Retinal synapses of the cat medial interlaminar nucleus and ventral lateral geniculate nucleus differ in size and synaptic organization

The retinal terminals of the medial interlaminar nucleus (MIN) and ventral lateral geniculate nucleus (VLG) have been examined quantitatively to determine if there are morphological differences in their synaptic ultrastructure which reflect their distinctive physiologies. The cross-sectional area and density (number per unit area) of synaptic contact zones with conventional and presynaptic dendrites (F2 profiles) were measured for each retinal terminal. The densities of F2 presynaptic dendrites and F1 flattened vesicle axon terminals were also measured. Retinal terminals in MIN were often large (mean size= 2.7 μm² area) and had a high density of synaptic contacts (0.14 per μm surface area) with conventional dendrites, presynaptic dendrites, and dendritic spines. A high density of F2 presynaptic dendrites (0.08 per μm² area) was found in MIN. F1 axon terminals were also found frequently (0.04 per μm²). MIN retinal terminals were often organized in glomeruli like those of the dorsal lateral geniculate nucleus. The retinal terminals in VLG were almost always small (mean size= 0.94 μm² area), although they also had a high density of synaptic contacts (0.17 per μm surface area). They frequently synapsed on small dendrites and dendritic spines and less frequently on large dendrites. Unlike MIN, retinal terminals in VLG rarely contacted F2 presynaptic dendrites which were much less frequent in VLG (0.01 per μm² area). Like MIN, VLG contained numerous F1 axon terminals (0.06 per μm² area). No typical retinal glomeruli were found in VLG. These results show that MIN, which contains many Y cells, has a population of large retinal terminals and many F2 presynaptic dendrites. VLG, which apparently has only W cells, contains only small retinal terminals and has fewer F2 presynaptic dendrites. Both have a high density of F1 flat vesicle axon terminals.     

Synaptology of retinal terminals in the dorsal lateral geniculate nucleus of the cat

We have made a fine structural investigation of the synaptic patterns made by axon terminals of retinal ganglion cells in the dorsal lateral geniculate nucleus of the cat. We compared the retinal input to dendritic processes that bear clusters of large appendages with the retinal input to relatively smooth dendritic segments that have only a few isolated spines. The study was restricted to the portion of laminae A and A1 that receive central visual field input. We were able to completely reconstruct 33 individual terminal boutons from long series of consecutive thin sections. Retinal terminals that were presynaptic to dendritic appendages tended to occupy the central position in the complex synaptic zones of geniculate fine structure called glomeruli. These terminals were surrounded by significantly more profiles than retinal terminals that were presynaptic to dendritic stems and averaged twice as many synaptic contacts per terminal bouton. The retinal input to dentritic appendages was heavily involved in a specific synaptic pattern called the triadic arrangement while retinal input to dendritic stems was only lightly involved in triads. Dendritic appendages in triads received greater synaptic input from profiles with flattened vesicles than did the dendritic stems that were found in triads.     

Parabrachial innervation of the cat's dorsal lateral geniculate nucleus: an electron microscopic study using the tracer Phaseolus vulgaris leucoagglutinin (PHA-L)

Ascending pathways from the brain stem play a key role, generally facilitatory, in controlling the transmission of retinal information through the lateral geniculate nucleus to the visual cortex (for reviews, see Singer, 1977; Burke and Cole, 1978; Sherman and Koch, 1986). In order to characterize the morphological basis of this brain-stem control, we used the electron microscope to study synaptic terminals labeled anterogradely from injections of the tracer Phaseolus vulgaris leucoagglutinin into the parabrachial region of the brain stem. The labeled axons, which are fine and unmyelinated in our material, form conventional synaptic contacts onto both relay cells and interneurons. These connections are surprisingly selective for certain postsynaptic elements such as the dendritic shafts and appendages of relay cells and the presynaptic dendritic terminals of interneurons. That is, the morphology of contacts made from parabrachial axons varies with the specific postsynaptic profile. Even a single axon can form symmetrical contacts onto F2 terminals, which are synaptic terminals deriving from dendrites of interneurons, and dendritic shafts of relay X cells, and form asymmetrical contacts onto dendritic appendages of the same relay X cells. Reconstructions of the dendritic segments postsynaptic to the labeled terminals show that the dendritic appendages receive retinal and parabrachial input in triadic relationships with F2 terminals: a retinal or parabrachial axon contacts the F2 terminal, and the F2 terminal plus the retinal or parabrachial axon contact the dendritic appendage. This positioning of the parabrachial innervation is well suited for control of retinal transmission. Finally, the dual morphology of the parabrachial synaptic contacts suggests that their actions may differ depending on the postsynaptic target.     

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.     

Glutamate-like Immunoreactivity in Retinal Terminals of the Mouse Suprachiasmatic Nucleus

With a view to identifying the neurotransmitter content of retinal terminals within the mouse suprachiasmatic nucleus, a highly specific antiserum to glutaraldehyde-coupled glutamate was used in a postembedding immunogold procedure at the ultrastructural level. Retinal terminals were identified by cholera toxin–horseradish peroxidase transported anterogradely from the retina and reacted with tetramethyl benzidine/tungstate/H₂O₂, or by their characteristically pale and distended mitochondria with irregular cristae. Controls included model ultrathin sections containing high concentrations of various amino acids. Alternate serial sections were labelled with anti-glutamate and anti-γ-aminobutyric acid (GABA). Data were analysed by computer-assisted image analysis. Density of glutamate labelling (gold particles per μm²) on whole retinal terminals was > 3 times higher than that on postsynaptic dendrites, and > 5 times higher than that on miscellaneous non-retinal non-glutamatergic terminals in the suprachiasmatic nucleus. The overall density of gold particles over retinal terminals was ～ 3 times higher than that over GABAergic terminals, in which glutamate-like immunoreactivity was mainly mitochondrial. Labelling of vesicles in retinal terminals was almost 5 times greater than the apparent labelling of vesicles in GABAergic terminals, underscoring the location of transmitter glutamate within synaptic vesicles in retinal terminals. In the retino-recipient region of the suprachiasmatic nucleus there was also a small population of non-retinal glutamatergic terminals. Their overall immunoreactivity was similar to or exceeded that of retinal terminals, but morphological features clearly distinguished between these two types of glutamate-containing terminals. The present results indicate that the vast majority of retinal terminals may use glutamate as a transmitter, in keeping with electrophysiological and neuropharmacological data from other sources. The possibility of cotransmitters within retinal terminals, suggested by the presence of dense-core vesicles among the glutamate-containing synaptic vesicles, has still to be addressed.     

Ultrastructural organization of GABA in the rabbit superior colliculus revealed by quantitative postembedding immunocytochemistry

We have studied the organization of gamma-aminobutyric acid (GABA)ergic profiles in the superior colliculus of the rabbit to determine whether the synaptic types found in cat and monkey also exist in a mammalian species whose visual system has a different organization. Ultrastructure of GABAergic profiles was examined by use of a polyclonal antibody to GABA and quantitative postembedding immunocytochemistry. Three distinct types of vesicle-containing profiles were labeled by the GABA antibody in the rabbit superior colliculus. One type was a putative presynaptic dendrite (PSD profile) that received synaptic input from other profiles and contained pleomorphic synaptic vesicles scattered throughout the profile. These PSD profiles frequently received retinal input and formed dendrodendritic synapses. A second type of profile was a large caliber dendrite, often horizontal in orientation (H profile), that had one or more discrete clusters of pleomorphic synaptic vesicles at sites of synaptic contact with conventional dendrites. These H profiles received few synaptic contacts. A third profile type was a putative axon terminal (F profile) with smaller, more flattened synaptic vesicles that densely and uniformly filled the profile. Quantitative analysis of gold particle density revealed that F profiles had a significantly higher gold particle density (14.3/ μm²) than did PSD or H profiles (10.4 and 10.2/ μm²), suggesting that GABAergic profile types contain different concentrations of GABA. The vesicle density of these profile types also differed, but no obvious relationship between vesicle and particle distributions was observed. We conclude that the profiles labeled by GABA in rabbit superior colliculus are similar to those in cat and monkey and must represent a phylogenetically conserved organization common to many mammals, and that particle density analysis of postembedding immunocytochemistry can distinguish different GABAergic profile types.     

Synaptic interactions between GABA-immunoreactive profiles and the terminals of functionally defined myelinated nociceptors in the monkey and cat spinal cord.

This study analyzes the synaptic interactions between the central terminals of A delta high threshold mechanoreceptors (A delta HTMs) and GABA-immunoreactive profiles. A delta HTM primary afferents from three monkeys and one cat were electrophysiologically identified and intracellularly labeled with HRP, and their terminal arborizations in laminae I and II of the sacrocaudal spinal cord were studied at the ultrastructural level. GABA-immunoreactive profiles in relation to A delta HTM terminals were demonstrated using postembedding colloidal gold techniques. Monkey A delta HTM terminals (n = 131) usually constituted the central element of synaptic glomeruli; they established large asymmetric synaptic contacts with 1-13 dendrites (modal value 2-4) and were surrounded by 0-6 peripheral axon terminals (modal value 2-3). The large majority (around 85%) of the peripheral axon terminals were GABA immunoreactive. They were found presynaptic to the A delta HTM terminal and/or to dendrites postsynaptic to the primary afferent terminal. Furthermore, all peripheral axon terminals found presynaptic to the A delta HTM terminals showed GABA immunoreactivity. Within a single A delta HTM fiber, this synaptic arrangement was found in 20-60% of its boutons. In addition, 28% of the postsynaptic dendritic profiles displayed weak GABA immunoreactivity. Some of them contained vesicles; however, only in a few cases did we observe synapses between a GABA-immunoreactive vesicle-containing dendrite and a dendritic profile postsynaptic to an A delta HTM terminal. Similar synaptology and interactions with GABA-immunoreactive profiles were displayed by the terminals of the single cat A delta HTM fiber studied. Our data support the hypothesis that GABA-containing neurons use both presynaptic and/or postsynaptic mechanisms to exert a powerful control, presumably inhibitory, over the transmission of nociceptive information between A delta HTM afferents and second-order neurons in monkey and cat spinal cord. Our results also imply that GABA may be released within the synaptic glomeruli formed by A delta HTM terminals either by local dendrites or by axon terminals. We discuss the possibility that these GABAergic synapses can be driven by inputs from both primary afferents and/or descending systems to modulate the transmission of nociceptive sensory information.     

Ultrastructural studies of retinal, visual cortical (area 17), and parabigeminal terminals within the superior colliculus of Galago crassicaudatus.

The morphology and synaptic relationships of anterogradely labeled retinal, visual cortical (area 17), and parabigeminal terminals have been analyzed within the superficial gray (stratum griseum superficiale) of Galago crassicaudatus. Our data regarding the retinocollicular projection reveal two populations of terminals based upon size. The population of smaller terminals are found in clusters, while the larger occur in isolation. Both populations of retinocollicular terminals form synapses primarily with dendritic spines, but synapses upon pale vesicle filled (PVF) profiles and dendritic shafts also occur. Corticotectal terminals contain round vesicles and make asymmetrical synapses, primarily onto dendritic spines; few form synapses with PVF profiles. Our findings suggest the possibility that there are two populations of corticotectal terminals based upon differences in size and morphology. Parabigeminotectal profiles contain densely packed round vesicles and make asymmetrical synapses. These terminals, which are exclusively cholinergic in Galago, are presynaptic to dendrites of various sizes. Convergence of retinal and cortical terminals has been observed. This convergence occurs on distinctly separate regions of the postsynaptic membrane. In contrast, convergence of retinal and parabigeminal terminals occurs on the same region of the postsynaptic cell(s).     

Characteristics of the synaptic apparatus of the parietal associative cortex (area 5b) in the cat brain

An electron microscopic examination of the associative cerebral cortex (area 5b) in cat was performed. The average density of axonal terminal profiles in this area was 263 +/- 16 terminals per 1000 micron2 of the slice area. 75.5% of axonal terminals contained synaptic vesicles and had asymmetric or symmetric contacts with visible active zones. 8.4% of axonal terminals had contacts without visible active zones. 24.5% of axonal terminals contained synaptic vesicles, but had no visible contacts with neighbouring structures. 84.9% of axonal terminals contained round or slightly elongated vesicles, 7.8% --a mixture of round and elongated vesicles and 7.3%--thin elongated vesicles. Among the axonal terminals with visible synaptic contacts 46.6% were of the axo-spine type, 50%--of axo-dendritic type and 3.4%--of axo-somatic type. 77% of axo-somatic terminals contained elongated vesicles and had symmetric contacts and 23% contained round vesicles and had asymmetric contacts.     

Ultrastructural synaptic organization of axon terminals in the ventral part of the cat oral pontine reticular nucleus

In an attempt to contribute to the current knowledge of the brainstem reticular formation synaptic organization, the ultrastructure and distribution of synaptic terminal profiles on neurons in the ventral part of the oral pontine reticular nucleus (vRPO), the rapid eye movement (REM) sleep-induction site, were studied quantitatively. Terminals with asymmetric contacts and rounded vesicles were classified according to vesicle density as type I or II (high or low density, respectively). The area, apposed perimeter length, and mitochondrial area of type I terminals, on average, were significantly smaller than those of type II terminals. Type III and IV terminals had symmetric contacts and oval and/or flattened vesicles; type III terminals formed synapses between them and on initial axons. Type V and VI terminals showed characteristics intermediate to those of asymmetric and symmetric synapses. Interestingly, some terminal features were related to both terminal area and postsynaptic dendritic diameter. The percentages of different synapses sampled on somata were as follows: asymmetric synapses (usually formed by type II terminals; mean +/- S.D.), 26.4% +/- 3%; symmetric synapses, 46.7% +/- 5.2%; and intermediate synapses, 26.9% +/- 6.1%. The percentages of different synapses sampled on dendrites were asymmetric synapses, 62.1% +/- 9%; symmetric synapses, 25.6% +/- 8.1%; and intermediate synapses, 12.3% +/- 1.7%. Comparison between large- and small-diameter dendrites revealed that the percentages of symmetric synapses and type II terminals decreased, whereas the percentages of type I terminals increased as postsynaptic dendritic diameters became smaller. Synaptic density was approximately four times lower on somata than on dendrites. The vRPO synaptic organization reflects some patterns that are similar to those found in other regions of the central nervous system as well as specific synaptic patterns that are probably related to its functions: the generation and maintenance of REM sleep and the control of eye movement or limb muscle tone.     

Quantitative comparison of retinal synapses in the dorsal and ventral (parvicellular) C laminae of the cat dorsal lateral geniculate nucleus

Three physiological classes of retinal ganglion cell project to the cat dorsal lateral geniculate nucleus (DLGN). The dorsal laminae A, A1, and magnocellular C receive X and Y retinal input, whereas the ventral parvicellular laminae C1 and C2 receive predominantly W input. We have compared quantitatively the retinal synaptic terminals of the dorsal and ventral laminae to determine whether there are morphological differences in the terminals that correspond to their different response properties. Anterogradely labeled retinal synaptic terminals in all laminae contained pale mitochondria and large, round synaptic vesicles. However, retinal terminals with pale mitochondria varied in size and synaptic organization in different laminae. The terminals in the A laminae were, on average, quite large and made numerous contacts with conventional dendritic profiles and with profiles that themselves contained synaptic vesicles (F2 profiles). The terminals in lamina C that contained pale mitochondria had a smaller overall mean area. Terminals with pale mitochondria in C1 and C2 were almost all small and synapsed with F2 profiles less frequently than did terminals in the A laminae or in lamina C. These results provide quantitative evidence that visual areas receiving W-type retinal input contain smaller retinal terminals and have a different synaptic organization from that of laminae receiving X and Y input.     

Light and electron microscopic localization of calcitonin gene-related peptide immunoreactivity in lamina II of the feline trigeminal pars caudalis/medullary dorsal horn: A qualitative study

Calcitonin gene-related peptide (CGRP) is a neuropeptide that is associated with a subset of primary afferent fibers and appears to have a role in nociception. The purpose of the present study was to perform a qualitative light, and especially electron microscopic (LM and EM), examination of CGRP-immunoreactivity (IR) within lamina II (substantia gelatinosa) of the feline pars caudalis/medullary dorsal horn (PC/MDH) of the spinal trigeminal nucleus. The LM investigation revealed massive CGRP-IR within lamina II outer, with fewer fibers that traversed lamina II inner. The EM preparations showed CGRP-IR in small, thinly myelinated and unmyelinated axons, preterminal axons, and in axon terminals that formed asymmetric synaptic contacts onto small dendritic profiles. The terminals with CGRP-IR were often the central element within glomeruli, where the terminal usually formed 2 or more asymmetric synaptic specializations onto 1 or more dendrites. Many of these postsynaptic dendrites contained synaptic vesicles. Other profiles were seen forming presynaptic contacts onto the terminal with CGRP-IR, and these profiles most likely represent presynaptic dendrites and/or other axon terminals of intrinsic origin. The synaptic association of terminals showing CGRP-IR with vesicle-containing dendrites, presynaptic dendrites, and/or other axon terminals suggests that terminals with CGRP-IR are especially susceptible to modulation. © 1993 Wiley-Liss, Inc.     

Fine structure of nigral and pallidal afferents in the thalamus: an EM autoradiography study in the cat

Ultrastructural characteristics and distribution of nigral and pallidal axon terminals on thalamic neurons were studied after injections of tritiated leucine into substantia nigra and entopeduncular nucleus respectively. Adult cats received 0.1–0.2-μl injections of 2, 3, 4, 5, ³H-leucine in a concentration 60 μCi/μl and were allowed to survive for 4–5 days. The brain tissue was processed for electron (EM) and light microscopic (LM) autoradiography. EM samples were obtained from the ventral medial and ventral anterior thalamic nuclei. Ultrastructural features of labelled nigral and pallidal boutons were analyzed both qualitatively and quantitatively. Ultrastructural characteristics of nigral and pallidal boutons appeared similar. Their length along postsynaptic membrane ranged from 0.8 to 10 μm, with average length of apposition around 2 μm. Both types of bouton contained small clear vesicles of extremely variable shape and formed symmetrical type contacts. Mean diameter of synaptic vesicles profiles (n = 500) was 32.5 nm and 33.3 nm in nigral and pallidal terminal respectively, and mean vesicle profile areas were 846 nm² in nigral terminals and 878m² in pallidal. Both parameters showed normal distribution in percentage distribution histograms. The mean ratios of longest and shortest diameters was 1.6 for synaptic vesicles in both types of boutons. Thus, no significant differences in morphological parameters of nigral and pallidal axon terminals were discovered except that pallidal afferents often formed “en passant”-type synapses while nigral did not. However, this feature alone is not sufficient for distinction between the two types of termi-nals in unlabelled tissue. Analysis of distribution of synaptic sites showed that only pallidal bou-tons formed axosomatic synapses on thalamocortical projection neurons (TCPN), which comprised 21% of total number of pallidal terminals studied. On primary dendritic trunks of TCPN the proportion of nigral boutons was larger (28.8%) as compared to pallidal (19%). The percentage of both types of bouton contacting secondary TCPN dendrites was similar (36% pallidal, 30.6% nigral), while the proportion of nigral terminals on tertiary TCPN den-drites was larger (23.6% versus 13%). Both afferents revealed a tendency to synapse preferentially at the branching points of TCPN dendrites with sev-eral boutons often found along the perimeter of the branching site. Small but equal proportions (8%) of both types of axon terminal were found to synapse on vesicle-containing dendrites of local circuit neurons. Nigral boutons were also found in complex synaptic arrangements in glomeruli. It is concluded that the organizations of pallidal and nigral afferent in-puts in the thalamus are rather similar. Both occupy strategic positions which would allow them to exert strong influence on the firing pattern of TCPN.     

Fine structure of calcitonin gene-related peptide immunoreactive synaptic contacts in the thalamus of the rat

Recent studies have shown a prominent calcitonin gene-related peptide immunoreactive (CGRP-ir) pathway extending from the external medial and external lateral para-brachial nuclei to the area surrounding and including the gustatory nuclei in the thalamus, and the cortex and amygdala. The function of the CGRP-ir pathway is not completely understood, but may be involved with the processing of both nociceptive and gustatory information in the thalamus. The purpose of this study was to characterize the nature of the CGRP-ir synaptic contacts in the gustatory nucleus. Electron microscopic examination of CGRP-ir synaptic contacts revealed two classes of CGRP-ir terminals. One class, which was large, formed asymmetric synaptic contacts on dendritic appendages, had many small, round synaptic vesicles, and heavy patches of reaction product which obscured any underlying organelles. Since similar terminals in unstained tissue contained large numbers of dense-cored vesicles, it was concluded that CGRP-ir was contained predominantly in dense-cored vesicles. A second class of CGRP-ir terminals was smaller and made either asymmetric or symmetric synaptic contacts. Both symmetric and asymmetric small terminals contained small, round synaptic vesicles and fewer patches of dense reaction product. Several of the CGRP-ir terminals making symmetric contacts also contained pleomorphic vesicles. There were very few contacts on cell bodies. There were no contacts on other CGRP-ir elements, somal or dendritic, or on axon terminals. None of the CGRP-ir terminal elements were postsynaptic to unlabeled terminals. Axons containing CGRP-ir were primarily unmyelinated, but a few myelinated axons were also seen. © 1993 Wiley-Liss, Inc.     

Synaptic organization of the nucleus gracilis of the cat. Experimental identification of dorsal root fibers and cortical afferents

The synaptic organization throughout the nucleus gracilis has been investigated in unoperated cats. Axon terminals of variable size can establish synaptic contacts with neuronal somata, dendritic processes, initial segment of axons or with other axon terminals at “complex synaptic arrangements.” Large boutons with rounded vesicles are regularly associated with smaller boutons containing flattened vesicles; the latter type of bouton forms frequently a double synapse being presynaptic to the large bouton and to the element postsynaptic to this (“complex synaptic arrangements”). Medium-sized to small axon terminals of the “isolated” type contain primarily either rounded or flattened vesicles. These boutons are surrounded by a thin glial process which also wraps the postsynaptic element, mostly represented by a small dendritic profile. The “isolated” type of bouton seems to be more abundant in the rostral than in the caudal part of the nucleus. In all unoperated control animals altered axons and axon terminals are present. They are enlarged and display hyperplasia and dilatation of tubular profiles of smooth endoplasmic reticulum as well as proliferation of microtubules and various aspects of mitochondrial degeneration. In cats sacrificed 48 hours after section of lumbo-sacral dorsal roots a high number of “dark” boutons are observed in various stages of degeneration. These terminals are identifiable with the large boutons containing rounded vesicles and postsynaptic to the smaller boutons with flattened vesicles. The morphology of dorsal root terminals in the nucleus gracilis is discussed in relation to that of primary afferent terminals in other central structures and to the functional aspects of axo-axonic contacts. The sensori-motor cortex was removed in another series of animals which were sacrificed after one to four days. As a consequence of such lesions cortical fiber terminals in the nucleus gracilis may undergo either the “dark” or the “light” type of degeneration. These terminals are of smaller size than those of primary afferents, they usually synapse on dendritic profiles of small diameter, are not involved in axo-axonic contacts and seem to contain rounded vesicles. Therefore they can be identified with at least some of the small and medium-sized boutons of the “isolated” type.     

Ultrastructural localization of dynorphin in the dentate gyrus in human temporal lobe epilepsy: a study of reorganized mossy fiber synapses

Substantial reorganization of mossy fibers from granule cells of the dentate gyrus occurs in a high percentage of humans with medically intractable temporal lobe epilepsy. To identify these fibers and determine their ultrastructural features in human surgical specimens, we used preembedding immunoperoxidase labeling of dynorphin A, an opioid peptide that is abundant in normal mossy fibers. In electron microscopic preparations, dynorphin A immunoreactivity was highly associated with dense core vesicles and was localized predominantly in axon terminals in the inner molecular layer of the dentate gyrus, although some dynorphin-labeled dense core vesicles were also observed in dendritic shafts and spines. The labeled terminal profiles were numerous, and, whereas they varied greatly in size, many were relatively large (2.3 microm in mean major diameter). The terminals contained high concentrations of clear round vesicles and numerous mitochondrial profiles, formed distinct asymmetric synapses, often had irregular shapes, and, thus, exhibited many features of normal mossy fiber terminals. The dynorphin-labeled terminals formed synaptic contacts primarily with dendritic spines, and some of these spines were embedded in large labeled terminals, suggesting that they were complex spines. The labeled terminals frequently formed multiple synaptic contacts with their postsynaptic elements, and perforated postsynaptic densities, with and without spinules, were present at some synapses. These findings suggest that the reorganized mossy fiber terminals in humans with temporal lobe epilepsy form abundant functional synapses in the inner molecular layer of the dentate gyrus, and many of these contacts have ultrastructural features that could be associated with highly efficacious synapses.     

Ultrastructure of chemically defined neuron systems in the dorsal horn of the monkey. I. Substance P immunoreactivity

The ultrastructural localization of substance P-like immunoreactivity (SPLI) in lamina I (marginal zone) and lamina II₀ (outer substantia gelatinosa) of the dorsal horn of the macaque monkey was examined by the indirect antibody peroxidase-antiperoxidase method. SPLI was found in small unmyelinated and finely myelinated axons and a variety of terminal types. The majority of SPLI terminals contained a few to many large granular vesicles (mean diameter 90 nm) in addition to a population of small clear round vesicles. A very few terminals contained mainly small round vesicles. SPLI terminals were presynaptic in axosomatic, axodendritic and axospinous contacts forming, in all but the axosomatic junctions, asymmetrical synapses. Some axosomatic junctions were symmetrical. SPLI terminals also formed the center of glomeruli with unlabeled dendrites and dendritic spines; some of the unlabeled dendrites contained a few small scattered vesicles and large dense-core vesicles. In more complex formations 2 to 4 SPLI terminals were associated with one another and linked by desmosomal contacts. The individual terminals in the complexes or ‘congregate terminals’ were simple large granular vesicle containing terminals (LGV), LGV-central terminals of associated glomeruli, or terminals containing mainly small round vesicles. In the apical region of lamina I an unlabeled terminal was found occasionally in contact with an SPLI terminal, which in turn synapsed onto a dendrite. These contacts have some synaptic characteristics and the SPLI terminal was possibly postsynaptic. Most of the types of SPLI terminals resemble closely terminal types shown to be of primary afferent origin. These terminals which make direct contact with dorsal horn dendrites may be the morphological substrate for postsynaptic excitation of dorsal horn neurons by substance P. The contacts of unlabeled terminals with SPLI terminals may represent a morphological substrate by which other neurochemical substances such as enkephalin or serotonin may modulate the substance P effects on dorsal horn neuronal activity.     

Inhibitory circuitry involving Y cells and Y retinal terminals in the C laminae of the cat dorsal lateral geniculate nucleus

We previously established (Datskovskaia et al. [2001] J Comp Neurol 430:85-100) that roughly 40% of Y retinal terminals contact interneurons in the A lamina of the dorsal lateral geniculate nucleus (dLGN) of the cat. However, we did not establish whether the dendritic terminals of interneurons postsynaptic to Y retinal terminals subsequently contact Y thalamocortical cells. To begin to address this issue, we examined the synaptic targets of Y retinal terminals in the magnocellular C lamina of the dLGN, which is populated almost exclusively by Y thalamocortical cells and interneurons. We utilized material generated from our previous work, in which we injected the superior colliculus with biotinylated dextran amine to backfill the geniculate branches of Y retinogeniculate axons in the dLGN. Sections prepared for electron microscopy were stained for gamma aminobutyric acid (GABA) to distinguish interneurons from thalamocortical cells. We found that the majority of profiles postsynaptic to Y retinal axons were the GABA-negative dendrites of thalamocortical cells (116/200, 58%). The remainder were GABA-positive dendrites of interneurons (84/200, 42%), many of which contained vesicles (F2 profiles; 54/200, 27%). In addition, we examined the synaptic targets of F2 profiles and found that almost all contacts of F2 profiles in the magnocellular C lamina were made onto the GABA-negative dendrites of thalamocortical cells (199/200, 99.5%). Thus, Y retinogeniculate axons contact interneurons and interneurons contact Y thalamocortical cells in the magnocellular C lamina of the dLGN. This indicates that interneurons are involved in modulation of the Y pathway.     

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.