New synapses associated with the granule-containing cells of rat sympathetic ganglia

The synapses of the rat superior cervical sympathetic ganglion were studied with both conventional and ultrastructural histochemical methods. Besides the cholinergic synapses polarized from preganglionic fibers to sympathetic ganglion neurons, two morphologically and functionally different types of synapses were observed in relation to the small granule-containing (catecholamine-containing) cells of the rat superior cervical ganglion. The first type is an efferent adrenergic synapse polarized from granule-containing cells to the dendrites of the sympathetic ganglion neurons. This type of synapse might mediate the inhibitory effects (slow inhibitory postsynaptic potentials) induced by catecholamines on the sympathetic neurons. The second type is a reciprocal type of synapse between the granule-containing cells and the cholinergic preganglionic fibers. Through such synapses, these cells could exert a modulating effect on the excitatory preganglionic fibers. Therefore, we propose that these cells, through their multiple synaptic connections, exhibit a local modulatory feedback system in the rat sympathetic ganglia and may serve as interneurons between the preganglionic and postganglionic sympathetic neurons.     

A light and fluorescence cytochemical and electron microscopic study of granule-containing cells in the intrapulmonary ganglia of Pseudemys scripta elegans

In the lung of the red-eared turtle, large numbers of intramural ganglia located in the intraparenchymal connective tissue are demonstrated. Numerous cells in close proximity to the principal ganglionic neurons displayed a bright blue-white formaldehyde-induced fluorescence. Microspectrofluorometric analysis revealed the presence of dopamine (DA) in all cells measured. Subsequent light histochemical staining of the fluorescent sections showed the DA-containing cells to display argentaffinity. Electron microscopy of serial sections revealed cells characterized by dense-cored vesicles corresponding to the intensely formaldehyde-induced fluorescent cells. The argentaffin technique performed directly on ultrathin sections selectively stained the dense-cored vesicles. After fixation with glutaraldehyde followed by dichromate, x-ray microanalysis showed the chromium to be incorporated into the dense granules. Cholinergic-type nerve endings formed axosomatic synaptic contacts with the DA-containing cells, which can therefore be considered as intrinsic postganglionic elements. No efferent synapses from the granule-containing cells to the principal ganglionic neurons could be observed. The granule-containing cells occurred solitarily and in clusters, partially invested with satellite cells, and usually located near fenestrated capillaries; they displayed cytoplasmic processes and indicated emiocytotic granule release. Adjacent granule-containing cells were separated by spaces about 20 nm wide, gradually widening to form intercellular channels with apically projecting microvilli and primary cilia. It is concluded that the intrapulmonary granule-containing cells of the red-eared turtle belong to the APUD system. Furthermore, morphologically these cells appeared to possess a special sensory apparatus which designates them as paraneurons. The possible physiological significance of these intrapulmonary granule-containing cells is discussed.     

Synapses of the superior cervical ganglion of the white rat

In the complexe system of synapses in the white rat's superior cervical ganglion, authors show the existence of: 1 - somato-somatic neuronal synapses 2 - somatic synapses between S.I.F. cells 3 - recurrent auto-synapses They moreover mention widening of synaptic cleft of the synapses between S.I.F. cells and their processes.     

Decentralisation of neurones in the pelvic ganglion of the guinea-pig: Reinnervation by adrenergic nerves

An electron-microscopic study has been made of adrenergic and cholinergic nerve fibres and synapses in the pelvic ganglion of the guinea-pig at intervals of up to 60 days following section of the hypogastric and pelvic nerves. Transection of the hypogastric nerves led to degeneration of 80-90% of the cholinergic nerve profiles and synapses in the ganglion. The small number of adrenergic nerves and synapses did not change, but 30-60 days after section, this number increased 8-10 times. Transection of the pelvic nerves led to degeneration of about 15% of the cholinergic nerve terminals, but no change in adrenergic terminals. After transection of both hypogastric and pelvic nerves, only about 1% of cholinergic nerves survived, but after 30-60 days, the number of adrenergic nerves increased 8-10 times. It is concluded that following cholinergic nerve degeneration in the ganglion, adrenergic nerves, probably originating as collateral sprouts from postganglionic neurones and granule-containing cells, can replace them to some extent.     

The granule-containing cell somata in the superior cervical ganglion of the rat,as studied by a serial sampling method for electron microscopy

Summary The surface of 4 granule-containing cells, in a cluster within the rat superior cervical ganglion, was studied by a serial sampling technique for electron microscopy. The result shows that all the 4 cells receive one, or three afferent synaptic boutons from the preganglionic fibers impinging upon their somata, and a somatic efferent synapse exists at two locations on each soma of the 2 of these cells. The postsynaptic element of the efferent synapse is observed to be represented by non-vesiculated and vesiculated segments of dendrites, soma and a possible axon collateral of the adrenergic principal neuron of the ganglion. There is a remarkably constant development of the attachment plaque between the granule-containing cells themselves, representing 1.7–2.3% of surface area for each cell. The surface area exposed to the extracellular space (covered only by a basal lamina) varies from 0.1 to 2.3% of the total perikaryal surface of the 4 cells. A tendency is noted that those cells without efferent synapses possess a more extensive area exposed to extracellular space than those forming somatic efferent synapse to the postganglionic elements.It is a pleasure to acknowledge the advice and encouragement of Professor A. Yamauchi throughout this work. I thank Mr. K. Kumagai and Miss K. Tsushida for their technical assistance.     

A light and electron microscopic study on the embryonic development of the rat carotid body.

The embryonic development of the rat carotid body was studied with electron microscopy. In the 11 mm embryo a cell aggregation consisting of undifferentiated cells and unmyelinated nerve fibers appears on the anterior wall of the third branchial artery. Granule-containing cells appear in the 12 mm embryo and continue to increase in number as the cellular aggregation increases in size and becomes separated from the wall of the third branchial artery. Synapse formation and the appearance of fenestrated capillaries occur almost simultaneously at the 17 mm stage. There are two types of synapses, one with membrane densification and vesicles clustered inside the nerve endings, the other with dense material and vesicles inside the granule-containing cells. At the 20 mm stage the undifferentiated cells send enveloping cytoplasmic processes toward adjacent granule-containing cells and the carotid body anlage displays rudimentary lobules.     

Novel processes invaginate the pre-synaptic terminal of retinal bipolar cells

Mixed-rod cone bipolar (Mb) cells of goldfish retina have large synaptic terminals (10 mum in diameter) that make 60-90 ribbon synapses mostly onto amacrine cells and rarely onto ganglion cells and, in return, receive 300-400 synapses from gamma-aminobutyric acid (GABA)-ergic amacrine cells. Tissue viewed by electron microscopy revealed the presence of double-membrane-bound processes deep within Mb terminals. No membrane specializations were apparent on these invaginating processes, although rare vesicular fusion was observed. These invaginating dendrites were termed "InDents". Mb bipolar cells were identified by their immunoreactivity for protein kinase C. Double-label immunofluorescence with other cell-type-specific labels eliminated Müller cells, efferent fibers, other Mb bipolar cells, dopaminergic interplexiform cells, and somatostatin amacrine cells as a source of the InDents. Confocal analysis of double-labeled tissue clearly showed dendrites of GABA amacrine cells, backfilled ganglion cells, and dendrites containing PanNa immunoreactivity extending into and passing through Mb terminals. Nearly all Mb terminals showed evidence for the presence of InDents, indicating their common presence in goldfish retina. No PanNa immunoreactivity was found on GABA or ganglion cell InDents, suggesting that a subtype of glycine amacrine cell contained voltage-gated Na channels. Thus, potassium and calcium voltage-gated channels might be present on the InDents and on the Mb terminal membrane opposed to the InDents. In addition to synaptic signaling at ribbon and conventional synapses, Mb bipolar cells may exchange information with InDents by an alternative signaling mechanism.     

Ultrastructural evidence for two-cell and three-cell neural pathways in the tentacle epidermis of the sea anemone Aiptasia pallida.

Sensory and ganglion cells in the tentacle epidermis of the sea anemone Aiptasia pallida were traced in serial transmission electron micrographs to their synaptic contacts on other cells. Sensory cell synapses were found on spirocytes, muscle cells, and ganglion cells. Ganglion cells, in turn, synapsed on sensory cells, spirocytes, muscle cells, and other neurons and formed en passant axo-axonal synapses. Axonal synapses on nematocytes and gland cells were not traced to their cells of origin, i.e., identified sensory or ganglion cells. Direct synaptic contacts of sensory cells with spirocytes and sensory cells with muscle cells suggest a local two-cell pathway for spirocyst discharge and muscle cell contraction, whereas interjection of a ganglion cell between the sensory and effector cells creates a local three-cell pathway. The network of ganglion cells and their processes allows for a through-conduction system that is interconnected by chemical synapses. Although the sea anemone nervous system is more complex than that of Hydra, it has similar two-cell and three-cell effector pathways that may function in local responses to tentacle contact with food.     

Morphological characteristics of chemosensory,visual, and statocyst pathways in the snailHelix lucorum

The following structural characteristics of the chemosensory, visual, and vestibular pathways of the snail (Helix lucorum) were demonstrated by using a variety of histological techniques. Large and small neurons of the tentacle ganglion, the bipolar cells of the olfactory nerve, and a proportion of optic tentacle bulb chemoreceptors within the olfactory nerve all send their processes to the CNS of the mollusk. Here they are divided up into numerous bundles of fibers in the neuropil of the ipsilateral cerebral ganglion. They are joined by processes from the central nervous system put out by all neurons of the protocerebrum and the cluster of cells of the commissural section of the metacerebrum. Ocular receptors do not send processes down below the enlargement of the upper optic nerve. This enlargement is also the site where processes from cells within the CNS and the nerve itself terminate. An area of arborization of processes from the visual pathway cells is located in the neuropil of the pleural portion of the metacerebrum. Hair cells of statocysts put out processes to the cerebral ganglion, whence axons of small metacerebral neurons extend towards the organ of balance. Some processes from vestibular pathway cells form an arborization zone at the ipsilateral cerebral ganglion, while others pass through the cerebral commissure to form their area of arborization in the contralateral ganglion. Processes from vestibular and visual pathway cells arborize in exactly the same area.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 18, No. 1, pp. 7–16, January–February, 1986.     

The major ganglion in the pelvic plexus of the male rat

Summary To further evaluate the role of autonomic ganglia in the regulation of pelvic visceral activity, the neural elements in the major pelvic ganglion of the male rat have been studied with histochemical and electron microscopic techniques. The principal findings are that the ganglion is composed of cholinergic and adrenergic ganglion cells as well as small intensely fluorescent (SIF) cells. Polarity in the ganglion is indicated by clustering of small ganglion cells which stain intensely for acetylcholinesterase (AChE) along the pelvic nerve while larger cells, with weak to moderate AChE activity, collect near small branches of the hypogastric nerve. Some cholinergic ganglion cells are enclosed by a plexus of adrenergic terminals. SIF cells appear to be in contact with both cholinergic and adrenergic cells, although many of the fluorescent beads around adrenergic neurons may be short dendrites of ganglion cells, rather than processes of SIF cells. Two types of SIF cells may be distinguished on the basis of size and morphology of their granulated vesicles. Afferent synapses of the cholinergic type were common on SIF cells of the large granule and small granule type. Portions of SIF cells with large granules occur within the capsule of ganglion cells. Contacts seen here were interpreted as efferent synapses from SIF cells to the dendrites of ganglion cells.     

Electron microscopy of the paracervical (Frankenhäuser) ganglion of the adult rat

Summary The ultrastructure of the paracervical (Frankenhäuser) ganglion in the rat was studied after immersion or perfusion fixation with glutaraldehyde followed by post-osmification. This ganglion is located at the uterovaginal junction in the vicinity of arteria uterina and contains three neuronal cell types. (1) Principal neurons have a fine structure mainly similar to the ganglion cells of other autonomic ganglia. (2) Small granule-containing cells occur in clusters often close to fenestrated capillaries. They are divided into two subgroups according to the size of their cytoplasmic granules; those containing only small granulated vesicles of 800 to 1400 Å in diameter and those having also large granulated vesicles of 2000 to 3000 Å in diameter. (3) Vacuolated nerve cells are large cells that resemble the principal neurons in their cytoplasmic components, except that they contain one to ten vacuoles with corpuscles of different size and shape. The possible physiological significance of the small, granule-containing cells in the uterine function is discussed.     

Ultrastructure of sympathetic ganglion cells and granule-containing cells in the paracervical (Frankenhäuser) ganglion of the newborn rat

Summary A study was made of the ultrastructure of the paracervical (Frankenhäuser) ganglion of the newborn rat, using immersion fixation by glutaraldehyde (2.5%) followed by OsO₄ (1%), or KMnO₄ (3%) fixation. The cells containing dense—core vesicles were divided into three groups: (1) primitive sympathetic cells, (2) cells containing some dense-core vesicles 700–1100 Å in size and structurally resembling sympathetic neurons, called principal neurons, and (3) cells containing many dense-core vesicles with a larger, darker dense core, 800–2000 Å in diameter, called granule-containing cells. Using glutaraldehyde-osmium fixation, the principal neurons were further divided into dark and light cells on the basis of electron opacity of the cytoplasmic matrix. The granule-containing cells were believed to correspond to the small, intensely fluorescent cells (SIF-cells) previously described using the formaldehyde-induced fluorescence technique. On the basis of the amount of granules, the granulecontaining cells were classified as mature or maturing SIF-cells and as more primitive SIF-cells, and developing sympathicoblasts. The development of synapses in autonomic ganglia was discussed.Grant: The Finnish Medical Foundation.     

Growth and synapse formation among major classes of adult salamander retinal neurons in vitro

Adult neurons, isolated from the salamander retina, were maintained in low-density cell culture and examined for synapse formation by electrophysiological and electron microscopic techniques. Morphologically identifiable rod, cone, horizontal, bipolar, and amacrine/ganglion cells survived for many months, grew processes, and formed numerous cell contacts. Intracellular recordings showed the presence of a variety of voltage- and time-dependent conductances and both electrical and chemical transmission among these cells. At the ultrastructural level, gap junctions, monad ribbon synapses, and conventional synapses, like those present in the intact retina, were observed in sibling cultures. Thus, all major classes of adult retinal neurons, in addition to ganglion cells, are able to regenerate processes and reform synapses. The regenerated synaptic contacts are functional and structurally diverse.     

Electrophysiological properties of rat nodose ganglion neurons co-transplanted with carotid bodies into the chick chorioallantoic membrane

The electrophysiological properties of nodose ganglion neurons were evaluated immediately after removing nodose ganglia from young adult rats and 3 to 10 days after nodose ganglia implantation -either alone or co-implanted with carotid bodies- onto the chick chorioallantoic membrane. Implanted and co-implanted nodose neurons were less excitable than acutely recorded nodose neurons. Co-implanted neurons also showed reduced amplitudes for both action potentials and spike after-hyperpolarizations relative to those found in. acutely recorded nodose ganglion neurons and a smaller time constant (T) than that found in implanted neurons. In addition, no spontaneous activity was recorded from nodose ganglion neurons co-implanted with carotid bodies during 3-9 days, which suggests that functional synapses between carotid glomus cells and nodose neurons were not yet established. Results indicate the feasibility of obtaining viable nodose neurons for up to 10 days grafted onto the chick chorioallantoic membrane, where they can conserve most of their passive and active membrane properties and also are susceptible to carotid bodies trophic influences. They also suggest that nodose neurons would need more time for the development of functional synapses when grafted with carotid body glomus cells.     

Ultrastructural study of small granule containing (SGC) cells in the lower mesenteric ganglion in drug treated guinea-pigs

Small granule containing cells (SGC or SIF cells) in the lower mesenteric ganglion of the guinea-pig were studied in control and experimental animals. After reserpine administration a decrease of dense population of granular vesicles was found. After injection of nialamide and dopamine to the animals in SGC cells significantly increased the number and density of granular vesicles. No real efferent synapses were found on SGC cell processes, although sites with accumulated synaptic-like vesicles, mostly with a dense core, were often observed.     

Studies of cardiac ganglia in pre- and postnatal rabbits

Summary This investigation was undertaken to describe the ultrastructure of cardiac ganglia in rabbits from day 18 of gestation to day 35 postpartum. Special attention was directed to the types of synaptic contacts made with the principal neurons and with the small granule-containing cells. The cardiac ganglia in all animals consisted mainly of parasympathetic postganglionic neurons, supporting cells, and small granule-containing (small intensely fluorescent) cells. The neurons received afferent synaptic terminals of two types. One type contained mainly small clear vesicles typical of most cholinergic terminals. The second type contained mainly small dense-core vesicles (these were most prominent after treatment of the animal with 5-hydroxydopamine), and were considered to be adrenergic terminals. These adrenergic terminals are probably part of an inhibitory system in the ganglia. The small granule-containing cells received typical afferent synaptic terminals of the cholinergic type, and also formed specialized contacts with certain axonal terminals. These latter specializations are considered to be reciprocal synapses which probably have a role in modulating ganglionic transmission.Supported by the Kentucky Heart Association and the Heart Association of Louisville and Jefferson County     

A dividing granule-containing cell in the pelvic ganglion of the guinea-pig

Summary A dividing granule-containing cell is described in the pelvic ganglion of the guinea-pig two days after pelvic nerve section. This appears to be the first report of a dividing granule-containing cell in adult tissue.     

Synaptic relationships in the plexiform layers of carp retina

Summary The synaptic contacts made by carp retinal neurons were studied with electron microscopic techniques. Three kinds of contacts are described: (1) a conventional synapse in which an accumulation of agranular vesicles is found on the presynaptic side along with membrane densification of both pre- and postsynaptic elements; (2) a ribbon synapse in which a presynaptic ribbon surrounded by a halo of agranular vesicles faces two postsynaptic elements; and (3) close apposition of plasma membranes without any vesicle accumulation or membrane densification.In the external plexiform layer, conventional synapses between horizontal cells are described. Horizontal cells possess dense-core vesicles about 1,000 Å in diameter. Membranes of adjacent horizontal cells of the same type (external, intermediate or internal) are found closely apposed over broad regions.In the inner plexiform layer ribbon synapses occur only in bipolar cell terminals. The postsynaptic elements opposite the ribbon may be two amacrine processes or one amacrine process and one ganglion cell dendrite. Amacrine processes make conventional synaptic contacts onto bipolar terminals, other amacrine processes, amacrine cell bodies, ganglion cell dendrites and bodies. Amacrine cells possess dense-core vesicles. Ganglion cells are never presynaptic elements. Serial synapses between amacrine processes and reciprocal synapses between amacrine processes and bipolar terminals are described. The inner plexiform layer contains a large number of myelinated fibers which terminate near the layer of amacrine cells.This work was supported by an N.I.H. grant NB 05404-05 and a Fight for Sight grant G-396 to P.W. and N.I.H. grant NB 05336 to J.E.D. The authors wish to thank Mrs. P. Sheppard and Miss B. Hecker for able technical assistance. P.W. is grateful to Dr. G. K. Smelser, Department of Ophthalmology, Columbia University, for the use of his electron microscope facilities.     

Fine structure of cardiac ganglion trunk in prawn, Penaeus japonicus bates.

The cardiac ganglion trunk of prawn, Peneaus japonicus Bates, is on the middle line of ventral wall of the cardiac tube and consists of nine ganglion cells, many nerve fibers and neuropils. These neuronal elements are insulated by supporting cells and connective tissue fibers. The peripheral area of the perikaryal cytoplasm of the ganglion cell is separated into many compartments by deep invaginations of the cell membrane. Each compartment is packed with a tight network of the agranular endoplasmic reticulum. Among nerve fiber bundles are many small areas of neuropil. Most of the synapses in the ganglion trunk are observed in the neuropil, but there are a few nerve terminals which form synapses with the somata of ganglion cells.     

Fate of ganglionic synapses and ganglion cell axons during normal and induced cell death

In order to understand the significance of cell death in the formation of neural circuits, it is necessary to determine whether before cell death neurons have (a) sent axons to the periphery; (b) reached the proper target organs; and (c) have established synaptic connections with them. Axon counts demonstrated that, after sending out initial axons, ciliary cells sprouted numerous collaterals at the time of peripheral synapse formation. Subsequently, large numbers of axons were lost from the nerves, slightly later than the onset of ganglion cell death. A secondary loss of collaterals later occurred unaccompanied by cell death. Measurements of conduction velocity and axon diameters indicated that all ganglion cell axons grew down the proper pathways from the start, but it was not possible to determine whether all axons had actually formed proper synapses. This was ascertained, however, in the ganglion itself where preganglionic fibres were shown to synapse selectively with all ganglion cells before cell death. During this period, degenerating preganglionic synapses were observed on normal cells. It can therefore be inferred that at least some preganglionics established proper synapses before dying and that a single synapse is not sufficient to prevent cell death. In this system neither preganglionic nor ganglionic cell death seems designed to remove improper connections but rather to remove cells that have not competed effectively for a sufficient number of synapses, resulting in a quantitative matching up of neuron numbers.