Rothermel, Andree ; Burghardt, Mirjam ; Volpert, Katja ; Robitzki, Andrea ; Layer, Paul G. ; Lantzsch, Christina (2006)
Knock-down of GFR-alpha-4 expression by RNA interference affects the development of retinal cell types in three-dimensional histotypic retinal spheres.
In: Investigative ophthalmology & visual science, 47 (6)
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
Abstract PURPOSE:
To determine the role of glial cell line-derived neurotropic factor family receptor alpha 4 (GFRalpha4) during retinogenesis in a three-dimensional histiotypic in vitro model of the embryonic chicken retina. METHODS:
Retinal spheres were cultured from dissociated 6-day-old chicken retina under permanent rotation and transfected with GFRalpha4 siRNA at culture day 2. Alterations on proliferation, apoptosis, and differentiation were determined by semiquantitative RT-PCR, in situ hybridization, and immunohistochemistry after 24, 48, and 72 hours. RESULTS:
In contrast to control cultures, retinal spheres transfected with GFRalpha4 siRNA showed reduced GFRalpha4 mRNA expression of only 38% after 24 hours, 3% after 48 hours, and 5% after 72 hours. Based on the suppression of GFRalpha4, a decline in proliferating cells from 10% to 4.8% even after 24 hours and a reduction of sphere size by up to 25% at later culture stages were observed. Moreover, the number of Pax 6-positive amacrine, ganglion, and horizontal cells was significantly decreased from 36% to 16% in GFRalpha4 siRNA-transfected retinal spheres 72 hours after transfection. Additionally, GFRalpha4 gene silencing affected the development of different types of photoreceptors, as revealed by a significant decrease of blue opsin mRNA expression from 29% to 2%, whereas green opsin mRNA and the number rho4D2-positive photoreceptors were significantly increased. CONCLUSIONS:
These data showed for the first time that GFRalpha4 plays an essential role in regulating, at least in vitro, the development and differentiation of various cell types during retinogenesis.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2006 |
| Autor(en): | Rothermel, Andree ; Burghardt, Mirjam ; Volpert, Katja ; Robitzki, Andrea ; Layer, Paul G. ; Lantzsch, Christina |
| Art des Eintrags: | Bibliographie |
| Titel: | Knock-down of GFR-alpha-4 expression by RNA interference affects the development of retinal cell types in three-dimensional histotypic retinal spheres |
| Sprache: | Englisch |
| Publikationsjahr: | 2006 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Investigative ophthalmology & visual science |
| Jahrgang/Volume einer Zeitschrift: | 47 |
| (Heft-)Nummer: | 6 |
| Kurzbeschreibung (Abstract): | Abstract PURPOSE: To determine the role of glial cell line-derived neurotropic factor family receptor alpha 4 (GFRalpha4) during retinogenesis in a three-dimensional histiotypic in vitro model of the embryonic chicken retina. METHODS: Retinal spheres were cultured from dissociated 6-day-old chicken retina under permanent rotation and transfected with GFRalpha4 siRNA at culture day 2. Alterations on proliferation, apoptosis, and differentiation were determined by semiquantitative RT-PCR, in situ hybridization, and immunohistochemistry after 24, 48, and 72 hours. RESULTS: In contrast to control cultures, retinal spheres transfected with GFRalpha4 siRNA showed reduced GFRalpha4 mRNA expression of only 38% after 24 hours, 3% after 48 hours, and 5% after 72 hours. Based on the suppression of GFRalpha4, a decline in proliferating cells from 10% to 4.8% even after 24 hours and a reduction of sphere size by up to 25% at later culture stages were observed. Moreover, the number of Pax 6-positive amacrine, ganglion, and horizontal cells was significantly decreased from 36% to 16% in GFRalpha4 siRNA-transfected retinal spheres 72 hours after transfection. Additionally, GFRalpha4 gene silencing affected the development of different types of photoreceptors, as revealed by a significant decrease of blue opsin mRNA expression from 29% to 2%, whereas green opsin mRNA and the number rho4D2-positive photoreceptors were significantly increased. CONCLUSIONS: These data showed for the first time that GFRalpha4 plays an essential role in regulating, at least in vitro, the development and differentiation of various cell types during retinogenesis. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie ?? fb10_zoologie ?? 10 Fachbereich Biologie > Developmental Biology and Neurogenetics |
| Hinterlegungsdatum: | 20 Nov 2008 08:21 |
| Letzte Änderung: | 20 Feb 2020 13:24 |
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