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眼科学报 2006年第4期22卷 244-251页

作者：王岚 Martin Rudolf christine a.curcio 彭大伟刘杏曾淑君中山大学中山眼科中心中山大学眼科学国家重点实验室 Department of Ophthalmology University of Alabama School of Medicine Birmingham Alabama 35233 USA 中山大学基础医学院药理学教研室

目的:检测大鼠视网膜色素上皮细胞系(RPE-J)是否表达载脂蛋白的基因及其是否具有合成和分泌中性脂质的功能。方法:将RPE-J细胞培养于组织培养板及Transwell上,用RT-PCR检测载脂蛋白mRNA的表达。将荧光标记颗粒加入细胞中培养以检测细胞... 详细信息

目的:检测大鼠视网膜色素上皮细胞系(RPE-J)是否表达载脂蛋白的基因及其是否具有合成和分泌中性脂质的功能。方法:将RPE-J细胞培养于组织培养板及Transwell上,用RT-PCR检测载脂蛋白mRNA的表达。将荧光标记颗粒加入细胞中培养以检测细胞吞噬功能。细胞中加入光感受器细胞外节,培养不同时间点后提取脂质,用薄层层析法和酶学荧光法分析细胞中脂质含量及变化。用放射性标记的OleicAcid([3H]-oleate)检测细胞新合成和分泌中性脂质的功能。结果:RPE-J细胞有载脂蛋白A-I,A-II,B,E,C-I,C-II,C-III,以及微粒体甘油三酯酸转移蛋白基因的表达。RPE-J细胞具有吞噬功能。吞噬光感受器细胞外节后,RPE-J细胞中甘油三酯含量增加5.8倍,磷脂含量增加2.6倍,非酯化胆固醇增加1.5倍,酯化胆固醇增加0.3倍。RPE-J细胞和培养基内可以检测到新合成的放射标记的中性脂质。细胞内脂质含量随时间延长而增加;培养基中只有磷脂随时间延长而增多。结论:RPE-J细胞有多种载脂蛋白及微粒体甘油三酯酸转移蛋白的基因表达。吞噬光感受器细胞外节后,RPE-J细胞中脂质增加。培养基中加入长链脂肪酸后,RPE-J细胞可以新合成和分泌中性脂质。

关键词：视网膜色素上皮细胞 RPE-J 脂蛋白载脂蛋白脂质光感受器细胞基因

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Hyperspectral autofluorescence characterization of drusen and sub-RPE deposits in age-related macular degeneration

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Annals of Eye Science 2021年第1期6卷 20-27页

作者： Yuehong Tong Thomas Ach christine a.curcio RTheodore Smith Department of Ophthalmology Icahn School of Medicine at Mount SinaiNew YorkNYUSA Department of Ophthalmology University Hospital BonnGermany Department of Ophthalmology and Visual Sciences University of Alabama at BirminghamALUSA

Background:Soft drusen and basal linear deposit(BLinD)are two forms of the same extracellular lipid rich material that together make up an Oil Spill on Bruch’s membrane(BrM).Drusen are focal and can be recognized *** contrast BLinD is thin and diffusely distributed,and invisible clinically,even on highest resolution OCT,but has been detected on en face hyperspectral autofluorescence(AF)imaging ex *** sought to optimize histologic hyperspectral AF imaging and image analysis for recognition of drusen and sub-RPE deposits(including BLinD and basal laminar deposit),for potential clinical ***:Twenty locations specifically with drusen and 12 additional locations specifically from fovea,perifovea and mid-periphery from RPE/BrM flatmounts from 4 AMD donors underwent hyperspectral AF imaging with 4 excitation wavelengths(λex 436,450,480 and 505 nm),and the resulting image cubes were simultaneously decomposed with our published non-negative matrix factorization(NMF).Rank 4 recovery of 4 emission spectra was chosen for each excitation ***:A composite emission spectrum,sensitive and specific for drusen and presumed sub-RPE deposits(the SDr spectrum)was recovered with peak at 510-520 nm in all tissues with drusen,with greatest amplitudes at excitationsλ_(ex)436,450 and 480 *** RPE spectra of combined sources Lipofuscin(LF)/Melanolipofuscin(MLF)were of comparable amplitude and consistently recapitulated the spectra S1,S2 and S3 previously reported from all tissues:tissues with drusen,foveal and extra-foveal ***:A clinical hyperspectral AF camera,with properly chosen excitation wavelengths in the blue range and a hyperspectral AF detector,should be capable of detecting and quantifying drusen and sub-RPE deposits,the earliest known lesions of AMD,before any other currently available imaging modality.

关键词： Age-related macular degeneration sub-RPE deposits drusen hyperspectral autofluorescence(AF)

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Characteristics of normal human retinal pigment epithelium cells with extremes of autofluorescence or intracellular granule count

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Annals of Eye Science 2021年第1期6卷 11-19页

作者： Katharina Bermond Andreas Berlin Ioana-Sandra Tarau Christina Wobbe Rainer Heintzmann christine a.curcio Kenneth R.Sloan Thomas Ach Department of Ophthalmology University Hospital WürzburgWürzburgGermany Department of Ophthalmology Ludwigshafen HospitalLudwigshafenGermany Leibniz Institute of Photonic Technology JenaGermany Institute of Physical Chemistry and Abbe Center of Photonics Friedrich-Schiller University JenaJenaGermany Department of Ophthalmology University of Alabama at BirminghamBirminghamALUSA Department of Ophthalmology University Hospital BonnBonnGermany

Background:Cells of the retinal pigment epithelium(RPE)accumulate different kinds of granules(lipofuscin,melanolipofuscin,melanosomes)within their cell bodies,with lipofuscin and melanolipofuscin being autofluorescent after blue light *** amounts of lipofuscin granules within the RPE have been associated with the development of RPE cell death and age-related macular degeneration(AMD);however,this has not been confirmed in histology so ***,based on our previous dataset of RPE granule characteristics,we report the characteristics of RPE cells from human donor eyes that show either high or low numbers of intracellular granules or high or low autofluorescence(AF)***:RPE flatmounts of fifteen human donors were examined using high-resolution structured illumination microscopy(HR-SIM)and laser scanning microscopy(LSM).Autofluorescent granules were analyzed regarding AF phenotype and absolute number of *** addition,total AF intensity per cell and granule density(number of granules per cell area)were *** the final analysis,RPE cells with total granule number below 5th or above the 95th percentile,or a total AF intensity±1.5 standard deviations above or below the mean were included,and compared to the average RPE cell at the same *** are presented as mean±standard ***:Within 420 RPE cells examined,42 cells were further analyzed due to extremes regarding total granule *** addition,20 RPE cells had AF 1.5 standard deviations below,28 RPE cells above the mean local AF *** granules predominate in RPE cells with low granule content and low AF *** cells with high granule content have nearly twice(1.8 times)as many granules as an average RPE ***:In normal eyes,outliers regarding autofluorescent granule load and AF intensity signals are rare among RPE cells,suggesting that granule deposition and subsequent AF follows intrinsic control mechanisms at a cellul

关键词： Retinal pigment epithelium(RPE) granules autofluorescence(AF) lipofuscin melanolipofuscin melanosomes

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