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Biomedical subjects

G Raviola

Publications and source records attributed to G Raviola.

At least 37 records · Page 2Linked to original sources
PubMed1982

Freeze-fracture analysis of the interendothelial junctions in the blood vessels of the iris in Macaca mulatta.

The interendothelial cell junctions in the blood vessels of the rhesus monkey iris were examined by both conventional electron microscopy and the freeze-fracturing technique. Endothelial cells of iris blood vessels were joined by two types of intercellular junctions: zonular tight junctions and gap junctions. The zonulae occludentes were represented by a complex network of branching and anastomosing strands that remained preferentially associated with the E fracture face. They varied in complexity from a single strand to eight or more but usually consisted of from two to four strands. Small gap junctions were inserted within the tight junctional network. Junctions similar to those typical of muscular and visceral venules were absent. Thus the intercellular clefts of all iris vessels are closed by zonulae occludentes that exhibit a degree of complexity intermediate between the endothelial junctions of the cerebral cortex and those of striated muscles and viscera. This finding strongly suggests that the blood vessels of the iris participate only minimally in aqueous humor dynamics.

Animals↗
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

Rod-shaped bodies and crystalloid inclusions in ocular vascular endothelia of adult and developing Macaca mulatta.

In addition to the usual organelles, endothelial cells of ocular blood vessels and Schlemm's canal in adult and fetal Macaca mulatta, contain two types of inclusions: rod-shaped bodies and crystalloids. The rod-shaped bodies are similar to those first described in arterial endothelia of the rat. They are between 0.2 to 0.35 micrometer in diameter, up to 2.5 micrometers in length, and are membrane bounded. The crystalloid inclusions are up to 1.2 micrometers in diameter and are associated with the granular endoplasmic reticulum. Their crystalline lattice consists of subunits, 28 nm in diameter, which are either arranged in rows separated by amorphous matrix or tightly packed in a honeycomb-like lattice. The rod-shaped bodies are much more numerous than the crystalloids; however, both kinds of inclusions are consistently found in both adult and fetal vessels. The represent normal components of ocular vascular endothelia which appear very early in the development of the eye.

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↗
PubMed1980

A simple staining method for blood vessels in flat preparations of ocular tissues.

We describe a technique for light microscopic visualization of the vasculature of the retina, choroid, ciliary body, and iris in flat preparations. The technique is based on the intravenous injection of horseradish peroxidase, the histochemical demonstration of this enzyme's activity, and the bleaching of the melanin that hinders the visualization of the injected vessels. This technique is reproducible and much simpler than other available methods in which dyes suspended in different media are injected directly into the ocular vessels.

Animals↗
PubMed1978

Degeneration and regeneration of autonomic nerve endings in the anterior part of rhesus monkey ciliary muscle.

The autonomic nerve plexus of the ciliary muscle was examined with the electron microscope in normal rhesus monkeys of different ages. In the anterior region of the muscle, at the boundary with the poorly innervated scleral spur and trabecular meshwork, 3.8-7.1% of the axons exhibit either degenerative or regenerative features. The cytoplasm of degenerating axons contains lamellated, dense and multivesicular bodies, vesicles, whorls of filaments, and membranous debris. The plasma membrane is often discontinuous and, on occasion, axonal debris and degenerative organelles are freely dispersed in the connective tissue spaces of the muscle. Degenerating axons contain a granular reaction product when stained for acid phosphatase activity. Regenerating axons are characterized by tightly packed mitochondria, glycogen particles, and aggregates of synaptic vesicles; they synapse with muscle cells and are negative to the acid phosphatase reaction. A quantitative analysis showed that in the anterior region of the ciliary muscle degenerating and regenerating axons increase in number with age, although the total number of axonal profiles remains constant. In the age groups examined, degenerating axons occurred with the same frequency as regenerating axons, thus, the age-dependent increase in axonal degeneration is accompanied by a parallel increase in axonal regeneration. We conclude that autonomic nerve endings in the anterior part of the ciliary muscle undergo a continuous process of renewal that is more prominent in old age.

Acid Phosphatase↗
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↗
PubMed1976

Giant nerve fibers in the ciliary muscle and iris sphincter of Macaca mulatta.

Unusually large nerve processes, containing numerous mitochondria, glycogen particles, and synaptic vesicles are described in both the ciliary muscle and the iris sphincter muscle of the rhesus monkey. The striking similarity of these axonal profiles to the dendritic enlargements observed by Sotelo and Palay (1968) is noted and the possibility that they represent growing ends of peripheral nerve fibers is suggested.

Animals↗
PubMed1976

Blood-aqueous barrier can be circumvented by lowering intraocular pressure.

Rhesus monkeys were injected intravenously with hypertonic urea (9 ml/kg body weight of 30% urea in 10% invert sugar) and the intraocular pressure was measured with an applamatic tonometer. When this pressure reached its minimum (20% of the normal value) horseradish peroxidase (molecular weight 40,000; radius of an equivalent hydrodynamic sphere about 2.5 nm; 0.5 g/kg body weight), was injected intravenously. Twenty minutes following peroxidase administration, either aqueous humor was sampled from the anterior chamber for biochemical determination of peroxidase activity, or one eyeball was enucleated and processed for light and electron microscopic localization of the enzymatic tracer. This experiment showed that: (1) therapeutic doses of hypertonic urea do not cause a breakdown of either the blood-retina or the blood-aqueous barriers; (2) as intraocular pressure decreases, peroxidase-containing blood flows back from the episcleral veins into the Schlemm canal; (3) macromolecules up to the dimensions of horseradish peroxidase leak through the intercellular clefts of the endothelium of the Schlemm canal, permeate the juxtacanalicular connective tissue and trabecular meshwork, and finally enter the anterior chamber. Thus, blood-borne substances can circumvent the blood-aqueous barrier when intraocular pressure is decreased, and administration of a hypertonic agent may represent a simple pharmacological device to cause penetration into the ocular chambers by drugs that are normally excluded from the interior of the eye.

Animals↗