title: Identification of an Alternative Splicing Product of the Otx2 Gene Expressed in the Neural Retina and Retinal Pigmented Epithelial Cells.
abstract: To investigate the complexity of alternative splicing in the retina, we sequenced and analyzed a total of 115,706 clones from normalized cDNA libraries from mouse neural retina (66,217) and rat retinal pigmented epithelium (49,489). Based upon clustering the cDNAs and mapping them with their respective genomes, the estimated numbers of genes were 9,134 for the mouse neural retina and 12,050 for the rat retinal pigmented epithelium libraries. This unique collection of retinal of messenger RNAs is maintained and accessible through a web-base server to the whole community of retinal biologists for further functional characterization. The analysis revealed 3,248 and 3,202 alternative splice events for mouse neural retina and rat retinal pigmented epithelium, respectively. We focused on transcription factors involved in vision. Among the six candidates suitable for functional analysis, we selected Otx2S, a novel variant of the Otx2 gene with a deletion within the homeodomain sequence. Otx2S is expressed in both the neural retina and retinal pigmented epithelium, and encodes a protein that is targeted to the nucleus. OTX2S exerts transdominant activity on the tyrosinase promoter when tested in the physiological environment of primary RPE cells. By overexpressing OTX2S in primary RPE cells using an adeno associated viral vector, we identified 10 genes whose expression is positively regulated by OTX2S. We find that OTX2S is able to bind to the chromatin at the promoter of the retinal dehydrogenase 10 (RDH10) gene.
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Christo
authors.initials: C
authors.lastname: Kole
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Naomi
authors.initials: N
authors.lastname: Berdugo
authors.affiliation: CEA-Institut de Génomique, GENOSCOPE, Centre National de Séquençage, 2 rue Gaston Crémieux, CP5706, 91057 Evry Cedex, France.
authors.forename: Corinne
authors.initials: C
authors.lastname: Da Silva
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Najate
authors.initials: N
authors.lastname: Aït-Ali
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Géraldine
authors.initials: G
authors.lastname: Millet-Puel
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Pagan
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Blond
authors.affiliation: Université de Strasbourg CNRS-ICube UMR 7357-LBGI-Faculté de Médecine-4, rue Kirschleger, F- 67085, Strasbourg, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Poidevin
authors.affiliation: Université de Strasbourg CNRS-IGBMC UMR7104-1 rue Laurent Fries-BP10142-F67404 Illkirch, France.
authors.forename: Raymond
authors.initials: R
authors.lastname: Ripp
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Fontaine
authors.affiliation: CEA-Institut de Génomique, GENOSCOPE, Centre National de Séquençage, 2 rue Gaston Crémieux, CP5706, 91057 Evry Cedex, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Wincker
authors.affiliation: Wilmer Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, United States of America.
authors.forename: Donald J
authors.initials: DJ
authors.lastname: Zack
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: José-Alain
authors.initials: JA
authors.lastname: Sahel
authors.affiliation: Université de Strasbourg CNRS-ICube UMR 7357-LBGI-Faculté de Médecine-4, rue Kirschleger, F- 67085, Strasbourg, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Poch
authors.affiliation: INSERM, U968, Paris, F-75012, France.|Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France.|CNRS, UMR_7210, Paris, F-75012, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Léveillard
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D015723:Gene Library; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000235:genetics; D051857:Otx Transcription Factors / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D012160:Retina / Q000166:cytology* / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism
chemical_list: D002843:Chromatin; D018076:DNA, Complementary; D051857:Otx Transcription Factors; C497660:Otx2 protein, mouse; C000609576:Otx2 protein, rat; D020033:Protein Isoforms; D012333:RNA, Messenger; D000429:Alcohol Oxidoreductases; C113992:trans-retinol dehydrogenase; D014442:Monophenol Monooxygenase