Ocular lesions of 6-aminonicotinamide toxicosis in rabbits

6-Aminonicotinamide, an antimetabolite of nicotinamide, given by intraperitoneal injection produced diarrhea, ascending paresis/paralysis, death, and bilateral ocular alterations in both sexes of New Zealand white and Dutch belted rabbits. Ocular vascular lesions consisted of iridal congestion with iridal hemorrhage and associated acute iritis and aqueous flare in some rabbits. Cytoplasmic swelling, vacuolation, and loss of staining affinity that represented hydropic change were present in both layers of ciliary epithelium and the inner layer of iridal epithelium. Cells in the outer layer of the iridal processes and the ciliary ridge, were most severely affected. Vacuoles were also present in the retinal pigment epithelium and scattered throughout the outer plexiform layer of the retina with a few in the inner and outer nuclear layers. Ocular alterations were prevented by simultaneous administration of nicotinamide and their development appeared related to nicotinamide deficiency. No ocular alterations were caused by nicotinamide administration alone.     

Fine structure of the retinal pigment epithelium, Bruch's membrane and choriocapillaris in the ostrich (Struthio camelus)

The fine structure of the retinal pigment epithelium (RPE), Bruch's membrane and choriocapillaris in the ostrich (Struthio camelus) was investigated by using light microscopy and transmission electron microscopy. In this species, the RPE consisted of a single layer of low columnar cells. The epithelial cells were joined laterally by two type junctions, zonulae occludentes and zonulae adherentes located in the midregion of the cells. These cells displayed numerous deep basal infoldings and thick extensive apical processes, which enclosed the outer segments of the rods. The epithelial cell nuclei were large, vesicular and located basally within the epithelial cells. Smooth endoplasmic reticulum was very abundant, while rough endoplasmic reticulum was scarce. Mitochondria of various shapes were abundant basally while polysomes were plentiful and widespread. In the light-adapted state melanosomes were located in the apical region and in apical processes of the epithelial cells. Myeloid bodies were large, numerous and often showed ribosomes on their outer surface. Bruch's membrane (complexus basalis) was typical pentalaminate throughout the retina, as noted in the majority of other vertebrates. The endothelium of the choriocapillaris facing Bruch's membrane was extremely thin but only moderately fenestrated. Some of the fenestrations displayed a double-layered diaphragm while the majority showed the more typical single-layered diaphragm noted in most species.     

Fine Structure of the Choriocapillaris, Bruch''s Membrane and Retinal Epithelium of the Cow

The fine structure of the choriocapillaris, Bruch's membrane and retinal epithelium was investigated in both the tapetal and non-tapetal fundus of the bovine eye. In ail locations the retinal epithelium consists of a single layer of cuboidal cells. The epithelial cells are joined laterally by apically located tight junctions and throughout the retina display numerous basal infoldings and fine apical processes which enclose rod outer segments. All retinal epithelial cells are rich in smooth endoplasmic reticulum and mitochondria and contain phagosomes. Although not as abundant, profiles of rough endoplasmic reticulum and polysomes are also noted in all locations. In non-tapetal areas, melano-somes are numerous whereas over the central tapetum fibrosum they are absent. The absence of melanosomes over a functional tapetum is to be expected. While lysosomes are present throughout the epithelial layer, over the tapetal region they appear to be more numerous. The apparent increase in lysosomal numbers in this location may indicate an enhanced shedding of outer segment material over the tapetum. Although some retinal epithelial cells are modified to accomodate a tapetum lucidum their morphology is basically similar throughout the retina and probably indicates that ail regions of the retinal epithelium are capable of the normal functions of this indispensible retinal layer. The choriocapillaris is heavily fenestrated on the border facing the retina and overlying the tapetum also displays fenestrae on its choroidal edge. Bruch's membrane ( complexus basalis ) is pentalaminate throughout the retina and is slightly thicker in the posterior fundus.     

Fine structure of the retinal pigment epithelium, Bruch's membrane and choriocapillaris in the horse

The fine structure of the retinal pigment epithelium (RPE), Bruch's membrane and choriocapillaris was investigated by light and transmission electron microscopy in both the tapetal and non-tapetal fundus of the horse eye. In all locations, the RPE consisted of a single layer of low cuboidal cells. The epithelial cells were joined laterally by apically located tight junctions. These cells displayed numerous basal infoldings and abundant thin apical processes which enclosed the rod outer segments. The epithelial cell nuclei were large and located basally. Within the epithelial cells, smooth endoplasmic reticulum was very abundant, while rough endoplasmic reticulum was scarce, polysomes and mitochondria, which often display a ring-shaped structure, were abundant. Melanosomes were abundant in the non-tapetal area but absent in the tapetal area. Bruch's membrane was pentalaminate throughout the retina. The endothelium of the choriocapillaris was heavily fenestrated.     

Development of the Retina in the Porcine Fetus A Light Microscopic Study

Morphogenesis of the porcine retina was studied using light microscopy from 4 weeks of gestation until birth (18 to 310 mm crown-rump length), and compared with the adult stage (6 months). Tissue samples were examined from the posterior and peripheral parts of the retina. At 18 mm the retina consists of an inner marginal layer and an outer layer of neuroblastic cells. At 18-40 mm the latter layer is divided into an inner and an outer neuroblastic layer by the transient layer of Chievitz. Subsequently, the development of the different retinal layers begins at the inner retinal border and moves progressively outwards; it also spreads from the posterior to the peripheral part of the neural retina. Many cells of the inner neuroblastic layer are prospective ganglionic cells which migrate inwards, thus forming the ganglion cell layer and the inner plexiform layer at 90 mm. At 120 mm, primitive horizontal cells appear within the outer neuroblastic layer. Separation of this layer into the inner nuclear, outer plexiform and outer nuclear layers is first evident at 180 mm. At this stage all retinal layers are present, except the layer of the photoreceptor cells which is not widespread until at 220 mm. The inner and outer segments of the photoreceptor cells lengthen considerably during the last month of gestation. During the late fetal stage the nerve fiber layer, the inner and outer plexiform layers and the layer of rods and cones all continue to increase in thickness. Concurrently, the ganglion cell layer and the inner and outer nuclear layers have reached their maximal thickness and become thinner. After the total thickness of the neural retina amounts to approximately 180 microns at two to three weeks before birth, it then thins to approximately 160 microns in the adult stage.     

Cytologic appearance of retinal cells included in a fine‐needle aspirate of a meningioma around the optic nerve of a dog

A 6‐year‐old Wirehair Dachshund had a meningioma around the optic nerve that caused exophthalmos. A benign mesenchymal tumor was suspected based on the cytologic pattern of a fine‐needle aspirate, and a meningioma was diagnosed by histopathologic examination. In addition to the meningioma cells, the cytologic smears included groups of cells from apparently 4 layers of normal retina. In particular, uniform rod‐shaped structures in the cytologic sample could suggest rod‐shaped bacteria, but these structures were identified as cylindrical outer segments of photoreceptor rod cells. Other retinal structures recognized included pigmented epithelial layer cells with their uniquely formed pigment granules, the characteristic bi‐lobed, cleaved nuclei from the outer nuclear layer, and nerve tissue likely from the outer plexiform layer of the retina.     

Fine structure of the retinal pigment epithelium, Bruch's membrane and choriocapillaris in the camel

The morphology of the retinal pigment epithelium (RPE) and closely associated Bruch's membrane and choriocapillaris was investigated by light and transmission electron microscopy in the camel (Camelus dromedarius). The study showed that RPE is composed of a single layer of hexanocuboidal cells that were joined laterally by a series of apically located tight junctions. In addition, adjacent from internal side of cell membrane at the level of tight junctions, an undefined structure which resembled the myofibrillar organization of skeletal muscles in appearance was located. These cells displayed numerous short basal infoldings and abundant thin apical processes which enclosed the rod outer segments. The epithelial cell nuclei were large, vesicular and eccentrically located. Within the epithelial cells, smooth endoplasmic reticulum was very abundant, while rough endoplasmic reticulum was present only in small amounts. Polysomes were also numerous and the mitochondria often displayed a ring-shaped structure. Lipofuscin granules were plentiful in all locations. Bruch's membrane (complexus basalis) was typically pentalaminate throughout the retina. The endothelium of the choriocapillaris facing Bruch's membrane was extremely thin and heavily fenestrated. These fenestrations displayed typical single-layered diaphragm as noted in most species.     

Multifocal retinopathy of Great Pyrenees dogs

Forty-four related Great Pyrenees dogs were examined ophthalmoscopically. Focal retinal elevations, multiple gray-tan-pink subretinal patches, and discrete areas of tapetal hyper-reflectivity were seen in 19 dogs, ranging from 13 weeks to 10 years of age. These lesions varied in size from focal spots that were barely visible with the indirect ophthalmoscope to areas that were larger than the optic disc. Complete blood cell counts, serum biochemical profiles, urinalyses, and blood pressure measurements were completed on four affected dogs and all were within normal reference ranges. Photopic and scotopic electroretinography was completed and the a-wave and b-wave amplitudes and latencies were similar for affected and age-matched nonaffected Great Pyrenees and other normal dogs. Electroretinograms that were examined twice during a 3-year period on three affected adult dogs did not reveal significant progressive deterioration of the a or b-wave parameters. Fluorescein angiography was completed on four affected dogs of ages 1 (n = 2), 5, and 6 years. These angiograms were repeated in three of these dogs 1 year later. The blood ocular barrier was intact in these dogs but there was blocked choroidal fluorescence. Postmortem examination, light microscopy, scanning and transmission electron microscopy were performed on three affected puppies and two affected adult dogs. These examinations revealed that the lesions in the puppies were limited to bilateral multiple areas of retinal pigment epithelial vacuolation, hypertrophy, and apparent separation from Bruch's membrane, and multiple serous retinal detachments. The affected adult dogs had focal retinal degeneration and retinal pigment epithelial hypertrophy, hyperplasia and pigmentation. Pedigree analysis and test mating confirm that this condition is inherited, probably as an autosomal recessive trait. This condition develops at approximately 13 weeks of age and the focal areas of retinal detachment and retinal pigment epithelial vacuolation progress to permanent and stable focal areas of retinal degeneration, and retinal pigment epithelial hypertrophy and pigmentation.     

Immunohistochemical study of caveolin-1 and -2 in the rat retina

The expression of caveolin-1 and -2 in the retina was examined; Western blot analysis showed that both were present. Immunohistochemistry indicated that caveolin-1 was expressed in the majority of retinal layers, including the ganglion cell layer, inner plexiform layer, outer plexiform layer, and in the vascular endothelial cells of the retina. Caveolin-2 was primarily immunostained in the vessels, but in a few other elements as well. This is the first demonstration of caveolin differential expression in the retina of rats, and suggests that caveolin plays an important role in signal transduction in glial cells and neuronal cells.     

Angiostatin and integrin alphavbeta3 in the feline, bovine, canine, equine, porcine and murine retina and cornea

PURPOSE: Angiogenesis is tightly controlled in the ocular tissues of domestic animals but its mechanisms are not fully understood. This is largely because of insufficient data on the expression of molecules that impact angiogenesis. Because angiostatin and one of its receptors integrin alphavbeta3 inhibit and promote angiogenesis, respectively, we hypothesized that the normal retina and cornea of domestic animals would express angiostatin but not integrin alphavbeta3. PROCEDURE: Normal eyes of the cat, cow, dog, horse, pig and rat were evaluated for angiostatin and integrin alphavbeta3 by light and electron immunocytochemistry and estern blots. RESULTS: Angiostatin was detected in the corneal epithelium of the cat, dog, horse, pig and rat, but was not found in cow corneal epithelium. Angiostatin was localized in the nerve fiber layer, ganglion cell layer, inner and outer plexiform layers, and the photoreceptor layer of the cat, cow, dog and rat. Horse and pig retinas showed additional staining in the matrix of the inner nuclear layer. Immunogold electron microscopy further confirmed angiostatin in cat retina. Western blots showed angiostatin in corneal and retinal homogenates. Integrin alphavbeta3 was absent in cornea and retina of all the species studied. CONCLUSION: These data show that angiostatin, an inhibitor of angiogenesis, is present while integrin alphavbeta3, which promotes angiogenesis, is absent in normal cornea and retina of the domestic animals in this study with the exception being angiostatin absence in cow corneal epithelium. Therefore, angiostatin may contribute to the anti-angiogenic environment in the normal domestic animal eye while its absence in the cow may contribute to greater propensity for corneal vascularization. Because integrin alphavbeta3 is one of the receptors for angiostatin, its absence may prevent angiostatin from killing normal retinal and corneal cells.     

Abnormal prion accumulation associated with retinal pathology in experimentally inoculated scrapie-affected sheep

The purpose of this study was to characterize the patterns of PrP(Sc) immunoreactivity in the retinae of scrapie-affected sheep and to determine the extent of retinal pathology as indicated by glial fibrillary acidic protein immunoreactivity (GFAP-IR) of Müller glia. Sections from the retina of 13 experimentally inoculated scrapie-affected and 2 negative control sheep were examined with immunohistochemical staining for PrP(Sc), GFAP, and PrP(Sc)/GFAP double staining. GFAP-IR of Müller glia is suggestive of retinal pathology in the absence of morphologic abnormality detected by light microscopy. Sheep with the least amount of PrP(Sc) in the retina have multifocal punctate aggregates of prion staining in the outer half of the inner plexiform layer and rarely in the outer plexiform layer. In these retinae, GFAP-IR is not localized with prion accumulation, but rather is present in moderate numbers of Müller glia throughout the sections of retina examined. The majority of sheep with retinal accumulation of PrP(Sc) have intense, diffuse PrP(Sc) staining in both plexiform layers, with immunoreactivity in the cytoplasm of multiple ganglion cells and lesser amounts in the optic fiber layer and between nuclei in nuclear layers. This intense PrP(Sc) immunoreactivity is associated with diffuse, intense GFAP-IR that extends from the inner limiting membrane to the outer limiting membrane. This is the first report of a prion disease in a natural host that describes the accumulation of PrP(Sc) in retina associated with retinal pathology in the absence of overt morphologic changes indicative of retinal degeneration.     

Sarcocystis neurona retinochoroiditis in a sea otter (Enhydra lutris kenyoni)

Sarcocystis neurona is an important cause of fatal disease in sea otters in the USA. Encephalitis is the predominant lesion and parasites are confined to the central nervous system and muscles. Here we report retinochoroiditis in a sea otter (Enhydra lutris kenyoni) found dead on Copalis Beach, WA, USA. Salient lesions were confined to the brain and eye. Multifocal nonsuppurative meningoencephalitis was present in the cerebrum and cerebellum associated with S. neurona schizonts. The retina of one eye had a focus of inflammation that contained numerous S. neurona schizonts and merozoites. The focus extended from the retinal pigment epithelium inward through all layers of the retina, but inflammation was most concentrated at the inner surface of the tapetum and the outer retina. The inner and outer nuclear layers of the retina were disorganized and irregular at the site of inflammation. There was severe congestion and mild hemorrhage in the choroid, and mild hemorrhage into the vitreous body. Immunohistochemistry with S. neurona-specific polyclonal rabbit antibodies stained schizonts and merozoites. To our knowledge this is the first report of S. neurona-associated retinochoroiditis in any naturally infected animal.     

Nodavirus associated with pathological changes in adult spotted coralgroupers (Plectropomus maculatus) in Thailand with viral nervous necrosis

The present study characterized viral nervous necrosis in sea cage-reared adult spotted coralgroupers (Plectropomus maculatus). Histopathological study showed extensive vacuolation and neuronal necrosis of the olfactory bulb and the optic lobe of the forebrain and the inner and outer nuclear layer of retina. Mild necrosis was observed in the spinal cord. Homogeneous intranuclear inclusion bodies were noted in the hyperplastic and hypertrophic glandular epithelial cells of the swim bladder suggesting viral etiology. Etiological diagnosis of VNN was confirmed by RT-PCR, immunohistochemistry and in situ hybridization. The immunohistochemistry and in situ hybridization gave strongly positive staining in the same area of the infected cells of the brain, spinal cord and retina correlating with histopathological changes. No positive reaction was detectable in the affected gas glandular epithelium and other organs, confirming the consistent neurotropism of this nodavirus. Nodavirus was mainly detected in the olfactory bulb of the brain. The result suggests nasal transmission was the major route of infection.     

Distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens in nervous and non-nervous organs of European seabass (Dicentrarchus labrax) during the course of an experimental challenge

The distribution of red-spotted grouper nervous necrosis virus (RGNNV) antigens was examined by immunohistochemistry in the nervous and non-nervous organs of juvenile European seabass (Dicentrarchus labrax) during the course of an intramuscular infection. Histological changes resulting from the infection were evaluated from 3 days to 2 months post-infection. The specific antibody response was also studied 2 months post-challenge. Viral proteins were present throughout the experimental period in the retina (inner nuclear layer, ganglion layer, outer limiting membrane, and outer plexiform layer), brain (cerebellum and tectum opticum), and liver (hepatocytes and endothelial cells). These proteins were also observed in the renal tubular cells, white pulp of spleen, and in fibroblasts and cartilage of caudal fin. This is the first report of RGNNV proteins appearing in these organs, where the immunostaining was only detected at certain sampling times after the onset of mortality. Brain and retina of virus-exposed fish showed high levels of vacuolation, while accumulation of fat vacuoles was observed in the liver. RGNNV infection also induced a specific antibody response as measured by an ELISA. In summary, this is the first study demonstrating the presence of viral proteins in cells of caudal fin, kidney and spleen of European seabass.     

Retinal Photoreceptor Fine Structure in the Great Blue Heron (Ardea herodias)

The morphology of the retinal photoreceptors of the great blue heron (Ardea herodias) has been investigated by light and electron microscopy. They consist of rods, single cones and double (unequal) cones present in a ratio of about 2:1:1 respectively. The rods are slender elongated cells with outer segments that reach to the retinal epithelial (RPE) cells and are surrounded by pigment-rich apical processes of the RPE cells in the light-adapted state. The rod inner segment displays an ellipsoid of mitochondria, an hyperboloid of glycogen, much rough ER, numerous polysomes, Golgi zones and autophagic vacuoles. The rod nucleus is located deep in the outer nuclear layer and the rod synaptic pedicle displays both invaginated and superficial synaptic sites. Single cones display a slightly tapered outer segment, a large electron lucent oil droplet and an ellipsoid of mitochondria in the apex of the inner segment. Double cones consist of a long thin chief member which shows an electron dense oil droplet and a shorter, stouter accessory cone with no oil droplet but a paraboloid of glycogen below the ellipsoid. As in the single cone, polysomes, RER and Golgi zones are present in the myoid region of both members of the double cone. All photoreceptor types have a connecting cilium joining inner and outer segments. Near the external limiting membrane, the chief and accessory cones show intercellular junctions. All cone photoreceptors are relatively small in diameter and hence tightly packed. While rods are felt to undergo retinomotor movements, cones are felt to move minimally or not at all. Both single and double cones display several invaginated (ribbon) synapses as well as numerous superficial (conventional) synaptic sites.     

Microvasculature of the hamster eye: scanning electron microscopy of vascular corrosion casts

The architecture of the retina, choroid and anterior eye segment was investigated in 12 Syrian hamsters using scanning electron micrographs of methylmethacrylate corrosion casts. The hamster eye receives its primary blood supply from the long posterior ciliary artery, which directly enters the optic nerve head, and divides into three branches: the central retinal artery and medial and lateral long posterior ciliary arteries. In the retina the central retinal artery divides into six radiating branches. Retinal arterioles form a slender and long course to capillaries. Retinal capillaries are extremely thin in diameter and form a sparse capillary network. The choroid is supplied by the long posterior ciliary arteries. Choroidal arterioles exhibit a thick and short course to the choriocapillaris. The choriocapillaris is thick and sinusoid-like, forming a dense network in the choroid. The ciliary body, iris and bulbar conjunctiva are supplied by the anterior ciliary arteries, which are branches of the long posterior ciliary arteries. Capillaries supplying the anterior margin of the ciliary process are large in diameter with an irregular bore, forming a thoroughfare channel draining blood in the ciliary arterioles into the pars plana vessels. Blood from the retina is drained by the central retinal veins. Venules from the anterior eye segment empty into the vortex veins via the pars plana vessels. Venous blood from the choroid is drained only by vortex veins via the choroidal veins. The functional significance of the vascular architecture and species differences are discussed.     

Congenital ocular anomalies in purebred and crossbred Rocky and Kentucky Mountain horses in Canada

Multiple congenital ocular anomalies in purebred and crossbred Rocky and Kentucky Mountain horses in Canada are frequently diagnosed with biomicroscopic and indirect ophthalmoscopic examination. In order of frequency detected, these include temporal ciliary epithelial cysts; iridal hypoplasia; prominent corneas; focal temporal retinal degeneration related to ciliary cysts; and, rarely, retinal detachment. A pedigree analysis confirms a dominant mode of inheritance with incomplete penetrance and with a linkage to coat color.     

Experimental udder infection with Escherichia coli. 2. Morphological changes of the epithelium in the area of the glands of the milk cistern and the large milk ducts

Optical light, transmission, and scanning electron microscopy were used in investigations of epithelia in the glandular region of the milk cistern and greater lactiferous ducts and yielded the following findings, four and six hours from infection: degeneration and necrosis of epithelial cells, intraepithelial foreign cell infiltration (neutrophilic granulocytes, lymphocytes, macrophages), intra-epithelial oedema and locally delimited epithelial loss. The lesions differed by intensity in various regions, so that a distinction could be made among five froms of epithelial alterations. The epithelial cells revealed their capability of absorbing pathogenic material and storing it in cytoplasmic vacuoles. This was considered to reflect active involvement of epithelium in antibacterial defence, and consequently, to reflect the role played by epithelium as a defence barrier in the milk cistern and greater lactiferous ducts.     

Drusen and lipid-filled retinal pigment epithelium cells in a rhesus macula

A middle-aged rhesus monkey with detailed clinical history exhibited progression of a macular abnormality with a variety of clinical drusen and pigment changes typical of the Cayo Santiago phenotype. Numerous frozen sections of one sample of the macular retina/retinal pigment epithelium (RPE)/choroid showed a single classical druse but extensive single and clustered lipid-filled RPE cells. The monkey exhibited functional outer retinal decline and an insignificant number of 'window defects' as found among signs of relatively benign human macular aging. The clinical and histologically defined results agree if lipid-filled RPE cells are included among the clinically apparent signs of drusen.     

An ultrastructural study of the equine lower respiratory tract

The surface features of the lower respiratory tract of 20 clinically normal horses of different ages and types were studied with scanning electron microscopy. Parallel light microscopical and transmission electron microscopical studies were also carried out. The ciliary carpet was virtually complete from the trachea to the lobar bronchi. In small bronchi, ciliation was less complete allowing numerous non-ciliated mucous cells to become obvious. The terminal bronchioles, populated mainly by non-ciliated bronchiolar epithelial cells, had an abrupt junction with alveolar ducts. Interalveolar pores were common particularly in older horses.