Interneuronal synapses in the local ganglia of the cat urinary bladder.

The interneuronal synapses of the urinary bladder in the cat were studied by electron microscopy. The great majority of the fibres containing vesicles are found within the ganglia occurring in the trigonum area. Morphologically differentiated synaptic contacts could be observed on the surface of the local neurons and between the different nerve processes. The presynaptic terminals can be divided into three types based on a combination of synaptic vesicles. Type I terminals, presumably cholinergic synaptic terminals, contain only small clear vesicles of 40-50 nm in diameter. Type II terminals, presumably adrenergic terminals, are characterized by small granulated vesicles of 40-60 nm in diameter. Type III terminals, probably of local origin, contain a variable number of large granulated vesicles of 80-140 nm in diameter. Occasionally, a single nerve fibre contacted several (two or four) other nerve processes forming a typical synapse. In other cases, on one nerve cell soma or on other nerve processes there are two or three different-type nerve terminals establishing synapses. It might be inferred from these observations that convergence and divergence can occur in the local ganglia and that cholinergic and adrenergic synaptic terminals can modulate the ganglionic activity. However, a local circuit also can play an important role in coordinating the function of the bladder.     

Ultrastructure of nerve plexus in flatworms

Summary Synaptic components from the peripheral nervous system of the polyclad flatworm, Notoplana acticola, are described from electron microscopic observations. Quasineuropile, defined as clusters of neurites containing synaptic vesicles, occurs as scattered islands along the peripheral nerve cords of the plexus. Some neurite clusters only contain one type of synaptic vesicle but others are mixed. The most usual synaptic configuration consists of a single presynaptic element and a pair of postsynaptic neurites sharing a common synaptic cleft. These synapses are polarized and contain clear, 420 Å vesicles. GABA-type synapses are also found. At least two kinds of solid-core vesicles also occur.     

The role of the synaptic vesicles in transmission at the insect nerve-muscle junction

The ultrastructure of nerve-muscle synapses on red and white fibres of locust ($\text{Schistocerca}$$\text{gregaria}$) retractor unguis muscle fibres was examined before and after stimulation. At a stimulation frequency of 15Hz, the synapses on the white fibres fatigued completely; those on the red fibres also fatigued, but during prolonged stimulation they showed intermittent periods of recovery. Synaptic vesicles at both types of fatigued synapses were reduced in volume compared with controls and at fatigued synapses on white fibres the vesicles had irregular outlines. Aggregation of vesicles were observed at fatigued synapses on red fibres and the synaptic cleft width at these synapses was less than at control synapses on red fibres. These results are discussed in the light of the vesicle hypothesis for synaptic transmission.     

Fine structure of nerve-melanophore contacts in the angelfish Pterophyllum scalare

Contacts between small unmyelinated nerve fibres and dermal melanophores of the angelfish, Pterophyllum scalare, exhibit several features characteristic of synapses, including small synaptic vesicles and dense core vesicles, a narrow synaptic cleft, electron-dense material at the postsynaptic membrane (cell membrane of the melanophore) and, occasionally, presynaptic densities. An analysis of serial thin sections shows that the synapses described here represent varicosities of an otherwise more or less straight nerve fibre. A single axon thereby may form several en passant synapses with a single melanophore. It is suggested that the synaptic contacts described here not only represent sites of transmitter release but also play a role as sites of firm attachment between nerves and melanophores which guarantee a stable arrangement of nerve fibres and melanophores.Supported by the Deutsche Forschungsgemeinschaft     

Synaptic connections of substance P-immunoreactive nerve terminals in the substantia nigra of the rat

Summary A monoclonal antibody that recognises the C-terminal part of substance P was used to study immunoreactive structures in the substantia nigra by the unlabeled antibody, peroxidase-antiperoxidase procedure. Immunoreactivity was present in nerve fibres in all parts of the substantia nigra, particularly in the pars reticulata and pars lateralis. Electron microscopically two types of bouton immunoreactive for substance P were found: Type 1 contained large electron-lucent vesicles, occasional large granulated vesicles and formed symmetrical synapses with dendrites. Type 2 boutons contained smaller, round electron-lucent vesicles, many large granular vesicles and formed asymmetrical synapses (having prominent postjunctional dense bodies) with dendrites and perikarya.Immunoreactive fibres with varicosities that had been identified light microscopically were studied in serial sections in the electron microscope. Each identified varicosity contained synaptic vesicles and formed a single synapse. An individual fibre formed boutons of only one kind (type 1 or type 2) and could form multiple synapses with the same neuron. Thus, an identified fibre in the pars compacta had eight varicosities, each of which was in synaptic contacts (type 2) with the dendrites or soma of the same neuron.The results are consistent with the concept that substance P is a synaptic transmitter in the substantia nigra and indicate that neurons in this region may receive a significant input from substance P-containing afferents, and that there are at least two types of such afferent fibres.     

Innervation and cytochemistry of the neuroepithelial bodies in the ciliated epithelium of the toad lung (Bufo marinus)

Summary Neuroepithelial bodies (NEB) were identified in the lung of Bufo marinus. The characteristics of the cells and their innervation were studied with electron and fluorescence microscopy before and after close vagosympathetic denervation. The bodies consist of low columnar cells which rest on the epithelial basal lamina. The majority of the cells do not reach the lumen of the lung (basal cells); the few which do (apical cells) are bordered by microvilli and possess a single cilium. The neuroepithelial cell cytoplasm contains a variety of organelles the most characteristic of which are dense cored vesicles. Microspectrofluorometry and electron microscopic cytochemistry indicate significant quantities of 5-hydroxytryptamine in these cells. The neuroepithelial bodies could be divided into three groups on the basis of their innervation: 1) About 60% of the NEBs are innervated solely by nerve fibres containing agranular vesicles which form reciprocal synapses; 2) about 20% are innervated solely by adrenergic nerve fibres which form distinct synaptic contacts; and 3) the remaining 20% are innervated by both types of nerve fibres. It is proposed that the NEBs are receptors monitoring intrapulmonary P_{CO 2} and so leading to modulation of activity in afferent nerve fibres (type containing agranular vesicles). The presence of NEBs solely with an adrenergic (efferent) innervation poses a problem with this interpretation.     

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 anterior sensory neuropile in the metathoracic ganglion of the locust Locusta migratoria]

The study of serial sections of the metathoracic ganglion of Locusta migratoria showed that the fibres of the tympanal nerve terminate in the dorsal half of the anterior sensory neuropile (ASN). Three types of synaptic endings were found in the ASN. Endings type I contain dense-core vesicles 600-850 A in diameter, more or less uniformly distributed in the axoplasm. They do not form specialized contacts with postsynaptic fibres and are localized only in the most ventral part of the ASN. Endings type II contain clear round vesicles 400-450 A in diameter (rare 250-300 A) and form typical synapses with dense pre-and postsynaptic membranes and synaptic cleft 150-200 A. Four types of contacts formed by these endings with postsynaptic fibres were found: 1 : 1 synapses; convergent, divergent and serial. All of them are well presented in the auditory neuropile. Endings type III contain both dense-core and clear vesicles in different relation. Only clear vesicles of these endings are connected with the active sites of the membrane.     

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.     

Neural Organization of the Median Ocellus of the Dragonfly : II. Synaptic structure

Two types of presumed synaptic contacts have been recognized by electron microscopy in the synaptic plexus of the median ocellus of the dragonfly. The first type is characterized by an electron-opaque, button-like organelle in the presynaptic cytoplasm, surrounded by a cluster of synaptic vesicles. Two postsynaptic elements are associated with these junctions, which we have termed button synapses. The second synaptic type is characterized by a dense cluster of synaptic vesicles adjacent to the presumed presynaptic membrane. One postsynaptic element is observed at these junctions. The overwhelming majority of synapses seen in the plexus are button synapses. They are found most commonly in the receptor cell axons where they synaptically contact ocellar nerve dendrites and adjacent receptor cell axons. Button synapses are also seen in the ocellar nerve dendrites where they appear to make synapses back onto receptor axon terminals as well as onto adjacent ocellar nerve dendrites. Reciprocal and serial synaptic arrangements between receptor cell axon terminals, and between receptor cell axon terminals and ocellar nerve dendrites are occasionally seen. It is suggested that the lateral and feedback synapses in the median ocellus of the dragonfly play a role in enhancing transients in the postsynaptic responses.     

Ultrastructure of the nerve cells and fibres in the urinary bladder wall of the cat.

The nerve elements in the urinary bladder of the cat were studied by electron microscopy. According to their ultrastructure, nerve cell somata can be classified into three types: the large cells with a cytoplasm rich in organelles, several processes and numerous synaptic contacts on their surface; the cytoplasm contained 80- 120-nm granulated vesicles. The second type is poor in cytoplasmic organelles and has very few processes and virtually no synaptic contacts on the soma. The third type contains numerous large 160- to 220-nm 'neurosecretory' vesicles in the cytoplasm. According to the morphology of the vesicle population, four types of nerve processes could be distinguished: Type a, with a dominant population of small (40-60 nm) agranular vesicles. These are thought to be sacral parasympathetic fibres. Type b, with small (40-60 nm) granular vesicles, which may be the noradrenergic sympathetic fibres. Type c, with 80- to 120-nm granulated vesicles, probably of local origin. Typed d, with large 160- to 220-nm 'neurosecretory' vesicles also of local origin. Different types of nerve fibres are converging on the local nerve cells. This suggests that the local circuits can play an important role in coordinating the function of the bladder. Therefore, ganglia may be considered as an elementary functional unit.     

Ultrastructure of the neurohypophysis of the teleost Poecilia latipinna in relation to neural control of the adenohypophysial cells

Summary The structure of the neurohypophysis of Poecilia latipinna (green molly, sailfin molly) was studied with the electron microscope. Profile diameters of neurosecretory granules in the non-myelinated neurohypophysial nerve fibres were measured and mathematically corrected for error due to section thickness. Six different types of nerve fibres could be distinguished by statistical classification of their granules and by other ultrastructural features. One fibre-type (type B) contained granules with a mean diameter of 85 nm, and the other five types (types Ala, Alb, A2, A3 and A4) all contained granules with mean diameters greater than 100 nm. Synaptic contacts were observed between type B fibres and all the adenohypophysial cell-types, although in the case of the ACTH cells the synapses were separated from the cell membrane by a continuous double basement membrane. Type A fibres were observed to contact the cells of the proximal pars distalis and pars intermedia, but did not form synapses. However, synapses occurred between type A fibres and pituicytes, and between type A fibres and the pericapillary basement membrane in the interior of the neurohypophysis. The possible roles of the different types of nerve fibres in controlling the adenohypophysial cells are discussed in the context of evidence from other teleosts.We thank Mr. W.A. Thomson and Mr. D.I. Hollingworth for technical assistance, and Dr. D.I.C. Pearson (Department of Physics, University of Nancy, Nancy, France) for advice on mathematical analysis and computer programs. The work was carried out during the tenure of an S.R.C. Research Studentship by T.F.C.B.     

Synaptogenesis in the neural ganglia of the heart in the human embryo

Electron microscopy was used to study synapse development in the cardiac ganglia of human fetuses ranging from 8 to 27 weeks of ovulation time. Staining with ethanolic phosphotungstic acid was used for analysis of synaptic active zones. Specialization of interneuronal links begins with the appearance of electron dense material on plasmalemmas of nerve cells in the places of simple contacts. First synapses with single synaptic vesicles and short osmiophilic zones were found in cardiac ganglia in 8-week-old fetuses. Large granular vesicles and mitochondria vesicles are formed from cisternae of agranular endoplasmic reticulum in the preterminal parts of axons and moved by axoplasmic transport to the osmiophilic zones of future synapses. Axodendritic synapses appeared earlier in the cardiac ganglia than axosomatic ones, the latter were observed from the middle of gestation. Transient neuroglial synapse-like contacts were found in the cardiac ganglia. Staining with phosphotungstic acid made it possible to distinguish the degree of synapse maturation according to active synaptic zones. The peculiarities of synaptic development in cardiac ganglia in comparison with that in the central nervous system may be accounted for by different origins of the neural tube and of neural crest and by the level of their phylogenic development.     

Ultrastructural observations on the central innervation of the guinea-pig pineal gland

Summary In the present study the central innervation of the guinea-pig pineal gland was investigated. The habenulae and the pineal stalk contain myelinated and non-myelinated nerve fibres with few dense-cored and electron-lucent vesicles. Some myelinated fibres leave the main nerve fibre bundles, lose their myelin-sheaths and terminate in the pineal gland. Although direct proof is lacking, the non-myelinated fibres appear to end near the site where the bulk of the myelinated fibres are located. Here a neuropil area exists where synapses between non-myelinated fibre elements are abundant. Neurosecretory fibres were also seen. The results support the concept of functional interrelationships between hypothalamus, epithalamus and the pineal gland.     

The Ultrastructure of the Nervous System of Gyratrix hermaphroditus (Turbellaria, Rhabdocoela)

The peripheral nervous system and the synapses of G. hermaphroditus are studied with the electron microscope. There is a submuscular as well as a subepithelial plexus. The subepithelial plexus is found among the muscles and between the muscles and the basement membrane. It consists of fibres containing large lucent and lysosome-like vesicles and fibres with only small lucent (synaptic) vesicles. In the deeper lying submuscular plexus also dense and dense-cored vesicles occur in the fibres. Cell bodies are not observed in the plexuses. The separate nerve supplies of the pharynx and the gonads contain nerve cells of the neurosecretory type. Fibres of the same kinds as in the brain are also seen here. The synapses in the neuropile are of two kinds. 1. Symmetrical synapses with an additional presynaptic network are most common. 2. Synapses without thickenings of membranes are observed between lateral membranes of neurites. In the peripheral nervous system are two other kinds of synapses also observed. 1. Asymmertical synapses with a denser and wider postsynaptic thickening and 2. neuromuscular junctions. Neurites containing accumulations of small vesicles against the basement membrane are also described. The organization of the peripheral nervous system is described and discussed in relation to the systematic position of G. hermaphroditus.     

The neurohypophysis of the crested newt

Summary In the median eminence of the newt a medial region and two lateral regions are described.In cross section, the medial region appears to be made up of 1) an outer or glandular zone (Zone I) containing aldehyde-thionine-positive and negative nerve fibres and blood capillaries. Nerve fibres appear aligned in palisade array along the capillaries. 2) An inner zone (Zone II) made up of a) a layer of aldehyde-thionine-positive nerve fibres (fibrous layer) belonging to the preoptic hypophyseal tract and b) a layer of ependymal cells lining the infundibular lumen and reaching the blood vessels with their long processes.The lateral regions display a less pronounced stratification and aldehyde-thionine positive nerve fibres are nearly absent.A slender lamina (ependymal border) containing mainly aldehyde-thionine-positive nerve fibres and ependymal cells connects the median eminence to the pars nervosa.At the ultrastructural level, in the outer zone of the medial region at least 4 types of nerve fibres and nerve endings are identified:Type I nerve fibres containing granular vesicles of 700–1000 Å and clear vesicles (250–400 Å).Type II nerve fibres containing granular vesicles and polymorphous granules of 900–1300 Å and clear vesicles (250–400 Å).Type III nerve fibres containing dense granules of 1200–2000 Å and clear vesicles of 250–400 Å.Type IV nerve fibres containing only clear vesicles of 250–400 Å. In the inner zone too, all these nerve fiber types are found among ependymal cells, while the fibrous layer consists of nerve fibres containing granules of 1200–2000 Å in diameter.In the lateral regions Type I, Type II and Type IV nerve fibres and their respective perivascular terminals are found; axons containing dense granules (1200–2000 Å) are scanty. In these regions typical synapses between Type I nerve fibres and processes rich in microtubules are visible.The classification and functional significance of nerve fibres in the median eminence are still unsolved, but it may be assumed that nerve fibres of the medial region belong to both the preoptic hypophyseal and tubero hypophyseal tract, while the lateral regions are characterized by nerve fibres of the tubero hypophyseal tract. Peculiar specializations of the ependymal cells in the median eminence of the newt are also discussed.Work supported by a grant from the Consiglio Nazionale delle Ricerche.The authors are indebted to Mr. G. Gendusa and P. Balbi for technical assistance.     

Structure and innervation of the pineal gland of the rabbit,Oryctolagus cuniculus (L.)

In the rabbit pineal gland two types of postganglionic nerve endings were found which are characterized by the presence of small dense-core vesicles or small clear vesicles. Pharmacological and cytochemical experiments showed then to be noradrenergic and cholinergic, respectively. Both types were often present in the same nerve bundle, occasionally in close opposition. Intrapineal neurons were only rarely observed. They showed cholinergic synapses on their perikaryon and dendrites as well as noradrenergic axo-dendritic close contacts. Bilateral extirpation of the superior cervical ganglia revealed the postganglionic sympathetic origin of the pineal noradrenergic nerve fibres. Moreover, it appeared that these ganglia are hardly, if at all, involved in the pathway of pineal cholinergic innervation. The results obtained from lesions of both facial nerves, taken together with the results reported in the literature, led to the conclusion that the postganglionic cholinergic nerve fibers in the pineal are of parasympathetic origin. A model for the sympathetic and parasympathetic pineal innervation is proposed.     

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.     

Pattern of synaptic connections in the pineal organ of the ayu,Plecoglossus altivelis (Teleostei)

Summary Synaptic connections were studied by means of electron microscopy in the sensory pineal organ of the ayu, Plecoglossus altivelis, a highly photosensitive teleost species. Three types of specific contacts were observed in the pineal end-vesicle: 1) symmetrically organized gap junctions between the basal processes of adjacent photoreceptor cells; 2) sensory synapses endowed with synaptic ribbons, formed by basal processes of photoreceptor cells and dendrites of pineal neurons; 3) conventional synapses between pineal neurons, containing both clear and dense-core vesicles at the presynaptic site. Based on these findings, the following interpretations are given: (i) The gap junctions may be involved in an enhancement of electric communication and signal encoding between pineal photoreceptor cells. (ii) The sensory synapses transmit photic signals from the photoreceptor cells to pineal nerve cells. (iii) The conventional synapses are assumed to be involved in a lateral interaction and/or summation of information in the sensory pineal organ. A concept of synaptic relationships among the sensory and neuronal elements in the pineal organ of the ayu is presented.Fellow of the Alexander von Humboldt Foundation, Federal Republic of Germany     

The release sites and targets of nerve cells immunoreactive to RFamide — an ultrastructural study ofMicrostomum lineare andDiphyllobothrium dendriticum (Plathelminthes)

Summary A detailed study of the ultrastructure and RFamide immunoreactivity (RF-IR) in the flatwormsMicrostomum lineare andDiphyllobothrium dendriticum has been made with the immunogold technique. The present ultrastructural study confirms the localization of RF-IR cells observed by light microscopic immunocytochemistry. The RF-IR is demonstrated in small, electron-dense vesicles in neuronal cell bodies and processes. RF-IR is not detected in the rough endoplasmatic reticulum or the Golgi system of the nerve cells. The targets of RF positive fibres are nerve fibres, muscles and glands. Gold labelling occurs in classic synapses, which points to a role in neurotransmission. RF-IR is also observed in nerve terminals lacking the characteristics of synapses. These release sites occur close to muscle fibres in the intestine and body musculature ofM. lineare. Thus, an additional, paracrine action of the neuropeptide is suggested.