Organotypic Culture of Neural Retina as a Research Model of Neurodegeneration of Ganglion Cells

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O. S. Gancharova^1,2,3*, V. N. Manskikh^1,3, A. A. Zamyatnin, Jr.^2,4, and P. P. Philippov^1,2

¹Lomonosov Moscow State University, Institute of Mitoengineering, 119991 Moscow, Russia; fax: (495) 939-5945; E-mail: info@mitotech.ru; olgancharova@gmail.com

²Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology, 119991 Moscow, Russia; fax: (495) 939-3181; E-mail: fxb@genebee.msu.su

³Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology, Faculty of Bioengineering and Bioinformatics, 119991 Moscow, Russia; fax: (495) 939-3181; E-mail: fxb@belozersky.msu.ru

⁴Institute of Molecular Medicine, Sechenov First Moscow State Medical University, 119991 Moscow, Russia; fax: (495) 622-9632

^* To whom correspondence should be addressed.

Received June 19, 2013

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Organotypic models deserve special attention among the large variety of methods of vertebrate retina cultivation. The purpose of this study was to make a detailed qualitative and quantitative characterization of a model employing roller organotypic cultivation of the neural retina of rat eye posterior segment, with special attention to morphological and functional characteristics of retinal ganglion cells. The study included morphological analysis of retina histological preparations as well as estimation of RNA synthesis and evaluation of neuron survival by the Brachet and TUNEL methods, respectively. Retina has been shown to display normal morphofunctional characteristics for the first 12 h of cultivation. After 24 h, a substantial number of ganglion cells underwent pyknosis and stopped RNA synthesis. Almost all the cells of the retinal ganglion layer became apoptotic by 3-4 days in vitro. In the course of cultivation, neural retina is detached from the underlying layers of the posterior eye segment and undergoes significant cytoarchitectonic changes. The causes of ganglion cell death during organotypic cultivation of eye posterior segment are discussed. This method can serve as a suitable model for the screening of new retinoprotectors and for research on ganglion cell death resulting from retina degenerative diseases, e.g. glaucoma.

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KEY WORDS: retina, ganglion cells, apoptosis, organotypic roller culture

DOI: 10.1134/S0006297913110084

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