Ocular albinism and hypopigmentation defects in Slc24a5-/- mice

As part of a high-throughput mutagenesis and phenotyping process designed to discover novel drug targets, we generated and characterized mice with a targeted mutation in Slc24a5, a gene encoding a putative cation exchanger. Upon macroscopic examination, Slc24a5-/- mice were viable, fertile, and indistinguishable by coat color from their heterozygous and wild-type litter mates. Ophthalmoscopic examination revealed diffuse retinal hypopigmentation, and a histologic examination of the eye confirmed the presence of moderate-to-marked hypopigmentation of the retinal pigmented epithelium (RPE), ciliary body, and iris pigment epithelium (IPE). Hypopigmentation was most severe in the anterior layer cells of the IPE, where melanosomes were smaller, paler, and more indistinct than those of the anterior stroma and posterior IPE. The pigment granules of the posterior IPE appeared to be nearly as dark as those in stromal melanocytes; however, both cell layers were thinner and paler than corresponding layers in wild-type mice. Ultrastructural analysis of the RPE, IPE, and ciliary body pigmented cells confirmed that mutation of Slc24a5 results in marked hypopigmentation of melanosomes in optic cup-derived pigmented neuroepithelium in the eyes. Milder reductions in melanosome size and pigmentation were noted in neural crest-derived melanocytes. The severe hypopigmentation of neuroepithelium-derived cells in the eyes resulted in a novel form of ocular albinism in Slc24a5-/- mice. Our findings suggest that SLC24A5 may be a candidate gene for some forms of ocular albinism and for the BEY1/EYCL2 locus previously associated with central brown eye color in humans.     

Management of intraocular inflammatory disease

The uvea of the eye is divided into the anterior uvea (iris and ciliary body) and posterior uvea (choroid). Clinical signs of anterior uveitis include conjunctival hyperemia, corneal edema, aqueous flare, miosis, and a decrease in intraocular pressure. Inflammation of the posterior uvea often involves the retina as well resulting in a chorioretinitis. Clinical signs of chorioretinitis may include multifocal lesions in tapetal or nontapetal fundus, retinal detachment, and a decrease in vision. The etiology of uveitis is complex and includes numerous infectious, neoplastic, immune-mediated, and other diseases. Treatment is directed at the underlying systemic disease, if present, as well as symptomatic treatment for the eye. The diagnostic and therapeutic approach to a patient with uveitis is discussed.     

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.     

A novel immunohistochemical marker of normal and neoplastic melanocytes in formalin-fixed, paraffin-embedded tissues of albino rats

In albino rats, spontaneous occurrence of melanocytic tumors is rare, with diagnosis difficult. This study evaluated immunoreactivity for PNL2 in normal and neoplastic melanocytes in formalin-fixed and paraffin-embedded tissues of albino rats. The samples consisted of 11 (1.57%) amelanotic melanomas in 700 rats (2 studies), 23 non-melanocytic tumors, and a wide variety of normal tissues. In normal albino rats, PNL2 stained the melanocytes in the iris and choroid of the eyeball and the hair bulb and basal cell layers of the epidermis of the whole body. In amelanotic melanoma, the tumor cells consisted of spindle cells with eosinophilic cytoplasm without melanin granules. PNL2 consistently stained cytoplasm in all amelanotic melanoma cells. In contrast, the nonmelanocytic tumor cells were not labeled. Electron microscopically, neoplastic, and normal melanocytes showed numerous cytoplasmic premelanosomes (stage II melanosome). In conclusion, PNL2 is direct against a fixative- and decalcific-resistant melanocyte-associated antigen, and has high specificity against normal and neoplastic melanocytes of albino rats.     

Immunohistochemical investigation of the distribution of immunoglobulins G, A and M within the anterior uvea of the normal equine eye

Distribution of the immunoglobulin (Ig) classes G, A and M within the anterior uvea of eight clinically normal equine eyes was examined using indirect immunoperoxidase labelling. Increased staining intensity of stromal IgG and IgA was observed within the ciliary processes, the iris stroma being relatively free of immunoglobulin. This may reflect anatomical variation in the permeability of the uveal microvasculature to lipid insoluble plasma macro-molecules. Intracellular IgG and IgA were observed within the non-pigmented ciliary epithelium in seven and four of the eight eyes respectively, although wide variation between numbers of Ig bearing cells in each eye was noted. It was suggested that these cells may have a role in the removal of Ig from the posterior chamber aqueous. Plasma cells, of IgG isotype, were observed in only one eye, suggesting that intraocular production of Ig is not a feature of the normal equine eye.     

Microvascular anatomy of the pig eye: scanning electron microscopy of vascular corrosion casts

The microvasculature of the eye of 10 pigs was investigated using scanning electron micrographs of corrosion casts. The ciliary body, iris and bulbar conjunctiva were supplied by the iridociliary ring artery via the long posterior ciliary artery. Capillaries of the ciliary process were of large diameter (23.2-27.5 microm) with an irregular bore, forming a thoroughfare channel draining blood in the ciliary arterioles into the pars plana venous vessels. Arterioles and venules in the iris exhibited a zigzag or spiral features. The third palpebra was supplied by the anterior ciliary artery. The capillary bed of the third palpebra was dense and was formed by many rows of fine hair-pin loops. Capillaries in the bulbar conjunctiva formed a sparse network disposing approximately parallel to the epithelium and formed a well-developed venous plexus, draining into the vortex veins. Retinal arterioles formed a slender and long course to capillaries. Retinal capillaries were extremely thin (3.0-4.0 microm in diameter). The choroid was supplied by the short posterior ciliary arteries. Choroidal arterioles exhibited a thick and short course to the choriocapillaris. The choriocapillaris was flat and sinusoid-like (8.9-13.9 microm in diameter), forming a dense sheet-like network. Blood from the choroid emptied into the episcleral vein via the vortex vein. Blood from the retina was drained by the posterior ciliary veins. The functional significance of this vascular architecture was discussed.     

Microvascular architecture of the rabbit eye: a scanning electron microscopic study of vascular corrosion casts

The microvasculature of the eyes of 5 rabbits was investigated using scanning electron microscopy on corrosion casts. The study revealed that the pars plana vessels draining blood from the iris and ciliary body coursed directly into the anterior vortex venous system constituting the scleral venous plexus (the venous circle of Hovius). The episcleral vasculature was found to possess a specialized morphology, with channels draining the aqueous humor. The capillaries of the third palpebral, bulbar and palpebral conjunctiva formed a single-layered capillary network approximately parallel to the epithelium and formed a well-developed venous plexus in the stroma. The retina was found to be merangiotic, meaning that vessels were present only in a small part of the retina, extending in a horizontal direction to form bands on either side of the optic disc. Channels representing the aqueous veins that drained blood mixed with aqueous humor were found to derive directly from the suprachoroidal space and communicate with the scleral venous plexus via the anterior vortex veins. The functional significance of the microvasculature of the iris, cilia, retina and choroid is discussed in this report as well. The elaborate microvasculature of the conjunctiva may be a prerequisite for the exchange of nutrients and gasses between the cornea and the vessels across the conjunctival epithelium when the eyelids are shut during sleep, and possibly for the dynamics of eye drop delivery. The scleral venous plexus in rabbits may be analogous to the scleral venous sinus (Schlemm's canal) in rats, primates and humans.     

Ultrastructural ocular lesions of 6-aminonicotinamide toxicosis in rabbits

6-Aminonicotinamide, given by intraperitoneal injection to male and female Dutch belted rabbits, produced swelling and vacuolation of ciliary and iridal epithelium plus vacuolation of the retinal pigment epithelial and outer plexiform layers of the retina. By transmission electron microscopy, inner and outer ciliary epithelial cells and inner iridal epithelial cells contained numerous coalescing, membrane-bound vacuoles of the cytocavitary network. These vacuoles were viewed as numerous interconnecting, intracytoplasmic cavities in scanning electron micrographs. Swelling of vacuolated epithelial cells and the presence of fibrin and proteinaceous fluid in the ciliary stroma resulted in thickening of the anterior ciliary processes with the formation of surface alterations detectable by scanning electron microscopy. In transmission electron micrographs the vacuoles in the retinal pigment epithelium were large, electron-lucent spaces and the vacuoles in the outer plexiform layer of the retina appeared to be intracytoplasmic spaces in axons of photoreceptor cells. Distention of cytocavitary structures has been reported in glial cells of animals given 6-aminonicotinamide and this change was apparently due to alterations in ion and water movement across cellular membranes that resulted in intracellular edema.     

Ocular lesions in Watanabe heritable hyperlipidemic rabbits

Hyperlipidemic ocular lesions are described for Watanabe heritable hyperlipidemic (WHHL) rabbits. Male WHHL rabbits 8 months old exhibited serum hyperlipidemia and ophthalmoscopically yellowish-white lesions along the corneoscleral junction and in the iris. Histopathologically, foamy macrophages aggregated in the stroma of the cornea, iris, and ciliary body were observed. These findings have been interpreted as lipid keratopathy. In addition, multiple clusters of a large number of foamy macrophages occurred throughout the choroid and sclera in association with the blood vessels. The lesions in the choroid and sclera could not be detected ophthalmoscopy, yet were much more prominent than those in the cornea, iris, and ciliary body, suggesting greater involvement and earlier onset of lipidosis at these sites associated with hyperlipidemia in WHHL rabbits.     

'D-shaped' and 'reverse-D-shaped' pupil in a cat with lymphosarcoma

A 7-year-old female spayed European shorthair cat was presented because of visual reduction and a green reflex in the right eye. Examination revealed a 'D-shaped' pupil in the right eye and a 'reverse-D-shaped' pupil in the left eye. Both eyes showed fibrinous anterior uveitis with exudates in the anterior chamber and precipitates on the posterior surface of the cornea. In both eyes a chorioretinitis was diagnosed which caused partial retinal detachment and peripapillary oedema in the left eye. Histopathology revealed intestinal and ocular lymphosarcoma. The aberrant pupil shapes which persisted after death were due to infiltration of the iris stroma and were not caused neurologically. The observation that in both eyes only the medial parts of the iris were affected could not be explained.     

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

Methylmethacrylate castings of the eye microvasculature were prepared from 10 Spargue-Dawley rats and studied by electron microscopy. The choroidal arterioles are larger in diameter than retinal arterioles, and have a shorter course to choroidal capillaries than retinal arterioles to retinal capillaries. Retinal capillaries are extremely thin in diameter and form a sparse retinal capillary network. The choriocapillaris is large and sinusoid-like, forming a compactly arranged network in the choroid. These differences in the microvasculature between the choroid and retina help explain the differences in ocular hemodynamics; that is, the blood flow in the choroid is faster than that in the retina. Capillaries of the iris show a zigzag configuration, which may be an accommodation for dilation and constriction of the pupil. Capillaries of the ciliary body are of large diameter forming leaf-like configurations, presumably to contribute to the secretion of the aqueous humor. Capillaries of the conjunctiva exhibit a somewhat coiled configuration, the arrangement of which reduce tension of the conjunctiva vessels caused by eyeball movement. Intra-arterial cushions, which control blood flow at the branching sites, are found in both choroidal or retinal arterioles.     

Comparative ocular anatomy of the western lowland gorilla

OBJECTIVE: To examine the lowland gorilla (Gorilla gorilla gorilla) eye and determine similarities to and differences between the mountain gorilla (Gorilla gorilla beringei) and the human eye. In addition, we compare our findings of G. g. gorilla to previous reports on the eye of this subspecies. PROCEDURES: A 13-year-old deceased male lowland gorilla and a 34-year-old deceased female lowland gorilla were included in the study. Gross and microscopic examinations of the formalin-fixed right eyeball of each gorilla were carried out. RESULTS: Globe dimensions of G. g. gorilla were similar to G. g. beringei and to humans. The limbal conjunctival epithelium and the choroid were densely pigmented. However, the distribution of the conjunctival pigment ring was different to that of G. g. beringei and the melanocytes of the choroid were unusually round. There were deep crypts in the anterior border layer of the iris, and the epithelium of the pars plana was uniquely irregular. Vertical corneal diameter was observed to be equal or greater than horizontal diameter in G. g. gorilla, which is in contrast to humans and to previous findings for G. g. beringei. Corneal thickness was closer to that of humans than to G. g. beringei. Posterior lens capsule thickness was noticeably greater than that of humans. CONCLUSIONS: Although some variation between the ocular anatomy of G. g. gorilla and G. g. beringei does exist, the gross and microscopic findings closely resemble each other in these two subspecies. In addition, the eye of Gorilla appears remarkably similar to the human eye. However, comparison of measurements with those in humans is somewhat limited because formalin-fixation can introduce tissue shrinkage and artifact.     

Characteristics of structures and lesions of the eye in laboratory animals used in toxicity studies

Histopathology of the eye is an essential part of ocular toxicity evaluation. There are structural variations of the eye among several laboratory animals commonly used in toxicity studies, and many cases of ocular lesions in these animals are related to anatomical and physiological characteristics of the eye. Since albino rats have no melanin in the eye, findings of the fundus can be observed clearly by ophthalmoscopy. Retinal atrophy is observed as a hyper-reflective lesion in the fundus and is usually observed as degeneration of the retina in histopathology. Albino rats are sensitive to light, and light-induced retinal degeneration is commonly observed because there is no melanin in the eye. Therefore, it is important to differentiate the causes of retinal degeneration because the lesion occurs spontaneously and is induced by several drugs or by lighting. In dogs, the tapetum lucidum, a multilayered reflective tissue of the choroid, is one of unique structures of the eye. Since tapetal cells contain reflecting crystals in which a high level of zinc has been demonstrated chemically, drug-induced tapetum degeneration is possibly related to zinc chelation. The eye of the monkey has a macula similar to that of humans. The macula consists only of cones with a high density, and light falls directly on the macula that plays an important role in visual acuity. Macular degeneration occurring in monkeys resembles histopathologically that of humans. Hence, the eye of the monkey is a suitable model to investigate macular degeneration and to assess drug-induced macular lesions.     

Ocular manifestations of a metastatic pulmonary adenocarcinoma in a cat

A primary pulmonary bronchogenic adenocarcinoma originating from an intramediastinal accessory lung was diagnosed in a 14.5-year-old cat. The cat had been admitted because of a cloudy right eye. Physical examination revealed a thin cat with severe iritis, aqueous flare, and a fibrin clot in the anterior chamber of the right eye. Right fundic examination revealed bullous retinal detachment superior to the optic disc. Euthanasia and necropsy were requested when FeLV test results were positive. Metastatic neoplastic cells similar to those of the primary tumor were detected in the choroid, ciliary body, and ciliary processes of the right eye. Ciliary and iridic stromal necrosis attributable to neoplastic embolization of uveal vessels had led to severe uveitis. Foci of metastasis were also in the heart, kidney, and cerebral meninges.     

Spontaneous amelanotic melanomas of the uveal tract in F344 rats.

Five intraocular amelanotic melanomas were identified in the National Toxicology Program's database consisting of records from more than 60,000 female and 60,000 male F344 rats, which were used as control and treated animals in 2-year carcinogenicity studies. The five spontaneous melanomas were grossly observed as white or yellow, unilateral nodules, which originated in the region of the iris and ciliary body, often also involving the choroid. These amelanotic melanomas were composed predominantly of spindle cells arranged in a whorled pattern often with perivascular orientation. Mitotic figures were common in five tumors. The spindle cells had a positive immunoreactivity for S-100 protein but were negative for desmin. Electron microscopic studies provided clear evidence that these tumors originated from the uveal melanocytes. Ultrastructurally, the spindle cells contained numerous cytoplasmic premelanosomes (stage II melanosomes) that were not associated with melanin. Special histochemical studies showed that the spindle cells had a negative reaction for melanin. Although electron microscopic features are critical in the diagnosis of amelanotic melanomas of the uveal tract, the whorled pattern of spindle cells is a useful histologic criterion in differential diagnosis of this tumor in F344 rats.     

成黑色素细胞生成过程中信号调节的研究进展

动物体内的黑色素细胞合成了黑色素从而决定其肤色和毛色。全身各处的黑色素细胞有2种来源:①由神经管上皮细胞分化而来的视网膜色素上皮细胞;②由神经嵴细胞在胚胎发育早期分化为成黑色素细胞进而发育成熟为黑色素细胞从而行使其功能。通过对小鼠和人类的发育遗传学研究,结果表明神经嵴细胞衍生的黑色素细胞的发育分化是一个非常复杂的过程,受到多重信号因子的调控。其中影响黑色素细胞发育过程的转录因子有SOX10、MITF和PAX3,信号通路有KIT及其配体KITL信号通路、WNT/β-catenin信号通路、EDN3及其受体EDNRB信号通路。MITF被认为是黑色素细胞发育过程中非常关键的调控因子,而3条通路也被认为与神经嵴细胞来源的黑色素细胞的发育有极为密切的关系且能调节MITF功能及其活性。作者总结了这个领域的研究进展并指出早期神经嵴细胞的发育调控可能是乌骨鸡和乌骨绵羊乌质性状形成的根本原因。     

Histiocytic chorioretinitis in a dog

A 3‐year‐old castrated male mixed‐breed dog presented with an acute bullous retinal detachment and thickened choroid of the right eye. Subretinal cytology revealed an atypical cell proliferation suggestive of neoplasia. The eye was enucleated, and the original diagnosis was a histologically benign choroidal melanocytic tumor. Further diagnostics revealed no other systemic abnormalities other than a nonhealing shoulder wound. Six months later, the left eye developed a bullous retinal detachment. This eye responded well to systemic steroids and the dog regained vision within a few weeks of initiating therapy. Results of immunohistochemistry with Melan‐A and CD204 of the previously enucleated right eye caused a revision of the histologic diagnosis from melanocytic tumor to histiocytic chorioretinitis. This case highlights the subtle and sometimes confusing distinction between neoplastic and inflammatory processes on both cytology and histopathology.