Reidel Boris, Orisme Wilda, Goldmann Tobias, Smith W Clay, Wolfrum Uwe
Institute of Zoology, Department of Cell and Matrix Biology, University of Mainz, Germany.
Vision Res. 2006 Dec;46(27):4464-71. doi: 10.1016/j.visres.2006.07.019. Epub 2006 Sep 18.
Vertebrate photoreceptor cells are polarized neurons highly specialized for light absorption and visual signal transduction. Photoreceptor cells consist of the light sensitive outer segment and the biosynthetic active inner segment linked by a slender connecting cilium. The function of mature photoreceptor cells is strictly dependent on this compartmentalization which is maintained in the specialized retinal environment. To keep this fragile morphologic and functional composition for further cell biological studies and treatments we established organotypic retina cultures of mature mice and Xenopus laevis. The organotypic retina cultures of both model organisms are created as co-cultures of the retina and the pigment epithelium, still attached to outer segments of the photoreceptor cells. To demonstrate the suitability of the culture system for physiological analyses we performed apoptotic cell death analyses and verified photoreceptor viability. Furthermore, light-dependent bidirectional movements of arrestin and transducin in photoreceptors in vivo and in the retinal cultures were indistinguishable indicating normal photoreceptor cell-biologic function in organotypic cultures. Our established culture systems allow the analysis of mature photoreceptor cells and their accessibility to treatments, characteristic for common cell culture. Furthermore, this culturing technique also provides an appropriate system for gene delivery to retinal cells and will serve to simulate gene therapeutic approaches prior to difficult and time-consuming in vivo experiments.
脊椎动物的光感受器细胞是高度特化的极化神经元，专门用于光吸收和视觉信号转导。光感受器细胞由光敏外段和通过细长连接纤毛相连的生物合成活跃的内段组成。成熟光感受器细胞的功能严格依赖于这种在特殊视网膜环境中维持的区室化。为了保持这种脆弱的形态和功能组成以用于进一步的细胞生物学研究和治疗，我们建立了成熟小鼠和非洲爪蟾的视网膜器官型培养物。这两种模式生物的视网膜器官型培养物均作为视网膜与色素上皮的共培养物创建，色素上皮仍附着于光感受器细胞的外段。为了证明该培养系统适用于生理学分析，我们进行了凋亡细胞死亡分析并验证了光感受器的活力。此外，体内和视网膜培养物中光感受器中抑制蛋白和转导蛋白的光依赖性双向运动没有区别，这表明器官型培养物中光感受器细胞生物学功能正常。我们建立的培养系统允许对成熟光感受器细胞进行分析，并使其能够接受常见细胞培养特有的处理。此外，这种培养技术还为向视网膜细胞进行基因递送提供了合适的系统，并将用于在困难且耗时的体内实验之前模拟基因治疗方法。
