Eldred Megan K, Charlton-Perkins Mark, Muresan Leila, Harris William A
Department of Physiology, Development and Neuroscience, Cambridge University, Cambridge CB2 3DY, UK.
Department of Physiology, Development and Neuroscience, Cambridge University, Cambridge CB2 3DY, UK
Development. 2017 Mar 15;144(6):1097-1106. doi: 10.1242/dev.142760. Epub 2017 Feb 7.
To investigate the cell-cell interactions necessary for the formation of retinal layers, we cultured dissociated zebrafish retinal progenitors in agarose microwells. Within these wells, the cells re-aggregated within hours, forming tight retinal organoids. Using a Spectrum of Fates zebrafish line, in which all different types of retinal neurons show distinct fluorescent spectra, we found that by 48 h in culture, the retinal organoids acquire a distinct spatial organisation, i.e. they became coarsely but clearly laminated. Retinal pigment epithelium cells were in the centre, photoreceptors and bipolar cells were next most central and amacrine cells and retinal ganglion cells were on the outside. Image analysis allowed us to derive quantitative measures of lamination, which we then used to find that Müller glia, but not RPE cells, are essential for this process.
为了研究视网膜层形成所需的细胞间相互作用，我们在琼脂糖微孔中培养了解离的斑马鱼视网膜祖细胞。在这些孔内，细胞在数小时内重新聚集，形成紧密的视网膜类器官。使用命运光谱斑马鱼品系，其中所有不同类型的视网膜神经元都显示出不同的荧光光谱，我们发现培养48小时后，视网膜类器官获得了独特的空间组织，即它们变得粗糙但清晰地分层。视网膜色素上皮细胞位于中心，光感受器和双极细胞次之，无长突细胞和视网膜神经节细胞位于外侧。图像分析使我们能够得出分层的定量测量值，然后我们用这些测量值发现，米勒胶质细胞而非视网膜色素上皮细胞对这一过程至关重要。
