Caffé A R, Söderpalm A, van Veen T
Department of Anatomy, Erasmus University Rotterdam, The Netherlands.
Curr Eye Res. 1993 Aug;12(8):719-26. doi: 10.3109/02713689308995767.
Previously we have presented the morphological features of a neonatal mouse retinal explant kept in culture for 3 to 4 weeks. To further evaluate the organotypic parameters of the tissue we have examined the presence of opsin, S-antigen, and interphotoreceptor retinoid-binding protein (IRBP) in the same experimental paradigm, using light microscopic immunocytochemistry. In vitro, opsin and S-antigen staining is found in photoreceptor somata from genetically normal explants and those derived from mice with the rd or the rds mutation. When present, inner and outer segments label more intensely. No IRBP staining has been found in cell bodies of any genotype. However, some labeling is found in the plexiform layers and in the inner segments. The results indicate that photoreceptor proteins are continuously produced in vitro. This further establishes the organotypic nature of the retinal explant in culture. The administration of growth factors to these explants has been investigated. Neither basic fibroblast growth factor nor nerve growth factor alone has affected the explants phenotype. However, the combination of these proteins has significantly retarded rd cell loss in vitro.
此前我们已展示了在培养中保存3至4周的新生小鼠视网膜外植体的形态学特征。为了进一步评估该组织的器官型参数，我们使用光学显微镜免疫细胞化学技术，在相同的实验模式下检测了视蛋白、S抗原和光感受器间类视黄醇结合蛋白（IRBP）的存在情况。在体外，在基因正常的外植体以及来自rd或rds突变小鼠的外植体的光感受器细胞体中发现了视蛋白和S抗原染色。当存在时，内段和外段的染色更为强烈。在任何基因型的细胞体中均未发现IRBP染色。然而，在丛状层和内段发现了一些标记。结果表明光感受器蛋白在体外持续产生。这进一步证实了培养中的视网膜外植体的器官型性质。已研究了向这些外植体施用生长因子的情况。单独使用碱性成纤维细胞生长因子或神经生长因子均未影响外植体的表型。然而，这些蛋白质的组合在体外显著延缓了rd细胞的损失。
