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Journal of Ocular Biology, Diseases, and Informatics

, Volume 2, Issue 1, pp 1–11

First Online: 27 January 2009Received: 08 December 2008Accepted: 12 January 2009

Abstract

Here, roles of pigment epithelial-derived factor PEDF and glial-derived neurotrophic factor GDNF on photoreceptor development in three-dimensional reaggregates from the retinae of the E6 chick embryo rosetted spheroids was investigated.
Fully dispersed cells were reaggregated under serum-reduced conditions and supplemented with 50 ng-ml PEDF alone or in combination with 50 ng-ml GDNF.
The spheroids were analyzed for cell growth, differentiation, and death using proliferating cell nuclear antigen, terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling, and other immunocytochemical stainings and semi-quantitative reverse transcription polymerase chain reaction RT-PCR methods.
PEDF strongly promoted synthesis of the messenger RNAs for blue and violet cone opsins and to a lesser extent on the red and green cone opsins.
This correlated with an increase in the number of cone photoreceptors, as determined by the cone cell marker CERN906.
Likewise, PEDF nearly completely inhibited rod differentiation, as detected by immunostaining with anti-rho4D2 and RT-PCR.
Furthermore, PEDF accelerated proliferation of cells in the spheroids and inhibited apoptosis.
As negative effects, PEDF inhibited the normal histotypic tissue formation of retinal aggregates and reduced the frequency of photoreceptor rosettes and IPL-like areas.
Noticeably, supplementation of PEDF-treated cultures with GDNF reversed the effects of PEDF on spheroid morphology and on rod differentiation.
This study establishes that PEDF strongly affects three-dimensional retinogenesis in vitro, most notably by inhibiting rod development and supporting proliferation and differentiation of cones, effects which are partially counteracted by GDNF.

KeywordsCytokines Muller glial cells Photoreceptor degeneration Retinal spheroids Tissue formation  Download fulltext PDF



Author: Katja N. Volpert - Joyce Tombran-Tink - Colin Barnstable - Paul G. Layer

Source: https://link.springer.com/



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