonists has been shown to result in synergistic analgesic effects. To determine the cellular basis for these functional associations, we examined the electron microscopic immunocytochemical localization of DOR and MOR in single sections through the superficial layers of the dorsal horn in the adult rat spinal cord (C2–C4). From a total of 270 DOR-labeled profiles, 49% were soma and dendrites, 46% were axon terminals and small unmyelinated axons, and 5% were glial processes. 6% of the DOR-labeled soma and dendrites, and <1% of the glial processes also showed MOR-like immunoreactivity (MOR-LI). Of 339 MOR-labeled profiles, 87% were axon terminals and small unmyelinated axons, 12% were soma and dendrites, and 2% were glial processes. 21% of the MOR-labeled soma and dendrites, but none of the axon terminals also contain DOR-LI. The subcellular distributions of MOR and DOR were distinct in axon terminals. In axon terminals, both DOR-LI and MOR-LI were detected along the plasmalemma, but only DOR-LI was associated with large dense core vesicles. DOR-labeled terminals formed synapses with dendrites containing MOR and conversely, MOR-labeled terminals formed synapses with DOR-labeled dendrites. These results suggest that the synergistic actions of selective MOR- and DOR-agonists may be attributed to dual modulation of the same or synaptically linked neurons in the superficial layers of the spinal cord.     

Chandelier cells in rat visual cortex

Golgi-impregnated chandelier cells in rat visual cortex have been examined by both light and electron microscopy. All of the chandelier cells impregnated have their cell bodies within layer II/III and although they occur throughout area 17, there are increased numbers at the area 17/18a border and to a lesser extent at the area 17/18 border. Most of the chandelier cells are bitufted neurons, with groups of dendrites extending from the upper and lower poles of an elongate cell body, but some cells have a more multipolar configuration. The perikaryal cytoplasm is rich in rough endoplasmic reticulum and both the cell body and the sparsely spinous dendrites receive axon terminals forming symmetric and asymmetric synapses. The axons of these neurons arise from either the lower pole of the cell body or the base of one of the dendrites in the lower tuft, and the axons form laterally spread plexuses which terminate in vertical strings of boutons. The boutons in each string synapse with axon initial segments of layer II/III pyramidal cells, the uppermost bouton in each string being 7 to 14 μm distant from the pyramidal cell body. Some layer II/III pyramidal cells seem to receive boutons from more than one chandelier cell, others from a single chandelier cell, and still others appear to receive no chandelier cell terminals. The axon terminals of the chandelier cells are irregular in shape, contain pleomorphic synaptic vesicles, and form symmetric synapses. Evidence is presented to show that axon terminals exhibiting the same morphological features and site of synaptic termination as those of the chandelier cells contain glutamic acid decarboxylase (GAD), the enzyme which synthesizes GABA. Hence the chandelier cells are probably GABAergic, inhibitory neurons. Other GAD-positive axon terminals synapse with the cell bodies, axon hillocks, and proximal portions of the axon initial segments of the layer II/III pyramidal cells, and these terminals are probably derived from the smooth and sparsely spinous stellate cells.     

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.     

Postembedding immunocytochemistry demonstrates directly that both retinal and cortical terminals in the cat superior colliculus are glutamate immunoreactive

Although the excitatory neurotransmitter glutamate is known to be present in the cat superior colliculus (SC), the types of synapses that contain glutamate have not been examined. We, therefore, studied the ultrastructure of synaptic profiles labeled by a glutamate antibody by using electron microscopic postembedding immunocytochemistry. In addition, unilateral aspiration lesions of areas 17–18 were made at 5–28 days before death in order to determine whether degenerating terminals from visual cortex were glutamate immunoreactive (Glu-ir). Three types of axon terminal were glu-ir: 1) those containing large, round synaptic vesicles and pale mitochondria, characteristic of retinal terminals (RT profiles); 2) those containing small, round synaptic vesicles and dark mitochondria (RSD profiles); and 3) those containing large, round synaptic vesicles and dark mitochondria (RLD profiles). Measures of mean gold particle density revealed that RT, RSD, and RLD profiles had similar average grain densities (11.3–12.7 particles/unit area). Other labeled profile types included cell bodies, large-calibre dendrites, and myelinated axons. Axon terminals containing flattened synaptic vesicles and vesicle-containing presynaptic dendrites, both of which contain γ-aminobutyric acid (GABA), had many fewer gold particles (3.6 and 4.8 mean particles/unit area, respectively). Following unilateral removal of visual cortex, normal RSD terminals were observed infrequently in the SC ipsilateral to the lesion. Synaptic terminals in the initial stages of degeneration were heavily labeled by the glutamate antibody, as were axon terminals and myelinated axons undergoing hypertrophied or neurofilamentous degeneration. These results show that both major sensory afferents to the superficial layers of cat SC contain glutamate—RT terminals from the retina and RSD terminals from visual cortex. The origin of RLD terminals is unknown. © 1996 Wiley-Liss, Inc.     

Synaptic patterns of the superficial layers of the superior colliculus of the rat

An electron microscopic study has been undertaken of the upper layers of the superior colliculus, into which run fibers from the retina and visual cortex. Study of the normal synaptic patterns shows large numbers of serial synapses, particularly near the surface of the colliculus. The presynaptic components usually contain round vesicles and the intermediate profiles always contain flattened vesicles. The latter may be dendrites, dendrite-like or dilatations of such profiles. Degeneration studies indicate that optic terminals are especially dense near the surface and that they are often the presynaptic terminal of a serial synapse. Very few optic terminals occur in the stratum opticum. Cortical afferents from the visual area mostly end deep in the stratum griseum superficiale, and in superficial stratum opticum, usually on small clear profiles. Studies of the progress of degeneration indicate that optic terminals first show a predominantly neurofilamentous and associated glycogen reaction, and later a predominantly electron dense reaction. Cortical terminals show an initial dense reaction, with occasional neurofilamentous endings. These time course fit well with light microscope data from Nauta and Glees stained material. The synaptic patterns described here closely resemble patterns described in retina and dorsal lateral geniculate body. How these reflect functional similarities has yet to be resolved.     

An electron microscopic observation of the vesicular acetylcholine transporter-immunoreactive fibers in the rat dorsal raphe nucleus

By using immunocytochemistry with an antibody directed against the vesicular acetylcholine transporter, many cholinergic neuronal processes were found to be immunopositive in the dorsal raphe nucleus. At the electron microscopic level, most of these processes were found to be axons. The immunopositive axon terminals made synapses on immunonegative dendrites and their spines whereas rare synapses were found between the immunopositive axon terminals and the immunonegative neuronal perikarya. Occasionally, the dendrites postsynaptic to an immunopositive axon terminal also received a synapse from an immunonegative axon terminal. The synapses made by the immunopositive axon terminals were usually symmetric and had a short active zone. Fewer immunostained dendrites were found, and they usually received asymmetric synapses from nonimmunostained axon terminals. The existence of cholinergic axon terminals and the synapses made by these terminals support the physiological data indicating that acetylcholine plays a role in the pain inhibition system in the dorsal raphe nucleus.     

Proportion of parvalbumin-positive basket cells in the GABAergic innervation of pyramidal and granule cells of the rat hippocampal formation

Recent studies have indicated that hippocampal GABAergic neurons in both the dentate gyrus and Ammon's horn contain immunoreactivity for the calcium-binding protein parvalbumin (PARV). Although the distribution of PARV-positive neurons has been previously described, detailed quantitative electron microscopic studies of the PARV-positive axon terminals in the hippocampal formation are lacking. In the present study, immunocytochemical methods were used to localize PARV-positive neurons and axon terminals to determine their similarity to GABAergic neurons. The PARV-positive cells and axon terminals are associated closely with the pyramidal and granule cell layers. In agreement with previous studies, the morphology of PARV-positive neurons is similar to that of GABAergic cells, including the basket cells of both the dentate gyrus and Ammon's horn. The PARV-positive axon terminals form exclusively symmetric synapses with somata, dendrites, dendritic spines, and axon initial segments. However, these terminals represent only a portion of the total number of terminals that form symmetric synapses. Quantitative results indicate that only 32-38% of the total number of terminals forming symmetric axosomatic synapses with principal cells of the dentate gyrus and Ammon's horn are PARV positive. Together with previous findings from light microscopic double-labeling studies, these data indicate that the PARV-positive terminals arise from a subpopulation of GABAergic hippocampal neurons. Finally, it is important to note that the terminal plexus of PARV-positive hippocampal axons overlaps at all postsynaptic sites with a plexus of PARV-negative axons.