The fine structure of the rat subthalamic nucleus: an electron microscopic study

The normal ultrastructure of the rat subthalamic nucleus (STH) was studied. The STH consisted of tightly packed neurons distributed within a neuropil filled with large numbers of blood vessels and thinly myelinated fibers. The somata of STH neurons (diameters, D, between 10 and 25 micron) contained abundant organelles but had only a small amount of both smooth and rough endoplasmic reticulum. The nuclei had deeply invaginated nuclear envelopes and pale nucleoplasm with little heterochromatin. STH neurons often were tightly apposed without any intervening glial membranes. Similar appositions were also found between somata and dendrites, dendrites and dendrites, and dendrites and initial axon segments. Although puncta adhaerentia were often observed, no gap junctions were found on any of these membrane appositions. In the neuropil, the dendrites were mostly smooth and thin (D between 0.5 and 1 micron) with an occasional stubby spine or thin dendritic appendage. At least two types of axon terminals were identified. Type 1 terminals (D up to 1 micron) contained medium-sized round vesicles (D about 45 nm) and formed asymmetrical synapses. Type 2 terminals were often large (D up to 5 micron) and contained both round and slightly flattened vesicles (D up to 50 nm). The type 2 terminals frequently formed adherens junctions with their postsynaptic targets in addition to forming relatively symmetrical synaptic junctions. The remaining axon terminals included a small number of terminals with various morphological characteristics and possibly some tangentially sectioned type 1 and type 2 terminals. Therefore they have not been classified as individual types in this study. A quantitative analysis indicated that the type 1 terminals formed synapses mainly with thin dendrites whereas the type 2 terminals formed synapses mainly with somata and larger dendrites.     

The fine structures of the suprachiasmatic nucleus of the golden hamster

The fine structure of the suprachiasmatic nucleus of the golden hamster was studied with special reference to the synaptic endings. The somata of SCN neurons contained well developed cytoplasmic organelles including the Golgi complex, mitochondria and polysomes. The nuclei had deeply invaginated nuclear membrane. Some neurons were characterized by the presence of a large number of granulated vesicles and an abundance of cytoplasmic organelles. Several kinds of synapses and gap junctions were also observed. Axo-somatic synapses were identified and could be differentiated based upon possessing two types of presynaptic elements. The first contained clear round synaptic vesicles (40-60 nm in diameter), the second contained clear round vesicles (40-60 nm in diameter) and dense cored vesicles (70-120 nm in diameter). Asymmetrical synaptic membrane thickening were observed on both the pre- and postsynaptic sides of most of the axosomatic synapses. Axo-dendritic synapses could also be divided into two sub-types according to their membrane specializations. In Type 1, the axon terminals contained both clear round vesicles and dense cored vesicles and formed asymmetrical synapses. Terminals in the second group (type 2) were characterized by symmetrical synapses that contained clear, round vesicles as well as dense cored vesicles. The type 1 terminals were evenly distributed throughout the SCN, but the type 2 terminals were encountered more frequently in the ventral SCN. These observations indicate that the ventral and dorsal components of the SCN may possess different functional roles.     

Ultrastructural single- and double-label immunohistochemical studies of substance P-containing terminals and dopaminergic neurons in the substantia nigra in pigeons.

The vast majority of striatonigral projection neurons in pigeons contain substance P (SP), and the vast majority of SP-containing fibers terminating in the substantia nigra arise from neurons in the striatum. To help clarify the role of striatonigral projection neurons, we conducted electron microscopic single- and double-label immunohistochemical studies of SP+ terminals and/or dopaminergic neurons (labeled with either anti-dopamine, DA, or anti-tyrosine hydroxylase, TH) in pigeons to determine: (1) the synaptic organization of SP+ terminals, (2) the synaptic organization of TH+ perikarya and/or dendrites, and (3) the synaptic relationship between SP+ terminals and TH+ neurons in the substantia nigra. Tissue single-labeled for SP revealed numerous SP+ terminals contacting thin unlabeled dendrites in the substantia nigra, but few SP+ terminals were observed contacting perikarya or large-diameter dendrites. SP+ terminals contained round, densely packed, clear vesicles, and often contained one or more dense-core vesicles. Synaptic junctions between SP+ terminals and their targets were more often symmetric (86%) than asymmetric. In tissue single-labeled for DA, we observed few terminals contacting DA+ perikarya, whereas terminals contacting DA+ dendrites were more abundant. Terminals contacting DA+ structures comprised at least four different morphologically distinct types based on the morphology of the clear synaptic vesicles and the type of synaptic junction. One type of terminal contained round clear vesicles and made symmetric synapses, and thus resembled the predominant type of SP+ terminal. The second type contained round clear vesicles and made asymmetric synapses, the third type contained medium-size pleomorphic clear vesicles and made symmetric synapses, and the fourth type contained small pleomorphic clear vesicles and made symmetric synapses. The presence of contacts between SP+ terminals and dopaminergic dendrites in the substantia nigra was directly demonstrated in tissue double-labeled for SP (by the peroxidase-antiperoxidase procedure, or PAP, with diaminobenzidine) and TH (by either the silver-intensified immunogold procedure or the PAP procedure with benzidine dihydrochloride). SP+ terminals commonly contacted thin TH+ dendrites in the substantia nigra, but few SP+ terminals contacted large-diameter TH+ dendrites or perikarya. Synapses between SP+ terminals and TH+ neurons were always symmetric. TH+ dendrites also were contacted by terminals not labeled for SP, which were more abundant than were SP+ terminals. Non-TH+ neurons were also contacted by both SP+ terminals and non-SP+ terminals.(ABSTRACT TRUNCATED AT 400 WORDS)     

Ultrastructural analysis of choline acetyltransferase-immunoreactive sympathetic preganglionic neurons and their dendritic bundles in rat thoracic spinal cord

We have used a monoclonal antibody against choline acetyltransferase (ChAT) to aid in the identification of sympathetic preganglionic neurons (SPNs) and to examine their ultrastructure in rat thoracic spinal cord. The clusters of ChAT-immunoreactive (ChAT-IR) preganglionic cell bodies and their distinctive bundles of dendrites give rise to a ladder-like appearance in horizontal light microscopic sections. This organization also produced a characteristic appearance of preganglionic neuropil when viewed electron microscopically. The intermediolateral (IML) nucleus contained numerous rostrocaudally oriented ChAT-IR dendrites. In addition, mediolaterally oriented ChAT-IR dendrites extended between the IML and the central autonomic region. Both the ChAT-IR dendrites and somata of preganglionic neurons were intimately associated with astroglial processes. The cell bodies were typically covered over a large proportion of their surface by a thin astrocytic sheath, and this was associated with a paucity of axon terminals forming axosomatic synapses. Instead, the vast majority of synapses upon SPNs were of the axodendritic type. The most frequently observed type of axon terminal contained numerous round, clear vesicles along with several dense-core vesicles (DCVs). In addition, some boutons contained a predominance of DCVs. Serial section analysis revealed that these apparently diverse morphological classes of synaptic boutons may simply represent variability of structure throughout a single terminal, with a greater proportion of DCVs being located distal to the synaptic specialization and a greater number of round, clear vesicles being present adjacent to the synapse. Analysis of the dendritic bundles revealed that individual dendrites, like the cell bodies, were often isolated from each other and the surrounding neuropil by astrocytic processes. This arrangement usually was interrupted only at regions of synaptic contact where astrocytic processes surrounded the synaptic complex as a whole. Thus, astroglial ensheathment of SPNs seems designed to minimize cross-talk between the bundled dendrites, as well as to isolate or segregate the diverse afferent inputs known to impinge on these cells.     

Fine structure and synaptic connections of the common spiny neuron of the rat neostriatum: A study employing intracellular injection of horseradish peroxidase

Medium-sized spiny neurons of the rat neostriatum, identified by intracellular injection of horseradish peroxidase, were examined at both light and electron microscopic levels. These neurons were characterized by their heavy investment of dendritic spines, beginning about 20 μm from the soma and continuing to the tips of the dendrites. Their axons arose from the soma or from a large dendritic trunk very near the soma, and tapered rapidly to form a main axonal branch from which arose several smaller initial collaterals. These arborized extensively throughout an area of about the same size as, and highly overlapping with, the dendritic field of the cell, while the main axon could be followed for distances of up to 1 mm in the direction of the globus pallidus. Three major synaptic types were seen in contact with spiny neurons. Boutons containing small round synaptic vesicles formed synapses exclusively with spiny regions of the dendrites, and most of these were axo-spinous. Small, very pleomorphic synaptic vesicles characterized a second bouton type of unknown origin, which made contacts with somata, initial segments, and dendrites, but not dendritic spines. Boutons containing large pleomorphic synaptic vesicles had the most widespread distribution, contacting all regions including dendritic spines. Spines receiving these contacts also were postsynaptic to boutons containing small round vesicles. Axon collaterals of spiny cells formed synapses with large pleomorphic vesicles and made synapses with somata, initial segments of axons, dendrites, and dendritic spines of striatal neurons, including other spiny cells.     

Fine structure of the medullary lateral line area of Chelon labrosus (order perciformes), a nonelectroreceptive teleost

The ultrastructure and synaptic organization of the nucleus medialis and cerebellar crest of the teleost Chelon labrosus have been investigated. The nucleus medialis receives projections from the anterior and posterior lateral line nerves. This nucleus consists of oval neurons and large crest cells (“Purkinje-like” cells) whose apical dendrites branch in the overlying molecular layer, the cerebellar crest. In the dorsal region of the nucleus medialis, the perikarya and smooth primary dendrites of the crest cells are interspersed among myelinated fibers and nerve boutons. The ventral layer of the nucleus medialis contains crest cell perikarya and dendrites as well as oval neurons. The cerebellar crest lacks neuronal bodies, but the apical dendrites of crest cells receive synapses from unmyelinated and myelinated fibers. In the cerebellar crest, two types of terminals are presynaptic to the crest cell dendrites: boutons with spherical vesicles that from asymmetric synapses with dendritic spines and boutons containing pleomorphic vesicles that from symmetric synapses with dendritic spines and boutons containing pleomorphic vesicles that from symmetric synapses directly on the dendritic shaft. Most axon terminals found on the somata and primary dedrites of crest cells in the nucleus medialis have pleomorphic vesicles and form symmetric contacts, though asymmetric with spherical vesicles and mixed synapses can be observed; these mixed synapses exhibit gap junctions and contain spherical vesicles. Unlike crest cells, the oval neuron perikarya receive three types of contacts (symmetric, asymmetric, and mixed). The origins and functions of these different bouton types in the nucleus medialis are discussed. © 1995 Willy-Liss, Inc.     

Immunocytochemical localization of choline acetyltransferase within the rat neostriatum: a correlated light and electron microscopic study of cholinergic neurons and synapses

Monoclonal antibodies to choline acetyltransferase (ChAT) were used in an immunocytochemical study to characterize putative cholinergic neurons and synaptic junctions in rat caudate-putamen. Light microscopy (LM) revealed that ChAT-positive neurons are distributed throughout the striatum. These cells have large oval or multipolar somata, and exhibit three to four primary dendrites that branch and extend long distances. Quantitative analysis of counterstained preparations indicated that ChAT-positive neurons constitute 1.7% of the total neuronal population. Electron microscopy (EM) of immunoreactive neurons initially studied by LM revealed somata characterized by deeply invaginated nuclei and by abundant amounts of organelle-rich cytoplasm. Surfaces of ChAT-positive neurons are frequently smooth, but occasional somatic protrusions and dendritic spines occur. Although infrequently observed, axons of ChAT-positive neurons branch, receive synapses, and become myelinated. Unlabeled boutons make both symmetrical and asymmetrical synapses with ChAT-positive somata and proximal dendrites, but are more numerous on distal dendrites. In addition, some unlabeled terminals form asymmetrical synapses with ChAT-positive somata and dendrites that are distinguished by prominent subsynaptic dense bodies. Light microscopy demonstrated a dense distribution of ChAT-positive fibers and punctate structures in the striatum, and these structures appear to correlate, respectively, with labeled preterminal axons and presynaptic boutons identified by EM. ChAT-positive boutons contain pleomorphic vesicles, and make symmetrical synapses primarily with unlabeled dendritic shafts. Furthermore, they establish synaptic contacts with somata, dendrites and axon initial segments of unlabeled neurons that ultrastructurally resemble medium spiny neurons. These observations, together with the results of other investigations, suggest that medium spiny GABAergic projection neurons receive a cholinergic innervation that is probably derived from ChAT-positive striatal cells. The results of this study also indicate that cholinergic neurons within caudate-putamen belong to a single population of cells that have large somata and extensive sparsely spined dendrites. Such neurons, in combination with dense concentrations of ChAT-positive fibers and terminals, are the likely basis for the large amounts of ChAT and acetylcholine detected biochemically within the neostriatum.     

An electron microscopic study of the origins of the projections of the first auditory area of the cortex (AI) to the medial geniculate body in cats

An electron microscope study of retrogradely labelled neurons in layer VI of the primary auditory cortex (AI) after injection of the horseradish peroxidase to the medial geniculate body was carried out in cats. Three-eight synapses (4.6 +/- 0.6 at an average) were revealed on the somata profiles of these retrogradely labelled cortico-geniculate neurons. Synapses occupied 10.8 +/- 1.0% of the somatic profile of cortico-geniculate neurons. Almost all (98.7%) of these axosomatic synapses had symmetrical contacts and were formed by axonal terminals with small elongated synaptic vesicles. HRP retrogradely labelled axonal terminals of geniculo-cortical fibres were revealed in neuropil of layer VI. They contained large round synaptic vesicles and formed asymmetrical synapses, mainly on spines. The role of axo-somatic synapses in regulation of the activity of cortico-geniculate neurons was discussed.     

Cytology and synaptology of the lateral reticular nucleus of the cat

The organization of lateral reticular nucleus (LRN) of the cat was investigated using electron microscopy and Golgi techniques. Golgi-Cox preparations revealed that the LRN consists of allodendritic and, especially, isodendritic neurons. The latter have been associated with neural centres that have important roles integrating signals from distant sources. Several forms of spines were identified with the Golgi method, and their ultrastructural correlates were determined. Somatic spines resembled stubby protrusions, while dendritic spines, where were usually observed on distal dendrites, appeared as pedunculated spines, racemose appendages and spine-crowned appendages. Ultrastructural examination of this nuclease revealed various synaptic relationships. The majority of the synaptic terminals were small (1.5--2.5 micrometer in diameter), contained round vesicles and usually contacted dendrites and spines. Other small terminals contained pleomorphic vesicles and contacted distal dendrites and spines. Large terminals (greater than 2.5 micrometer in diameter) with round or pleomorphic vesicles contacted the somata or proximal dendrites. Three types of "synaptic configurations," which consisted of discrete aggregations of neuronal processes invested by astrocytic lamellae, were also identified. These structural arrangements likely provide a basis for the integration of inputs to the LRN from spinal and supraspinal centres.     

GABAergic neural elements in the rat basilar pons: electron microscopic immunochemistry

Previous light microscopic immunoperoxidase studies of glutamic acid decarboxylase (GAD)-immunoreactive neural elements in the rat basilar pontine nuclei revealed immunocytochemical reaction product in neuronal somata and axon terminals. In the present study, pre-embedding immunoperoxidase labeling of GAD or gamma-aminobutyric acid (GABA) and postembedding immunogold labeling of GABA allowed the ultrastructural visualization of these neural elements in the basilar pontine nuclei of colchicine-treated animals. At the electron microscopic level, immunolabeled neuronal somata exhibited smoothly contoured nuclei, whereas some dendrites also contained reaction product after immunocytochemical treatment and were postsynaptic to both immunoreactive and nonimmunoreactive axon terminals. Synaptic boutons immunoreactive for GAD or GABA exhibited cross-sectional areas that ranged from 0.1 to 3.8 microns 2 and generally appeared round or elongated in most sections. The majority (95%) of immunolabeled boutons contained pleomorphic synaptic vesicles and formed symmetric synapses at their postsynaptic loci; however, boutons exhibiting round vesicles and boutons forming asymmetric synapses (5%) were also immunopositive. Small (less than 1.5 microns 2) GAD- or GABA-labeled axon terminals formed synaptic contact mainly with small dendritic profiles, dendritic spines, and neuronal somata, whereas large labeled boutons (greater than 1.5 microns 2) formed synapses with all sizes of dendritic profiles. Occasionally, a single immunolabeled bouton formed synaptic contact with two separate postsynaptic dendrites. It is suggested that the immunolabeled neuronal somata and dendrites observed in the rat basilar pontine nuclei represent a population of pontine local circuit neurons; however, it is known that GABAergic cell groups extrinsic to the pontine gray provide afferent projections to the basilar pons, and therefore at least some immunoreactive axon terminals present in the pontine nuclei are derived from these extrinsic sources. The ultrastructural observation of GABAergic neural elements in the rat basilar pontine nuclei confirms previous light microscopic findings and provides an anatomical substrate through which GABAergic neurons, whether arising from an intrinsic or extrinsic source, might exert an inhibitory influence on target cells within the pontine nuclei.     

Fine structural survey of the rat's brainstem sensory trigeminal complex

The fine structural organization of the principal sensory trigeminal nucleus was compared with that of the spinal trigeminal nucleus (subnuclei oralis, interpolaris, and the deep layers of caudalis) in adult albino rats. Direct comparisons indicate similarities between all of the subdivisions of the brainstem trigeminal complex both in the major morphological classes of neurons present and in basic patterns of synaptic connections. Major differences between the several subdivisions occur in the relative numbers and distribution of the different cell types. The spinal trigeminal nucleus is distinguished by more numerous large (22-40 micron) polygonal neurons which give rise to long straight primary dendrites. Both the perikaryal surface and the thick primary dendrites of many of these cells are densely innervated by synaptic terminals. Especially large cells of this type are a prominent feature of subnucleus oralis. By contrast, the principal sensory nucleus is distinguished by its high density of small to medium-sized (8-20 micron) round or ovoid neurons. These smaller neurons tend to receive a sparse axosomatic innervation. In addition to these differences the spinal trigeminal neuropil is distinguished by the striking manner in which it is broken up by large rostrocaudally oriented bundles of myelinated axons. Proximal dendrites of polygonal and fusiform neurons often wrap around these large axon bundles. Morphologically heterogeneous populations of synaptic terminals with round vesicles (R terminals) and terminals with predominantly flattened vesicles (F terminals) occur in all of the subdivisions of the trigeminal complex. Both types of terminal make primarily axodendritic synapses, but both also make axosomatic synapses, and axospinous synapses with somatic as well as dendritic spines. In addition, axoaxonic synaptic contacts from F terminals onto large R terminals are seen in all subdivisions. Convincing examples of presynaptic dendrites were not observed in any of the brainstem subdivisions. Synaptic glomeruli, characteristic groupings of dendrites and synaptic terminals, are found throughout the brainstem trigeminal complex. The dendritic elements in these glomeruli tend to be small-diameter dendrites, spines, and large, spinelike appendages. Within the glomerulus these elements are postsynaptic to a single large R terminal and may also be postsynaptic to smaller F terminals. In addition, axoaxonic synaptic contacts from the F terminals onto the R terminal are a consistent feature of trigeminal synaptic glomeruli.(ABSTRACT TRUNCATED AT 400 WORDS)     

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.     

Ultrastructure of the dorsal motor nucleus of the vagus nerve in monkey with a comparison of synaptology in monkey and cat

Neurons of the dorsal motor nucleus of the vagus nerve were studied following injections of horseradish peroxidase into the vagus nerve in a monkey (Macaca fascicularis). In frozen sections, the dorsal motor nucleus appeared to be completely filled by labeled medium-sized (20-30 micron in long axis) neurons. Labeled dendrites from these neurons often extended outside the borders of the nucleus into the nucleus of the tractus solitarius. In 1 micron thick plastic sections and ultrathin sections of the dorsal motor nucleus, two distinct types of neurons were observed with the light and electron microscope. Medium-sized neurons with abundant cytoplasm and an oval nucleus were retrogradely labeled with HRP, while small (10-15 micron in long axis) neurons with a paucity of organelles and an invaginated nucleus remained unlabeled. Medium-sized neurons outnumbered the small neurons by approximately five to one. The synaptic organization of the dorsal motor nucleus in monkey was studied and compared with that in cat. The porportions of different types of axosomatic synapses were similar in both species. Terminals containing round vesicles and making symmetrical or asymmetrical contact with the postsynaptic structure were more common than synaptic terminals containing pleomorphic vesicles. In both species, there was a slightly greater synaptic density on the medium-sized neurons than on the small neurons. The synaptic density in the monkey dorsal nucleus was greatest on the smallest dendrites in the neuropil and least on the somata.     

Neuronal associations in the rat suprachiasmatic nucleus demonstrated by immunoelectron microscopy.

The synaptic associations of neurons in the suprachiasmatic nucleus (SCN) of rats were examined by single immunolabeling for somatostatin (SRIH) and arginine vasopressin (AVP), and double immunolabeling for SRIH plus AVP and vasoactive intestinal polypeptide (VIP) plus AVP. Single immunolabeling showed that SRIH neurons, which displayed some somatic and dendritic spines, formed synaptic contacts with immunonegative and positive axon terminals. AVP neurons also formed synaptic contacts with both immunonegative and positive axon terminals. The immunonegative terminals contained small, spherical clear vesicles or flattened clear vesicles. A few immunopositive AVP fibers made synapses with immunonegative somatic or dendritic spines. Double immunolabeling showed synaptic associations between SRIH axons and AVP cell bodies or dendritic processes, and between AVP axons and the somata or dendrites of SRIH neurons. These findings suggest a reciprocal relation between the two types of neurons. Synaptic contacts between AVP neurons and VIP axon terminals were also demonstrated. Previously, we found synapses between SRIH axons and VIP neurons. Thus SRIH neurons appeared to regulate AVP and VIP neurons. On the basis of these findings, two possible oscillation systems of the SCN are proposed.     

An electron microscope study of the red nucleus in the cat, with special reference to the quantitative analysis of the axosomatic synapses.

The synaptic organization of the red nucleus in the cat was investigated using the electron microscope and the axosomatic synapses were analyzed quantitatively using serial sections. The bouton covering ratios were found to be 61.5, 16.6 and 6.1% in large, medium-sized and small neurons, respectively. In a vast majority of axosomatic terminals, the synaptic apposition length ranged from 1.2 to 1.4 mum. There were 15-17 axon terminals on each 100 sq. mum of perikaryal surface of a magnocellular neuron. Seventy-four per cent of axosomatic terminals on the magnocellular neuron were filled with spherical vesicles and 22% had flattened vesicles. No clear correlation appears to exist between the shape of synaptic vesicles and the type of the postsynaptic differentiation. Somatic thorns were observed rather frequently on the magnocellular neurons. Axo-dendrodendritic serial synapses were occasionally observed to be present in the red nucleus. All postsynaptic components of these serial synapses contained pleomorphic vesicles. The possible existence of the Golgi type II cells in the red nucleus is discussed in relation to the components consituting the serial synapses.     

A light and electron microscopic study of the dorsal motor nucleus of the vagus nerve in the cat

The morphology of the dorsal motor nucleus of the vagus nerve (DMV) in the cat was studied with the aid of light and electron microscopy. In frozen sections stained by the Kluver-Barrera method or stained to show retrograde labeling in the DMV following injections of horseradish peroxidase (HRP) in the cervical vagus nerve and the stomach wall a range of sizes of DMV neurons was observed but it was not possible to distinguish separate types. In contrast, two distinct types of neurons, one medium-sized and the other small, were identified with the light microscope in Golgi-Cox and 1-μm Epon sections and with the electron microscope in ultrathin sections. The medium-sized neurons had a range of sizes but generally measured 18 × 25 μm and possessed three to four proximal dendrites which branched two or three times. Spines were observed occasionally on the soma and on dendrites. These neurons contained a well-developed cytoplasm and a noninvaginated round to oval nucleus. The small neurons generally measured 9 × 14 μm and were round or slightly elongated in shape. Their dendritic processes were fewer and thinner than those of the medium-sized neurons and extended for shorter lengths. Their soma contained scanty cytoplasm and an invaginated nucleus. The medium-sized neurons outnumbered the small neurons by more than three to one but both neuronal types were distributed evenly throughout the nucleus. The medium-sized neurons seemed to correspond in size to the parasympathetic efferent neurons of the viscera as indicated by the HRP studies. Axosomatic synapses on both types of neurons and axodendritic synapses were observed in the DMV. Terminals containing mainly small clear round vesicles and making asymmetrical contact with the postsynaptic membrane were involved in the majority of synapses on both the soma and dendrites. Terminals containing predominantly pleomorphic vesicles and making symmetrical contact with the postsynaptic membrane were also common, comprising up to one-third of all synapses observed. Serial sections revealed that most synaptic terminals contained varying numbers of large (75–110 nm) dense-cored vesicles. Smaller dense-cored vesicles (45–55 nm) were sometimes observed, often close to the area of synaptic contact. Terminals 1–2 μm in diameter which contacted dendrites 1–3 μm in diameter formed the most common synaptic combination throughout the rostral to caudal extent of the DMV. No distinct regional differences were observed with respect to distribution of synaptic types.     

Direct synaptic contacts on the myenteric ganglia of the rat stomach from the dorsal motor nucleus of the vagus

The myenteric ganglia regulate not only gastric motility but also secretion, because a submucous plexus is sparsely developed in the rodent stomach. We have examined whether the neurons of the dorsal motor nucleus of the vagus (DMV) have direct synaptic contacts on the myenteric ganglia and the ultrastructure of the vagal efferent terminals by using wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). The myenteric ganglia of the rat were composed of four types of neurons, i.e., small, medium-sized, large, and elongated neurons. The average numbers of axosomatic terminals per profile were 2.0 on the small neurons, 3.1 on the medium-sized neurons, 1.2 on the large neurons, and 4.2 on the elongated neuron. More than half of the axosomatic terminals contained round vesicles and formed asymmetric synaptic contacts on the small, medium-sized, and large neurons. About 80% of the axosomatic terminals on the elongated neurons contained pleomorphic vesicles and formed asymmetric synaptic contacts. When WGA-HRP was injected into the DMV, many anterogradely labeled terminals were found around the myenteric neurons. The labeled terminals were large (3.16 +/- 0.10 microm) and contacted exclusively the somata. Most of them (about 90%) contained round vesicles and formed asymmetric synaptic contacts. Serial ultrathin sections revealed that almost all neurons in a ganglion received projections from the DMV. The vagal axon terminals generally contacted the medium-sized or the elongated neurons, whereas a few labeled terminals contacted the small and the large neurons. The present results indicate that the DMV projects to all types of neurons and that their axon terminals contain mostly round synaptic vesicles and form asymmetric synaptic contacts.     

Ultrastructure and synaptic organization of axon terminals from brainstem structures to the mediodorsal thalamic nucleus of the rat.

The ultrastructural characteristics and synaptic organization of afferent terminals from the brainstem to the mediodorsal thalamic nucleus (MD) of the rat have been studied with the electron microscope, by means of anterograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Labeled fibers were seen predominantly in the lateral portion of MD after the injections of WGA-HRP into the substantia nigra pars reticulata (SNr), the superior colliculus (SC), and the dorsal tegmental region (DT). The boutons arising from the SC were relatively small (less than 1.5 microns in diameter), formed asymmetric synaptic contacts with small dendrites and dendritic spines, and contained round synaptic vesicles. The axon terminals from the DT were mostly large boutons (2-4.5 microns) with asymmetric synaptic specializations and round vesicles. These boutons and their postsynaptic targets formed synaptic glomeruli that were entirely or partially ensheathed by glial lamellae. The ultrastructural features are almost identical to those of boutons in the medial and central segments of MD that were previously shown to originate from the basal amygdaloid nucleus and the piriform cortex. The boutons from the SNr had a wide range in size, but the majority were medium-sized to large (1.5-4 microns). The nigral boutons established symmetric synaptic contacts with dendritic shafts and occasionally with somata, and contained pleomorphic vesicles. However, like the DT terminals, they participated in glomerular formations. The nigral terminals closely resemble previously described terminals in the medial part of MD from the ventral pallidum, except that the nigral terminals formed en passant and axosomatic synapses as well as axodendritic synapses. A combined immunohistochemistry and WGA-HRP tracing study revealed that the nigral inputs were immunoreactive for glutamic acid decarboxylase and the axon terminals from the DT were immunoreactive for choline acetyltransferase. In a separate study, the colliculothalamic fibers have been shown to take up and transport the transmitter specific tracer [3H]-D-aspartate, and are therefore putatively glutamatergic and/or aspartatergic. Taken together with this, the present results suggest that the collicular afferents are excitatory and glutamatergic and/or aspartatergic, that the inputs from the DT are also excitatory and cholinergic, while the nigral inputs are inhibitory and GABAergic.     

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.