Calbindin 28kD and parvalbumin immunoreactive neurons receive different patterns of synaptic input in the cat superior colliculus.

Recent evidence suggests that neurons containing the calcium binding proteins calbindin 28kD (CB) and parvalbumin (PV) have differing distributions which match respectively the distribution of W and Y retinal ganglion cell inputs to the cat superior colliculus (SC). In this study we have used electron microscope immunocytochemistry to study directly the synaptic inputs to neurons containing CB and PV. Aspiration lesions of areas 17-18 of visual cortex were made 4 days prior to sacrifice in order to identify degenerating cortical terminals (CT). Retinal terminals (RTs) were identified by their characteristic morphology including large round synaptic vesicles and pale mitochondria. We photographed RTs and CTs that were in contact with immunoreactive profiles sampled in both the superficial gray and optic layers (ol) of SC. CB immunoreactive (ir) dendrites were usually of small to medium caliber and were found to receive synaptic input from RTs. These RTs were all small profiles forming a single synaptic contact with asymmetric densifications. CBir profiles also received other synaptic input, including from terminals with dark mitochondria that contained flattened synaptic vesicles (F profiles). No CBir dendrites were found to receive CT input even though degenerating CTs were found in the vicinity of CBir profiles. By contrast, both RT and CT were found to contact PVir dendrites. RT terminals contacting PVir dendrites were both small and larger profiles with round synaptic vesicles and asymmetric synaptic densifications. CT were undergoing electron dense degeneration but still sometimes formed asymmetric synaptic densifications with PV neurons. PV cells also received F profile synaptic input. We conclude that CB neurons receive small RT synapses that are probably of W origin, while PV neurons receive both RT and CT synapses which are likely related to the Y pathway.     

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.     

Synaptic organization of thalamocortical axon collaterals in the perigeniculate nucleus and dorsal lateral geniculate nucleus

We examined the synaptic targets of large non-gamma-aminobutyric acid (GABA)-ergic profiles that contain round vesicles and dark mitochondria (RLD profiles) in the perigeniculate nucleus (PGN) and the dorsal lateral geniculate nucleus (dLGN). RLD profiles can provisionally be identified as the collaterals of thalamocortical axons, because their ultrastrucure is distinct from all other previously described dLGN inputs. We also found that RLD profiles are larger than cholinergic terminals and contain the type 2 vesicular glutamate transporter. RLD profiles are distributed throughout the PGN and are concentrated within the interlaminar zones (IZs) of the dLGN, regions distinguished by dense binding of Wisteria floribunda agglutinin (WFA). To determine the synaptic targets of thalamocortical axon collaterals, we examined RLD profiles in the PGN and dLGN in tissue stained for GABA. For the PGN, we found that all RLD profiles make synaptic contacts with GABAergic PGN somata, dendrites, and spines. In the dLGN, RLD profiles primarily synapse with GABAergic dendrites that contain vesicles (F2 profiles) and non-GABAergic dendrites in glomerular arrangements that include triads. Occasional synapses on GABAergic somata and proximal dendrites were also observed in the dLGN. These results suggest that correlated dLGN activity may be enhanced via direct synaptic contacts between thalamocortical cells, whereas noncorrelated activity (such as that occurring during binocular rivalry) could be suppressed via thalamocortical collateral input to PGN cells and dLGN interneurons.     

The fine structure of the perigeniculate nucleus in the cat

The fine structure of the cat's perigeniculate nucleus has been analyzed and compared to that of dorsal thalamic relay nuclei. Golgi preparations and electron micrographs of perigeniculate cells commonly show somatic spines. The most common presynaptic elements for these spines and for the adjacent perikaryal surfaces are relatively large axon terminals containing round synaptic vesicles and making multiple asymmetric contacts. These "RLD" terminals (so termed for their round vesicles, large average size of the terminals, and dark mitochondria) are also presynaptic to dendritic spines and shafts of proximal and secondary dendrites. Comparisons with adjacent parts of the dorsal lateral geniculate nucleus show that these RLD terminals are cytologically distinct from retinogeniculate terminals and that small numbers of RLD terminals also occur in the geniculate A laminae. Three other major classes of perigeniculate synaptic terminals, resemble major classes of terminals in the dorsal lateral geniculate nucleus. These include two types of terminal with flat or ovoid synaptic vesicles and dark mitochondria, "FD1" and "FD2" terminals, and a class of small terminal with densely clustered round vesicles and dark mitochondria, "RSD" terminals. RSD terminals, which resemble corticogeniculate axon terminals, represent the only class of perigeniculate terminal that does not contact perikarya. FD2 terminals resemble lateral geniculate presynaptic dendrites and participate in serial and triadic synaptic contacts, being both pre- and postsynaptic; however, in contrast to the arrangement characteristic of thalamic relay nuclei, these contacts do not occur within synaptic glomeruli. A fifth major class of perigeniculate presynaptic terminal has large flat or polymorphic synaptic vesicles and pale mitochondria. These "FP" terminals are seen infrequently in the lateral geniculate A laminae. Similarities between perigeniculate and lateral geniculate fine structure may relate in part to common sources of afferent input to the two nuclei.     

Projections from auditory cortex to the cochlear nucleus in rats: Synapses on granule cell dendrites

Previous work has demonstrated that layer V pyramidal cells of primary auditory cortex project directly to the cochlear nucleus. The postsynaptic targets of these centrifugal projections, however, are not known. For the present study, biotinylated dextran amine, an anterograde tracer, was injected into the auditory cortex of rats, and labeled terminals were examined with light and electron microscopy. Labeled corticobulbar axons and terminals in the cochlear nucleus are found almost exclusively in the granule cell domain, and the terminals appear as boutons (1–2 μm in diameter) or as small mossy fiber endings (2–5 μm in diameter). These cortical endings contain round synaptic vesicles and form asymmetric synapses on hairy dendritic profiles, from which thin (0.1 μm in diameter), nonsynaptic “hairs” protrude deep into the labeled endings. These postsynaptic dendrites, which are typical of granule cells, surround and receive synapses from large, unlabeled mossy fiber endings containing round synaptic vesicles and are also postsynaptic to unlabeled axon terminals containing pleomorphic synaptic vesicles. No labeled fibers were observed synapsing on profiles that did not fit the characteristics of granule cell dendrites. We describe a circuit in the auditory system by which ascending information in the cochlear nucleus can be modified directly by descending cortical influences. © 1996 Wiley-Liss, Inc.     

Fine structure of the visual dorsolateral anterior thalamic nucleus of the pigeon (Columba livia): a hodological and GABA-immunocytochemical study

The ultrastructure of the lateroventral subcomponent of the visual dorsolateral anterior thalamic nucleus of the pigeon (DLLv) was analyzed using hodological techniques and GABA-immunocytochemistry. Two types of GABA-immunonegative hyperpalliopetal neurons and a single type of strongly GABA-immunoreactive (-ir) interneuron were identified, the latter displaying long dendrites with some containing synaptic vesicles (DCSV). Ten types of axon terminal were identified and divided into two categories. The first, GABA-immunonegative and making asymmetrical synaptic contact, contain round (RT1, RT2, RT3) or pleiomorphic synaptic and many dense-core vesicles (DCT). RT1 terminals are retinothalamic and RT2 terminals hyperpalliothalamic; both mainly contact dendrites of projection neurons (72% and 78% respectively), less frequently dendrites of interneurons and sometimes DCSV; RT1 terminals are rarely involved in synaptic triads. The second category are consistently GABA-immunopositive. Four types (PT1-4), distinguished by their pleiomorphic synaptic vesicles, make symmetrical synaptic contact essentially with dendrites of projection neurons, more rarely on dendrites of interneurons (PT2). PT1 terminals are very probably those of interneurons, whereas the rare PT4 terminals are of retinal origin. A fifth type (RgT) contains round synaptic vesicles and makes asymmetrical synaptic contact with dendrites of projection neurons and interneurons. PT2 and RgT terminals occasionally contact DCSV of interneurons, which are sometimes involved in synaptic triads. Two final subcategories (DCgT1-2) contain many dense-core vesicles. Our findings are compared with those of previous studies concerning the fine structure and neurochemical properties of the GLd of reptiles and mammals, with special reference to the origin of the extraretinal and extracortical projections to this structure.     

The GABAergic neurons and axon terminals in the lateralis medialis-suprageniculate nuclear complex of the cat: GABA-immunocytochemical and WGA-HRP studies by light and electron microscopy

In order to get more detailed information on the neural circuit of the lateralis medialis-suprageniculate nuclear (LM-Sg) complex of the cat, the GABAergic innervation of this complex was studied by GABA immunohistochemical techniques. Small immunoreactive cells were found throughout the LM-Sg complex. On the basis of their ultrastructural features, these GABAergic cells were identified as Golgi type II interneurons. The neuropil of this nucleus displayed a conspicuous granular immunoreactivity. Ultrastructurally, the immunoreactive neural profiles in the neuropil were identified as the presynaptic dendrites of interneurons, myelinated axons, or axon terminals. The GABAergic dendritic profiles, containing pleomorphic synaptic vesicles, were involved in synaptic glomeruli. Additionally, GABAergic axon terminals containing pleomorphic synaptic vesicles formed symmetric axodendritic synaptic contacts mainly in the extraglomerular neuropil. They appeared to correspond to either axon terminals from the thalamic reticular nucleus (TRN) or the axon terminals of interneurons. The projections from the TRN to the LM-Sg complex were studied by using wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). Following injection of WGA-HRP into the LM-Sg complex, a number of retrogradely labeled cells were observed in the TRN. The connections between the TRN and the LM-Sg complex appeared to be topographically organized, the dorsal TRN being connected mainly with the dorsomedial portion of the LM-Sg complex, and the ventral TRN being connected chiefly with the ventrolateral portion of the LM-Sg complex. Following injection of the tracer into the TRN, ultrastructural examination of anterograde labeling in the LM-Sg complex revealed that labeled terminals contain pleomorphic vesicles and make symmetric synaptic contacts mainly with small to medium-sized dendrites. The labeled terminals were not involved in synaptic glomeruli. The present results provide anatomic support for the contention that the projection cells of the LM-Sg complex may be inhibited by both the TRN axons and interneurons, probably through the mediation of GABA.     

Effects of monocular deprivation on geniculocortical synapses in the cat

In monocularly deprived (MD) cats, many cells in the lateral geniculate nucleus (LGN) but few cells in the visual cortex respond to input from the deprived eye, suggesting that the connections to visual cortex from the deprived geniculate laminae may have been disrupted. I have examined these connections in MD cats by using electron microscopic autoradiography of visual cortex after injections of tritiated lysine into single laminae of LGN. After injections into either deprived or experienced laminae, there was label over terminals that contained mitochondria and round synaptic vesicles and that made asymmetric contacts with dendritic profiles. However, the terminals of deprived afferents differed from those of experienced afferents: They were 25% smaller, contained 33% fewer mitochondria, were more likely to make synapses that were presynaptically convex (and thus, perhaps, immature), and synapsed onto smaller spines. These morphological changes were greater for afferents to upper layer IV than for afferents to lower layer IV. The geniculocortical synapses from deprived laminae were also reduced in number. To correct for variations in injection size and for a probable reduction in protein synthesis by cells in the deprived laminae, I computed the ratio of labeled synaptic terminals to labeled myelinated axons. Injections into the deprived laminae labeled 43% fewer synaptic terminals per labeled myelinated axon than did injections into the experienced lamina. The finding that the synaptic terminals of deprived afferents are both abnormal morphologically and fewer in number can help to explain the reduced effectiveness of the deprived eye in driving cortical cells.     

A note on the synapses between vesicle containing profiles in the pontine nuclei of the cat

In the cat synapses between vesicle containing profiles were observed in ventral and dorsolateral pontine nuclei. The presynaptic elements consisted of two types of axon terminals: axon terminals characterized by a population of small (38-40 nm) round synaptic vesicles (SSV) and axon terminals containing pleomorphic synaptic vesicles (PSV). The postsynaptic pale elements (PP) had pleomorphic vesicles and some features attributed to dendrites. In the dorsolateral pontine nucleus most of PP profiles took part in serial synapses, usually as an intermediate component, they were rarely observed in triads. On the basis of their electron microscopical appearance and synaptic relations they might be considered to represent a dendritic part of putative interneurons.     

Ultrastructure of the mammillotegmental projections to the ventral tegmental nucleus of Gudden in the rat.

This study examines the termination pattern of axons from the medial mammillary nucleus within the ventral tegmental nucleus of Gudden (TV) in rats by using anterograde transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) and visualized with tetramethylbenzidine. The neuropil of TV contains three classes of axodendritic terminals, that is, terminals containing round, flat, and pleomorphic synaptic vesicles. These types make up 55.6%, 26.1%, and 18.3%, respectively, of all normal axodendritic terminals. Injection of WGA-HRP into the medial mammillary nucleus permits ultrastructural recognition of anterogradely labeled terminals within the TV. More than 80% of the labeled terminals contain round synaptic vesicles and form asymmetric synaptic contacts, whereas about 16% contain flat synaptic vesicles with symmetric synaptic contacts. There are a few labeled terminals with pleomorphic vesicles and only a few axosomatic terminals. Almost all labeled terminals are small, having diameters of less than 1.5 microns. Compared with the distributions of normal and labeled terminals with round vesicles, there is an increase of the percentage of labeled terminals with round vesicles on the intermediate dendrites (1-2 microns diameter) and a decrease on the distal dendrites (less than 1 micron diameter). Anterogradely labeled axon terminals often contact retrogradely labeled dendrites. These results suggest that the medial mammillary neurons send mainly excitatory as well as a few inhibitory inputs to the dendrites of TV and have direct reciprocal contacts with the TV neurons.     

Immunoelectron microscopic study of glutamate inputs from the retrosplenial granular cortex to identified thalamocortical projection neurons in the anterior thalamus of the rat

We have carried out an ultrastructural study to determine the characteristics and distribution of glutamate-containing constituents of the anterodorsal (AD) and anteroventral (AV) thalamic nuclei in adult rats. We used a polyclonal antibody to glutamate and a postembedding immunogold detection method in animals in which the neurons of AD/AV projecting to the cortex had been retrogradely labelled and the terminals of corticothalamic afferents anterogradely labelled by injection of cholera toxin-horseradish peroxidase (HRP) into the retrosplenial granular cortex. The heaviest immunogold labelling was over axon terminals 0.42 to 2.2 microm in diameter containing round synaptic vesicles and establishing Gray type 1 (asymmetric) synaptic contact (type 1 terminals) on HRP-labelled or non-labelled dendrites. Mean gold particle densities over such terminals were 3-4 times higher than the densities over the dendrites to which they were presynaptic and 5-6 times higher than over terminals establishing Gray type 2 (symmetric) synaptic contacts (type 2 terminals). Gold particle densities over neuronal cell bodies and dendrites and over a subpopulation of myelinated axons were intermediate between the densities over type 1 and type 2 terminals. In adjacent serial sections immunoreacted for gamma aminobutyric acid, type 2 terminals were heavily immunolabelled whereas type 1 terminals and other profiles with moderate gold particle densities after glutamate immunoreaction displayed very low labelling. A subpopulation of small type 1 axon terminals (up to 1 microm diameter) contained HRP reaction product identifying them as cortical in origin; they contacted small dendritic profiles (most <1 microm diameter) many of which also contained HRP reaction product. We conclude that terminals of the corticothalamic projection from retrosplenial granular cortex to AD/AV are glutamatergic and innervate predominantly distal dendrites of thalamocortical projection neurons.     

Electron microscopic identification of axon terminals of retinopretectal fibers in the cat by a combined horseradish peroxidase and tritiated amino acids tracing method

The terminal nucleus of the optic tract in the pretectum was observed electron microscopically in the cat after intravitreous injection of a mixture of HRP and tritriated amino acids. Autoradiographic silver grains, and often also HRP granules, were seen in large axon terminals containing round synaptic vesicles and pale mitochondria. These terminals occasionally made synaptic contacts with the presynaptic dendrites and were involved in the formation of the synaptic triad.     

Serotonin-containing structures in the nucleus raphe dorsalis of the cat: an ultrastructural analysis of dendrites, presynaptic dendrites, and axon terminals

In the nucleus raphe dorsalis of the cat, an electron microscopic immunocytochemistry method was used to identify the fine structure of serotoninergic dendritic profiles and axon terminals analyzed in serial sections. Two classes of serotoninergic dendrites were distinguished in the nucleus. The first class was constituted by conventional serotonin (5-HT) dendrites that were contacted by unlabeled axon terminals containing differing populations of synaptic vesicles. The second class consisted of serotoninergic dendrites that contained vesicles in their dendritic shafts. Such 5-HT dendrites were further subdivided into two groups according to their synaptic contacts. In some 5-HT vesicle-containing dendrites, the vesicles were densely packed in small clusters and were associated with a well-defined synaptic specialization. These dendrites were classified as serotoninergic presynaptic dendrites and established synaptic contacts with unlabeled and labeled dendrites and were contacted by unlabeled axon terminals. In other 5-HT vesicle-containing dendrites, extensive serial section examination showed that the vesicles could be observed near the membrane but were never found to be associated with any synaptic membrane specialization. Serotoninergic axon terminals that were presumed to be recurrent collaterals of 5-HT neurons were present in the nucleus. Some of them were observed in synaptic contact with dendrites or dendritic protrusions whereas others did not exhibit synaptic specializations. The existence of serotoninergic dendrodendritic synaptic contacts and axon terminals suggests direct local interactions between serotoninergic neurons within the nucleus raphe dorsalis.     

Fine structure of the lateral mammillary projection to the dorsal tegmental nucleus of Gudden in the rat.

The synaptic organization of projections from the lateral mammillary neurons within the dorsal tegmental nucleus of Gudden is studied in the rat with the aid of anterograde transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) and visualized with tetramethylbenzidine. The dorsal tegmental nucleus consists of the pars ventralis (TDV) and the pars dorsalis (TDD). The normal neuropil of the dorsal tegmental nucleus contains three classes of axodendritic terminals, that is, terminals containing round, flat, and pleomorphic vesicles. They make up 44%, 5%, and 51%, respectively, of all axodendritic terminals in the TDV, and 62%, 1%, and 37% in the TDD. Injection of WGA-HRP into the lateral mammillary nucleus permits ultrastructural recognition of many anterograde labeled terminals within both the TDV and TDD. In the TDV, 81% of the labeled terminals contain round synaptic vesicles and make asymmetric synaptic contacts. A few of the labeled terminals contain pleomorphic vesicles and make symmetric synaptic contacts. More than 50% of the labeled terminals contact intermediate dendrites (1-2 microns diameter). In the TDD, almost all labeled terminals are small, contain round vesicles, and make asymmetric synaptic contacts. These terminals mainly contact intermediate as well as distal (less than 1 micron diameter) dendrites. There are only a few labeled terminals with pleomorphic vesicles and no terminals with flat vesicles. The termination pattern of the lateral mammillary neurons in the TDV is similar to that in the TDD. Anterograde labeled axon terminals often contact retrograde labeled dendrites in the TDV. No reciprocal connections are present in the TDD. These results suggest that the TDV and the TDD receive mainly excitatory and a few inhibitory inputs from the lateral mammillary nucleus. The TDV neurons also have direct reciprocal connections with the lateral mammillary neurons.     

Synaptic interrelationships between the optic tectum and the ipsilateral nucleus isthmi in Rana pipiens

The nucleus isthmi is reciprocally connected to the ipsilateral optic tectum. Ablation of the nucleus isthmi compromises visually guided behavior that is mediated by the tectum. In this paper, horseradish peroxidase (HRP) histochemistry and electron microscopy were used to explore the synaptic interrelationships between the optic tectum and the ipsilateral nucleus isthmi. After localized injections of HRP into the optic tectum, there are retrogradely labeled isthmotectal neurons and orthogradely labeled fibers and terminals in the ipsilateral nucleus isthmi. These terminals contain round. Clear vesicles of medium diameter (40–52 nm). These terminals make synaptic contact with dendrites of nucleus isthmi cells. Almost half of these postsynaptic dendrites are retrogradely labeled, indicating that there are monosynaptic tectoisthmotectal connections. Localized HRP injection into the nucleus isthmi labels terminals primarily in tectal layers B, E, F, and 8. The terminals contain medium-sized clear vesicles and they form synaptic contacts with tectal dendrites. There are no instances of labeled isthmotectal terminals contacting labeled dendrites. Retrogradely labeled tectoisthmal neurons are contacted by unlabeled terminals containing medium-sized and small clear vesicles. Fifty-four percent of the labeled fibers connecting the nucleus isthmi and ipsilateral tectum are myelinated fibers (average diameter approximately 0.6 μm). The remainder are unmyelinated fibers (average diameter approximately 0.4 μm). © 1994 Wiley-Liss, Inc.     

Serotonin-containing axon terminals in the hypoglossal nucleus of the rat. An immuno-electronmicroscopic study

The morphology and distribution of serotonin-containing axon terminals in the rat hypoglossal nucleus (XII) was investigated immunocytochemically at the electron microscopic level. Serotonin-positive profiles were found throughout all regions of XII and included unmyelinated axons, varicosities and axon terminals. Most labeled profiles (68.1%) were nonsynaptic unmyelinated axons and varicosities, while synaptic profiles, ending on dendrites and somata, were seen less frequently (28.7%). The majority of labeled axon terminals (76.9%) ended on small-to-medium-sized dendrites. Most axodendritic terminals contained small, round agranular vesicles (20-55 microns), several large (60-100 microns) dense core vesicles, and were associated with a pronounced asymmetric postsynaptic specialization. By contrast, labeled axosomatic terminals were seen less often than those ending on dendrites (23.0%). Axosomatic terminals typically contained small, round, agranular and large dense core vesicles and were associated with a symmetric or no postsynaptic specialization. These results provide the structural substrates for elucidating the functional role of serotonin in tongue control.     

Corticotectal terminals in the superior colliculus of the rabbit: a light- and electron microscopic analysis using horseradish peroxidase (HRP)-tetramethylbenzidine (TMB)

The projection from the striate cortex to the superior colliculus was studied light- and electron microscopically by means of anterogradely transported horseradish peroxidase and tetramethylbenzidine histochemistry. Labeled boutons were found in the stratum zonale (SZ) and in the stratum griseum superficiale (SGS), not in stratum opticum (SO). There are two maxima of frequency of labeled boutons, one in middle SGS at about 500 microns depth, and a smaller one in upper SGS at about 200 microns depth. Such a bimodal distribution of corticotectal terminals has not been described in any species before. Labeled myelinated axons were found in SGS and SO with a maximal frequency in middle SGS at about 400 microns depth. The myelinated axons in SZ, which are commonly considered to be of cortical origin, were not labeled. The labeled cortical terminals contained numerous round synaptic vesicles and predominantly dark mitochondria. They formed usually asymmetrical synapses and contacted dendrites, some of which contained synaptic vesicles. Occasionally, labeled boutons were observed which definitely did not belong to the type that is generally considered to be of cortical origin.     

Identification of signal substances in synapses made between primary afferents and their associated axon terminals in the rat trigeminal sensory nuclei

The relationships between primary afferent terminals (PATs) and their associated presynaptic terminals in the rat trigeminal sensory nuclear complex (TSNC) were examined with special reference to amino acid transmitters glutamate (Glu) and gamma-aminobutyric acid (GABA). Primary afferent terminals anterogradely labeled from the trigeminal ganglion with the B subunit of cholera toxin conjugated to horseradish peroxidase (CTB-HRP) were sectioned for electron microscopy. Serial sections from the principal nucleus (Vp), dorsomedial parts of the oral and interpolar nuclei (Vdm), and lamina III/IV of caudal nucleus (Vc) were immunostained for Glu and GABA by using a postembedding immunogold technique. The tracer, CTB-HRP to the trigeminal ganglion, preferentially labeled myelinated primary afferents. Sections immunostained with Glu antiserum showed that most labeled PATs were enriched with immunoreactivity (IR) for Glu. The Glu-IR PATs contained clear, round, synaptic vesicles and formed asymmetric synaptic contacts with somata or dendrites. They were frequently postsynaptic to, unlabeled axon terminals filled with a mixture of clear, round, oval, and flattened vesicles (p-endings), with symmetric synaptic junctions. The frequency of synapses onto somata or primary dendrites per Glu-IR PAT was higher in the Vdm than in either the Vp or Vc lamina III/IV. The frequency of contacts of the p-endings per Glu-IR PAT was higher in the Vp than in the Vdm and Vc lamina III/IV. Sections immunostained with GABA antiserum showed that most axon terminals presynaptic to PATs were enriched with GABA in the three nuclei. The GABA-IR axon terminals and their postsynaptic PATs had a similar ultrastructural character to p-endings and their postsynaptic Glu-IR PATs, respectively. The present study suggests that primary afferent neurons with large-caliber fibers use glutamate as a neurotransmitter and are subject to presynaptic modulation by GABAergic fibers.     

Synaptic organization of projections from basal forebrain structures to the mediodorsal thalamic nucleus of the rat.

The synaptic organization of the mediodorsal thalamic nucleus (MD) in the rat was studied with the electron microscope, and correlated with the termination of afferent fibers labeled with wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). Presynaptic axon terminals were classified into four categories in MD on the basis of the size, synaptic vesicle morphology, and synaptic membrane specializations: 1) small axon terminals with round synaptic vesicles (SR), which made asymmetrical synaptic contacts predominantly with small dendritic shafts; 2) large axon terminals with round vesicles (LR), which established asymmetrical synaptic junctions mainly with large dendritic shafts; 3) small to medium axon terminals with pleomorphic vesicles (SMP), which formed symmetrical synaptic contacts with somata and small-diameter dendrites; 4) large axon terminals with pleomorphic vesicles (LP), which made symmetrical synaptic contacts with large dendritic shafts. Synaptic glomeruli were also identified in MD that contained either LR or LP terminals as the central presynaptic components. No presynaptic dendrites were identified. In order to identify terminals arising from different sources, injections of WGA-HRP were made into cortical and subcortical structures known to project to MD, including the prefrontal cortex, piriform cortex, amygdala, ventral pallidum and thalamic reticular nucleus. Axons from the amygdala formed LR terminals, while those from the prefrontal and insular cortex ended exclusively in SR terminals. Fibers labeled from the piriform cortex formed both LR and SR endings. Based on their morphology, all of these are presumed to be excitatory. In contrast, the axons from the ventral pallidum ended as LP terminals, and those from the thalamic reticular nucleus formed SMP terminals. Both are presumed to be inhibitory. At least some terminals from these sources have also been identified as GABAergic, based on double labeling with anterogradely transported WGA-HRP and glutamic acid decarboxylase (GAD) immunocytochemistry.     

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.