Purchase this article with an account.
X Luo, C Romano; Adult Wild-type and Transgenic Mouse Retinal Cells in Dissociated Cultures: Characterization and Effects of Excitotoxins. . Invest. Ophthalmol. Vis. Sci. 2002;43(13):723.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: In vitro systems provide unparalled control of experimental conditions, but most primary cell culture systems use embryonic or neonatal tissue. Since the properties of developing and adult neurons differ greatly, we would like to study adult retinal neurons in dissociated culture. This will provide insights into mechanisms of adult-onset visual diseases such as glaucoma. The mouse is an attractive species to use because of the availability of genetically modified strains. Methods: Mixed retinal cultures were prepared from mice 5-6 weeks old following a procedure modified from that of Luo and Hicks (IOVS, 42:1096, 2001) used for pig. Cell types were identified using immunolabeling of specific markers. Strains expressing CFP in ganglion cells (Neuron, 28:51, 2000) were also employed. Excitotoxins were added to the medium for specified times, and cell populations were counted at specified later times. Results: Robust survival of, and expression of markers by, virtually all cell types were apparent. For example, ganglion cells extended neurofilament-positive processes for hundreds of microns. Ganglion cells from CFP-expressing mice retained fluorescence in culture. When subject to excitotoxic conditions, mouse amacrine cells were more susceptible to excitotoxic damage than those from pig. Ganglion cells were resistant to KA damage. Conclusion: Dissociated cultures from adult mice provide a novel and promising in vitro experimental system in which to investigate the cellular biology and pathology of adult mammalian neurons.
This PDF is available to Subscribers Only