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E Raviola

Publications and source records attributed to E Raviola.

At least 19 recordsLinked to original sources

Synaptic connections of the narrow-field, bistratified rod amacrine cell (AII) in the rabbit retina.

The synaptic connections of the narrow-field, bistratified rod amacrine cell (AII) in the inner plexiform layer (IPL) of the rabbit retina were reconstructed from electron micrographs of continuous series of thin sections. The AII amacrine cell receives a large synaptic input from the axonal endings of rod bipolar cells in the most vitreal region of the IPL (sublamina b, S5) and a smaller input from axonal endings of cone bipolar cells in the scleral region of the IPL (sublamina a, S1-S2). Amacrine input, localized at multiple levels in the IPL, equals the total number of synapses received from bipolar cells. The axonal endings of cone bipolar cells represent the major target for the chemical output of the AII amacrine cell: these synapses are established by the lobular appendages in sublamina a (S1-S2). Ganglion cell dendrites represent only 4% of the output of the AII amacrine and most of them are also postsynaptic to the cone bipolars which receive AII input. The AII amacrine is not presynaptic to other amacrine cells. Finally, the AII amacrine makes gap junctions with the axonal arborizations of cone bipolars that stratify in sublamina b (S3-S4) as well as with other AII amacrine cells in S5. Therefore, in the rabbit retina 1) the rod pathway consists of five neurons arranged in series: rod-->rod bipolar-->AII amacrine-->cone bipolar-->ganglion cell; 2) it seems unlikely that a class of ganglion cells exists that is exclusively devoted to scotopic functions. In ventral, midperipheral retina, about nine rod bipolar cells converge onto a single AII amacrine, but one of them establishes a much higher proportion of synaptic contacts than the rest. Conversely, each rod bipolar cell diverges onto four AII amacrine cells, but one of them receives the largest fraction of synapses. Thus, within the pattern of convergence and divergence suggested by population studies, preferential synaptic pathways are established.

Animals

Synaptic connections of rod bipolar cells in the inner plexiform layer of the rabbit retina.

We have reconstructed from electron micrographs of a continuous series of thin sections the synaptic connections of the axonal arborizations of all the rod bipolar cells contained in a small region of the retina of the rabbit. We observed that all rod bipolars share the same pattern of connectivity and are probably functionally equivalent. As a rule, they do not contact ganglion cells. Their prevalent synaptic output is on narrow-field, bistratified, and indoleamine-accumulating amacrine cells. Their dominant inputs are the reciprocal synapses from the indoleamine-accumulating amacrines, but they also receive a sizable number of synaptic contacts from other, non-reciprocal, amacrine cells. The lateral spread of scotopic signals at the synapse between rod bipolars and narrow-field, bistratified amacrines is small. Finally, in the rabbit, as in the cat, a narrow-field, bistratified amacrine is inserted in series along the rod pathway.

Animals

Neural control of eye growth and experimental myopia in primates.

Macaque monkeys become myopic when raised with fused lids to expose the retina to formless shadows during the period of postnatal eye development. The effect of the abnormal visual input is an excessive expansion of the posterior segment of the eye, a process that seems to be controlled by the nervous system. The mechanism by which the nervous system influences eye growth appears to be different in the stumptailed macaque (Macaca arctoides) and the rhesus macaque (M. mulatta). Lid-fused arctoides monkeys do not develop myopia when the ciliary muscle is paralysed or the optic nerve is cut, suggesting that the abnormal growth is caused by excessive accommodation. In contrast, paralysis of the ciliary muscle or optic nerve section does not prevent the development of myopia in the rhesus macaque, suggesting that in this species the axial growth is controlled by the retina. In both species neonatal lid fusion causes a marked increase in retinal vasoactive intestinal polypeptide (VIP). VIP is contained in a single type of amacrine cell whose dendrites spread in the middle of the inner plexiform layer. It remains to be determined whether the increase in the level of VIP is related to the abnormal axial elongation caused by lid fusion. At present we are also exploring the effects of accommodation on the growth of the eye by training juvenile arctoides monkeys to work on complex visual discrimination paradigms. Preliminary results show that performing a visual task at close range may influence the axial length and refraction in this macaque species.

Accommodation, Ocular

Axonless horizontal cells of the rabbit retina: synaptic connections and origin of the rod aftereffect.

An axonless horizontal cell (AHC) of the rabbit retina was penetrated with a microelectrode and stained with horseradish peroxidase after recording its light responses. The cell was then serially sectioned and its connections examined with the electron microscope. Physiologically, the cell exhibited cone-dominated responses and a minor rod influence known as rod aftereffect. Electron microscopy showed that this AHC was only connected to cones. Therefore, the rod aftereffect could only invade the cell through the gap junctions between the synaptic endings of rod and cone photoreceptors. In the synaptic invaginations of the cone pedicles contacting the cell, only one of the lateral elements was stained. This suggests that the two lateral elements of each cone-invaginating synapse belong to two different horizontal cells. By staining intracellularly adjacent AHCs, we showed that the two lateral processes may originate from two horizontal cells belonging to the same morphological type.

Animals

Connections of indoleamine-accumulating cells in the rabbit retina.

To study the connections of the neurons of the rabbit retina that accumulate indoleamines, we injected 5,7-dihydroxytryptamine into the vitreous body. It accumulated within a subset of amacrine cells and could be visualized there by aldehyde-induced fluorescence. The fluorescent labeling was photo-converted to an insoluble, osmiophilic product by irradiation in the presence of diaminobenzidine, and the tissue was examined by electron microscopy. Preservation of the structure of the tissue after photoconversion was satisfactory and the dendrites of the indoleamine-accumulating cells could easily be identified. They form a dense plexus near the junction of the inner plexiform and ganglion cell layers, where they exhibit large synaptic endings that occupy a substantial fraction of the surface of rod bipolar terminals. The dendrites of the indoleamine-accumulating cells receive input from rod bipolars at dyad synapses, where the other postsynaptic partner is a dendrite of a narrow-field, bistratified amacrine cell; in addition, they receive amacrine cell input throughout the inner plexiform layer. The only outputs we observed are reciprocal synapses onto the rod bipolar endings. Thus, these amacrine cells appear to exert an important effect on the transmission of scotopic information through the retina.

5,7-Dihydroxytryptamine

Increase in retinal vasoactive intestinal polypeptide after eyelid fusion in primates.

Lids were fused in six neonatal and one adult macaque monkey (Macaca mulatta and Macaca arctoides) and were kept fused for 1 to 18.5 months. The juvenile macaques, but not the adult one, developed myopia due to excessive elongation of the eye. In all animals, the immunohistochemical reactivity of the retina for vasoactive intestinal polypeptide (VIP) was much higher in the closed than in the open eyes. The neuropeptide was localized to the perikaryon and dendrites of amacrine cells. No difference was observed in substance P immunoreactivity between open and closed eyes, suggesting that the observed effect is selective. The change in VIP immunoreactivity could be the result of an increase in peptide synthesis, a decrease in peptide release, or a combination of the two. These results indicate that VIP may play a part in the regulation of postnatal ocular growth.

Animals

Rod photoreceptors dissociated from the adult rabbit retina.

Rod photoreceptors have been isolated from the adult rabbit retina using enzymatic and mechanical dissociation procedures; their fine structure, synaptic activity, and long-term viability were examined using conventional electron-microscopic, quick-freezing, and cell culture techniques. Freshly dissociated photoreceptors were well-preserved compared to their counterparts in the intact retina. About half of the cells, however, exhibited broad continuity between inner and outer segments. Quick-frozen, freeze-substituted rods differed from chemically fixed cells in 3 respects: (1) there was an increased amount of granular matrix in the cytoplasm, mitochondria, and rough endoplasmic reticulum; (2) branching and anastomosing profiles of smooth endoplasmic reticulum had disappeared from the inner segment; and (3) the number of synaptic vesicles within the spherule was highly variable, in some cases leaving synaptic ribbons completely denuded of their halo of vesicles. Light-adapted, solitary rod cells continued to be synaptically active: their endings were capable of endocytosis when placed in the dark in the presence of extracellular ferritin and tracer was incorporated into vesicles and vacuoles; this uptake was much reduced when the cells were incubated with the tracer in the light. Thus, synaptic vesicle regeneration was stimulated in the dark, suggesting that vesicles underwent exocytosis in the dark. Isolated rod cells adhered poorly to most standard substrates; without proper adhesion, cells deteriorated in 2-4 hr. However, photoreceptors did adhere to glutaraldehyde-fixed Vitrogen gels and could be maintained for over 48 hr on this substrate if kept in a complete medium at 22 degrees C. In contrast, Müller cells adhered quickly to a laminin substrate with their endfoot processes. The differential adhesion properties of Müller and photoreceptor cells may be useful in obtaining pure populations of glial cells or neurons from the adult mammalian retina.

Animals

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

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

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

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

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