Neurons and their interconnections in the frontal cortex of monkeys

It was shown by the Golgi and Golgi-Kopsch method that pyramidal cells of layers II–IV in the frontal cortex of the monkeyMacaca rhesus have numeruous, mainly recurrent axon collaterals by means of which they form vertical connections. Pyramidal cells with ascending axons are found. Axons of stellate basket neurons unite pyramidal cells in both horizontal (modules) and vertical (micromodules) directions; depending on the direction of the axon collaterals, two groups of stellate neurons can be distinguished. Groups of 14 to 16 pyramidal cells whose apical dendrites are connected into bundles were found. Axons of pyramidal cells in layers II–IV descend in the composition of the pyramidal tract and give off collaterals which run toward the bodies and dendrites of neighboring pyramidal cells, united into the same group, forming terminal and en passant junctions. Besides bundles, special kinds of "local" cell groups with U-shaped axons are found.A. A. Zhdanov Leningrad State University. Translated from Neirofiziologiya, Vol. 15, No. 2, pp. 115–120, March–April, 1983.     

Cell contacts and smooth muscle bundle formation in tissue transplants into the anterior eye chamber

Summary Segments of the taenia coli from guinea-pig were transplanted into the anterior chamber of the eye. Depending on such factors as the total volume of the transplant and the presence or absence of ganglion cells degeneration was either very extensive (90% or more of the total number of muscle cells) or localized (alternating regions of degenerating and normal structure). During days 1–2 muscle cells lost their plasma membranes so that their cytoplasmic contents were dispersed into the intercellular spaces. Many cells produced numerous small processes which were pinched off and dispersed in a similar manner. Following a period of intense mitotic activity (3–8 days) numerous cells with the characteristics of embryonic smooth muscle cells were evident. Within 10–14 days these differentiating cells produced bulbous protrusions and assumed more irregular outlines than at 3–8 days. The protrusions formed close contacts (50–100Å intercellular space) and tight junctions between adjacent muscle cells. Aggregation of muscle cells into bundles was under way between 14–28 days. At approximately 4–6 weeks these developing muscle groups were invaded by nerve fiber bundles. The pattern of the innervation and the form and size of the muscle bundles simulated the normal. These findings are discussed in relation to the possible functions of the intercellular contacts and cellular protrusions which characterise various periods of regeneration.This work was supported by the Australian Research Grants Committee. The transplants were carried out by Dr. T. Malmfors assistant, Miss Ulla Enberg.     

Neurons and their synaptic organization in the visual cortex of the rat

Summary Cells in the visual cortex (area 17) of adult rats were impregnated by the rapid Golgi method and characterized by light microscopy. Selected cells were then sectioned for electron microscopy and their cytological characteristics and the pattern of synapses on their cell bodies and dendrites were studied Twelve classical pyramidal cells from layers II–VI, two pyramid-like cells from layer VI, two inverted pyramidal cells from layers V and VI, ten spine-free non-pyramidal cells from layers II–VI and two spinous non-pyramidal cells from layer IV were examined.The cytoplasmic features of the identified cells, where these could be discerned, corresponded to those previously reported for the different cell types in conventionally prepared tissue. Pyramidal Cells received exclusively type 2 synaptic contacts on their cell bodies, type 1 contacts on their dendritic spines and a mixture of synaptic types (type II predominating) on their shafts, where synaptic density was relatively low. This pattern of synaptic contacts was consistent for all portions of the dendritic tree; inverted pyramidal cells and pyramid-like cells showed the same synaptic organization as classical pyramids. The axon collaterals of pyramidal cells established type I contacts with dendritic spines (or, rarely, shafts) of unknown origin. Non-Pyramidal Cells received both type 1 and type 2 contacts (the former predominating) on their cell bodies and dendrites. The spinous variety also received type I contacts on their dendritic spines. Axon terminal of spine-free non-pyramidal cells established type II synaptic contacts with dendritic shafts of unknown origin. The similarity in synaptic organization between the spine-free and spinous non-pyramidal cells examined in this study suggest that the latter correspond to the sparsely spinous stellate cells rather than to the spinous stellate cells of cat and monkey visual cortex.We thank the Medical Research Council for financial support     

Nerve-purkinje fiber relationship in the moderator band of bovine and caprine heart

Summary The moderator band in the heart of the ox and goat contains bundles of Purkinje fibers and nerve fibers separated by connective tissue. The axons are mostly unmyelinated and embedded in the cytoplasm of Schwann cells.Small bundles of axons run close to the Purkinje fibers. The axons dilate into varicosities 0.5 to 1.6 in diameter (mean 0.95 ), containing three types of vesicles: 1) agranular vesicles with a diameter of 400–500 Å, 2) large dense-cored vesicles with a diameter of 800–1200 Å, 3) small dense-cored vesicles with a diameter of 500 Å. Most varicosities contain agranular vesicles together with a few large dense-cored vesicles.The gap between the varicosities and the nearest Purkinje fiber is unusually wide and normally varies between 0.3 and 0.8 . No intimate nerve-Purkinje fiber contacts, with a cleft of 200 Å, were observed.     

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.     

Three-dimensional organization of the collagen fibrils in the rat sciatic nerve as revealed by transmission- and scanning electron microscopy

Summary The organization of collagen fibrils in the rat sciatic nerve was studied by scanning electron microscopy after digestion of cellular elements by sodium hydroxide treatment, and by conventional transmission electron microscopy. The epineurium consisted mainly of thick bundles of collagen fibrils measuring about 10–20 m in width; they were wavy and ran slightly obliquely to the nerve axis. Between these collagen bundles, a very coarse meshwork of randomly oriented collagen fibrils was present. In the perineurium, collagen fibrils occupied the interspaces between the concentrically arranged perineurial cells; in each interspace, they formed a sheet of characteristic lacework elaborately interwoven by thin (about 3 m or less in width) bundles of collagen fibrils. In the subperineurial region, there was a distinct sheet of densely woven collagen fibrils between the perineurium and underlying endoneurial fibroblasts. In the endoneurium, collagen fibrils surrounded individual nerve fibers in two layers as scaffolds: the inner layer was made up of a delicate meshwork of very fine collagen fibrils, and the outer one consisted of longitudinally oriented bundles of about 1–3 m in width. The collagen fibril arrangement described above may protect the nerve fibers against external forces.     

Ultrastructural evidence for the presence of actin filaments in mouse eggs at fertilization

Summary Mouse eggs at fertilization were permeated with glycerol solutions and then reacted with heavy meromyosin to show actin filaments by electron microscopy. The meiotic area of the egg surface is devoid of microvilli and is supported by a thick layer (0.6–0.8 m in width) of submembranous filaments. A much thinner layer (less than 0.3 m) is present in the remaining non-meiotic microvillous area and underlying its membrane is a very thick layer of cross-filaments and filament bundles.     

Morphology of the Respiratory Trees in the Holothurians Apostichopus japonicus and Cucumaria japonica

The morphology of the respiratory trees in the holothurians Apostichopus japonicus and Cucumaria japonica was studied using histochemical and electron microscopic techniques. The epithelium of the respiratory tree cavity in A. japonicus consists of columnar cells about 17–20 m high. In C. japonica, this epithelium is composed of two cell types: bulbous cells embedded in the connective tissue layer and secretory cells; cells are 1–12 m high. A characteristic feature of cells of the respiratory tree cavity epithelium in these species is the presence of numerous phagosomes and coated vesicles in the apical cytoplasm. The cell surface has many microvilli and a single cilium. Cells of the coelomic epithelium contain vacuoles in the apical part and myofibrils in the basal region; the thickness of this epithelium in A. japonicus and C. japonica is 9–16 and 10–30 m, respectively. Based on the different structure of the respiratory trees and the absence of hemocytes in A. japonicus, it is suggested that the holothurian species studied have different routes of oxygen transport from the environment to the internal organs.     

The fine structure of smooth muscle cells and their relationship to connective tissue in the rabbit portal vein

Summary The smooth muscle of rabbit portal vein was studied by electron microscopy with particular emphasis on the mechanical linkage between the muscle cells and on the distribution of connective tissue.The media of this vein is composed of inner circular and outer longitudinal muscle layers which are orientated almost perpendicularly to each other. The muscle of the inner circular layer shows very irregular contours with much branching and anastomosing of the cytoplasmic processes, which often make membrane contacts with neighbouring cells to form an extensive network of cytoplasmic processes. The muscle cells of the outer longitudinal layer are arranged in densely packed bundles and are spindle-shaped, with no branching processes. Opposing dense areas from neighbouring cells, with variable gap distances (30–100 nm) and close membrane contacts (intermediate junctions) with a gap of 11 nm were observed in both circular and longitudinal muscle layers.In the terminal regions of muscle cells in both circular and longitudinal layers a specialized anchoring structure was present which was closely related to extracellular elastic tissue. Muscle cells in the longitudinal layer showed the most elaborate structure, the tapering end of the muscle cell showing a honeycomb-like structure penetrated by columns of connective tissue compounds. The functional implications of these structures are discussed.     

Electron microscopic observations on the renal caliceal wall in the rat

Summary The fine structure of the rat caliceal wall at its attachment to the renal parenchyma is described. Particular attention is paid to the smooth muscle cells and their associated nerves. A single overlapping layer of epithelial cells lines the renal papilla which changes abruptly to a layer of 3–5 cells where the calix gains attachment to the renal substance. In this region there is an associated increase in the underlying connective tissue which contains smooth muscle cells. These cells possess filaments, are surrounded by a basal lamina, and occur scattered among large bundles of collagen fibres. The muscle cells possess numerous branching processes as well as shorter projections which make close contacts with adjacent cells. Large numbers of axons and their associated Schwann cells are also observed in this region. The axons possess swellings, some of which lie within 800 Å of smooth muscle cells, and contain large and small granulated vesicles and agranular vesicles. They are therefore considered to be adrenergic effectors.Further out in the caliceal wall typical spindle-shaped smooth muscle cells are observed lying parallel to one another to form closely packes bundles and are associated with relatively few nerves.The significance of these observations is discussed.     

Structure of cells and nerve endings in abdominal vagal paraganglia of the rat

Summary The abdominal vagal paraganglia of the rat consist of small groups of cells, interspersed by blood vessels and nerve bundles and lying close to, or within, the vagus nerve or its branches. Each cell group consists of 2–10 Type I cells incompletely invested by 1–3 satellite cells. Type I cells are characterised by the presence of numerous dense-cored vesicles in their cytoplasm and may exhibit synaptic-like contact with each other.Small efferent nerve endings make synaptic contacts with Type I cells. Larger cup-shaped afferent nerve endings also make synaptic contacts of two kinds with Type I cells. Nerve-nerve synapses are often seen within or close to paraganglia.Attention is drawn to the close similarity of fine structure of abdominal vagal paraganglia, carotid body and small intensely fluorescent cells of the superior cervical ganglion in rats. Possible functional implications of this morphological similarity are discussed.     

Electron microscopy of neurosecretory nerve fibres in the neural lobe of the embryonic mouse

Summary Nerve fibres of the neurosecretory hypothalamo-hypophyseal tract were studied in embryonic C3H mouse neural lobes; at least four glands at each gestational day 15–19 were examined.Single axons and small bundles of fibres are visible at gestational days 15 and 16. By day 17 large fibre bundles penetrate between glial cells. They increase in number during the next two days.Electron-lucent and electron-dense vesicles are seen in the fibres of the 15th and 16th gestational days. In the 17–19 day-old embryos development is characterized by a successive rise in the number of the two types of vesicles. The mean diameter of the electron-lucent vesicles is approximately unchanged in all the stages examined (50 nm). The electron-dense vesicles increase in size from approximately 80–90 nm at days 15–16 to 140 nm at the 19th gestational day.By day 19 contacts between neurosecretory fibre terminals and the outer basement membrane of internal and peripheral capillaries are occasionally observed. The possibly adrenergic nature of a few terminals contacting peripheral vascular structures in 17 and 18 day-old embryos is suggested.This investigation was supported by grant No. 2180-020 from the Swedish Natural Science Research Council. The skilful technical assistance of Mrs. Ulla Wennerberg is gratefully acknowledged.     

Electron microscopic study of the intercellular junctions during the ciliary epithelial differentiation of the eye of the common frog]

Intercellular junctions in the eye ciliary epithelium were studied in Rana temporaria by means of transmission electron microscopy beginning from the stage of appearance of the light sensitivity in the larval eye and until the completion of metamorphosis. In the ciliary epithelium inner layer the apical parts of adjacent cells are tied by contacts forming the junction complex already at a very early developmental stage. The most apical position in this complex is occupied by the focal junction which appears to be an early stage of zonula occludens; this complex includes also zonula adherens and macula adherens which may follow in any succession focal junction or zonula occludens. No definite order was found in the localization of cell contacts between the outer and inner layers of ciliary epithelium, as well as between the side surfaces of the cells within each layer. Contacts were found everywhere termed as "lengthy" which may be considered as gap junctions. Regional differences were found in the ultrastructure of the ciliary epithelium cells with respect to unequal distribution of functional loads.     

Cellular and paracellular pathway resistances in the “tight” Cl−-secreting epithelium of rabbit cornea

Summary The high transverse resistance of the isolated rabbit cornea (6–12 k·cm²) is associated with the corneal epithelium, a Cl^–-secreting tissue which is modulated by -adrenergic and serotonergic receptors. Three methods were employed to determine the resistances for the apical membrane, basolateral membrane, and paracellular conductive pathways in the epithelium. In the first method, the specific resistance of the apical membrane was selectively and reversibly changed. Epinephrine was used to increase apical Cl^– conductance and Ag⁺ was used to increase apical cation permeability. The second method utilized a direct measure of the spontaneous cellular ionic current. The third method obtained estimates of shunt resistance using transepithelial electrophysiological responses to changes in apical membrane resistance. The results of the first method were largely independent of the agent used. In addition, the three methods were in general agreement, and the ranges of mean values for apical membrane, basolateral membrane, and shunt resistances were 23–33, 3–4, and 12–16 k·cm², respectively, for the normal cornea. The apical membrane was the major, physiologically-modulated barrier to ion permeation. The shunt resistance of the corneal epithelium was comparable to that found previously for other tight epithelia. Experiments using Ag⁺ in tissues that were bathed in Cl^– and HCO₃-free solutions indicated that under resting conditions the apical membrane is anion-selective.     

Barrier membranes at the outer surface of the brain of an elasmobranch,Raja erinacea

Summary This report gives the results of the first electron-microscopic examination of the cell layers covering the outer brain surface and the inner surface of the cartilaginous skull in the skate, Raja erinacea. The perivascular glial blood-brain barrier — a characteristic of elasmobranchs — extends to the outer surface of the brain. This outer barrier layer is surrounded, in turn, by a subarachnoid compartment (depth: 30–40 m), containing loose connective tissue and blood vessels; by an arachnoid-like epithelium (10–15 cell layers), impermeable to horseradish peroxidase; and, by perimeningeal fluid, a fluid with a slow turnover rate and a protein composition different from plasma. The inside of the skull, facing the perimeningeal fluid, is covered by a multilayered (10–15 layers) cuboidal epithelium, also impermeable to horseradish peroxidase. Closely apposed cells in the luminal layer of this epithelium have apical microvilli and numerous vesicular profiles, containing material of moderate electron density. These observations may explain, in terms of structure, the regulated protein content of perimeningeal fluid and the restricted exchange of solutes between brain and perimeningeal fluid in elasmobranchs.     

Role of hippocampal neurons in theta-wave generation

Temporal relations between hippocampal unit activity and phases of the theta-waves were studied in unanesthetized rabbits immobilized with tubocurarine. When spontaneous thetarhythm potentials were recorded layer by layer from the dorsal hippocampus, a change in their polarity was observed 0.15–0.2 mm deeper than the pyramidal layer (i.e., in the radial layer). Most hippocampal neurons fired synchronously with the extracellular thetawaves. The numerous pyramidal cells, in which the spontaneous activity was inhibited during stimulation of the contralateral hippocampus and sciatic nerve and a hyperpolarization potential developed, were excited mainly during the positive phase of the theta-waves. Intracellular recording showed that their membrane potential decreased during the positive phase and increased during the negative phase. The less numerous basket cells, whose response to stimulation of the contralateral hippocampus consisted of a short high-frequency volley of spikes, and whose response to sciatic nerve stimulation was marked by a prolonged increase of frequency of the spontaneous discharges, were activated mainly during the negative phase of the theta-waves. It is concluded from these findings that inhibitory mechanisms play a role in the theta-activity of the hippocampus. It is postulated that the extracellular theta-waves are integral EPSPs of the basal dendrites and of the proximal segments of the apical dendrites, and that IPSPs of the pyramidal cell bodies play the principal role in the generation of the intracellular theta-rhythm.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 4, No. 5, pp. 531–539, September–October, 1972.     

Fine structure study on the development of trabeculae in the siphonous green algaCaulerpa simpliciuscula C. Ag.

Summary Bundles of fibrils and tubular structures were found to be associated with growing trabeculae ofCaulerpa simpliciuscula. In the rhizome tips, the bundles had an average diameter of 0.1 to 0.3 m, and a length greater than 10 m. The fibrils in the bundles were oriented in a strictly parallel fashion, with an individual thickness of 3–8 nm. The development of trabeculae started with the apposition of material of low electron density onto the bundles, which in this way became the inner skeleton of the trabeculae.Although fibre bundles with the same internal structure also occurred in the frond tip, these rarely contributed to trabecula formation. In the frond tips a different type of bundle with paracrystalline structure was found associated with the trabecular surface, forming a temporary connection between adjacent trabeculae. Permanent connection was achieved by deposition of further layers of trabecular material. These bundles in the frond tip consisted of two layers of tubular elements with a wall thickness of 80 Å and an inner diameter of 20–25 nm.Both fibre bundles and tubular bundles appear to contribute to trabecula formation. The similarity of these structures to the vacuolar inclusions observed in other siphonous algae is discussed.     

Wall texture in the spore of a vesicular-arbuscular mycorrhizal fungus

Summary InGlomus epigaeum Daniels and Trappe, a vesicular-arbuscular mycorrhizal fungus, the mature spore has a complex multi-layered wall containing a regular pattern of wall subunits.The outer wall (2–4 m thick) consists of a simple layer of parallel microfibrils. The inner wall (5–6 m thick) is built from two layers possessing different organization. The innermost layer, near the plasmalemma has a texture of apparently dispersed fibrils, whereas the second layer is regularly organized with an arced texture. Ten to twelve bundles of fibrils connected by apparently bow-shaped fibrils are consistently observed. The appearance of this arced organization depends on the section plane and on the angle of observation in the electron microscope as confirmed by tilting experiments. Wall subunits are evident as straight electron transparent fibrils; particularly well-defined in negatively stained frozen sections: their diameter is about 3.5nm.The regular pattern of wall subunits in this fungal cell wall is compared with the textures shown by cellulose fibrils in algae or higher plants and by chitin fibrils in arthropod cuticle.Research work supported by CNR, Italy. Special grant I.P.R.A.—Sub-project 1. Paper No. 55.     

The postnatal development of lamina V pyramidal cells in the sensomotor cortex of the albino rat. 2. Quantitative studies on Golgi preparations]

Using Golgi-Cox preparates the postnatal development (1st to 90th day after birth) of the patterns of apical dendrite spines in layer V pyramids in albino rat sensorimotor cortex is represented. Especially it is reported about the evolution of the total amount of apical dendrite spines, of length of apical dendrites, of spine density (spine dendrite quotients, DQ), of interspinous distance (space between two adjacent visible spines, in the proximal dendrite with increasing spine distribution and in the district with crest of spine distribution separately) and finally of cell body length.     

Vascular Endothelial-Cadherin Stabilizes at Cell–Cell Junctions by Anchoring to Circumferential Actin Bundles through α- and β-Catenins in Cyclic AMP-Epac-Rap1 Signal-activated Endothelial Cells

Vascular endothelial (VE)-cadherin is a cell–cell adhesion molecule involved in endothelial barrier functions. Previously, we reported that cAMP-Epac-Rap1 signal enhances VE-cadherin–dependent cell adhesion. Here, we further scrutinized how cAMP-Epac-Rap1 pathway promotes stabilization of VE-cadherin at the cell–cell contacts. Forskolin induced circumferential actin bundling and accumulation of VE-cadherin fused with green fluorescence protein (VEC-GFP) on the bundled actin filaments. Fluorescence recovery after photobleaching (FRAP) analyses using VEC-GFP revealed that forskolin stabilizes VE-cadherin at cell–cell contacts. These effects of forskolin were mimicked by an activator for Epac but not by that for protein kinase A. Forskolin-induced both accumulation and stabilization of junctional VEC-GFP was impeded by latrunculin A. VE-cadherin, α-catenin, and β-catenin were dispensable for forskolin-induced circumferential actin bundling, indicating that homophilic VE-cadherin association is not the trigger of actin bundling. Requirement of α- and β-catenins for forskolin-induced stabilization of VE-cadherin on the actin bundles was confirmed by FRAP analyses using VEC-GFP mutants, supporting the classical model that α-catenin could potentially link the bundled actin to cadherin. Collectively, circumferential actin bundle formation and subsequent linkage between actin bundles and VE-cadherin through α- and β-catenins are important for the stabilization of VE-cadherin at the cell–cell contacts in cAMP-Epac-Rap1 signal-activated cells.