In vitro formation of neuroglial synapses

Neuroglial synapses have been systematically observed in histotypical cerebellar rotary tissue cultures. These axoglial contacts appear as excitatory-like synapses in which the presynaptic element contains rounded synaptic vesicles accumulated at the presynaptic active zone. The post-synaptic structures were always the somata as well as processes of astrocytic glial cells. The axoglial contacts appear in these cultures when they are prepared from 16-day-old chick embryos cerebellum and cultivated during 6 days. To explain these neuroglial interrelationship it has been suggested as a working hypothesis that glial and nerve cells develop plastic capacities in order to establish synaptic contact in the in vitro conditions.     

Morphological evidence for local microcircuits in rat vestibular maculae

Previous studies suggested that intramacular, unmyelinated segments of vestibular afferent nerve fibers and their large afferent endings (calyces) on type I hair cells branch. Many of the branches (processes) contain vesicles and are presynaptic to type II hair cells, other processes, intramacular nerve fibers, and calyces. This study used serial section transmission electron microscopy and three-dimensional reconstruction methods to document the origins and distributions of presynaptic processes of afferents in the medial part of the adult rat utricular macula. The ultrastructural research focused on presynaptic processes whose origin and termination could be observed in a single micrograph. Results showed that calyces had 1) vesiculated, spine-like processes that invaginated type I cells and 2) other, elongate processes that ended on type II cells pre- as well as postsynaptically. Intramacular, unmyelinated segments of afferent nerve fibers gave origin to branches that were presynaptic to type II cells, calyces, calyceal processes, and other nerve fibers in the macula. Synapses with type II cells occurred opposite subsynaptic cisternae (C synapses); all other synapses were asymmetric. Vesicles were pleomorphic but were differentially distributed according to process origin. Small, clear-centered vesicles, ˜40–60 nm in diameter, predominated in processes originating from afferent nerve fibers and basal parts of calyces. Larger vesicles ˜70–120 nm in diameter having ˜40–80 nm electron-opaque cores were dominant in processes originating from the necks of calyces. Results are interpreted to indicate the existence of a complex system of intrinsic feedforward (postsynaptic)-feedback (presynaptic) connections in a network of direct and local microcircuits. The morphological findings support the concept that maculae dynamically preprocess linear acceleratory information before its transmission to the central nervous system. J. Comp. Neurol. 379:333–346, 1997. © 1997 Wiley-Liss, Inc.     

Transformation of cells of astrocyte lineage into macrophage-like cells in organotypic cultures of mouse spinal cord tissue

Phagocytic cells on the surface of the explants and their relationships to the surface were examined morphologically and immunocytochemically in organotypic cultures of mouse spinal cord tissue. Phagocytic cells were rounded, had smooth cytoplasmic surfaces and were occasionally closely apposed to underlying cells by junctional complexes. These cells contained dense bodies, vacuoles, smooth and coated vesicles, a few microtubules and bundles of intermediate filaments similar to astroglial filaments. The superficial layer of the explant which usually consisted of astroglial cell bodies and their processes, sometimes contained immature neuroepithelial cells with numerous free ribosomes, centrioles, Golgi apparatus, microtubules and infrequently, intermediate filaments. Overall, the cells resembled poorly differentiated astrocytes. Numerous dense bodies and coated vesicles were observed in some of these immature cells as well as in astrocytes in the surface layer of the explant. Cytoplasmic bridges between immature cells within the explant and phagocytic cells on the surface were observed. Immunocytochemistry revealed the presence of glial fibrillary acidic protein within these surface phagocytic cells. It thus appears that immature neuroepithelial cells of astrocytic lineage are capable of transforming into macrophage-like cells in organotypic culture.     

Postnatal development of the cerebellar cortex in the rat. II. Phases in the maturation of Purkinje cells and of the molecular layer

The growth and synaptic maturation of Purkinje cells and of the molecular layer were studied in the cerebellar cortex of rats aged 0, 3, 5, 7, 10, 12, 15, 21 and 30 days with histological (including Golgi), histochemical, autoradiographic and electron microscopic techniques. Five phases were distinguished in the maturation of Purkinje cells. During the first phase, Purkinje cells become dispersed and aligned in a monolayer but as yet few or no synapses are formed. Next, two transient structures appear: a hypertrophied apical cone composed of ?reticular”? cytoplasm, and lateral perisomatic processes which establish conspicuous asymmetrical synapses with climbing fibers. During the third phase the perisomatic processes disappear; the ?reticular”? cytoplasm streams upward into the growing dendrites; the soma is invaded by permanent inconspicuous, symmetrical synapses of basket cells; and, finally, it is surrounded by glial processes, which marks the end of the synaptic maturation of the soma. During the fourth phase parallel fibers form synapses with dendritritic spines in the lower half of the molecular layer. During the fifth phase, which occurs after the disappearance of the external germinal layer, parallel fibers establish synapses with dendritic spines in the upper molecular layer. The ?march”? of synaptogenesis in the molecular layer from the bottom upward is characterized by three successive events: an initial gradient in the appearance and disappearance of coated vesicles, heralding synaptogenesis; a similar subsequent trend in the formation of synapses; and finally, the interposition in the same sequence of glial processes between Purkinje cell dendrities and parallel fibers, marking the cessation of synaptogenesis.     

Regeneration of dorsal root fibers into the adult rat spinal cord

Regeneration of dorsal root nerve fibers into the spinal cord of adult rat was studied with the electron microscope after crushing the roots. Regenerated dorsal root myelinated fibers were observed in the substantia gelatinosa and posterior funiculi around the tenth week after lesion. Cytoplasmic processes of oligodendrocytes were often found close to the young myelinated nerve fibers. The astrocytic response subsided as regeneration progressed. Clusters of small, circular profile of cellular processess were found in the neuropil about the sixth week and are considered to be regenerated unmyelinated axons. At this period, groups of segments of cellular processes containing clear vesicles were also encountered in the substantia gelatinosa and resembled growth cones described in peripheral regenerating nerves.     

Junctional specializations between growth cones and glia in the developing rat pyramidal tract: synapse-like contacts and invaginations

The ultrastructure of contacts between axonal growth cones and glial cells in the developing pyramidal tract was examined by serial sectioning at the third cervical spinal cord segment in 0-, 2-, and 4-day-old rats. Junctional specializations, composed of synapse-like contacts and invaginations, were frequently observed at the contact zone between growth cones and glial elements. The synapse-like contacts consist of clear, round vesicles of 43 +/- 6 nm in the presynaptic growth cone, a pre- and a postsynaptic density, separated by a cleft of 12.1 +/- 0.9 nm. The invaginations consist of small protrusions of the growth cone into the glial element. The invaginated glial membrane is coated. Within the glial element, close to the invagination, frequently organelles were observed that closely resemble endosomes and prelysosomes. Therefore, it is suggested that the invagination represents a stage in endocytosis or possibly phagocytosis of the protruding part of the growth cone by the glial cell. The junctional specializations are formed by growth cones and, less frequently, by axon shafts. The targets of these specialized contacts are, in general, immature glial cells located within the tract area. Occasionally, however, invaginations were also observed into myelinating oligodendrocytes, suggesting that the population of immature target cells includes oligodendrocyte precursors. With regard to the functional significance of these temporary growth cone-glial contacts, several possibilities are discussed, including the suggestion that outgrowing pyramidal tract axons provide immature glial cells with chemical messages, which may influence the timing of glial cell maturation in the tract.     

Cell types and synaptic organization of the medullary electromotor nucleus in a constant frequency weakly electric fish,Sternarchus albifrons

The medullary electromotor nucleus (EMN) of Sternarchus albifrons was studied at the light and electron microscopic levels. The EMN consists of a dense meshwork of myelinated axons and glial elements with interposed large neurons; it is provided with an abundant supply of capillaries. Two types of essentially adendritic nerve cells were distinguished on the basis of size: giant neurons (approx. 70 μm in diameter) and large neurons (approx. 30 μm in diameter). Their population ratio is 1:4. Only giant cells are labelled following the injection of retrograde tracer into the spinal cord; they are therefore identified with the so-called “relay cells” of other gymnotids. Tracer experiments further suggest that the descending axons of these relay cells give off collateral branches throughout the elongated spinal electromotor nucleus. In contrast, the large cells remain unlabelled and therefore lack spinal projections; they most likely correspond to “pacemaker cells”. The perikaryal surface, including axon hillock and proximal part of initial segment of both types of EMN cells, is contacted by clusters of synaptic terminals and astrocytic processes. Two main varieties of synaptic terminals occur: (1) large endings and (2) ordinary end feet with standard size (S-type) and variable size (Sv-type) clear, spherical vesicles. The junction between large endings and EMN cells is characterized by the combination of gap junctions and surrounding intermediate junctions whose freeze-fracture characteristics were morphometrically analyzed. The large endings were formed by nodes of Ranvier as well as by fiber terminations, and synchronization within the EMN may be achieved by presynaptic fibers. Some of the contacts occur directly on the initial segment, which could allow activity to bypass the soma. It is concluded that the electromotor system of Sternarchus is comprised of a rapid conduction pathway where medullary pacemaker and relay cells as well as spinal electromotor neurons are coupled by synapses with gap junctions. In contrast to the spinal electromotor neurons, the medullary EMN cells receive synapses with morphological characteristics of chemical transmission, and the S-type and Sv-type terminals may possibly correspond to Gray's Type I and Type II synapses, respectively. These synapses may be involved in modulation of the electric organ discharge frequency.     

The neural organization of the first optic ganglion of the principal eyes of jumping spiders (Salticidae)

The first optic ganglion (FOG) of the principal eyes of jumping spider has been studied by both light and electron microscopy. Each FOG receives projections from the ipsilateral principal eye in a compact optic nerve. Some of the retinal axons, as they pass to the ganglion, cross in a chiasm within the optic nerve. The FOG is composed of a rind of nerve cell bodies which are unipolar cells giving rise anteriorly to second order nerve fibers in the central neuropile of the ganglion and posteriorly to processes which pass into the second optic ganglion. The neuropil is composed of neural and glial processes and of scattered glial cell bodies and it can be divided into a terminal zone and a fibrous zone. The terminal zone is the site of termination for all of the retinal axons. A unique feature of the retinal axon terminals is the presence of numerous fine post-terminal processes extending away from the main portion of the retinal terminal. The fibrous zone is composed of second order nerve fibers with terminals in the terminal zone and it is continuous with a connective between the FOG and the second optic ganglion called the chiasm where the majority of the second order fibers cross. All of the synapses in the FOG are modified ribbon synapses and occur in the terminal zone within ensheathed synaptic glomeruli as dyads or triads. Large retinal terminals containing pleomorphic vesicles are presynaptic to smaller second order terminals containing spherical vesicles. Second order terminals are also presynaptic to retinal terminals in addition to forming serial-like synapses with each other.     

Electron microscopic study of cardiac ganglia in human fetuses

The early events in the development of the heart ganglia and nerves in human fetuses ranging from 5 to 12 weeks of gestation age were studied by transmission electron microscopy. The first neuroblasts in the atrial mesenchyme differ from surrounding cells in the presence of short cisternae of rough endoplasmic reticulum and in the absence of glycogen particles. The most valuable criterion for identification of neuroblasts is the presence of contacts with preganglionic nerve terminals. Only ganglia composed of compact aggregations of neuronal cells and nerve terminals have a complete glial sheath. The first signs of synapse formation were seen in 5-week-old fetuses; well developed synapses with many synaptic vesicles were found from the 8th week of gestation onwards. These were predominantly axodendritic synapses. It is proposed that synaptogenesis begins with the appearance of osmiophilic zones at the sites of interneuronal contacts, then synaptic vesicles move by axonal transport to reach the preformed specialized junctions and stop in axonal presynaptic varicosities.     

Acute transplantation of glial-restricted precursor cells into spinal cord contusion injuries: survival, differentiation, and effects on lesion environment and axonal regeneration

Transplantation of stem cells and immature cells has been reported to ameliorate tissue damage, induce axonal regeneration, and improve locomotion following spinal cord injury. However, unless these cells are pushed down a neuronal lineage, the majority of cells become glia, suggesting that the alterations observed may be potentially glially mediated. Transplantation of glial-restricted precursor (GRP) cells--a precursor cell population restricted to oligodendrocyte and astrocyte lineages--offers a novel way to examine the effects of glial cells on injury processes and repair. This study examines the survival and differentiation of GRP cells, and their ability to modulate the development of the lesion when transplanted immediately after a moderate contusion injury of the rat spinal cord. GRP cells isolated from a transgenic rat that ubiquitously expresses heat-stable human placental alkaline phosphatase (PLAP) were used to unambiguously detect transplanted GRP cells. Following transplantation, some GRP cells differentiated into oligodendrocytes and astrocytes, retaining their differentiation potential after injury. Transplanted GRP cells altered the lesion environment, reducing astrocytic scarring and the expression of inhibitory proteoglycans. Transplanted GRP cells did not induce long-distance regeneration from corticospinal tract (CST) and raphe-spinal axons when compared to control animals. However, GRP cell transplants did alter the morphology of CST axons toward that of growth cones, and CST fibers were found within GRP cell transplants, suggesting that GRP cells may be able to support axonal growth in vivo after injury.     

Development of axosomatic synapses of the Xenopus spinal cord with special reference to subsurface cisterns and C-type synapses

The relationships of highly flattened subsurface cisterns (SCCs) were investigated electron microscopically in the spinal cord at various devel- opmental stages of tadpoles and adult toads, Xenopus laeuis. In medial ventral motor cells (MVCs) of the adult, more than 90% of 156 SSCs examined were situated postsynaptically. Similarly, more than 909% of 540 SSCs in lateral motor column cells (LMCs) were postsynaptic. By contrast, in early developmental stages, the SSCs were initially formed by regional flattening of cisterns of rough-surfaced endoplasmic reticulum just beneath the cell surfaces opposite to glial processes. Then, the glial processes were displaced by nerve endings with an elongated bouton, and thus the C-type synapses were formed. The ratio of post- synaptic SSCs to the total SSCs reached the adult level at around Stage 60. This finding suggests that the SSCs in the MVCs and LMCs draw a certain type of nerve ending to form C-type synapses. Such a mechanism is totally lacking in the dorsal and lateral small nerve cells, since the SSCs in these cells were always situated under the surface opposite to glial processes throughout the developmental stages and in the adult. In mature C-type synapses, an aggregate of synaptic vesicles and a structural specialization of presynaptic membrane occurred only at the region where the postsynaptic membrane was associated with the SSC. The postsynaptic membrane itself of the C-type synapse showed no marked structural specialization at any stage of development or in the adult. The postsynaptic SSC in the mature C-type synapse seems to be involved in some way in the reception of synaptic transmission.     

Axonal projections and synaptogenesis by supraspinal descending neurons in the spinal cord of the chick embryo.

Following the injection of horeseradish peroxidase (HRP) into the brachial spinal cord of the chick on embryonic day (E)4.5, retrogradely labeled neurons can be found in the brainstem (Okado and Oppenheim: Journal of Comparative Neurology 232: 143-161, 1985). By contrast, following high cervical spinal transection, functional (behavioral) deficits are not observed until E10 (Oppenheim: Journal of Comparative Neurology 160: 37-50, 1975). To determine whether this temporal difference between projections and function reflects a delay in synaptogenesis, we looked for the presence of anterogradely HRP-labeled pre-synaptic terminals in brachial cord following injection of HRP into the boundary between brainstem and spinal cord at ages between E3.5 and E7. HRP-labeled fibers were observed in the branchial cord by E4.5 and were diffusely distributed in the ventral and lateral marginal zones (presumptive ventral and lateral funiculi, respectively). Although some axo-dendritic and axo-somatic synapses were observed in the brachial cord prior to E6, the presynaptic profiles were always unlabeled by HRP and thus must originate from propriospinal sources. The first HRP-labeled supraspinal synapses were found in the ventral and lateral funiculi on E6. They contained several clear spherical synaptic vesicles and were axo-dendritic in nature. The cells of origin of the postsynaptic dendrites were determined by injecting HRP into the wing-bud to label the brachial motoneurons retrogradely and the presynaptic component was identified as supraspinal by HRP injections into the brainstem/spinal cord boundary to orthogradely label the descending fibers. Several double-labeled axo-dendritic synapses were found in the ventral and lateral funiculi of E6 brachial cord. Therefore, at least some descending supraspinal fibers make synapses directly onto motoneuron dendrites. We conclude that 1) there is a delay of about 1.5 days between the arrival of supraspinal fibers and synapse formation in the brachial cord, 2) the earliest synapses are axo-dendritic in nature, 3) at least some supraspinal fibers make direct contact with motoneuron dendrites as early as E6, and 4) synaptogenesis from propriospinal sources precedes that from supraspinal descending axons. These observations provide evidence indicating that the temporal difference between the onset of projections of supraspinal descending fibers and the onset of their function may be partly owing to delayed synaptogenesis.     

Reciprocal synapses between inner hair cell spines and afferent dendrites in the organ of corti of the mouse

We provide, for the first time, ultrastructural evidence for the differentiation of reciprocal synapses between afferent dendrites of spiral ganglion neurons and inner hair cells. Cochlear synaptogenesis of inner hair cells in the mouse occurs in two phases: before and after the onset of hearing at 9-10 postnatal (PN) days. In the first phase, inner hair cells acquire afferent innervation (1-5 PN). Reciprocal synapses form around 9-10 PN on spinous processes emitted by inner hair cells into the dendritic terminals, predominantly in conjunction with ribbon afferent synapses. During the second phase, which lasts up to 14 PN, synaptogenesis is led by the olivocochlear fibers of the lateral bundle, which induce the formation of compound and spinous synapses. The afferent dendrites themselves also develop recurrent presynaptic spines or form mounds of synaptic vesicles apposed directly across inner hair cell ribbon synapses. Thus, in the adult 2-month mouse, afferent dendrites of spiral ganglion neurons are not only postsynaptic but also presynaptic to inner hair cells, providing a synaptic loop for an immediate feedback response. Reciprocal synapses, together with triadic, converging, and serial synapses, are an integral part of the afferent ribbon synapse complex. We define the neuronal circuitry of the inner hair cell and propose that these minicircuits form synaptic trains that provide the neurological basis for local cochlear encoding of the initial acoustic signals.     

Morphological changes in the cat cerebral cortex produced by superfusion of ouabain

The morphological changes following superfusion of the cat cerebral cortex with ouabain were studied. Autoradiography of [3H]ouabain was performed to study drug distribution. The resulting lesion consists of an upper vacuolated layer which is distinctly separated from an underlying region containing dark neurones. Ouabain is confined to the vacuolated layer. Swelling of apical dendrites and many presynaptic terminals are the main morphological changes occurring in the vacuolar layer. Depletion of synaptic vesicles, clustering of vesicles around the synaptic membrane, and the production of coated vesicles and cisternae are further changes found within presynaptic endings. Whilst swelling of fibrous astrocytes within the glialimitans occurs, this is not true of astrocytic processes elsewhere in ouabain exposed regions. Regions containing dark neurones are characterised by a general swelling of astroglial processes. The results strongly suggest that apical dendrites and presynaptic endings possess high activities of Na+, K+-ATPase whereas the activity on astroglial processes within the neuropil is relatively low. Astroglial swelling in areas of dark neurones is produced by some change in the chemical milieu surrounding the processes which appears unrelated to Na+, K+-ATPase inhibition.     

Glial process elongation and branching in the developing murine neocortex: a qualitative and quantitative immunohistochemical analysis

Cells of astroglial lineage in the murine cerebrum undergo a succession of transformations during prenatal and early postnatal development. The bipolar radial cell, the earliest astroglial form to appear, provides a radially aligned, parallel array of fibers that serves as a guide to neuronal migration. The multipolar astrocyte is the representative of this lineage that persists in the adult cerebrum. The processes of the multipolar astrocytes form a complex reticulum, which is considered critical to the development, function, and maintenance of neural circuits. A monopolar radial cell appears to be transitional between the two. The shift from the radial glial fiber system to a diffuse glial network is achieved largely in the E17-P2 interval in the mouse. This phenomenon has been studied qualitatively and quantitatively by staining cerebral tissue with monoclonal antibody RC2, a specific and sensitive ligand for cells of astroglial lineage in the mouse. Elongation and branching of glial processes contribute to the glial transformation. Elongation of radial fibers occurs under the guidance of other radial glial fibers (fasciculated elongation) or independently of other fibers (nonfasciculated elongation). Fasciculated elongation results in an increase in the density of radial glial fibers that span the cortical layers. Nonfasciculated elongation appears to be associated with process branching. This is the initial event in transformation of the bipolar radial cells to monopolar radial or multipolar cells. Only nonfasciculated elongation is characteristic of processes of the monopolar radial cells and multipolar astrocytes. Branching of the processes of all three cell forms appears to occur both by bifurcation at the elongating tip and by sprouting from the fiber shaft. Elongating fibers are tipped by growth cones that are relatively simple in shape as compared to those observed at the tips of elongating axons. Growth cones at the tips of nonfasciculated fibers are more complex in form than those at the tips of radial fibers elongating in contact with other radial fibers.     

Synaptogenesis in the rat suprachiasmatic nucleus: a light microscopic immunocytochemical survey

MabQ155, a monoclonal antibody against synaptophysin, has been used to conduct a light microscopic survey of synaptogenesis in the suprachiasmatic nucleus of the perinatal rat. Synaptophysin is an integral component of synaptic vesicle membranes which is expressed in growth cones and growth cone filopodia as well as in mature synapses. With the light microscope, mabQ155 immunoreactivity in growth cones can be distinguished from that in presynaptic terminals on the basis of the size of immunoreactive puncta. The current study presents a qualitative and quantitative analysis of synaptogenesis from the day of birth (P0) to postnatal day 10 (P10). In our quantitative analysis we have used daily intervals during the first postnatal week, distinguished between growth cones and presynaptic terminals, and divided the suprachiasmatic nucleus into sampling regions that are related to the progress of synaptogenesis. Our data demonstrate regional differences in synaptogenesis within the suprachiasmatic nucleus (SCN), document the temporal progression from the penetration of growth cones to the appearance of mature synapses, and provide information about gradients of synaptogenesis in the nucleus during development.     

Electron microscopic observations on neuron-like cells in the ground squirrel pineal gland

Summary Ultrastructural details of neuron-like cells as well as synaptic nerve endings in the pineal gland of the ground squirrel are described. The neuronlike cells are situated mainly in the distal portion of the gland. Since the neuron-like cells differ considerably from the pinealocytes and exhibit cytological features characteristic of nerve cells, they are presumably true neurons. The cell bodies or processes of the neuron-like cells receive many synapses. The nerve endings synapsing on these cells contain numerous small non-granulated and some large granulated vesicles but no small granulated vesicles. These synaptic nerve endings are rather abundant within the parenchyma but they are also found in perivascular spaces ensheathed completely or partially by the Schwann cell cytoplasm. Similar nerve endings occasionally make typical synaptic contacts with the cell bodies or processes of the pinealocytes. Since the adrenergic nerve endings do not form synapses on the pinealocytes in the ground squirrel, it is clear that nerve endings other than adrenergic ones also distribute within the pineal gland of this animal. These synaptic nerve endings may contribute, at least to some extent, to the innervation of the ground squirrel pineal gland, whether they are derived from the nerve fibers penetrating the pineal gland or from the neuron-like cells.     

Neuronal sprouting and synapse formation in response to injury in the mouse organ of corti in culture

The effect of mechanical injury on induction of regenerative phenomena within the neurosensory epithelium was investigated in cultures of neonatal mouse cochlea. The oldest examined culture in which new neuronal growth followed insult, was injured at 13 days in vitro and fixed 24 h later. By far, the most vigorous regenerative reaction was observed in a 3-day culture 4 h post-injury. The reaction included sprouting of nerve fibers injured directly, synapse formation between the surviving hair cells and sprouting neuronal growth cones, wrapping of growing nerve fibers by extending processes of hair cell cytoplasm, and collateral sprouting of synaptically-engaged nerve endings and of nerve fibers in passage.     

Postnatal development of axosomatic synapses in the rat visual cortex: morphogenesis and quantitative evaluation

Postnatal development of axosomatic synapses was studied in the rat visual cortex in order to obtain experimental data that may explain how the unequal distribution of asymmetric and symmetric synapses evolves on the soma of cortical neurons. Three types of synaptic junctions were identified: asymmetric or type 1 synapses, with postsynaptic densities greater than or equal to 20 nm, symmetric type 2 synapses, and symmetric synapses with an intermediate structure. The third synapse type had a structure similar to that of type 1 synapses, although the postsynaptic densities were thinner than 20 nm. Type 1 synapses developed in three phases. In phase 1, the first postnatal week, there were many free postsynaptic thickenings and immature synapses whereby a higher degree of postsynaptic differentiation was visible in comparison to the presynaptic elements. During the following 10 days, phase 2, type 1 synapses containing thin postsynaptic densities and intermediate synapses temporarily increased in number. Intermediate synapses are interpreted as precursors of type 1 synapses that have relatively immature postsynaptic elements. Toward the end of synaptogenesis, phase 3, the free postsynaptic thickenings reappeared while type 1 synapses containing well developed postsynaptic elements prevailed. Throughout the whole postnatal period, the numerical density of axosomatic type 1 synapses remained very low and the ratio of asymmetric to symmetric synapses at the neuronal somata was inversely proportional to that at the dendrites. Also, there was a significant decrease in the numerical density of type 1 synapses between postnatal days (P) 17 and 30. Data normalized according to cortical growth suggest that this is probably due to a decrease in the number of axosomatic type 1 synapses. This corresponds to the observation that in layers III and V a few type 1 synapses were found on pyramid-like cells up to P10 which then disappeared in later stages. Axosomatic type 2 synapses appear to be formed by two different presynaptic processes. The first presynaptic type contains flocculent material with glycogen granules and resembles axonal growth cones. These junctions contain multiple adhesion patches, intermediate junctions, one or more active zones, narrow synaptic clefts, and small pleomorphic vesicles. All of these are structural features of adult type 2 synapses. The growth-cone-like presynaptic elements disappeared after about 3 weeks. The second presynaptic type is smaller in size and also forms contacts with a structure similar to adult type 2 synapses.(ABSTRACT TRUNCATED AT 400 WORDS)     

HVEM serial-section analysis of rabbit foliate taste buds: I. Type III cells and their synapses

Serially sectioned rabbit foliate taste buds were examined with high voltage electron microscopy (HVEM) and computer-assisted, three-dimensional reconstruction. This report focuses on the ultrastructure of the type III cells and their synapses with sensory nerve fibers. Type III cells have previously been proposed to be the primary gustatory receptor cells in taste buds of rabbits and other mammals. Within rabbit foliate taste buds, type III cells constitute a well-defined, easily recognizable class and are the only taste bud cells observed to form synapses with intragemmal nerve fibers. Among 18 type III cells reconstructed from serial sections, 11 formed from 1 to 6 synapses each with nerve fibers; 7 reconstructed type III cells formed no synapses. Examples of both convergence and divergence of synaptic input from type III cells onto nerve fibers were observed. The sizes of the active zones of the synapses and numbers of vesicles associated with the presynaptic membrane specializations were highly variable. Dense-cored vesicles 80-140 nm in diameter were often found among the 40-60 nm clear vesicles clustered at presynaptic sites. At some synapses, these large dense-cored vesicles appeared to be the predominant vesicle type. This observation suggests that there may be functionally different types of synapses in taste buds, distinguished by the prevalence of either clear or dense-cored vesicles. Previous investigations have indicated that the dense-cored vesicles in type III cells may be storage sites for biogenic amines.