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Biomedical subjects

E Raviola

Publications and source records attributed to E Raviola.

At least 55 records · Page 3Linked to original sources
PubMed1981

Paracellular route of aqueous outflow in the trabecular meshwork and canal of Schlemm. A freeze-fracture study of the endothelial junctions in the sclerocorneal angel of the macaque monkey eye.

The intercellular junctions of the endothelial cells of the trabecular meshwork and canal of Schlemm were examined with the electron microscope in the macaque monkey eye by both thin-sectioned specimens and the freeze-fracturing technique. The endothelial cells that line the beams of the meshwork are joined by gap junctions and short, isolated strands of tight junction; zonulae occludentes are absent. Thus aqueous humor can freely traverse the patent endothelial clefts of the trabecular meshwork. The endothelial cells of the canal of Schlemm are joined by zonulae occludentes and a small number of minute gap junctions. In 57% of their length, the tight junctions consist of one or two strands; the strands are rarely more than four. They remain preferentially associated with the E-face of the membrane, run parallel to one another, and only exceptionally branch or anastomose. Thus they are provided with free endings and do not form a bidimensional network. As a result of this organization, the zonula occludens is traversed by meandering channels of extracellular space or split pores, which connect the open endothelial clefts on the luminal and tissue fronts of the junction. The frequency of slit pores is 0.134 per micrometer of zonula occludens. They occupy 0.87% of the intercellular boundary and 0.0015% of the area of the endothelium. Estimates of the fluid conductance of the zonulae occludentes indicate that the intercellular clefts of the endothelium of Schlemm's canal filter but a small fraction of the amount of aqueous humor that leaves the anterior chamber through the conventional route.

Animals↗
PubMed1980

Structure of rapidly frozen gap junctions.

The structure of gap junctions in the rabbit ciliary epithelium, corneal endothelium, and mouse stomach and liver was studied with the freeze-fracturing technique after rapid freezing to near 4 degrees K from the living state. In the ciliary epithelium, the connexons were randomly distributed, separated by smooth membrane matrix. In the corneal endothelium, both random and crystalline arrangements of the connexons were observed. In the stomach and liver, the connexons were packed but not crystalline. Experimental anoxia or lowered pH caused crystallization of the connexons within 20-30 min. In the ciliary epithelium, the effects of prolonged anoxia or low pH could not be reversed . In addition, invaginated or annular gap junctions increased in number, but their connexons were usually distributed at random. Rapid freezing thus demonstrates that gap junctions of different tissues are highly pleiomorphic in the living state, and this may explain their variations in structure after chemical fixation. The slow time-course and irreversibility of the morphological changes induced by prolonged anoxia or low pH suggest that connexon crystallization may be a long-term consequence rather than the morphological correlate of the switch to high resistance.

Animals↗
PubMed1979

Increase in axial length of the macaque monkey eye after corneal opacification.

The cornea of one eye was opacified in two young macaque monkeys by multiple stromal injections of a suspension of polystyrene particles (latex). Ultrasound measurements showed that the eye with opaque cornea grew at a faster rate, so that after 1 year it was more than 1 mm longer than the normal eye. This difference in axial length was due to elongation of the posterior segment, since lens thickness, depth of anterior chamber, and corneal curvature were identical in both eyes. At histological examination, no pathological changes were observed in the anterior segment of the latex-injected eye except for a scant vascularization of the corneal opacity. The result of this experiment demonstrates that opacification of the corneal has effects on axial length similar to, although less marked than, those on lid fusion and therefore supports our previous conclusion that the myopia caused by lid fusion is triggered by the abnormal visual impact and involves central visual pathways.

Animals↗
PubMed1978

Membrane recycling in the cone cell endings of the turtle retina.

The ultrastructural effects of dark, light, and low temperature were investigated in the cone cell endings of the red-eared turtle (Pseudemys scripta elegans). Thin sections revealed that in dark-adapted retinas maintained at 22 degrees C, the neural processes which contact the cone cells at the invaginating synapses penetrated deeply into the photoreceptor endings. When dark-adapted retinas were illuminated for 1 h at 22 degrees C, the invaginating processes were apparently extruded from the synaptic endings. On the other hand, 1-h exposure to a temperature of 4 degrees C in the dark caused the invaginating processes to become much more strikingly inserted than at room temperature. A morphometric analysis showed that the ratio between the synaptic surface density of the endings and their total surface density decreased in the light and increased in the dark and cold. Freeze-fracturing documented fusion of synaptic vesicles with the presynaptic membrane in all conditions tested. These observations suggest that the changes in configuration of the pedicles in the light, dark, and cold reflect a different balance between addition and retrieval of synaptic vesicle membrane from the plasmalemma; in the dark, the rate of vesicle fusion is increased, whereas in the cold, membrane retrieval is blocked. When the eyecups were warmed up and illuminated for 30-45 min after cold exposure, a striking number of vacuoles and cisterns appeared in the cytoplasm and coated vesicles were commonly seen budding from the plasmalemma. 60-90 min after returning to room temperature, the endings had reverted to their normal configuration, and the vast majority of vacuoles, cisterns, and coated vesicles had disappeared. When horseradish peroxidase was included in the incubation medium, very few synaptic vesicles were labeled at the end of the period of cold exposure. 30-45 min after returning to 22 degrees C, vacuoles and cisterns contained peroxidase, whereas most synaptic vesicles were devoid of reaction product. 2 h after returning to 22 degrees C, coated vesicles, vacuoles, and cisterns had disappeared and a number of synaptic vesicles were labeled. These experiments suggest that vacuoles, cisterns, and coated vesicles mediate the retrieval of the synaptic vesicle membrane that has been added to the plasmalemma during cold exposure.

Animals↗
PubMed1978

Intercellular junctions in the ciliary epithelium.

The fine structure of the intercellular junctions in the ciliary epithelium of rhesus monkeys and rabbits was studied with conventional electron microscopy of thin-sectioned specimens and the freeze-fracturing technique. In the rhesus monkey, a zonula occludens, zonula adhaerens, gap junctions, and desmosomes interconnect the nonpigmented cells, whereas gap junctions, puncta adhaerentia, and desmosomes connect pigmented to nonpigmented cells, and pigmented cells to one another. In the rabbit, desmosomes are absent between nonpigmented cells, and substituted for by puncta adhaerentia. The zonula occludens between nonpigmented cells greatly varies in its complexity in different regions of the cell perimeter, and in places, it may consist of very few intramembrane strands; this suggests that the ciliary epithelium is relatively leaky to ions and small molecules. Gap junctions are ubiquitous in the ciliary epithelium and particularly numerous at the interface between pigmented and nonpigmented layers; this finding indicates that the cells of the ciliary epithelium are joined in a metabolic syncytium. All gap junctions are characterized by the crystalline configuration which is typical of the uncoupled state; furthermore, in specimens fixed by immersion, they may be caused by uncoupling and take place in the time interval elapsing between interruption of the blood supply and arrival of the fixative fluid. Puncta adhaerentia resemble zonulae adhaerentes in their structural details but are macular in shape instead of encompassing the cell perimeter in a beltlike fashion. In contrast with desmosomes, the intercellular cleft of puncta adhaerentia has an irregular width and contains opaque material, but this never gives rise to the central band typical of desmosomes. On the inner aspect of the junctional membranes, there is a layer of fluffy material but no plaque of insertion for a bundle of tonofilaments. Finally, puncta adhaerentia have no representation in the interior of the plasmalemma and are intimately associated with cytoplasmic microfilaments. They probably anchor to the plasmalemma the contractile apparatus of the ciliary epithelial cells.

Animals↗
PubMed1978

Effect of dark-rearing on experimental myopia in monkeys.

When lids are surgically fused in rhesus monkeys before eye growth is completed, a high degree of myopia develops, which is caused by an elongation of the eye globe. The present study shows that in monkeys raised in the dark after monocular lid fusion, refraction and axial length were normal in both the closed and the open eye. Myopia, however, readily developed and the eye elongated when a monkey raised in the dark was transferred to illuminated quarters. These findings indicate that visual stimulation through the translucent lids was necessary for the development of this experimental ametropia.

Animals↗
PubMed1976

Ultrastructure of perfusion-fixed fetal capillaries in the human placenta.

The ultrastructure of human placental capillaries was investigated using perfusion fixation and the freeze-fracturing technique. The capillaries have a continuous endothelium especially rich in microfilaments, whereas micropinocytotic vesicles are exceedingly scarce. The endothelial cells are connected by three types of junctions: (1) zonulae occludentes characterized by 2 to 4 focal regions of membrane contact in thin-sectioned specimens and an equal number of ridges on the membrane E-face in freeze-fractured specimens; (2) small gap junctions associated with the zonula occludens. (3) attachment plaques resembling zonulae adhaerentes in their fine structure. Endothelial cells are provided with long, circularly oriented pseudopodial extensions, which may be responsible for intermittent constrictions of the vessel lumen. These findings indicate that diaplacental transport at the level of the fetal capillary is controlled by the cytoplasm of the endothelial cells and probably occurs only to a very limited extent by way of micropinocytotic vesicles.

Capillaries↗
PubMed1975

Intramembrane organization of specialized contacts in the outer plexiform layer of the retina. A freeze-fracture study in monkeys and rabbits.

Freeze-fracture analysis of the neural connections in the outer plexiform layer of the retina of primates (Macaca mulatta and Macaca arctoides) demonstrates a remarkable diversity in the internal structure of the synaptic membranes. In the invaginating synapses of cone pedicles, the plasma membrane of the photoreceptor ending contains an aggregate of A-face particles, a hexagonal array of synaptic vesicle sites, and rows of coated vesicle sites, which are deployed in sequence from apex to base of the synaptic ridge. The horizontal cell dendrites lack vesicle sites and have two aggregates of intramembrane A-face particles, one at the interface with the apex of the synaptic ridge, the other opposite the tip of the invaginating midget bipolar dendrite. Furthermore, the horizontal cell dendrites are interconnected by a novel type of specialized junction, characterized by: (a) enlarged intercellular cleft, bisected by a dense plate and traversed by uniformly spaced crossbars; (b) symmetrical arrays of B-face particles arranged in parallel rows within the junctional membranes; and (c) a layer of dense material on the cytoplasmic surface of the membranes. The plasmalemma of the invaginating midget bipolar dendrite is unspecialized. At the contact region between the basal surface of cone pedicles and the dendrites of the flat midget and diffuse cone bipolar cells, the pedicle membrane has moderately clustered A-face particles, but no vesicle sites, whereas the adjoining membrane of the bipolar dendrites contains an aggregate of B-face particles. The invaginating synapse of rod spherules differs from that of cone pedicles, because the membrane of the axonal endings of the horizontal cells only has an A-face particle aggregate opposite the apex of the synaptic ridge. Specialized junctions between horizontal cell processes, characterized by symmetrical arrays of intramembrane B-face particles, are also present in the neuropil underlying the photoreceptor endings. Small gap junctions connect the processes of the horizontal cells; other gap junctions probably connect the bipolar cell dendrites which make contact with each cone pedicle. Most of the junctional specializations typical of the primate outer plexiform layer are also found in the rabbit retina. The fact that specialized contacts between different types of neurons interacting in the outer plexiform layer have specific arrangements of intramembrane particles strongly suggests that the internal structure of the synaptic membranes is intimately correlated with synaptic function.

Animals↗
PubMed1973

Gap junctions between photoreceptor cells in the vertebrate retina.

In the outer plexiform layer of the retina the synaptic endings of cone cells make specialized junctions with each other and with the endings of rod cells. The ultrastructure of these interreceptor junctions is described in retinas of monkeys, rabbits, and turtles, in thin sections of embedded specimens and by the freeze-fracturing technique. Cone-to-rod junctions are ribbon-like areas of close membrane approximation. On either side of the narrowing of the intercellular space, the junctional membranes contain a row of particles located on the fracture face A (cytoplasmic leaflet), while the complementary element, a row of single depressions, is located on fracture face B. The particle rows are surrounded by a membrane region that is devoid of particulate inclusions and bears an adherent layer of dense cytoplasmic material. Cone-to-cone junctions in some places are identical to cone-to-rod junctions, while in other places they closely resemble typical gap junctions (nexus). Interreceptor junctions, therefore, represent a morphological variant of the gap junction, and probably mediate electrotonic coupling between neighboring photoreceptor cells.

Animals↗
PubMed1972

Evidence for a blood-thymus barrier using electron-opaque tracers.

In order to verify the existence of a blood-thymus barrier to circulating macromolecules, the permeability of the vessels of the thymus was analyzed in young adult mice using electron opaque tracers of different molecular dimensions (horseradish peroxidase, cytochrome c, catalase, ferritin, colloidal lanthanum). Results show that although blood-borne macromolecules do penetrate the thymus, their parenchyma] distribution is limited to the medulla of the lobe by several factors: (a) the differential permeability of the various segments of the vascular tree; (b) the spatial segregation of these segments within the lobe; (c) the strategic location of parenchymal macrophages along the vessels. The cortex is exclusively supplied by capillaries, which have impermeable endothelial junctions. Although a small amount of tracer is transported by plasmalemmal vesicles through the capillary endothelium, this tracer is promptly sequestrated by macrophages stretched out in a continuous row along the cortical capillaries and it does not reach the intercellular clefts between cortical lymphocytes and reticular cells. The medulla contains all the leaky vessels, namely postcapillary venules and arterioles. Across the walls of the venules, large quantities of all injected tracers escape through the clefts between migrating lymphocytes and endothelial cells; also the arterioles have a small number of endothelial junctions which are permeable to peroxidase, but do not allow passage of tracers of higher molecular weight. The tracers released by the leaky vessels penetrate the intercellular clefts of the medulla, but they never reach the cortical parenchyma, even at long time intervals after the injection. Therefore, a blood-thymus barrier to circulating macromolecules does exist, but is limited to the cortex. Medullary lymphocytes are freely exposed to blood-borne substances.

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