Cone photoreceptor recovery after experimental detachment and reattachment: An immunocytochemical, morphological, and electrophysiological study
Cone photoreceptor recovery after experimental detachment and reattachment: An immunocytochemical, morphological, and electrophysiological study
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DOI：
10.1167/iovs.02-0633
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发表时间：
2003-01-01
影响因子：
4.4
通讯作者：
Jacobs, GH
中科院分区：
文献类型：
--
作者：
Sakai, T;Calderone, JB;Jacobs, GH
目的.目的比较视锥细胞占优势的黄鼠视网膜脱离和复位后视网膜形态和功能的恢复。地松鼠（Spermophilus beecbeyi）视网膜脱离1天，重新连接7，35，或96天（n 2，每个时间点）。分别在脱离后和再附着后的不同时间记录闪烁ERGS。对比度响应函数进行了测量等色调制和选择性调制的短波长敏感（S）和中波长敏感（M）锥。在实验结束时，制备视网膜用于光学显微镜检查或用针对视杆视蛋白、S和M视锥视蛋白、细胞色素氧化酶、突触素、胶质细胞酸性蛋白（GFAP）、细胞视网膜色素脱结合蛋白（CRALBP）、光感受器间结合蛋白（IRBP）和花生凝集素凝集素（PNA）的抗体进行免疫细胞化学染色。从用生物素化PNA和抗S视锥视蛋白标记的全样品制备物产生感光器密度图。光感受器核和锥体外节（OS）的细胞计数与闪烁ERG数据进行了比较。通过TUNEL法检测细胞死亡。重新附着阻止了光感受器细胞死亡，并逆转了光感受器间基质的破坏以及穆勒细胞蛋白质的重新分布。抗GFAP染色显示，它还激活了一些星形胶质细胞。S-和M-cone OS显示在重新附着后长度逐渐恢复，并且这种恢复持续到检查的最长时间点。视网膜电图对比度增益也恢复后，重新附着，但这些达到渐近水平，约一周后重新附着。外核层（ONL）细胞计数与ERG对比度增益之间存在显著相关性。M锥和S锥的恢复指数无差异。视网膜电图可用于跟踪视网膜脱离和复位后发生的视网膜变化。
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 beecbeyi) retinas were detached for I day and reattached for 7, 35, or 96 days (n 2, each time point). Flicker ERGS were recorded I (lay 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 Muller 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.