Corticotropin-releasing factor-like immunoreactive nerve fibers in the rat superior cervical ganglion and their fine structures

The existence of nerve fibers containing corticotropin-releasing factor (CRF)-like immunoreactivity (CRFI) in the rat superior cervical ganglion (SCG) was demonstrated by using immunocytochemistry. They were found to be extrinsic in origin, because no CRFI neurons were seen in the SCG and decentralization resulted in the disappearance of CRFI fibers in the SCG on the operated side. These findings were also confirmed by immunoelectron microscopic analysis; CRFI fibers contained a number of small clear synaptic vesicles but were devoid of large granular and agranular vesicles. These morphological characteristics are identical to those of the preganglionic fibers. The present immunoelectron microscopic analysis revealed that most of the CRFI fibers in the SCG make synaptic contact predominantly with the dendrites of the principal cells, partly with their somas and rarely with a non-CRFI terminal. Thus, the present study provides direct morphological evidence that CRF directly influences the function of the principal cells of the SCG and that CRFI fibers are preganglionic.     

Immunocytochemical analysis of [Met5]enkephalin-Arg6-Gly7-Leu8 immunoreactive structures in the rat superior cervical ganglion

Indirect immunofluorescence and immunoelectron microscopy were employed to analyze the enkephalinergic systems in the rat superior cervical ganglion (SCG). These systems were identified using specific antiserum against [Met5]Enkephalin-Arg6-Gly7-Leu8 (ENK-8), a peptide which is derived only from proenkephalin A. Abundant ENK-8 like immunoreactive (ENK-8-LI) neurons and fibers were observed in the SCG, but their distribution patterns were heterogenous; ENK-8-LI neurons were localized preferentially in the caudal two-thirds of the SCG, while immunoreactive fibers were found to be distributed more densely in the rostral one-third than in the remaining part of the SCG. Most of the ENK-8-LI neurons were large and had ultrastructural features resembling those of principal cells, some were identified electron microscopically as small intensely fluorescent (SIF) cells. ENK-8-LI fibers were varicose in appearance and surrounded the perikarya of neurons. Since most of these fibers were not detected after experimental decentralization of the SCG and since ENK-8-LI terminals were seen to contain small lucent vesicles, most of the former were thought to be preganglionic fibers. Immunoreactive fibers mainly formed synaptic contacts with the dendrites of non-immunoreactive principal cells, but a small proportion of ENK-8-LI principal cells also received synaptic input from them. Occasionally, immunoreactive fibers formed synapses with the processes or the soma of both ENK-8-LI and non-immunoreactive SIF cells. On the basis of these findings, we conclude that: (1) preganglionic ENK-8-LI fibers terminate mainly on the principal cells, which are devoid of ENK-8-LI structures; (2) the majority of ENK-8-LI neurons are principal cells, while the remainder are SIF cells; (3) inputs to these cells mainly involve structures lacking ENK-8 immunoreactivity; and (4) there are, however, a small number of ENK-8-LI preganglionic fibers which terminate on ENK-8-LI principal cells and SIF cells.     

Immunocytochemical analysis of [Met]enkephalin-Arg-Gly-Leu immunoreactive structures in the rat superior cervical ganglion

Indirect immunofluorescence and immunoelectron microscopy were employed to analyze the enkephalinergic systems in the rat superior cervical ganglion (SCG). These systems were identified using specific antiserum against [Met⁵]Enkephalin-Arg⁶-Gly⁷-Leu⁸ (ENK-8), a peptide which is derived only from proenkephalin A. Abundant ENK-8 like immunoreactive (ENK-8-LI) neurons and fibers were observed in the SCG, but their distribution patterns were heterogenous; ENK-8-LI neurons were localized preferentially in the caudal two-thirds of the SCG, while immunoreactive fibers were found to be distributed more densely in the rostral one-third than in the remaining part of the SCG. Most of the ENK-8-LI neurons were large and had ultrastructural features resembling those of principal cells, some were identified electron microscopically as small intensely fluorescent (SIF) cells. ENK-8-LI fibers were varicose in appearance and surrounded the perikarya of neurons. Since most of these fibers were not detected after experimental decentralization of the SCG and since ENK-8-LI terminals were seen to contain small lucent vesicles, most of the former were thought to be preganglionic fibers. Immunoreactive fibers mainly formed synaptic contacts with the dendrites of non-immunoreactive principal cells, but a small proportion of ENK-8-LI principal cells also received synaptic input from them. Occasionally, immunoreactive fibers formed synapses with the processes or the soma of both ENK-8-LI and non-immunoreactive SIF cells. On the basis of these findings, we conclude that: (1) preganglionic ENK-8-LI fibers terminate mainly on the principal cells, which are devoid of ENK-8-LI structures; (2) the majority of ENK-8-LI neurons are principal cells, while the remainder are SIF cells; (3) inputs to these cells mainly involve structures lacking ENK-8 immunoreactivity; and (4) there are, however, a small number of ENK-8-LI preganglionic fibers which terminate on ENK-8-LI principal cells and SIF cells.     

Biogenic amine localization in cardiac ganglion intrinsic neurons: Electron microscopic histochemistry of SIF cells

The parasympathetic cardiac ganglion in the mudpuppy, N. maculosus, contains postganglionic nerve cells and intrinsic neurons, many of which are small intensely fluorescent (SIF) cells. Several bioactive substances have been localized in the intrinsic nerve cells which may have integrative effects at synapses within the ganglion. Ganglionic intrinsic neurons can be identified electron microscopically by the presence of numerous cytoplasmic granular vesicles 80-120 nm in diameter. Throughout the ganglion there are bundles of unmyelinated fibers some of which are filled with granular and agranular vesicles and axosomatic terminals with similar vesicles synapsing on principal parasympathetic nerve cells. To understand the aminergic contribution to ganglionic synaptic circuitry the chromaffin reaction was used. The intrinsic neurons (i.e., SIF cells) were readily identified by their characteristic intracellular granule population. All intrinsic nerve cells identified showed granules which were positively labelled by the chromaffin reaction. Granular vesicles in synaptic profiles on principal cells (P cells) were also labelled indicating a direct aminergic synaptic innervation to these cells. The cell bodies of intrinsic neurons, ensheathed with supportive glial-like cellular processes, rarely received synapses. Elemental microanalysis was used to verify the chromium content of the electron dense product within the granular vesicles. These studies demonstrated direct aminergic synaptic input to at least a subpopulation of principal parasympathetic cells in the cardiac ganglion of mudpuppy.     

The lateral reticular nucleus of the opossum (Didelphis virginiana). I. Conformation,cytology and synaptology

When viewed in Nissl preparations, the lateral reticular nucleus (LRN) of the opossum can be divided into three subgroups: a medial internal portion, a lateral external portion and a rostral trigeminal division. Neurons within the internal division measure 13-45 μ in their greatest dimension whereas those within the external and trigeminal portions measure 11-32 μ and 14-27 μ respectively. Golgi impregnations reveal that many neurons in all three subdivisions display a radial dendritic pattern although some of the nerve cells within the external division have dendrites which orient mainly in a ventromedial to dorsolateral direction. The cell bodies of LRN neurons are relatively spine-free. However, a small percentage of neurons exhibit clusters of sessile spines on proximal and more distal dendritic segments. No locally ramifying axons or axon collaterals were found within the LRN. Synaptic terminals within the LRN were divided into four categories: (1) small terminals measuring 2.5 μ or less containing agranular spherical vesicles; (2) small terminals (2.5 μ or less) with agranular pleomorphic synaptic vesicles, i.e., a mixture of spherical and elliptical synaptic vesicles; (3) small terminals (2.5 μ or less) containing agranular spherical or pleomorphic vesicles with a variable number (4-27) of dense core vesicles; and (4) large terminals (greater than 2.5 μ) which contain agranular spherical synaptic vesicles and a variable number of dense core vesicles (1-17). Dendritic diameters were measured from Golgi impregnations and correlated with cross-sectioned profiles in electron micrographs to help determine the post-synaptic distribution of synaptic endings. Small terminals containing agranular spherical or pleomorphic synaptic vesicles contact the soma and entire dendritic tree in each portion of the nucleus, whereas the small terminals containing dense core vesicles are usually located on distal dendrites or spines. Some large terminals make multiple synaptic contacts with a cluster of spines, others contact groups of small (distal) dendrites. In order to identify two of the major afferent systems to the LRN, 15 adult opossums were subjected to either a cervical spinal cord hemisection or a stereotaxic lesion of the red nucleus. Two days subsequent to spinal hemisection, large terminals in the caudal part of the ipsilateral LRN exhibit either an electron dense or filamentous reaction. Their postsynaptic loci are spines and shafts of proximal dendrites or a number of distal dendrites and spines. In addition, small terminals containing spherical agranular synaptic vesicles undergo an electron dense reaction in the same areas. Their postsynaptic loci are proximal or distal dendrites. Two days subsequent to rubral lesions, small terminals containing agranular spherical synaptic vesicles undergo a dark reaction in rostral portions of the contralateral nucleus. They contact intermediate or distal dendrites and occasionally spines.     

Direct observations of synapses between GABA-immunoreactive boutons and identified spinocervical tract neurons in the cat's spinal cord.

Three spinocervical tract neurons in adult cats were physiologically characterized and intracellularly labelled with horseradish peroxidase. The neurons were reconstructed and examined with the light microscope and were prepared for postembedding immunochemical analysis by using an antiserum which specifically recognizes GABA in glutaraldehyde-fixed tissue. Semithin sections were tested and examined with the light microscope. Somata, proximal, and distal dendrites of all three cells were associated with numerous punctate GABA-immunoreactive structures. Immunoreactive perikarya of small neurons in the vicinity of spinocervical tract cells were also observed. Ultrastructural analysis, with the immunogold technique, revealed that somata and proximal dendrites of all three neurons received synaptic contacts (about 37% of total synapses) from GABA-immunoreactive boutons and that distal dendrites were also associated with substantial numbers of immunoreactive structures (about 27% of synapses). Immunoreactive boutons were small (about 1 micron in diameter), contained irregularly shaped agranular vesicles, and formed symmetrical synaptic junctions with identified neurons. An additional group of immunoreactive boutons was observed to be associated with one of the cells only; these contained many large dense-core vesicles in addition to small agranular vesicles. Boutons containing round agranular vesicles and flattened agranular vesicles were not observed to be immunoreactive. The evidence supports the idea that much of the postsynaptic inhibition observed in spinocervical tract neurons is mediated by GABA and that even the most distal dendrites of these neurons receive inhibitory inputs.     

Immunocytoehemical localization of substance P in the spinal trigeminal nucleus of the rat: A light and electron microscopic study

The neuropeptide substance P is a transmitter candidate for certain primary afferent fibers which terminate in the substantia gelatinosa. In this study the light and electron microscopic localization of substance P in the substantia gelatinosa of the spinal trigeminal nucleus of the rat has been studied using immunocytochemical procedures. Substance P immunoreactive fibers were observed mainly in lamina I and outer lamina II. Ultrastructural analysis revealed immunoreactivity in unmyelinated fibers and in axon terminals which contained agranular spherical vesicles and large dense-cored vesicles and which made predominantly simple asymmetric axodendritic synaptic contacts. Immunoreactive terminals only rarely formed the central terminal of synaptic glomeruli and in only one example was a stained terminal possibly postsynaptic to an unstained terminal. The majority of synapses were onto small dendrites in outer lamina II and in some cases these dendrites were themselves presynaptic to other dendrites. Immunoreactive terminals also synapsed with the soma and proximal dendrites of large neurons on the border of laminae I and II. The results show that there are at least two distinct targets for substance P immunoreactive terminals in the substantia gelatinosa, namely the large lamina I neurons and lamina II probable intemeurons. Some of the former may be projection neurons while some of the latter may correspond to the inhibitory islet cells described by Gobel and colleagues in the cat. In addition the results indicate that few substance P immunoreactive terminals receive axoaxonic synapses and emphasize instead the role of postsynaptic interactions. In particular the results suggest several sites at which substance P might interact postsynaptically with the neuropeptide enkephalin.     

Fine structure of noradrenergic terminals and their synapses in the rat spinal dorsal horn: an immunohistochemical study

Noradrenergic fibers in the spinal dorsal horn originate from neurons in the A5-7 cell groups, and may participate in the modulation of pain. Here we studied the fine structure of noradrenergic terminals in the rat by immunohistochemistry using antiserum against dopamine-beta-hydroxylase (DBH). We also investigated the relationship between such terminals and primary afferent terminals. DBH-like immunoreactive terminals were found in lamina I and the outer layer of lamina II of the dorsal horn and they contained many clear round vesicles and some large granular vesicles. More than half of these terminals made synaptic contact with other neuronal elements with membrane specialization. Most of the postsynaptic structures of these terminals were small dendrites (69%); 28% were spines, and no synaptic contact was made with primary afferent terminals. These findings suggest that noradrenaline acts on the spinal dorsal horn neurons postsynaptically mainly via a direct synaptic mechanism.     

The morphological relationships between substance P immunoreactive processes and ventral horn neurons in the human and monkey spinal cord

The distribution of substance P (SP)-containing neural processes in the ventral horn region of the human and monkey spinal cord was studied by staining for SP by the indirect antibody peroxidase-antiperoxidase technique and by light and electron microscopic examination of stained material. Immunoreactive fibers and terminals were found in all major cell columns of the ventral horn in the human and monkey spinal cord, with the immunoreactive profiles in close apposition to motoneurons. The immunoreactivity was contained in synaptic knoblike (bouton) structures, in which the label was associated with small agranular vesicles, dense-core vesicles, and mitochondria. The immunoreactive synaptic knobs formed synaptic junctions predominantly with dendrites. The labeled knobs were similar to "S" (agranular spheroid vesicles) and "G" (dense-core vesicles) type bulbs described previously in the monkey ventral horn. Their location, mainly on the neuron dendrites and soma supports the physiological evidence that SP-produced membrane depolarization is mediated primarily by a direct action of the neurochemical on the neuron rather than by a transynaptic mechanism.     

Ultrastructural characterization of synaptic terminals formed on newly generated neurons in a song control nucleus of the adult canary forebrain

The fine structure of synaptic terminals contacting neurons generated in the forebrain of adult male canaries was investigated by autoradiography and electron microscopy. The procedure for labeling the new neurons included pretreating adult canaries with 3H-thymidine and sacrificing them 23-45 days later. Neurons were identified as newly generated by the presence of 3H-thymidine in the cell nucleus. The new neurons in the nucleus hyperstriatum ventralis, pars caudalis (HVc) were identified by autoradiography and light microscopy and examined with electron microscopy. Several types of synaptic terminals contacted the cell body and proximal dendrites of the newly formed neurons. Synaptic junctions were formed by terminals that contained spherical, agranular vesicles, large dense-core vesicles and spherical, agranular vesicles, and pleomorphic or flattened synaptic vesicles. Terminals that contained spherical vesicles were most often associated with asymmetric synaptic densities, and terminals that contained pleomorphic or flattened vesicles formed symmetric junctions. New neurons were also contacted by small terminals that contained few vesicles and had little pre- or postsynaptic density associated with the junction; these terminals may be a special type or may be in the process of developing their synaptic contact with the new neuron. In addition, rare terminals that appeared to be degenerating or to contain debris from other degenerating neural elements contacted new neurons. In summary, these data indicate that the new neurons, which are known to be inserted into existing neural networks, receive synaptic input from at least three different sources.     

An electron microscopic study on VIP-like immunoreactive nerve fibers in the celiac ganglion of guinea pigs

An immuno-electron microscopic study revealed that VIP-like immunoreactive nerve fibers in the celiac ganglion of guinea pigs were characterized by a conspicuously numerous large granular vesicles mixed with small clear vesicles. The immunoreactive materials were localized in the core of the large granular vesicles and a distinct halo was recognized between the core and the limiting membrane of the vesicles. These fibers made numerous axo-dendritic and a few axo-somatic synapses with the post-ganglionic principal neurons and also formed some synapses with vesicle-containing neuronal profiles which are presumably preganglionic axons. The immunoreactive fibers were presynaptic at all these synaptic sites. In addition, some synaptic contacts were found between two adjacent immunoreactive nerve fibers. These findings strongly suggests that VIP might be involved in the ganglionic transmission of the prevertebral ganglia.     

Principal neurons and small intensely fluorescent (SIF) cells in the rat superior cervical ganglion have distinct developmental histories.

Sympathetic ganglia contain 2 adrenergic derivatives of the neural crest: principal neurons and small, intensely fluorescent (SIF) cells. The developmental mechanisms responsible for the generation of these 2 cell classes in vivo are not well understood. To examine the possible developmental and lineage relationships between differentiating principal neurons and SIF cells, a fluorescence microscopic study utilizing antibodies against tyrosine hydroxylase (TH) and catecholamine histofluorescence has been combined with the ultrastructural examination of embryonic and postnatal rat superior cervical ganglia (SCG). On embryonic day 12.5, before neuroblasts had become postmitotic, the cells in the SCG possessed intense TH immunoreactivity and had weak to bright catecholamine histofluorescence, but no cells displayed the fine structure of mature SIF cells or neurons. At embryonic days 16.5 and 18.5, postmitotic principal neurons expressed more moderate levels of TH and catecholamines characteristic of the late embryonic and postnatal SCG. By contrast, a small number of cells containing intense TH or catecholamine fluorescence were present in embryonic day 16.5 and older ganglia. Almost all of the intensely fluorescent cells observed were found apposed to capillaries within the ganglion. These embryonic intensely fluorescent cells were larger than SIF cells seen postnatally. Ultrastructural examination of developing ganglia confirmed that cells containing numerous large, dense-cored vesicles (LDCVs) were a prominent feature of ganglia that also contained intensely fluorescent cells. In addition, some embryonic cells containing LDCVs were mitotic. From these and other studies, it seems likely that during development, neuron precursors, in response to differentiation factors such as fibroblast growth factor (FGF) and/or NGF, acquire overt neuronal traits and become postmitotic. Subsequently, cells resembling mature SIF cells appear next to blood vessels, where they may have received other instructional signals such as glucocorticoids. This developmental scheme suggests that the differentiation of principal neurons and SIF cells is independently regulated, and that the ability of SIF cells to convert into principal neurons observed in vitro cannot account for the generation of neurons in vivo.     

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.     

Ultrastructural evidence for communication between intramuscular vagal mechanoreceptors and interstitial cells of Cajal in the rat fundus.

To assess whether afferent vagal intramuscular arrays (IMAs), putative gastrointestinal mechanoreceptors, form contacts with interstitial cells of Cajal of the intramuscular type (ICC-IM) and to describe any such contacts, electron microscopic analyses were performed on the external muscle layers of the fundus containing dextran-labelled diaminobenzidin (DAB)-stained IMAs. Special staining and embedding techniques were developed to preserve ultrastructural features. Within the muscle layers, IMA varicosities were observed in nerve bundles traversing major septa without contact with ICC-IM, contacting unlabelled neurites and glial cells. IMA varicosities were encountered in minor septa in contact with ICC-IM which were not necessarily in close contact with muscle cells. In addition, IMA varicosities were observed within muscle bundles in close contact with ICC-IM which were in gap junction contact with muscle cells. IMAs formed varicosities containing predominantly small agranular vesicles, occasionally large granular vesicles and prejunctional thickenings in apposition to ICC-IM processes, indicating communication between ICC and IMA via synapse-like contacts. Taken together, these different morphological features are consistent with a hypothesized mechanoreceptor role for IMA-ICC complexes. Intraganglionic laminar ending varicosities contacted neuronal somata and dendrites in the myenteric plexus of the fundus, but no contacts with ICC associated with Auerbach's plexus were encountered.     

Met5-enkephalin-Arg6-Gly7-Leu8-like immunoreactivity in the pelvic ganglion of the male rat: a light and electron microscopic study

By using both light and electron microscopic immunocytochemical methods, Met5-Enkephalin-Arg6-Gly7-Leu8 (MEAGL)-like immunoreactive structures were detected in the pelvic ganglion of male rats. Denervation studies were carried out to determine the origin of these immunoreactive fibers and the projection of immunoreactive neurons within the pelvic ganglion. MEAGL-like immunoreactivity was found in numerous axon boutons, some small, intensely fluorescent (SIF) cells, and a few principal ganglion neurons. Most of the immunoreactive nerve fibers formed pericellular plexuses surrounding the ganglion cells. In addition, there were a few scattered varicose fibers. These fiber plexuses could be classified into two types: type I (approximately 90% of fibers), which consisted of 80-120 small boutons that synapsed on either the dendrites (80% of cases) or somata (20% of cases) of principal neurons; and type II (approximately 10% of fibers), which consisted of 20-40 larger boutons that formed axodendritic synapses exclusively. After transection of the hypogastric and pelvic nerves, virtually all of the pericellular fiber plexuses disappeared, whereas the scattered varicose fibers remained. According to their ultrastructure, these remaining fibers were considered to arise from SIF cells. Following the injection of Fast Blue into the bladder wall, some of the MEAGL-like immunoreactive principal neurons were retrogradely labeled. The results of this study indicate that there are two origins for the MEAGL-like immunoreactive fibers detected in the pelvic ganglion: most arise from preganglionic neurons in the spinal cord, and a small proportion may originate from intraganglionic MEAGL-like immunoreactive SIF cells or principal neurons. Some MEAGL-like immunoreactive principal neurons may project to the urinary bladder.     

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.     

Topography and synaptology of mamillary body projections to the mesencephalon and pons in the rat.

The anterograde and retrograde transport of horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) was used to study the anatomical organization of descending projections from the mamillary body (MB) to the mesencephalon and pons at light and electron microscopic levels. Injections of WGA-HRP into the medial mamillary nucleus resulted in dense anterograde and retrograde labeling in the ventral tegmental nucleus, while injections in the lateral mamillary nucleus resulted in dense anterograde labeling in the dorsal tegmental nucleus pars dorsalis and dense anterograde and retrograde labeling in the pars ventralis of the dorsal tegmental nucleus. Anterogradely labeled fibers in the mamillotegmental tract diverged from the principal mamillary tract in an extensive dorsocaudally oriented swath of axons which extended to the dorsal and ventral tegmental nuclei, and numerous axons turned sharply ventrally and rostrally to terminate topographically in the dorsomedial nucleus reticularis tegmenti pontis and rostromedial pontine nuclei. The anterograde labeling in these two precerebellar relay nuclei was distributed near the midline such that projections from the lateral mamillary nucleus terminated mainly dorsomedial to the terminal fields of projections from the medial mamillary nucleus. In the dorsal and ventral tegmental nuclei, labeled axon terminals contained round synaptic vesicles and formed asymmetric synaptic junctions primarily with small diameter dendrites and to a lesser extent with neuronal somata. A few labeled terminals contained pleomorphic vesicles and formed symmetric synaptic junctions with dendrites and neuronal somata. Labeled axon terminals were also frequently found in synaptic contact with retrogradely labeled dendrites and neuronal somata in the dorsal and ventral tegmental nuclei. These findings indicate that neurons in the dorsal and ventral tegmental nuclei are reciprocally connected with MB projection neurons. In the nucleus reticularis tegmenti pontis and medial pontine nuclei, labeled axon terminals contained round synaptic vesicles and formed asymmetric synaptic junctions primarily with small diameter dendrites. The present study demonstrates that projections from the medial and lateral nuclei of the MB are topographically organized in the mesencephalon and pons. The synaptic morphology of mamillotegmental projections suggests that they may have excitatory influences primarily on the distal dendrites of neurons in these brain regions.     

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.     

Immunohistochemical study on met-enkephalin-arg-gly-leu-like immunoreactive nerve fibers in the rat adrenal medulla

Met-enkephalin-Arg-Gyl-Leu-like immunoreactivity was revealed in many nerve fibers as well as in chromaffin cells in the rat adrenal medulla. The immunoreactive nerve fibers were seen among and around chromaffin cells in the forms of punctate structures or varicose processes and they were characterized by numerous small clear vesicles mixed with several large granular vesicles. The nerve fibers formed synaptic contacts with both Met-enkephalin-Arg-Gly-Leu-like immunoreactive and non-immunoreactive chromaffin cells. The immunoreactive material in the nerve was generally confined to the granular cores. The present findings suggest that preproenkephalin A and its derivatives are produced in adrenal efferents which may be preganglionic cholinergic, and that they may play a significant role in the function of adrenal chromaffin cells.     

Synaptic organization of GABAergic projections from the substantia nigra pars reticulata and the reticular thalamic nucleus to the parafascicular thalamic nucleus in the rat

The ventrolateral part of the parafascicular thalamic nucleus (PF), which is considered to take part in the control mechanism of orofacial motor functions, receives projection fibers not only from the dorsolateral part of the substantia nigra pars reticulata (SNr) but also from the ventral part of the reticular thalamic nucleus (RT) [Tsumori et al., Brain Res. 858 (2000) 429]. In order to better understand the influence of these fibers upon the PF projection neurons, the morphology, synaptology and chemical nature of them were examined in the present study. After ipsilateral injections of Phaseolus vulgaris-leucoagglutinin (PHA-L) into the dorsolateral part of the SNr and biotinylated dextran amine (BDA) into the ventral part of the RT, overlapping distributions of PHA-L-labeled SNr fibers and BDA-labeled RT fibers were seen in the ventrolateral part of the PF. At the electron microscopic level, the SNr terminals made synapses predominantly with the medium to small dendrites and far less frequently with the somata and large dendrites, whereas approximately half of the RT terminals made synapses with the somata and large dendrites and the rest did with the medium to small dendrites of PF neurons. Some of single dendritic as well as single somatic profiles received convergent synaptic inputs from both sets of terminals. These terminals were packed with pleomorphic synaptic vesicles and formed symmetrical synapses. After combined injections of PHA-L into the dorsolateral part of the SNr, BDA into the ventral part of the RT and wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the ventrolateral part of the striatum or into the rostroventral part of the lateral agranular cortex, WGA-HRP-labeled neurons were embedded in the plexus of PHA-L- and BDA-labeled axon terminals within the ventrolateral part of the PF, where the PHA-L- and/or BDA-labeled terminals were in synaptic contact with single somatic and dendritic profiles of the WGA-HRP-labeled neurons. Furthermore, the SNr and RT axon terminals were revealed to be immunoreactive for gamma-aminobutyric acid (GABA), by using the anterograde BDA tracing technique combined with immunohistochemistry for GABA. The present data suggest that GABAergic SNr and RT fibers may exert different inhibitory influences on the PF neurons for regulating the thalamic outflow from the PF to the cerebral cortex and/or striatum in the control of orofacial movements.