Cellular Characterization of OCT and Outer Retinal Bands Using Specific Immunohistochemistry Markers and Clinical Implications
Cellular Characterization of OCT and Outer Retinal Bands Using Specific Immunohistochemistry Markers and Clinical Implications
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DOI：
10.1016/j.ophtha.2017.09.016
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发表时间：
2018-03-01
期刊：
影响因子：
13.7
通讯作者：
Pinilla, Isabel
中科院分区：
文献类型：
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作者：
Cuenca, Nicolas;Ortuno-Lizaran, Isabel;Pinilla, Isabel
目的：光学相干断层扫描（OCT）在许多眼科疾病的诊断、治疗和随访方面取得了技术突破，特别是视网膜和神经眼科病理状况。到目前为止，在OCT中观察到的条带所代表的特定细胞类型上出现了一些争议，特别是在4个外视网膜条带上。设计：使用人视网膜切片和免疫细胞化学在中央凹水平将OCT中观察到的4个外高反射条带与组织学结构相关联。参与者：来自人类供体的眼睛。人视网膜的垂直冷冻切片用针对视锥细胞、双极细胞、线粒体、Muller细胞和视网膜色素上皮（RPE）细胞的特异性抗体进行免疫染色，并使用共聚焦显微镜观察。组织学和OCT之间的形态学相关性在fovealevel.Results：三重免疫标记允许区分细胞类型和不同的细胞室。鸟嘌呤核苷酸结合蛋白β 3（GNB3）和细胞视网膜色素结合蛋白（CRALBP）抗体的免疫染色显示，所有视网膜层在小凹，特别是外核层和Henle纤维层之间的分离。CRALBP和细胞色素C（Cyt C）的免疫标记显示，超反射带1和2，在OCT中观察到，分别对应于外界膜和锥体椭圆体，由锥体肌球分开。CRALBP、细胞色素C和GNB 3的研究表明，RPE的交错排列沿着视锥细胞的整个外节延伸，我们不认为它们是形成第三条带的结构。然而，在RPE内的视锥细胞外节的小片段的鉴定使我们将第三条带表征为位于RPE顶部的视锥细胞吞噬体。最后，我们提出第四条带对应于线粒体在RPE基底部分的积累，如细胞色素C免疫反应性所确定的，并且第3条带和第4条带之间的低反射带对应于RPE核和黑素体区。这项研究提出了一种新的解释外视网膜带，导致更准确的解释OCT图像，提供有关视锥细胞健康及其与RPE关系的信息，并有助于更好地了解视网膜疾病的诊断和预后。(C)2017年美国眼科学会
Purpose: OCT has been a technological breakthrough in the diagnosis, treatment, and follow-up of many ocular diseases, especially retinal and neuro-ophthalmologic pathologic conditions. Until now, several controversies have arisen over the specific cell types that the bands observed in the OCT represent, especially over the 4 outer retinal bands.Design: To correlate the 4 outer hyperreflective bands observed in the OCT with the histologic structures using human retinal sections and immunocytochemistry at the fovea level.Participants: Eyes from human donors.Methods: Vertical cryosections of human retinas were immunostained with antibodies specific for cones photoreceptors, bipolar cells, mitochondria, Muller cells, and retinal pigment epithelium (RPE) cells and were visualized using confocal microscopy.Main Outcome Measures: Morphological correlation between histology and OCT at the fovea level.Results: Triple immunolabeling allowed distinguishing between cells types and different cell compartments. Immunostaining with guanine nucleotide-binding protein beta 3 (GNB3) and cellular retinaldehyde-binding protein (CRALBP) antibodies showed all retinal layers at the foveola, especially the separation between the outer nuclear layer and the Henle fiber layer. CRALBP and cytochrome C (Cyt C) immunolabeling revealed that hyperreflective bands 1 and 2, observed in the OCT, correspond to the outer limiting membrane and the cone ellipsoids, respectively, separated by the cone myoids. CRALBP, cytochrome C, and GNB3 showed that the RPE interdigitations extend along the entire external segment of the cones, we do not believe them to be the structure responsible for forming the third band. However, the identification of small fragments of cone outer segments within the RPE led us to characterize the third band as the cone phagosomes located in the top of the RPE. Finally, we propose that the fourth band corresponds to the accumulation of mitochondria at the basal portion of the RPE, as identified by cytochrome C immunoreactivity, and that the hyporeflective band between bands 3 and 4 corresponds to the RPE nuclei and melanosomes zone.Conclusions: This study proposes a new interpretation of the outer retinal bands that leads to a more accurate interpretation of OCT images, providing information about the health of cones and their relationship with the RPE, and could help to form a better understanding of retinal disease diagnosis and prognosis. (C) 2017 by the American Academy of Ophthalmology