Ultrastructural characteristics of layer III pyramidal neurons in the cat primary auditory cortex (Al)

Electron microscope studies were made of retrogradely horseradish peroxidase-labeled pyramidal neurons forming transcallosal projections in layer III of the cat primary auditory cortex (Al). These showed a significant proportion of the somatic membrane to be covered with processes of astroglia, while synapses occupy 20% of the synaptic surface on average. Between 4 and 10 axosomatic synapses were identified on the profiles of callosal cell somata. All these were formed by axonal terminals containing small, flattened synaptic vesicles and had symmetrical contacts. Average length of these synaptic contacts equaled 1.6 µm. Numerous anterogradely horseradish peroxidase-labeled axonal terminals of callosal fibers were found in cortical area Al in amongst retrogradely HP-labeled neurons. The ultrastructural pattern of these is described.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 4, pp. 520–526, July–August, 1990.     

Synaptic organization of the cat parietal association cortex (area 5b)

An electron microscopy study was made of synaptic organization in the cat association cortex, area 5b. A total of 1635 axonal terminals were discovered over 6215 µm² (240 electronic imagings of slices of different association cortex layers); i.e., an average of 263±16 terminals per 1000 µm² expanse. It was found that 75.5% of axon terminals contained synaptic vesicles and formed either one- or two-sided contact with postsynaptic structures; 24.5% of axonal terminals contained synaptic vesicles but formed no distinct synaptic contacts with nearby neurons; 84.9% of terminals contained round-shaped or slightly oval synaptic vesicles; 7.8% had both rounded and elongated shapes, and vesicles were very elongated in the remaining 7.3%. Of the axonal terminals having synaptic contacts, axo(dendritic)-spinal terminals accounted for 46.6%, and axodendritic and axosomatic endings amounted to 50.0% and 3.4% respectively (in all 77% of axosomatic terminals contained elongated vesicles and maintained symmetrical contact, while 23% had round-shaped vesicles and formed asymmetrical contact). Calculations show that for each 1 mm³ an average of 258 million axonal terminals are found forming synaptic contacts in the cat association cortex as well as 84 million terminals containing synaptic vesicles but not forming contact.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 2, pp. 174–185, March–April, 1989.     

Morphology of the cat auditory cortex

The ultrastructural features of the primary auditory cortex of the cats and the character of the endings of geniculo-cortical afferent fibers in the early stages of experimental degeneration evoked by destruction of the medial geniculate body were studied. In all layers of the cortex asymmetrical synapses with round synaptic vesicles on dendritic spines and on thin dendritic branches of pyramidal and nonpyramidal neurons are predominant. Symmetrical synapses with flattened or polymorphic vesicles are distributed chiefly on the bodies of the neurons and their large dendrites. Because there are few symmetrical synapses which could be regarded as inhibitory it is postulated that inhibitory influences may also be transmitted through asymmetrical synapses with round vesicles. Other types of contacts between the bodies of neurons, dendrites, and glial processes also were found in the auditory cortex. Degenerating terminals of geniculo-cortical fibers were shown to terminate chiefly in layer IV of the cortex on pyramidal and nonpyramidal neurons. Degeneration was of the dark type in asymmetrical synapses with round vesicles. The results are dicussed in connection with electrophysiological investigations of the auditory cortex.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 5, No. 5, pp. 519–524, September–October, 1973.     

Distribution and ultrastructure of thalamic afferents in the cat auditory cortex

Ultrastructural features of thalamic afferent fibers were studied in the cat auditory cortex in the early stages (on the 4th day) of experimental degeneration produced by destruction of the medial geniculate body. A coordinate grid was used in conjunction with an electron micro-scope to study the topography of the degenerating elements over wide areas of sections, so that the density of degeneration could be determined quantitatively in different layers of the cortex. Degenerating axons were found in all layers. Most of the large (5–7 µ) degenerating axons are located in layer VI; their diameters were smaller in the upper layers of the cortex. Degenerative changes affecting synaptic terminals of thalamo-cortical afferents were of the "dark" type. Fibers of the geniculo-cortical tract were shown to terminate mainly in cortical layer IV. A few degenerating synapses were found in the molecular layer. Terminals with sperical synaptic vesicles are found mainly on the spines of dendrites where they form "asymmetrical" contacts. A few degenerating axo-somatic synapses were observed on stellate neurons in layer IV. The results are discussed in connection with electrophysiological investigations of the cat auditory cortex during stimulation of specific afferent fibers.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 4, No. 6, pp. 612–620, November–December, 1972.     

Ultrastructure and morphometric parameters of glutamatergic synapses on nigrothalamic and unidentified neurons of the reticular zone of theSubstantia nigra in cats

Nigrothalamic neurons were identified into thesubstantia nigra by their retrograde labelling with horseradish peroxidase. Axon terminals that contain glutamate (the excitatory transmitter) were revealed immunocytochemically with an immunogold electron microscopic technique. Ultrastructural parameters (the large and small diameters of axon terminals, area of their profiles, coefficient of form of profiles, large and small diameters of synaptic vesicles) were analyzed in all 240 synapses under study. Synaptic contacts localized on both nigrothalamic and unidentified neurons belonged to three morphologically specific groups. Synapses of the groups I and III, according to classification by Rinvik and Grofova, were characterized by a symmetric type of synaptic contact and contained polymorphic synaptic vesicles. Contacts in group-II synapses were asymmetric, and respective terminals contained round vesicles. Among the studied synapses, 65.8% were classified as group-I contacts, 25.0% belonged to group II, and 9.2% belonged to group III. Glutamate-positive axon terminals formed predominantly group-II synapses; such connections constituted 70% of this group's synapses. Sixty percent of glutamate-positive synapses were localized on the distal dendrites and 23% on the proximal dendrites, while 17% of such synapses were distributed on the somata of nigral neurons. Such a pattern of distribution of glutamate-positive synapses was observed on both nigrothalamic and unidentified nigral neurons. About 7% of glutamate-positive synapses were formed by very large axon terminals containing round synaptic vesicles; yet, the contacts of these terminals were of a symmetric type. Twenty percent of group-I synapses, i.e., synapses considered inhibitory connections, were found to manifest a weak immune reaction to glutamate.Neirofiziologiya/Neurophysiology, Vol. 28, No. 6, pp. 285–295, November–December, 1996.     

Ultrastructural characteristics of callosal neurons in deep layers of cat primary auditory cortex (AI)

We have carried out an electron microscopic investigation of retrogradely HRP-labeled nonpyramidal neurons in layers V and VI of the primary auditory cortex (AI), which are sources of transcallosal projections. We have established that on average 15.8±1.7% of the perikaryon surface of these cells is occupied by axo-somatic synapses. We detected in ultrathin sections from two to nine synapses on the profiles of the perikaryon of callosal neurons. All of these axo-somatic synapses are formed by axon terminals containing small flat synaptic vesicles and are characterized by symmetrical contacts. The length of the cross section of the contacts was on average 1.6±0.1 µm. The axon terminals of callosal fibers, antegradely labeled by the enzyme, form in the deep layers of the cortex asymmetrical synapses on the spines and stems of the dendrites. A possible functional significance of the axo-somatic synapses in the production of the impulse activity of callosal neurons in the deep layers of the AI region, is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 23, No. 5, pp. 549–556, May, 1991.     

Electron microscope study of sources in the cat primary auditory cortex (AI) projecting to the medial geniculate body

An electron microscope study of retrogradely labeled pyramidal neurons in layer VI of the primary auditory cortex (AI) after injecting horseradish peroxidase (HP) into the medial geniculate body was carried out in cats. Not less than 57.8±1.9% on average of the perimeter of perikaryon profiles of corticogeniculate neurons labeled with HP were found to be covered with astroglia processes. Between three and eight synapses occupying an average of 10.8±1.0% of the perimeter length were found on the perikaryon profiles of these neurons. Nearly all synapses (a total of 98.7%) at the soma of corticogeniculate neurons had symmetrical active zones, being made up of axonal terminals with flattened synaptic vesicles. Anterogradely HP-labeled axonal terminals of geniculocortical fibers were also found in the neuropil of layer VI in area AI, in addition to retrogradely labeled neurons. They contained large round synaptic vesicles and formed asymmetrical synapses. The potential role of axosomatic synapses in the shaping of corticogeniculate neuronal activity is discussed.A. A. Bogomolets Institute, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 2, pp. 171–178, March–April, 1990.     

Synaptic changes in the feline association cortex (area 5b) following lesion of the thalamic posterolateral nucleus

Numbers of intact and degenerating axonal terminals (AT) were determined in area 5b of the cat association cortex two, four, and 30 days after electrolytic destruction of the posterolateral thalamic nucleus (PL). The proportion of degenerating AT in area 5b two and four days after PL lesion was found to be 7.4 and 7.3%, respectively. Intact AT in the same field accounted for 83.6% of the total in intact animals four days after PL lesion. A 16.4% reduction occurred, consisting of AT with round vesicles and asymmetrical synaptic contacts (i.e., primarily excitatory AT); 40% of the damaged AT formed synapses on dendrites and 60% on spines. It follows that neurons of cortical association area 5b receive direct excitatory and mainly axonospinal afferents from the thalamic PL. Degenerating AT accounted for 4.6% 30 days after PL lesion, as compared with a decline of 31.2% and a rise of 7.0% in axospinal and axodendritic AT, respectively. No change occurred in axosomatic AT. The total number of AT had declined by 10.0%. Aspects of reinnervation of area 5b neurons following degeneration of the neuronal synapses induced by PL lesion are discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 5, pp. 661–667, September–October, 1989.     

Ultrastructural parameters of GABA-ergic and non-GABA-ergic synaptic contacts on neurons of the catsubstantia nigra

Nigrothalamic neurons were identified in the reticular part of thesubstantia nigra using labelling by the retrograde axonal transport of horseradish peroxidase. Nine parameters of the synaptic contacts (n=195) were analyzed, including the size and shape of terminals and size and type of synaptic vesicies. Sixty-six percent of axon terminals studied formed symmetric contacts and contained large polymorphic vesicles (group I). Two-thirds of these synapses were located on the distal dendrites, while one-third was distributed on the perikarya and proximal dendrites. Synapses of group II (29% of all synapses analyzed) exhibited asymmetric contacts and contained round agranular vesicles. Among these synapses, 79% were located on the distal dendrites, 19% were located on the proximal dendrites, and only 2% were located on the neuronal perikarya. Axon terminals of group III (5% of total population) exhibited symmetric contact and contained small polymorphic vesicles. High-resolution immunogold EM histochemistry indicated that 63% of the group-I axon terminals were GABA-positive. The majority of synapses on the labelled nigrothalamic neurons (21 contacts of 25) belonged to group I. Among these 21 synapses, 19 were axo-somatic and mostly GABA-positive. Within group II, 30% of synapses showed slightly expressed GABA-positivity.Neirofiziologiya/Neurophysiology, Vol. 27, No. 2, pp. 147–157, March–April, 1995.     

The time course of synapse formation of mouse neuroblastoma cells in monolayer cultures

Summary Neuroblastoma cells grown on substrates in culture develop long processes and assume the morphology of normal neurons as judged light microscopically. The development of synapses in the cultured tissue is studied by periodic electron microscopic examination of the areas of contact between cells. The initial expiants are free of any apparent synaptic contacts. After 48 h in culture, simple swellings or boutons are detected at the periphery of the cells or at the end of the fine processes. These initial synaptic profiles contain a few vesicles but lack mitochondria. The synaptic vesicles appear to originate from the smooth endoplasmic reticulum. Further expiants remain primitive, only the number of vesicles in the cytoplasmic swellings or boutons increases. These clusters of vesicles are 40–60 nm in diameter and morphologically distinguishable from the synaptic vesicles of normal neurons. There are no postsynaptic folds or membrane thickenings. Specialized cell contacts between cells are also present.     

Presynaptic Membrane Retrieval and Endosome Biology: Defining Molecularly Heterogeneous Synaptic Vesicles

The release and uptake of neurotransmitters by synaptic vesicles is a tightly controlled process that occurs in response to diverse stimuli at morphologically disparate synapses. To meet these architectural and functional synaptic demands, it follows that there should be diversity in the mechanisms that control their secretion and retrieval and possibly in the composition of synaptic vesicles within the same terminal. Here we pay particular attention to areas where such diversity is generated, such as the variance in exocytosis/endocytosis coupling, SNAREs defining functionally diverse synaptic vesicle populations and the adaptor-dependent sorting machineries capable of generating vesicle diversity. We argue that there are various synaptic vesicle recycling pathways at any given synapse and discuss several lines of evidence that support the role of the endosome in synaptic vesicle recycling.Chemical synapses contain discrete numbers of synaptic vesicles, which are capable of sustaining neurotransmitter release. Sustained neurotransmission occurs despite the secretory demands imposed by persistent and diverse patterns of neuronal electrical activity. Maintaining synaptic vesicle numbers requires local mechanisms to regenerate these vesicles to prevent their exhaustion, preserve plasma membrane surface area, and to maintain the molecularly distinct identity of a vesicle versus plasma membrane. Rizzoli and Betz (2005) eloquently draw a parallel between chemical neurotransmission with synapse chatter saying that some synapses “whisper,” whereas others “shout.” The “louder” the synapse, the more synaptic vesicles are required, extending from a few hundred vesicles (whisperers) to nearly thousands (shouters). This beautiful analogy implies that every synapse has just one “voice” or species of vesicle. Here we will present the case that synapses are more like choirs in which multiple vesicle species or “voices” contribute to the “pianissimo” or “fortissimo” parts of chemical neurotransmission.Synaptic terminals show a range of structural and functional differences in distinct regions of the brain, suggesting that the mechanisms for exocytosis/endocytosis coupling, as well as local vesicle recycling, may also be diverse. On one side, the Calyx of Held nerve terminal participates in fast and sustained synaptic transmission at high frequency (800 Hz), which is crucial for sound localization in the auditory brainstem (Taschenberger and von Gersdorff 2000; Borst and Soria van Hoeve 2012). The Calyx of Held houses ∼70,000 synaptic vesicles with nearly 3000 vesicles docked per Calyx terminal. These docked vesicles are distributed across the ∼500 active zones that exist per Calyx where vesicle fusion occurs (Satzler et al. 2002). On the other hand, hippocampal synapses fire action potentials at ∼0.5 Hz in bursts (Dobrunz and Stevens 1999). This synapse contains ∼200 synaptic vesicles and one active zone with ∼10 vesicles docked (Schikorski and Stevens 1997). With such a wide functional and structural gamut of synapses, it is reasonable to hypothesize that synaptic vesicles may differ in their retrieval mechanisms, not just at the rate at which the process occurs but also in the molecular pathways used.Two synaptic vesicle retrieval mechanisms, namely clathrin/AP-2/dynamin-dependent biogenesis and kiss-and-run, have been summarized in outstanding recent reviews (see, for example, Augustine et al. 2006; Rizzoli and Jahn 2007; Smith et al. 2008; Royle and Lagnado 2010; Ferguson and De Camilli 2012; Saheki and De Camilli 2012). Therefore, here we focus on the coupling of secretion and membrane retrieval, as well as endosome sorting. We will discuss new developments supporting the existence of diverse functional and molecular pools of synaptic vesicles and how endocytosis and endosome retrieval mechanisms may generate these vesicle pools.     

Serial synapses in Aplysia

Serial synapses occur between small profiles in the neuropil of Aplysia abdominal ganglion. Material was fixed in phosphate buffered OsO₄, embedded in epon, and sections were stained with uranyl acetate and lead citrate. A class of synapses had the following characteristics: (1) synaptic vesicles clustered against the presynaptic membrane, (2) a widened extracellular space of about 20 nm containing electron-dense material, (3) straightening of the pre- and postsynaptic membranes, and (4) no postsynaptic membrane specialization. Some density between the presynaptic membrane and the adjacent synaptic vesicles was occasionally observed. Synapses occurred between small profiles in the neuropil (typical profile diameters were 1–3 m?m). In this sample of approximately 100 synapses, four serial synapses were identified. The serial synaptic profiles were all small. In addition to the finding of serial synapses, 40% of the postsynaptic profiles contained vesicles similar to the synaptic vesicles seen in presynaptic profile. Serial synapses may be the anatomical substrate of presynaptic inhibition and facilitation and of dishabituation.     

How to make a synaptic ribbon: RIBEYE deletion abolishes ribbons in retinal synapses and disrupts neurotransmitter release

Synaptic ribbons are large proteinaceous scaffolds at the active zone of ribbon synapses that are specialized for rapid sustained synaptic vesicles exocytosis. A single ribbon‐specific protein is known, RIBEYE, suggesting that ribbons may be constructed from RIBEYE protein. RIBEYE knockdown in zebrafish, however, only reduced but did not eliminate ribbons, indicating a more ancillary role. Here, we show in mice that full deletion of RIBEYE abolishes all presynaptic ribbons in retina synapses. Using paired recordings in acute retina slices, we demonstrate that deletion of RIBEYE severely impaired fast and sustained neurotransmitter release at bipolar neuron/AII amacrine cell synapses and rendered spontaneous miniature release sensitive to the slow Ca²⁺‐buffer EGTA, suggesting that synaptic ribbons mediate nano‐domain coupling of Ca²⁺ channels to synaptic vesicle exocytosis. Our results show that RIBEYE is essential for synaptic ribbons as such, and may organize presynaptic nano‐domains that position release‐ready synaptic vesicles adjacent to Ca²⁺ channels.     

Possible mechanisms of synapse formation during ontogenesis

Electron microscopic investigation of synaptogenesis in the sensomotor cortex and in the caudate nucleus has been performed in the prenatal ontogenesis (16-22 days) and in newborn rats. The first immature synapses are demonstrated to appear beginning on the 16th day of embryogenesis. At the end of the prenatal development and especially in newborn animals desmosome-like, asymmetric and symmetric, mixed and complex forms of the synaptic contacts are revealed. As a result of the analysis performed on the ultrastructural organization of the contacts, a hypothesis explaining mechanisms of development of various elements of the synapses has been suggested. A part of the synaptic contacts of the asymmetric and symmetric types is supposed to be genetically programmed and membrane specialization of these contacts is formed earlier than synaptic vesicles appear. Other part of the synapses undergoes certain stages of differentiation before the functionally mature contact is formed. The initial stage in the synapses formation in formation of the desmosome-like junction. The second stage is appearance of synaptic vesicles in the area of this contact. The third stage includes development of pre- and postsynaptic membranous specialization and owing to this the contact acquires either asymmetric or symmetric appearance. For the ontogenetic periods investigated establishment of complex forms of the intercellular junctions (tangent, reciprocal, etc.) is specific; this evidently demonstrates certain plastic rearrangements in the synapses during the process of development.     

Ultrastructural organization of the neuropil in layer I of the cat cerebral cortex

The ultrastructure of layer I in the middle ectosylvian gyrus (area 22) of the cat's cerebral cortex was investigated. Beneath the subpial astrocytic layer most of the neuropil in layer I was shown to be occupied by nerve fibers and their terminals, terminal branches, dendritic spines, and astrocytic processes surrounding them. More than 90% of the presynaptic terminals contained spherical synaptic vesicles. The predominant types of interneuronal junctions are axo-spinous and axo-dendritic synapses of asymmetrical type. Presynaptic terminals, which contain flattened and pleomorphic synaptic vesicles, take part in the formation of all symmetrical junctions, accounting for 6% of the total number of synapses. Large polymorphic outgrowths filled with vacuoles — so-called multivacuolar sacs — are described. These structures were invaginated into varicose expansion of the terminal branches of apical dendrites of pyramidal neurons. They are shown to be outgrowths of presynaptic terminals. Dependence of synaptic function on the shape of the synaptic vesicles is examined.I. S. Beritashvili Institute of Physiology, Academy of Sciences of the Georgian SSR, Tbilisi. Translated from Neirofiziologiya, Vol. 15, No. 1, pp. 50–55, January–February, 1983.     

The Organum vasculosum laminae terminalis

Summary The Organum vasculosum laminae terminalis (OVLT) of the duck is lined innerly by specialized ependymal cells (tanycytes) and outwardly by a well-developed superficial vascular network, the capillaries of which often show a fenestrated endothelium. The OVLT also includes glial cells, internal non-fenestrated capillaries, bundles of fine nerve fibers and three groups of axonal swellings. One type contains granulations of 1000–1400 Å in diameter as well as 300–500 Å clear vesicles. The second type exhibits granulations and dense core vesicles of 500–800 Å in diameter along with small electron-lucent vesicles having diameters of 300–400 Å. In the third type, exclusively clear vesicles 300–600 Å in diameter are found. Asymmetrical synapses on dendrites and neuronal perikarya are found at every level of the organ. In the most external zone, the interposition of tanycyte endings sometimes allows neurosecretory axons to reach the parenchymal basement membrane (basal lamina).When tritiated molecules (amino acids or monoamines) are administered either in vitro by incubation or in vivo by intraventricular injections, radioautographic grains are observed over the tanycyte perikarya. Although this labeling is observed at every time point following the administration of the tracers, within three minutes only ³H-GABA appears to be concentrated in the cytoplasmic processes of the tanycytes. ³H-noradrenaline and ³H-serotonin are taken up and retained by some axons of the second type described above. Noradrenergic fibers are primarily localized in the inner zone of the OVLT where they display axodendritic synaptic contacts. Serotonergic fibers appear sparsely distributed in the OVLT but are more numerous in the lateral edges of the organ where synaptic differentiations on dendrites or on dendritic spines are also observed.It is concluded that the duck OVLT probably displays a neuroendocrine activity. Uptake and selective transport of exogenous molecules by tanycytes are also suggested by the present radioautographic observations. Finally, monoaminergic innervation is discussed at the OVLT level with special reference to the occurrence of serotonergic synapses.Supported by the Département de Biologie du C.E.A., and the I.N.S.E.R.M. (C.R.A.T., 74.1.438.45)     

Structural plasticity of synapses in hippocampal slices after oxygen-glucose deprivation

We analyzed structural rearrangements of synaptic contacts in the stratum radiatum of the CA1 area of cultured rat hippocampal slices under conditions of the development of potentiation of synaptic transmission induced by short-term (10 min) oxygen-glucose deprivation (OGD). Studies were carried out using electron microscopy and 3D reconstruction of cellular compartments. Within the 1st h after OGD, we observed increases in the volume of pre-synaptic terminals and post-synaptic spines and also in the area of postsynaptic densities (PSDs) in both asymmetric excitatory and symmetric inhibitory synapses, especially in the case were the PSD was perforated. We also observed significant activation of glial cells (increases in their volume and area of contacts of their processes with the components of synapses). Therefore, OGD results in activationassociated structural rearrangements of both excitatory and inhibitory synapses of the hippocampal CA1 area. Such rearrangements are accompanied by a clearly pronounced reaction of the glia, which correlates with an important role of the latter in modulation of the functioning of neurons.     

Electron Microscopic Analysis of the Inner Synaptic Layer in the River Lamprey (Lampetra fluviatilis)

Abstract Different types of synaptic contacts between bipolar, amacrine and ganglion cells were scored on random electron micrographs and on montages comprising the entire thickness of the inner synaptic layer. Currently accepted criteria were used when classifying the different cell processes. The percental distribution of dyads was estimated to 56 % amacrine-amacrine dyads, 34 % amacrine-ganglion dyads and 10 % ganglion-ganglion dyads. The ratio of amacrine conventional synapses to bipolar ribbon synapses was 6.8 : 1. The density per unit area of conventional synapses (0.035/μm²) and ribbon synapses (0.005/ μm²) was found markedly low as compared with other vertebrate species except the carp. The inner synaptic layer of the river lamprey is suggested to be of the intermediate type in which both simple and complex ganglion cell receptive fields may be expected.     

THREE-DIMENSIONAL ULTRASTRUCTURE OF THE CRAYFISH NEUROMUSCULAR APPARATUS

The synapse-bearing nerve terminals of the opener muscle of the crayfish Procambarus were reconstructed using electron micrographs of regions which had been serially sectioned. The branching patterns of the terminals of excitatory and inhibitory axons and the locations and sizes of neuromuscular and axo-axonal synapses were studied. Excitatory and inhibitory synapses could be distinguished not only on the basis of differences in synaptic vesicles, but also by a difference in density of pre- and postsynaptic membranes. Synapses of both axons usually had one or more sharply localized presynaptic "dense bodies" around which synaptic vesicles appeared to cluster. Some synapses did not have the dense bodies. These structures may be involved in the physiological activity of the synapse. Excitatory axon terminals had more synapses, and a larger percentage of terminal surface area devoted to synaptic contacts, than inhibitory axon terminals. However, the largest synapses of the inhibitory axon exceeded in surface area those of the excitatory axon. Both axons had many side branches coming from the main terminal; often, the side branches were joined to the main terminal by narrow necks. A greater percentage of surface area was devoted to synapses in side branches than in the main terminal. Only a small fraction of total surface area was devoted to axo-axonal synapses, but these were often located at narrow necks or constrictions of the excitatory axon. This arrangement would result in effective blockage of spike invasion of regions of the terminal distal to the synapse, and would allow relatively few synapses to exert a powerful effect on transmitter release from the excitatory axon. A hypothesis to account for the development of the neuromuscular apparatus is presented, in which it is suggested that production of new synapses is more important than enlargement of old ones as a mechanism for allowing the axon to adjust transmitter output to the functional needs of the muscle.     

Synaptic connections of fibers of the rubrospinal tract in the cat spinal cord

The ultrastructure of the lateral part of laminae VI and VII of the spinal gray matter (the location of most of the terminal branches of the rubrospinal tract) was investigated in cats under normal conditions and at various times after destruction of the red nucleus. The neuron population of this region is formed by cells fairly homogeneous in size (25–40µ). The structure of the dendritic profiles is simple and they carry only infrequent and small membranous appendages. Most synapses are axo-dendritic. The axon terminals are divided into three groups depending on the size and shape of the synaptic vesicles and the presence of post-synaptic specialization. A few glomerular axon terminals contacting with various structures are found. Small axon terminals located chiefly on dendrites and their appendages show degenerative changes 1–8 days after destruction of the red nucleus. As a rule the degenerating terminals contain round synaptic vesicles. The glomerular terminals do not degenerate.A. A. Bogomol'ets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 6, No. 6, pp. 610–618, November–December, 1974.