Symmetric Synapses Research Articles

A quantitative analysis of the synaptic development of the lobus parolfactorius of the chick (Gallus domesticus).

The lobus parolfactorius (LPO) of the chick has been shown to undergo an increase in the mean synaptic numerical density (Nvsyn) in response to one-trial passive avoidance learning (Stewart et al. 1987). The present study was undertaken in order to describe the pattern of normal development of synapses in the LPO, to further investigate the significance of this plastic response. The LPO's from each hemisphere of pre-hatch (16 days), and post-hatch (1 day, 9 day and 22 day old) chicks were processed for electron microscopy. Synapses were classified into asymmetric spine, asymmetric shaft, symmetric spine, and symmetric shaft synapses, on the basis of the density of the post-synaptic thickening and the nature of the post-synaptic target. A 3-dimensional stereological probe was used (the 'disector') to calculate Nvsyn, and mean projected height (Hsyn) of the post-synaptic density (PSD). Mean values for each age and hemisphere were compared with a 2-way analysis of variance test using paired samples. A six-fold increase in Nvsyn was seen between 16 days in ovo, and 9 days post-hatch. There was a hemispheric asymmetry at 9 days post-hatch, with the left hemisphere LPO containing 1.6 times as many synapses per micron-3. There was a subsequent period of reduction in synaptic density in the left hemisphere LPO between 9 and 22 days post-hatch. The Nv of all classes of synapse increased with age, but the proportions of the symmetrical synapses with respect to the total number of synapses, decreased with age. This decrease was of a similar magnitude for each hemisphere. A hemispheric difference was seen in post-hatch asymmetric synapses, with a greater proportion of asymmetric spine synapses in the left hemisphere. The magnitude of the hemispheric asymmetry was constant throughout the 3 week period of post-hatch development, but was not present in pre-hatch chicks. The PSD increased in length in each hemisphere by approximately 40% between post-hatch day 1 and post-hatch day 9. These data show that the LPO contains a synaptic population which undergoes substantial modification during the first week post-hatch. An asymmetry exists at post-hatch day 9 which is not present at the earlier ages investigated, nor indeed after 22 days post-hatch. This may have significance with regard to studies of passive avoidance learning in the one-day old chick, where an increase in both the size and number of synapses in the LPO has been demonstrated (Stewart et al. 1987).(ABSTRACT TRUNCATED AT 400 WORDS)

Direct synaptic contacts of 5-hydroxytryptamine-, neuropeptide Y-, and somatostatin-immunoreactive nerve terminals on the preganglionic sympathetic neurons of the guinea pig

Direct synapses from 5-hydroxytryptamine-, neuropeptide Y-, and somatostatin-immunoreactive axon varicosities to the pregnaglionic sympathetic neurons retrogradely labeled with horseradish peroxidase were observed in the thoracic intermediolateral nucleus of the guinea pig. Symmetric synapses were predominant and both axo-dendritic and axo-somatic contacts were observed.

An EM study of the dorsal nucleus of the lateral lemniscus: inhibitory, commissural, synaptic connections between ascending auditory pathways

The dorsal nucleus of the lateral lemniscus (DNLL) and its connections constitute one of the ascending auditory pathways to the inferior colliculus. One notable feature of this nucleus is the heavy commissural connections between DNLL on opposite sides of the midbrain. These commissural connections may have a significant impact on the ascending pathway. In this study, the fine structure of DNLL in the cat and its commissural connections were examined. Both anterograde and retrograde transport methods were used simultaneously at the EM level. Injections of 3H-leucine mixed with WGA-HRP were made in one DNLL. After axonal transport, EM autoradiographic methods were used to identify the anterogradely labeled axonal endings from the opposite DNLL. In the same location, retrogradely labeled neurons with crossed connections were identified with HRP histochemistry. Two types of axonal endings were found in DNLL, those with round synaptic vesicles forming asymmetrical synaptic junctions and those with pleomorphic vesicles and symmetrical synapses. Both types were equally common. However, only endings with pleomorphic vesicles were labeled after injections in the contralateral DNLL. The labeled endings from the opposite DNLL appeared to represent a homogeneous population, even though a number of variations in the 2 types of endings were found. Labeled endings were presynaptic to all parts of neurons in DNLL, but a large proportion of the synapses were on cell bodies and large dendrites. Two patterns of nuclear morphology and distribution of rough endoplasmic reticulum were identified and may represent different cell types. Examples of both cell types were observed to project to the contralateral side and received labeled synaptic endings. The major finding of this study is that the crossed connections between DNLL exhibit the morphology associated with inhibitory function. Since neurons in DNLL are thought to use GABA as a neurotransmitter, the crossed connections could provide inhibitory inputs to DNLL on each side. Since some neurons receive numerous axosomatic inputs from the contralateral DNLL and also project to the opposite side, they may participate in direct reciprocal, inhibitory connections between the nuclei. Crossed inhibitory connections in the DNLL pathway may be important in regulating the flow of ascending auditory information.

Chapter 3 Adrenergic neurons in the rostral ventrolateral medulla: ultrastructure and synaptic relations with other transmitter-identified neurons

The first part of this chapter demonstrates that the C1 adrenergic neurons have high mitochondrial content and a close proximity to capillaries and glia suggestive of a high metabolic activity and a possible chemosensory function. Adrenergic terminals arising primarily from these neurons (1) can influence sympathetic nerve discharge through direct contacts on sympathetic preganglionic neurons in the IML of the spinal cord; and (2) are one of the more prevalent synaptic inputs to the principally noradrenergic neurons in the locus coeruleus. In both the IML and locus coeruleus, adrenergic terminals may be either excitatory (asymmetric synapses) or inhibitory (symmetric synapses) depending on their distribution on the post-synaptic target. The second part of this chapter shows that C1 adrenergic neurons in the RVL are modulated by synaptic associations with a variety of transmitter systems (see schematic Fig. 8). Specifically, C1 adrenergic neurons receive (1) major inhibitory input (symmetric synapses) from GABA-ergic and opioid terminals as well as from unidentified (unlabelled) transmitter-containing terminals; (2) major excitatory input (asymmetric synapses) from terminals containing substance P as well as other unidentified terminals and (3) minor inputs from cholinergic, adrenergic and noradrenergic pathways. Moreover, cholinergic terminals in the RVL form symmetric synapses mainly on unidentified transmitter-containing neurons rather than the C1 neurons suggesting that the reported cardiovascular effects of cholinergic agents in the RVL are most likely mediated via inhibitory interneurons. Within the RVL, adrenergic and noradrenergic terminals innervate cholinergic and opioid neurons. Thus, these results not only provide direct evidence that a number of transmitters modulate the activity of C1 adrenergic neurons, but also suggest new directions for studies of functional interactions involving catecholaminergic regulation of other transmitter-containing neurons within the RVL.

Corticotropin-Releasing Factor Neurons Innervate Dopamine Neurons in the Periventricular Hypothalamus of Juvenile Macaques

Corticotropin-releasing factor (CRF) and dopamine (DA) are important integrators of the endocrine and autonomic response to stress. CRF neurons in the anterior portions of the periventricular nucleus (PV) and parvocellular paraventricular nucleus (pvPVN) occur close to A14 DA neurons in these same locations. Since CRF has been shown to act as an excitatory neurotransmitter, possible CRF interactions with the DA system were investigated using double-label immunocytochemistry. Coronal vibratome sections through the PV and pvPVN were obtained from colchicine-treated and nontreated juvenile female cynomolgus macaques. They were sequentially immunostained for tyrosine hydroxylase (TH) (to identify DA neurons) with PAP and DAB, and for CRF using 15 nm colloidal gold. By light microscopy, areas of coincidence of TH- and CRF-immunoreactive cell bodies in the PV and pvPVN were obvious, but double-stained elements were not observed. By electron microscopy, asymmetrical synapses frequently occurred between CRF axons and TH dendrites or somata. Symmetrical axosomatic synapses sometimes appeared adjacent to these CRF/TH synapses, while symmetrical axoaxonic synapses were rare. We conclude that CRF neuronal efferents synaptically activate A14 DA neurons in the primate PV and pvPVN. Parallel CRF/DA symmetrical synapses also suggest coexistence of a companion transmitter within some of these same CRF neurons. Our own previous work and recent independent studies indicate that this transmitter is probably GABA. Thus the CRF neuronal system, which is known to alter secretion of several pituitary hormones, may also act through hypothalamic periventricular DA neurons to mediate other responses to stress.

Synaptic organization of the striatum.

The striatum, the main component of the basal ganglia, is composed of mainly one type of neuron, the so-called medium spiny neuron. This neuron cell type, which constitutes over 90% of striatal neurons, is the major output neuron of the striatum. Combined ultrastructural neuroanatomical methods have elucidated the organization of afferent connectivity to these neurons. The major physiologic function of striatal efferent activity appears to be inhibition of tonically active GABAergic neurons in the globus pallidus and substantia nigra pars reticulata. Thus, the excitatory input from the cerebral cortex, whose afferents make asymmetric synapses with the spines of medium spiny neurons, appears to drive the efferent activity of the striatum. Other extrinsic and intrinsic afferent synapses are situated in a position to regulate the effect of the corticostriatal excitatory input to the medium spiny neurons. For example, dopaminergic afferents from the midbrain make mainly symmetric synapses with the spine necks and dendritic shafts of the medium spiny neurons. Medium spiny neurons themselves have local axon collaterals, in addition to their efferent axon that exits the striatum, which serve to link together local clusters of medium spiny neurons. These local axon collaterals, which contain either GABA, substance P, or enkephalin, also make mainly symmetric synapses with the necks of spines or dendritic shafts of medium spiny neurons. Other afferents with similar synaptic connections to these neurons arise from cholinergic or somatostatinergic striatal intrinsic neurons. Additionally, the patterns of extrinsic and intrinsic afferents to medium spiny neurons and their extrinsic projections are related to the organization of medium spiny neurons into two mosaically organized macroscopic compartments, the striatal patches and matrix.

Projection pathways, co-existence of peptides and synaptic organization of nerve fibers in the inferior mesenteric ganglion of the guinea-pig

The presence of immunoreactive enkephalin, dynorphin, vasoactive intestinal polypeptide, cholecystokinin, substance P and neuropeptide Y in nerve fibers that project to the guinea-pig inferior mesenteric ganglion was analysed, after different denervation and ligation procedures. A quantitative analysis demonstrates that enkephalin- and substance P fibers reach the ganglion mainly via lumbar splanchnic and partly via intermesenteric nerves. Dynorphin-, vasoactive intestinal polypeptide- and cholecystokinin fibers reach the ganglion mainly via colonic and partly via hypogastric or intermesenteric nerves. Neuropeptide Y fibers enter via intermesenteric, lumbar splanchnic and hypogastric nerves and pass through the ganglion. Analysis of serial 0.5 micron sections tends to confirm co-existence: of dynorphin, vasoactive intestinal polypeptide and cholecystokinin in fibers projecting from the colon; of dynorphin with substance P in the lumbar splanchnic nerves; and of neuropeptide Y with substance P in the hypogastric and colonic fibers. Synaptic contacts, predominantly axodendritic, onto the ganglion cells from enkephalin-, vasoactive intestinal polypeptide-, and substance P-containing terminals were revealed by electron microscopy. Enkephalin-immunoreactive axon varicosities are filled with small, clear vesicles with a few large, cored vesicles and form asymmetric synapses; dynorphin-, vasoactive intestinal polypeptide- and cholecystokinin-immunoreactive axon varicosities are rich in large, dense-cored vesicles and form symmetric synapses.

Morphological identification of an interneuron in the hypoglossal nucleus of the rat: A combined Golgi‐electron microscopic study

Hypoglossal small neurons of adult and juvenile (10-15-day-old) rats were examined by a combined Golgi-electron microscopic technique. In adult rats, Golgi-impregnated neurons were fusiform or ovoid (17 X 12 micron) and emitted a few primary dendrites with few branches and few spines and an axon mainly from the proximal portion of the primary dendrite. At the ultrastructural level, the soma displayed an invaginated nucleus with a conspicuous nucleolus and a relatively scanty cytoplasm in which cisternae of rough endoplasmic reticulum were not organized into extensive lamellar arrays. A moderate number of axon terminals, which contained spherical clear or pleomorphic vesicles approximately 25-40 nm in diameter, formed symmetric or asymmetric synapses on the soma, the dendrites, the axon hillock, and the initial segment. In a preparation from the juvenile rat, we could trace a full axon trajectory of the small neuron. In this sample, in which a small neuron and a motoneuron were simultaneously impregnated, the axon of the small neuron was found to receive an axoaxonic symmetric synapse with pleomorphic vesicles on a varicosity and to contact the motoneuron dendrites by means of another varicosity of the main axon and of two boutons from an axon collateral. The varicosity and the boutons contained pleomorphic synaptic vesicles and synapsed symmetrically with the motoneuron dendrites. We identified the small neuron in the rat hypoglossal nucleus morphologicaly as a Golgi type II interneuron. We discuss its function in relation to the GABAergic nature of small neurons (Takasu et al.: J. Comp. Neurol. 263:42-53, '87).

Fine structure of NPY-containing neurons in the lateral geniculate nucleus and their terminals in the suprachiasmatic nucleus of the rat

Fine structures of neuropeptide Y (NPY)-like immunoreactive neuronal perikarya in the lateral geniculate nucleus and their terminals in the suprachiasmatic nucleus were investigated by peroxidase-antiperoxidase immunocytochemistry using male Wistar rats. NPY-like immunoreactive preterminal axons from both axo-somatic and axo-dendritic synapses on the neurons mainly located in the ventral portion of the suprachiasmatic nucleus. Almost all of them appeared as symmetrical synapses. Immunoreactive neuronal perikarya in the lateral geniculate nucleus show good development of cell organelles such as rough-surfaced endoplasmic reticulum and mitochondria. NPY-like immunoreactivity was distributed throughout the perikarya. The functional role of NPY-like immunoreactive terminals in the suprachiasmatic nucleus were briefly discussed.

Selective staining of a subset of GABAergic neurons in cat visual cortex by monoclonal antibody VC1.1

VC1.1 is a monoclonal antibody generated against cat area 17, which selectively outlines subsets of cortical neurons (Arimatsu et al., 1987). This study was conducted to determine the ultrastructural distribution of the VC1.1 antigen and to identify the particular subclasses of cortical neurons that were labeled. In the light microscope, VC1.1 delineated the surfaces of neurons located mainly in layer IV but also in other layers. The staining surrounded neuronal cell bodies and dendrites in a periodic or meshwork pattern but did not label axons. VC1.1-labeled neurons were morphologically heterogeneous and included multipolar, bipolar, and bitufted classes. In the electron microscope, VC1.1 immunoreactivity surrounded presynaptic membranes of terminal boutons and intersynaptic sections of postsynaptic membranes, but was not present within terminal boutons or synaptic clefts. Both asymmetric and symmetric synapses were immunoreactive. Labeling was also observed intracellularly on VC1.1-outlined neurons, associated with perisynaptic portions of plasma membranes. Tract-tracing methods were used in conjunction with immunocytochemistry to determine whether VC1.1 identified projection neurons, local circuit neurons, or a combination of both types. Layer V and VI corticogeniculate and corticotectal projection neurons were retrogradely labeled with rhodamine fluorescent latex microspheres. In a large sample of retrogradely labeled neurons, none were VC1.1-positive, suggesting that VC1.1 stained a population of local circuit neurons. Additional immunocytochemical double-labeling studies with an antiserum to GABA and VC1.1, revealed that VC1.1-positive neurons were immunoreactive to GABA. These were a major subset of the GABAergic neurons in area 17 and tended to have medium to large cell bodies. It is concluded that VC1.1 identifies a new, immunologically distinct subset of GABAergic neurons in area 17. The restricted distribution of this antigen on perisynaptic portions of GABA-containing cells and surrounding terminal boutons onto these cells suggests that this antigen may play an important role in inhibitory cortical circuits.

Ultrastructural organization of normal and transplanted rat fascia dentata: II. A quantitative analysis of the synaptic organization of intracerebral and intraocular grafts.

As part of an ultrastructural analysis of the normal rat fascia dentata and intracerebral and intraocular dentate transplants the synapses in the dentate molecular layer were quantified. Hippocampal and dentate tissue from 21-day-old rat embryos were grafted into the brain of developing and adult rats and to the anterior eye chamber of adult rats. After 100 or 200 days of survival the recipient rat brains and the recipient eyes were processed for electron microscopy, and the graft dentate molecular layer with the adjacent granule cell layer selected for ultrastructural analysis. Tissue from the dentate molecular layer of normal adult rats served as controls. The dentate synapses were classified as asymmetric (Gray's type 1) or symmetric (Gray's type 2), and according to the postsynaptic element (cell body, dendritic shaft, dendritic spine). The spine synapses were further classified into simple and complex types according to the spine-terminal configuration. Also, the length of synaptic contacts of the individual synaptic types was measured in some grafts, just as the percentage of the cross sectional area of the neuropil covered by blood vessels. The results showed that the synaptic density, expressed as number per unit area of neuropil, to a large extent was the same within the different parts of the normal dentate molecular layer. Compared with this the synaptic density was reduced with 16.4% in dentate molecular layer of the intracerebral graft, primarily because of a 17.6% reduction of simple synapses on dendritic spines and almost halving of the symmetric synapses on dendritic shafts. The synaptic density was independent of the age of the recipient, the intracerebral location of the graft, and the survival time. Although the synaptic length of some of the individual synaptic types increased, this did not compensate for the loss of synapses. In the intraocular grafts the synaptic density was lower than in the intracerebral grafts. Despite the reduced synaptic density, which mainly involved two synaptic types, we conclude that grafted dentate granule cells can develop a remarkably normal, ultrastructural synaptic organization even in the absence of major afferent inputs. This outcome must accordingly be achieved by reorganization of the available intrinsic afferents.

Synaptic structure of the monoamine and peptide nerve terminals in the intermediolateral nucleus of the guinea pig thoracic spinal cord.

Synaptic organization of the intermediolateral nucleus of the guinea pig thoracic spinal cord was examined with particular focus on monoamine- and peptide-containing nerve terminals. Axon varicosities having flat synaptic vesicles constituted 17% of all axons in the nucleus and formed exclusively symmetric synapses. Enkephalin-, substance P-, somatostatin-, 5-hydroxytryptamine-, and catecholamine-immunoreactive nerve terminals were densely distributed, while neurotensin, vasoactive intestinal polypeptide-, oxytocin-, and cholecystokinin-8-immunoreactive nerves were sparse in the nucleus. Coexistence of 5-hydroxytryptamine and enkephalin was demonstrated, and coexistence of somatostatin and enkephalin as well as somatostatin and 5-hydroxytryptamine in the same axons was also shown by serial semithin sections. Catecholamine axons labelled by 5-hydroxydopamine formed axodendritic and axosomatic synapses and made direct synaptic contacts on the preganglionic sympathetic neurons identified by retrograde transport of horseradish peroxidase. Direct synaptic contacts from enkephalin- and substance P-immunoreactive axons to preganglionic sympathetic neurons were also revealed. Enkephalin-, substance P-, and 5-hydroxytryptamine-immunoreactive axons formed axodendritic and axosomatic synapses. Catecholamine axon varicosities constituted 19% of all axon varicosities in the nucleus and 30% of them showed synaptic specializations in a sectional plane. Axon varicosities immunoreactive to enkephalin, 5-hydroxytryptamine, and substance P constituted approximately 35, 19, and 13% of all axon varicosities, respectively, while those with synaptic contacts made up 27, 30, and 26%, respectively, in a sectional plane. Enkephalin-, 5-hydroxytryptamine-, and noradrenaline-immunoreactive axons showed mainly symmetric synaptic contacts.

Morphology and synaptic connections of unmyelinated primary axons in the border zone of rat trigeminal nucleus oralis

This anterograde horseradish peroxidase study examines the morphology and synaptic connections of a population of primary axons which terminate in the dorsal one-half of the border zone (BZ) of rat trigeminal nucleus oralis. Unmyelinated parent fibers in the spinal V tract enter BZ directly and each terminate by continuing as a sparsely branched, long caudally directed strand containing several axonal endings. Primary endings lie in glomeruli where each forms an asymmetrical synapse on a central dendrite. Other glomerular components include two types of non-primary endings. One contains flattened synaptic vesicles, and forms a symmetrical synapse on either the primary ending or the central dendrite, while the other contains pleomorphic synaptic vesicles and establishes a symmetrical synapse on the central dendrite.

Morphological characterization of cholinergic neurons in the horizontal limb of the diagonal band of Broca in the basal forebrain of the rat.

A monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine synthesizing enzyme, was used to identify cholinergic neurons in the nucleus of the horizontal limb of the diagonal band of Broca at the light and electron microscopic levels. ChAT-labelled somata were fusiform, triangular or round in shape and varied considerably in size. Depending on the type of the cell, one to four dendrites emerged from the soma, but an axon could rarely be seen. The nuclei of most cells were round or oval, showed invaginations and displayed prominent nucleoli. The karyoplasm of the larger fusiform and triangular neurons contained abundant organelles including parallel arrays of granular endoplasmic reticulum. The synaptic input to labelled perikarya and proximal dendrites was sparse. It consisted chiefly of asymmetrical synaptic contacts, sometimes with postjunctional densities, but a few symmetrical synapses were also noted. ChAT-positive axon terminals were not identified which suggests that axon collaterals are rare within the nucleus of the horizontal limb of the diagonal band of Broca.

GABAergic and catecholaminergic synaptic interactions in the Macaque hypothalamus: Double label immunostaining with peroxidase-antiperoxidase and colloidal gold

Immunogold staining (IGS) for glutamic acid decar☐ylase (GAD) was combined with the peroxidase-antiperoxidase (PAP) technique for tyrosine hydroxylase (TH) to analyze γ-aminobutyric acid-catecholaminergic neuronal interactions in the rhesus hypothalamus. At the light-microscopic level, TH-immunoreactive (-IR) perikarya and their fibers (brown) were observed in the anterior ventral periventricular area (AVPV), the arcuate nucleus (ARC) and the adjacent periventricular zone (ARC-PVZ). GAD-IR processes (light red) were also present throughout the hypothalamus and appeared to contact some TH-IR neurons. At the electron-microscopic level, PAP was present in perikarya, dendrites, axons and axon terminals of TH-IR neurons. Colloidal gold particles (15 nm) were found only in dendrites and axon terminals of GAD-IR neurons. Labeled GAD terminals typically contained small, clear synaptic vesicles, while TH terminals contained these and sometimes one or two dense-core vesicles. In the ARC and ARC-PVZ, asymmetrical (Gray I) axodendritic synapses occurred between GAD and TH-IR profiles, with TH/GAD directionality more prevalent. Symmetrical (Gray II) synapses were less common, with either TH or GAD presynaptic in axodendritic and dendrodendritic contacts. GAD/GAD interactions were not observed, but TH/TH contacts appeared to be mostly dendrodendritic. In the AVPV, only symmetrical synapses were encountered, and their directionality was difficult to determine. GAD- and TH-IR dendrites frequently established dendrodendritic synapses, but GAD/TH dendrosomatic synapses were seldom seen. These results illustrate the complex interactions of GAD- and TH-containing elements in the neuroendocrine hypothalamus.

Bulbospinal projections in the primate: a light and electron microscopic study of a pain modulating system.

The projections of the nucleus raphe magnus (NRM) and the immediately adjacent reticular formation were studied in the macaque monkey following injections of the rostroventral medulla with 3H-leucine and examination of the resultant labeled axons and terminals by light and electron microscopic autoradiography. Five monkeys had accurately placed injections, which resulted in fiber pathway labeling that coursed caudally, laterally, and dorsally to project to laminae I, II, and V of subnucleus caudalis of the trigeminal and then traveled in the dorsolateral funiculus of the cord and terminated in similar laminae of the spinal dorsal horn at cervical levels. The pathway was only lightly labeled caudal to the cervical enlargement and could not be readily discerned above background in the thoracic or lumbar cord. Electron microscopy revealed that axons and terminals serving this system constitute a heterogeneous population. Large-diameter myelinated axons (3-6-micron diameter), small myelinated axons (0.75-3-micron diameter), and clusters of nonmyelinated axons were labeled. Terminals in laminae I, II, and V contained mixtures of clear round and granular vesicles or clear pleomorphic and granular vesicles or formed the central element in synaptic glomeruli. The labeled profiles formed asymmetrical or symmetrical synapses on medium and small dendrites; labeled axosomatic synapses were not observed. In rare instances there were contacts between labeled profiles and vesicle-containing structures, which were probably dendritic, but whether the NRM axon was pre- or postsynaptic to such structures could not be determined. It was concluded that the NRM in the monkey is organized in a manner quite similar to that previously described in the cat. The wide variety of fiber types and synaptic terminals serving this system suggests that different classes of neurons participate in it, probably using several transmitter substances that result in varying postsynaptic effects on neurons located in the trigeminal complex and dorsal horn.

A Golgi and ultrastructural analysis of the centromedian nucleus of the cat

The morphology of neurons in the centromedian nucleus (CM) was studied in rapid Golgi preparations of the adult cat. The ultrastructure of the nucleus, particularly its synaptic organization, was also studied with electron microscopy. The CM contains three types of neurons referred to as principal neurons, Golgi type II neurons, and bushy neurons. Principal neurons are the most numerous, have long dendrites, which branch infrequently, and are divided into two subgroups: principal-A neurons with dendrites that arborize radially, whereas principal-B neurons display horizontal orientations. Both subgroups show a frontal orientation in their dendritic organization and give rise to myelinated axons. Golgi type II neurons with their characteristic sinuous dendrites and unmyelinated axons are thought to be interneurons. The occurrence of bushy neurons in the cat's CM is a new finding. These bushy neurons resemble those of thalamic specific relay nuclei and give rise to myelinated axons. In addition to these three cell types, neurons with intermediate features between these three neuronal types are also described. The ultrastructure of CM neurons resembles, in general, typical central nervous system neurons. Presynaptic profiles are classified into four main categories. SR (small round) boutons are small in size, contain clear, round vesicles, and form asymmetrical synaptic contacts with predominantly small-diameter dendrites. LR (large round) boutons are relatively large and contain both clear and dense-cored vesicles. They interdigitate and form multiple, moderately asymmetrical synapses with their postsynaptic targets. Pale profiles are identified by their relatively electron-light appearance. They contain round vesicles and are thought to be dendritic in origin. The last category of presynaptic profiles is pleomorphic boutons. They contain vesicles of different shapes and are further subdivided into two subtypes: pleomorphic-I ends on soma and dendritic trunks, whereas pleomorphic-II contacts small-diameter dendrites. Both subtypes form symmetrical synapses. The glomeruli of specific thalamic relay nuclei generally contain dendrites, LR boutons, and pale profiles. In addition to these, pleomorphic-II boutons also participate in the formation of the glomerulus of the cat's CM.

Ultrastructural analysis of synapses involving tyrosine hydroxylase-containing neurons in the ventral periventricular hypothalamus of the macaque

Immunocytochemical staining for tyrosine hydroxylase (TH) in the adult macaque brain revealed a network of catecholaminergic (CA) cell bodies and fibers in the arcuate (ARC), anterior ventral periventricular (APV)_and lateral suprachiasmatic nuclei (SCN). Coronal Vibratome sections immunostained with PAP or colloidal gold (15nm) were thin sectioned and examined by electron microscopy. We examined 280 TH-immunopositive processes in individual or in serial thin sections. Of these, 190 engaged in a total of 270 synapses identified as Gray Type I asymmetrical synapses (AS) with distinct postsynaptic densities or Gray Type II symmetrical synapses (SS) without such specializations. The majority (80%) of all synapses were axodendritic, 63% of which exhibited SS and 37% AS, representing almost all of the AS observed. In nearly every case, unlabeled axon terminals containing round, 45 nm, clear vesicles and occasional small dense core vesicles contacted TH-labeled dendrites. About 15% of the synapses were dendrodendritic, all of which were symmetrical. Rare contacts involving other elements (axosomatic, dendrosomatic) constituted only 5% of the total, and occurred predominantly as SS. The predominance of AS and the prevalence of SS almost exclusively on TH-containing dendrites indicates that these CA neurons receive extensive afferent input of from other neurotransmitters. TH-labeling of both neural elements in most dendrodendritic, and some axodendritic SS, also suggest that they modulate one another within the ARC, APV and SCN. The results suggest that these CA neurons performed an important role in local integration, and may act elsewhere to affect the common final pathway of the neuroendocrine system in primates.

Mitral cell differentiation and synaptogenesis of rat presumptive olfactory bulb in organ culture.

The ultrastructure of differentiating rat presumptive olfactory bulb in organ culture was investigated with particular reference to mitral cell differentiation and formation of synapses. The presumptive olfactory bulb and olfactory mucosa were dissected en bloc from rat embryos on the fifteenth day of gestation and cultured for 7 days, after which the explants were examined by electron microscopy. The presumptive olfactory bulb had differentiated into a laminated structure with layers corresponding to the glomerular, external plexiform and mitral cell layers. Mitral-like cells were identified by their location and large cell size. Ultrastructural observations indicated that they were relatively well-differentiated. Their dendrites extended into the glomerular layer in which they were postsynaptic to incoming olfactory axons. The distal part of these dendrites frequently contained coated vesicles. Both asymmetrical and symmetrical synapses were found. The symmetrical synapses involved dendrodendritic contacts between periglomerular cells. Synapses in reciprocal arrangements were not observed in the organ cultures.

Projection of olfactory bulb efferents to layer I GABAergic neurons in the entorhinal area. Combination of anterograde degeneration and immunoelectron microscopy in rat

Glutamic acid decarboxylase (GAD) immunoelectron microscopy in combination with anterograde degeneration was applied in rats to study the synaptic targets of olfactory bulb afferents to the lateral subdivision (LEA) of the entorhinal area (EA). Immunoreactive neurons and terminals are scattered throughout all layers of LEA. After olfactory bulb resection, terminal degeneration occurs in layer Ia of EA. Using the electron microscope we examined serial thin sections of 12 and 14 immunoreactive neurons sampled from layer Ia of the dorsal (DLEA) and ventral (VLEA) subdivisions of LEA, respectively. The morphology of all these neurons is similar: they are small (short axis 5-9 micron, long axis 7-12 micron) and possess eccentrically located, indented nuclei provided with filamentous nuclear rodlets. The immunoreactive neurons have thin, smooth dendrites which usually emerge abruptly from the somata. We observed a single cilium on 5 of the immunoreactive neurons. In layer Ia of both DLEA and VLEA, the somata of the immunoreactive neurons are contacted by degenerating, non-immunoreactive boutons showing asymmetric synaptic junctions. In addition to these boutons, 4 other categories of axo-somatic terminals can be distinguished: normal, non-immunoreactive boutons forming asymmetric synapses and containing spherical synaptic vesicles; normal, non-immunoreactive boutons with symmetric synapses and pleomorphic synaptic vesicles; normal, non-immunoreactive boutons with asymmetric synapses, containing dense-cored vesicles in addition to spherical synaptic vesicles; and normal, immunoreactive boutons with symmetric synapses and pleomorphic synaptic vesicles. It is suggested that the GAD-immunoreactive neurons which receive olfactory bulb input correspond to local circuit neurons with intralaminar axons which innervate each other as well as the distal segments of the apical dendrites of projection neurons with cell bodies in layers II and III. Thus, the olfactory input in EA seems to be wired not only for excitation of layers II and III pyramidal neurons but also for feed-forward inhibition using GABAergic intermediary neurons, strategically located in the area of termination of olfactory bulb fibers.