Glycosaminoglycans in components of the rabbit eye: synthesis and characterization

Purpose. To trace the eye components involved in proteoglycan synthesis and to characterize the sulfated glycosaminoglycans which are associated to these macromolecules.Methods. Sodium [S-35]-sulfate was injected intravitreally and the rabbits were killed at different time intervals after the injection. The glycosaminoglycans of choroid, ciliary body, cornea, iris, lens capsule, retina and sclera were extracted and processed for estimations of their specific activities, and for electrophoresis plus autoradiography with or without previous treatment with specific enzymes. In addition, methacrylate sections of the eyes were analysed by autoradiography.Results. The peak of specific activities of the glycosaminoglycans of all eye components occurred at 2 days after the intravitreal injection of [S-35]-sulfate. The autoradiography of the agarose gels revealed three types of glycosaminoglycans, namely, heparan-, chondroitin- and dermatan sulfate, only in the retina. The other eye components contained heparan sulfate and either chondroitin or dermatan sulfate. Tissue autoradiography together with the biochemical techniques contributed to unravel the origin of the glycosaminoglycans in the eye components.Conclusions. The results of the present investigation have shown that heparan sulfate, contrasting to chondroitin sulfate and dermatan sulfate, is synthesized in all eye components studied and that the glycosaminoglycan composition differs according to the tissue of origin.

ENDO-OLIGOPEPTIDASE-A, A PUTATIVE ENKEPHALIN-GENERATING ENZYME, IN THE VERTEBRATE RETINA

Endo-oligopeptidase A, EC 3.4.22.19, converts small enkephalin-containing peptides into the corresponding enkephalins in vitro. We investigated the presence of endooligopeptidase A in the retina and its possible colocalization with enkephalins in retinal neurons. The specific activity of endo-oligopeptidase A found in pigeon retinae (30.3 +/- 7.3 mU/mg, mean +/- standard deviation) was four times higher than in rabbit retinae (7.0 +/- 1.1 mU/mg). The enzyme activity was not modified by EDTA, but it was enhanced by dithiothreitol and inhibited by zinc and 5,5'-dithiobis(2-nitrobenzoic acid). Immunohistochemical experiments with a purified antiserum against rabbit endo-oligopeptidase A revealed labeled neurons in both the inner nuclear layer and the ganglion cell layer of pigeon and rabbit retinae. Double-labeling immunofluorescence experiments demonstrated that about 90% of neurons containing endo-oligopeptidase A-like immunoreactivity also contained [Leu5]-enkephalin-like immunoreactivity. These colocalization results may represent an important step toward the demonstration of the possible involvement of endo-oligopeptidase A in enkephalin generation in vivo.

Macro- and microstructural organization of the rabbit's celiac-mesenteric ganglion complex (Oryctolagus cuniculus)

The macro- and microstructures of the rabbit celiac-mesenteric ganglion complex are described in 20 young animals. We found ten celiac ganglia, twenty-seven cranial mesenteric ganglia and eleven celiac-mesenteric ganglia. The celiac ganglia had a rectangular shape in nine cases (90%) and a circular one in one case (10%). The cranial mesenteric ganglia presented triangular (66.7%), rectangular (11.1%), L-shape (18.5%) and semilunar (3.7%) arrangements. The celiac-mesenteric ganglia were organized in three patterns: a single left celiac-mesenteric ganglion having a caudal portion (72.7%); celiac-mesenteric ganglia without a caudal portion (18.2%) and a single celiac-mesenteric ganglion with two portions: left and right (9.1%).The microstructure was investigated in nine celiac-mesenteric ganglia. The results showed that the celiac-mesenteric ganglion is actually a ganglion complex constituted of an agglomerate of ganglionic units separated by nerve fibers, capillaries and septa of connective tissue. Using the semi-thin section method we described the cellular organization of the celiac-mesenteric ganglion complex. Inside of each ganglionic unit, there were various cell types: principal ganglion neurons (PGN), glial cells (satellite cells) and SIF cells (small intensely fluorescent cells or small granular cells), which are the cytologic basis for each ganglionic unit of the rabbit's celiac-mesenteric ganglion complex.

Effects of mercury intoxication on the response of horizontal cells of the retina of thraira fish (Hoplias malabaricus)

Methyl mercury (MeHg) is highly neurotoxic, affecting visual function in addition to other central nervous system functions. The effect of mercury intoxication on the amplitude of horizontal cell responses to light was studied in the retina of the fish Hoplias malabaricus. Intracellular responses were recorded from horizontal cells of fish previously intoxicated with MeHg by intraperitoneal injection (IP group) or by trophic exposure (T group). Only one retina per fish was used. The doses of MeHg chloride administered to the IP group were 0.01, 0.05, 0.1, 1.0, 2.0, and 6.0 mg/kg. The amplitudes of the horizontal cell responses were lower than control in individuals exposed to 0.01 (N = 4 retinas), 0.05 (N = 2 retinas) and 0.1 mg/kg (N = 1 retina), whereas no responses were recorded in the 1.0, 2.0, and 6.0 mg/kg groups. T group individuals were fed young specimens of Astyanax sp previously injected with MeHg corresponding to 0.75 (N = 1 retina), 0.075 (N = 8 retinas) or 0.0075 (N = 4 retinas) mg/kg fish body weight. After 14 doses, one every 5 days, the amplitude of the horizontal cell response was higher than control in individuals exposed to 0.075 and 0.0075 mg/kg, and lower in individuals exposed to 0.75 mg/kg. We conclude that intoxication with MeHg affects the electrophysiological response of the horizontal cells in the retina, either reducing or increasing its amplitude compared to control, and that these effects are related to the dose and/or to the mode of administration.

Evidence of reciprocal connections between the dorsal raphe nucleus and the retina in the monkey Cebus apella

Possible connections between the retina and the raphe nuclei were investigated in the monkey Cebus apella by intraocular injection of cholera toxin B subunit (CTb). CTb-positive fibers were seen in the lateral region of the dorsal raphe nucleus (DR) on the side contralateral to the injection, and a few labeled perikarya were observed in the lateral portion of the DR on the ipsilateral side. Our findings suggest that direct and reciprocal connections between the retina and DR may exist in Cebus apella. These connections might be part of an important pathway through which the light/dark cycle influences the Activity and/or functional status of raphe neurons, with potential effects on a broad set of neural and behavioral circuits. (c) 2007 Elsevier B.V. All rights reserved.

Comparison of synthetic hydroxyapatite and porous polyethylene implants in eviscerated rabbit eyes

Purpose: Synthetic hydroxyapatite and porous polyethylene (Polipore) spheres were placed in rabbits' eviscerated cavities to evaluate tissue reaction and volume maintenance.Methods. Fifty-six Norfolk albino rabbits underwent unilateral evisceration and implantation of synthetic hydroxyapatite (H group, 28 animals) or porous polyethylene spheres (P group, 28 animals). Postoperative reactions, animal behavior, and socket conditions were monitored. Light microscopy and morphometric evaluation with statistical analysis of the exenterated orbits were performed at 7, 15, 30, 60, 90, 120, and 180 days. Scanning electron microscopy was appraised 7, 60, and 180 days after surgery.Results: Two animals from the H group and 1 from the P group had extrusion 7 days after surgery. Throughout the experimental period, the synthetic hydroxyapatite caused more inflammation than the porous polyethylene material. Ingrowth in the sphere occurred 7 to 15 days after the surgery in both groups, and the tissue reaction became denser at approximate to60 to 90 days, when bony metaplasia began in the H group. Volume maintenance was better in the P group and with a smaller pseudocapsule surrounding the implanted sphere than in the H group.Conclusions: Clinical findings demonstrated mild inflammation inside the sphere and in the pseudocapsule surrounding it and better cavity volume maintenance in the P group animals. The authors consider porous polyethylene a more suitable material than synthetic hydroxyapatite for use in anophthalmic cavity reconstruction.

Effects of Inhaled Nitric Oxide on Oxidative Stress and Histopathological and Inflammatory Lung Injury in a Saline-Lavaged Rabbit Model of Acute Lung Injury

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

High-frequency oscillatory ventilation attenuates oxidative lung injury in a rabbit model of acute lung injury

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development of a rabbit's urethral sphincter deficiency animal model for anatomical-functional evaluation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Autologous transplantation of adult adipose derived stem cells into rabbit urethral wall

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reproductive responses of rabbit does to a supplemental lighting program

The objective was to investigate whether the productivity of rabbit does can be improved, when natural photoperiod is decreasing, by adopting a supplemental lighting program. Three experiments were conducted involving two groups: control, submitted to the natural decreasing photoperiod, and supplemented with a lighting program which provided 14 h light/24 h beginning at 10 weeks of age. In the first experiment, 20 nulliparous does, 10 from each group, were euthanized 8 h after being presented to a buck; the overall number of follicles, whose diameter exceeded I mm, was determined macroscopically. The right ovaries were collected, histologically analyzed, and electronically measured. In the second experiment, 30 nulliparous does, 15 from each group, were presented to a buck (day 1). Receptive does were euthanized on day 8 to evaluate embryonic survival (number of normal embryos/ovulation rate). In the third experiment, 48 nulliparous does, 24 from each group, were followed from the first presentation to the buck until the weaning of the first litter. The effect of treatment on reproductive and body weight traits of does, and litter performance traits, at birth and weaning, was evaluated. The average number of follicles whose diameter exceeded 1 mm was higher in the treatment group (12.05 +/- 1.07 vs. 8.63 +/- 1.00, P=0.03 7). Receptive does of the treatment group had heavier ovaries relative to those of the control group (790 +/- 59 vs. 470 +/- 64 mg, P=0.004), whereas no treatment difference regarding this trait was found for non-receptive ones. Treatment had a favorable effect on pregnancy rate of total exposed and of receptive does (80.0% vs. 33.3%, P=0.01, and 92.3% vs. 50.0%, P=0.02, respectively). The number of underdeveloped embryos was lower (0.067 +/- 0.380 vs. 2.500 +/- 0.455, P=0.004), embryonic survival up to day 8, and uterus weight was higher in the treatment group (0.839 +/- 0.075 vs. 0.534 +/- 0.087, P=0.033 and 13.83 +/- 0.72 vs. 10.99 +/- 0.84, P=0.037, respectively). Number of presentations tended to be lower (1.32 +/- 0.17 vs. 1.75 +/- 0.16, P=0.077) and adjusted litter size in the first reproductive cycle tended to be higher (7.09 +/- 0.89 vs. 5.22 +/- 0.68, P=0.091) in the treatment group relative to the control.