Zebrafish (Danio rerio) provide many advantages as a model organism for studying ocular disease and development, and there is great interest in the ability to non-invasively assess their photoreceptor mosaic. Despite recent applications of scanning light ophthalmoscopy, fundus photography, and gonioscopy to in vivo imaging of the adult zebrafish eye, current techniques either lack accurate scaling information (limiting quantitative analyses) or require euthanizing the fish (precluding longitudinal analyses). Here we describe improved methods for imaging the adult zebrafish retina using spectral domain optical coherence tomography (OCT). Transgenic fli1:eGFP zebrafish were imaged using the Bioptigen Envisu R2200 broadband source OCT with a 12-mm telecentric probe to measure axial length and a mouse retina probe to acquire retinal volume scans subtending 1.2 × 1.2 mm nominally. En face summed volume projections were generated from the volume scans using custom software that allows the user to create contours tailored to specific retinal layer(s) of interest. Following imaging, the eyes were dissected for ex vivo fluorescence microscopy, and measurements of blood vessel branch points were compared to those made from the en face OCT images to determine the OCT lateral scale as a function of axial length. Using this scaling model, we imaged the photoreceptor layer of five wild-type zebrafish and quantified the density and packing geometry of the UV cone submosaic. Our in vivo cone density measurements agreed with measurements from previously published histology values. The method presented here allows accurate, quantitative assessment of cone structure in vivo and will be useful for longitudinal studies of the zebrafish cone mosaics.

Previous studies on fish retina have suggested that a curved, non-fused embryonic fissure is associated with, and perhaps caused by, asymmetric growth along the retina's marginal germinal zone (where neurons and Miiller glia are added appositionally throughout life). In this report retinal growth was measured directly in adult green sunfish (Lepomis cyanellus), which has a curved, non-fused embryonic fissure. Growth was asymmetric in both small and large fish: ventral and nasal retina grew more than temporal and dorsal retina. This asymmetry was due to different net rates of cellular addition, rather than differential passive expansion. The absolute rates of retinal growth in the centroperipheral direction were roughly exponential functions of fish size—smaller fish grow faster than large fish—but the area of new retina added per unit time did not vary with fish size. Visual implications of asymmetric retinal growth are evaluated.

The present study examines the changes in ultraviolet (UV) photosensitivity that occur during the growth of rainbow trout (Salmo gairdneri). A comparison of the ocular media transmission of small (n = 3) and large (n = 3) trout eyes did not reveal large changes in the transmission of UV radiation through the eye. We used the heart-rate conditioning technique to measure spectral sensitivity in immobilized trout. Four trout, each weighing less than 30 g, exhibited a UV-sensitivity peak at 360 nm while four additional trout weighing more than 60 g each exhibited no evidence of UV sensitivity. Spectral-sensitivity measurements of two trout weighing 44 g and 60 g revealed UV sensitivity, but when measured one month later (after a 25% increase in body weight) both fish exhibited no UV-sensitivity peak. At this time their sensitivity appeared to conform to the known blue-sensitive cone mechanism.

The presence, density distribution, and mosaic regularity of cone types were studied in the retina of the diurnal agouti, Dasyprocta aguti. Longwave-sensitive (L-) and shortwave-sensitive (S-) cones were detected by antibodies against the respective cone opsins. L- and S-cones were found to represent around 90 and 10% of the cone population, respectively. There was no evidence for L- and S-opsin coexpression in agouti cones. L-cone densities were highest, up to 14,000/mm2, along a horizontal visual streak located about 2–3 mm dorsal to the optic nerve, and the L-cone distribution showed a dorsoventral asymmetry with higher densities in ventral (about 10,000/mm2) than in dorsal (about 4000/mm2) retinal regions. This L-cone topography parallels the agouti’s ganglion cell topography. S-cones had a peak density of 1500–2000/mm2 in the central retinal region but did not form a visual streak. Their distribution also showed a dorsoventral asymmetry with densities around 600/mm2 in dorsal and around 1000/mm2 in ventral retinal regions. The patterning of cone arrays was assessed by the density recovery profile analysis. At all eccentricities evaluated, the S-cone mosaic less efficiently packed than the L-cone mosaic. Rod densities ranged from 47,000/mm2 in peripheral to 64,000/mm2 in central retina, and rod:cone ratios were 4:1–9:1. The comparatively low rod density and high cone proportion appear well adapted to the diurnal lifestyle of the agouti.

The retina of salmonid fishes has two types of cone photoreceptors: single and double cones. At the nuclear level, these cones are distributed in a square mosaic such that the double cones form the sides of the square and the single cones occupy positions at the centre and at the corners of the square. Double cones consist of two members, one having visual pigment protein maximally sensitive to green light (RH2 opsin), the other maximally sensitive to red light (LWS opsin). Single cones can have opsins maximally sensitive to ultraviolet (UV) or blue light (SWS1 and SWS2 opsins, respectively). In Pacific salmonids, all single cones express UV opsin at hatching. Around the time of yolk sac absorption, single cones start switching opsin expression from UV to blue, in an event that proceeds from the ventral to the dorsal retina. This transformation is accompanied by a loss of single corner cones such that the large juvenile shows corner cones and UV opsin expression in the dorsal retina only. Previous research has shown that adult Pacific salmon have corner cones over large areas of retina suggesting that these cones may be regenerated and that they may express UV opsin. Here we used in-situ hybridization with cRNA probes and RT-PCR to show that: (1) all single cones in non-growth zone areas of the retina express blue opsin and (2) double cone opsin expression alternates around the square mosaic unit. Our results indicate that single cone driven UV sensitivity in adult salmon must emanate from stimulation of growth zone areas.