Study of the production, composition and outflow of intraocular media in standard and Black Moor goldfish (Carassius auratus)

The goldfish is extensively used in vision research due to several factors: it is a poikilothermic animal, which allows ex-vivo experiments using its retina for longer periods of time; its retinal organization is very similar to that of mammals; it has large neurons, which facilitates electrophysiological and morphological studies and, unlike mammals, it is endowed with constant neurogenesis of some retinal cell types. However, the production, composition and outflow of ocular media in this species, which are fundamental for image formation onto the retina, are poorly understood. The standard goldfish has normal eye development, while the Black Moor goldfish is extremely myopic due to excessive eye growth (buphthalmos), a condition that is present in certain types of mammalian glaucoma. The present study investigated morphophysiological aspects of the aqueous humor production and drainage system and vitreous humor composition in these two varieties of goldfish to better understand the causes underlying the eye growth of the Black Moor. To study the drainage of the aqueous humor, anterior chamber dye injections and histological processing were performed in animals of both varieties after predetermined time intervals (5, 15, 30, 60, 120 minutes and 1, 2, 3, 4, and 7 days, n <= 3 eyes of each variety for each experimental group). No differences were observed in the drainage of aqueous humor between the two varieties. In both, the aqueous humor is drained mainly by the trabecular meshwork contained in the ventral portion of the drainage angle, between the iris and the annular ligament. Two secondary drainage routes were found in the two goldfish varieties studied: (i) the iridic route and (ii) drainage through epiretinal vessels. These pathways are apparently slower than the trabecular route, since the dye was found in these regions 4 days after the injections. An ultrastructural study of the retina and ciliary epithelium was also carried out in both varieties through transmission electron microscopy. The retinal pigment epithelium of the Black Moor presented more scarce melanin granules and we observed some disorganization of the photoreceptor layer, as well as a reduction in its density. Through an invasive device constructed by Joselevitch and Smith (1997), the intraocular pressure (IOP) of 14 eyes of each variety was measured. The Black Moor presented slightly increased IOP values compared to the standard animal (3.6 ± 0.9 mmH2O for demekin and 1.9 ± 0.6 mmH2O for the standard goldfish). The protein composition of the vitreous body was studied by polyacrylamide gel electrophoresis using a vitreous humor pool of 4 animals of the standard variety and 2 of the Black Moor variety. Low molecular weight proteins (15 to 25 kDa) were expressed in both - although with greater expression in the standard variety - and others of higher molecular mass, expressed only in the vitreous humor of the Black Moor (50-75 kDa and 150 kDa). These results suggest, therefore, that there are important differences in the retinal ultrastructure, the protein composition of the vitreous humor and the intraocular pressure of the Black Moor. To better understand the ocular growth in this variety, it is necessary to carry out a proteomic study, identifying which are the vitreous humor proteins evidenced through electrophoresis, as well as morphological studies in animals of different sizes aiming the quantification of the retinal damages caused by its ocular growth (AU)