Fundus Autofluorescence and Photoreceptor Cell Rosettes in Mouse Models
Fundus Autofluorescence and Photoreceptor Cell Rosettes in Mouse Models
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DOI：
10.1167/iovs.14-14136
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发表时间：
2014-09-01
影响因子：
4.4
通讯作者：
Sparrow, Janet R.
中科院分区：
文献类型：
--
作者：
Flynn, Erin;Ueda, Keiko;Sparrow, Janet R.
目的.本研究旨在探讨眼底自发荧光（autofluorescence，AF）、视网膜色素上皮（RPE）脂褐素积聚和感光细胞变性之间的相关性，并探讨眼底AF斑的结构基础。使用共焦扫描激光检眼镜（cSLO）在有色Rdh 8（-/-）/Abca 4（-/-）小鼠（1-9月龄）中采集眼底AF图像（558透镜; 488 nm激发）和光谱域光学相干断层扫描（SD-OCT）扫描。对于定量眼底AF（qAF），将灰度级（GL）校准为内部荧光参考。通过定量HPLC测量视网膜双维甲酸。进行外核层（ONL）厚度的组织计量学分析，并通过荧光显微镜检查视网膜的冷冻切片。Rdh 8（-/-）/Abca 4（/）小鼠的量化A2 E和qAF强度增加，直至4个月大。A2 E水平在4月龄后下降，但qAF强度值继续上升。Rdh 8（-/-）/Abca 4（-/-）小鼠中A2 E水平的下降导致感光细胞活力降低，如ONL变薄所反映的。眼底AF图像中的超自发荧光斑点对应于SD-OCT图像和苏木精和伊红染色的组织切片中的感光细胞凋亡。通过荧光显微镜检测到玫瑰花结的内节/外节核心发出自发荧光。当神经视网膜病变时，来自感光细胞的AF可以有助于无创性眼底AF图像。在眼底AF图像中的超自发荧光斑点至少在某些情况下可归因于感光细胞凋亡。
PURPOSE. This study was conducted to study correlations among fundus autofluorescence (AF), RPE lipofuscin accumulation, and photoreceptor cell degeneration and to investigate the structural basis of fundus AF spots.METHODS. Fundus AF images (558 lens; 488-nm excitation) and spectral-domain optical coherence tomography (SD-OCT) scans were acquired in pigmented Rdh8(-/-) / Abca4(-/-) mice (ages 1-9 months) with a confocal scanning laser ophthalmoscope (cSLO). For quantitative fundus AF (qAF), gray levels (GLs) were calibrated to an internal fluorescence reference. Retinal bisretinoids were measured by quantitative HPLC. Histometric analysis of outer nuclear layer (ONL) thicknesses was performed, and cryostat sections of retina were examined by fluorescence microscopy.RESULTS. Quantified A2E and qAF intensities increased until age 4 months in the Rdh8(-/-) / Abca4 (/) mice. The A2E levels declined after 4 months of age, but qAF intensity values continued to rise. The decline in A2E levels in the Rdh8(-/-) / Abca4(-/-) mice paralleled reduced photoreceptor cell viability as reflected in ONL thinning. Hyperautofluorescent puncta in fundus AF images corresponded to photoreceptor cell rosettes in SD-OCT images and histological sections stained with hematoxylin and eosin. The inner segment/outer segment-containing core of the rosette emitted an autofluorescence detected by fluorescence microscopy.CONCLUSIONS. When neural retina is disordered, AF from photoreceptor cells can contribute to noninvasive fundus AF images. Hyperautofluorescent puncta in fundus AF images are attributable, in at least some cases, to photoreceptor cell rosettes.