Effects of crystallin-?-b2 on stressed RPE in vitro and in vivo.

Böhm MR, Melkonyan H, Oellers P, Thanos S

Forschungsartikel (Zeitschrift) | Peer reviewed

Zusammenfassung

Crystallins are thought to play a cytoprotective role in conditions of cellular stress. The aim of this study was to determine the effects of crystallin-?-b2 (cry?-b2) and crystallin-?-b3 (cry?-b3) on ARPE-19 cells in vitro and on the retinal pigment epithelium (RPE) in vivo.The influence of cry?-b2 and cry?-b3 on the viability, proliferation and dying of ARPE-19 was measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium assay, bromo-2-deoxyuridine assay and life/death assay. The expressions of cry?-b2, cry?-b3, glial-derived neurotrophic factor (GDNF), and galectin-3 (Gal-3) in ARPE-19 cells were evaluated using immunohistochemistry (IHC), Western blotting (WB) and real-time-quantitative-PCR (qRT-PCR). To evaluate the response of cry?-b2 and cry?-b3 to stressed ARPE-19 cells, the cells were exposed to UV-light. In a rat model, cry?-b2-expressing neural progenitor cells (cry?-b2-NPCs) were injected intravitreally after retinal stress induced by optic nerve axotomy to examine whether they influence the RPE. Protein expression was examined 2 and 4 weeks postsurgery using IHC and WB.Detectable alterations of GDNF, and Gal-3 were found in ARPE-19 cells upon exposure to UV light. Adding the crystallins to the medium promoted proliferation and increased viability of ARPE-19 cells in vitro. The obtained data support the view that these crystallins possess epithelioprotective properties. Likewise, in vivo, intravitreally injected cry?-b2 and transplanted cry?-b2-NPCs protected RPE from indirectly induced stress.The data suggest that the RPE response to retinal ganglion cell denegeration is mediated via crystallins, which may thus be used therapeutically.

Details zur Publikation

FachzeitschriftGraefe's Archive for Clinical and Experimental Ophthalmology
Jahrgang / Bandnr. / Volume251
Ausgabe / Heftnr. / Issue1
Seitenbereich63-79
StatusVeröffentlicht
Veröffentlichungsjahr2013
Sprache, in der die Publikation verfasst istEnglisch

Autor*innen der Universität Münster

Böhm, Michael Rudolf Robert
Klinik für Augenheilkunde
Melkonyan, Harutyun
Klinik für Augenheilkunde