Photoreceptor vitality in organotypic cultures of mature vertebrate retinas validated by light-dependent molecular movements.
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.
Reidel
Boris
B
Institute of Zoology, Department of Cell and Matrix Biology, University of Mainz, Germany.
Orisme
Wilda
W
Goldmann
Tobias
T
Smith
W Clay
WC
Wolfrum
Uwe
U
D000818:Animals; D019393:Arrestin / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D009924:Organ Culture Techniques; D010775:Photic Stimulation; D010786:Photoreceptor Cells / Q000502:physiology*; D021381:Protein Transport; D012160:Retina / Q000033:anatomy & histology / Q000378:metabolism*; D015293:Transducin / Q000378:metabolism; D014714:Vertebrates / Q000378:metabolism*; D014785:Vision, Ocular / Q000502:physiology; D014981:Xenopus
D019393:Arrestin; D015293:Transducin