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Cone Photoreceptor Recovery after Experimental Detachment and Reattachment: An Immunocytochemical, Morphological, and Electrophysiological Study

¹From the Neuroscience Research Institute, the ⁴Departments of Molecular, Cellular, and Developmental Biology and ³Psychology, University of California, Santa Barbara; and the ²Department of Ophthalmology, Jikei University School of Medicine, Tokyo, Japan.

PURPOSE. To compare the morphologic and functional recovery of the retina after detachment and reattachment in an animal with a cone-dominant retina, the ground squirrel.

METHODS. Ground squirrel (Spermophilus beecheyi) retinas were detached for 1 day and reattached for 7, 35, or 96 days (n = 2, each time point). Flicker ERGs were recorded 1 day after the detachment and at various times after reattachment. Contrast-response functions were measured for isochromatic modulation and for selective modulation of short-wavelength–sensitive (S) and middle-wavelength–sensitive (M) cones. At the end of the experiment, retinas were prepared for light microscopy or immunocytochemical staining with antibodies to rod opsin, S and M cone opsins, cytochrome oxidase, synaptophysin, glial fibrillary acidic protein (GFAP), cellular retinaldehyde–binding protein (CRALBP), interphotoreceptor–binding protein (IRBP), and peanut agglutinin lectin (PNA). Photoreceptor density maps were created from wholemount preparations labeled with biotinylated PNA and anti–S cone opsin. Cell counts of photoreceptor nuclei and cone outer segments (OS) were compared with flicker ERG data. Cell death was examined by the TUNEL method.

RESULTS. Reattachment stopped photoreceptor cell death and reversed the disruption of interphotoreceptor matrix as well as the redistribution of Müller cell proteins. It also activated some astrocytes based on anti-GFAP staining. S- and M-cone OS showed a gradual recovery in length after reattachment, and this recovery continued to the longest time points examined. ERG contrast gains also recovered after reattachment, but these reached asymptotic levels by approximately a week after reattachment. There were significant correlations between outer nuclear layer (ONL) cell counts and ERG contrast gains. No differences were noted in the indices of recovery of M and S cones.

CONCLUSIONS. The ERG can be used to follow specifically the changes in the retina that occur after retinal detachment and reattachment.

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