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- Engelsberg, Karl, et al.
(författare)
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Early development of retinal subtypes in long-term cultures of human embryonic retina.
- 2008
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Ingår i: Current Eye Research. - : Informa UK Limited. - 0271-3683 .- 1460-2202. ; 33:2, s. 185-191
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Tidskriftsartikel (refereegranskat)abstract
- Purpose: To study early signs of neuronal and glial differentiation in the human embryonic retina. Materials and Methods: 6.5-to 8-week-old human embryos were obtained from elective abortions. The neuroretinas were kept in culture as full-thickness sheets for 7-42 days. Results: The control retinas consisted of a neuroblast cell layer and a thin marginal zone. Most explants displayed presence of retinal lamination, but also contained regions of disorganization. Vimentin labeling showed vertically arranged Müller cells in all explants. Recoverin-labeled photoreceptors appeared in explants kept 14 days and longer. By labeling with antibodies against PKC and parvalbumin, rod bipolar cells and amacrine cells could be seen in explants kept for 42 days in culture.Conclusions: We have shown that the embryonic full-thickness neuroretina can survive in a culture environment for at least 6 weeks, and can develop several types of the retina-specific neuronal and glial cells.
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- Petersen, Anne, 1962, et al.
(författare)
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The proteasome and intracellular redox status: implications for apoptotic regulation in lens epithelial cells.
- 2007
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Ingår i: Current eye research. - : Informa UK Limited. - 0271-3683 .- 1460-2202. ; 32:10, s. 871-82
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Tidskriftsartikel (refereegranskat)abstract
- Purpose: This study aimed to investigate redox regulation of the proteasome as well as the effect of proteasome inhibition on intracellular oxidative status and apoptosis. Methods: Oxidative stress was induced in cultured human lens epithelial cells (HLECs) and intact mouse lenses by 100 mu M H(2)O(2). HLECs were also exposed to the reduced and the oxidized forms of glutathione (GSH/GSSG) and the reducing agent dithiotreitol (DTT). The chymotrypsin-like, the trypsin-like, and the peptidylglutamyl peptidase activities of the proteasome were measured using synthetic fluorogenic substrates. Superoxide as well as peroxide production, mitochondrial membrane potential, and the level of GSH was measured in HLECs after proteasome inhibition by MG-132 or lactacystin. Apoptosis was determined by measuring caspase-3 activation and by studying apoptotic nuclei after staining with Hoechst 33342. Results: All three peptidase activities of the proteasome were inhibited by 100 mu M H(2)O(2) and by the oxidized form of glutathione (GSSG), whereas the reduced form (GSH) stimulated chymotrypsin-like and peptidylglutamyl peptidase activities in HLECs lysates. Intact mouse lenses exposed to 100 mu M H(2)O(2) exhibited loss of transparency and trends of decreased chymotrypsin-like proteasome activity as well as decreased GSH levels. Inhibition of the proteasome in cultured HLECs caused significant increase in apoptosis and disturbed intracellular redox balance. Simultaneous addition of exogenous GSH completely abolished the increased apoptosis seen after MG-132 treatment. Conclusions: This study supports the hypothesis that intracellular proteolytic and oxidative regulatory systems are tightly coupled. The current data also indicate that apoptosis by proteasome inhibition is mediated through oxidative mechanisms.
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