Visual cycle protein RPE65 persists in new retinal cells during retinal regeneration of adult newt

Adult newts can regenerate their entire retina through transdifferentiation of the retinal pigment epithelium (RPE). The objective of this study was to redescribe the retina regeneration process by means of modern biological techniques. We report two different antibodies (RPE‐No.112 and MAB5428) tha...

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Bibliographic Details
Published in:	Journal of comparative neurology (1911) Vol. 495; no. 4; pp. 391 - 407
Main Authors:	Chiba, Chikafumi, Hoshino, Akika, Nakamura, Kenta, Susaki, Kanako, Yamano, Yuka, Kaneko, Yuko, Kuwata, Osamu, Maruo, Fumiaki, Saito, Takehiko
Format:	Journal Article
Language:	English
Published:	Hoboken Wiley Subscription Services, Inc., A Wiley Company 01-04-2006
Subjects:	Amino Acid Sequence Animals Biomarkers - analysis Cell Differentiation - physiology Connexin 43 - metabolism Eye Proteins - genetics Eye Proteins - metabolism Freshwater Gene Expression - physiology Homeodomain Proteins - metabolism Immunoblotting Immunohistochemistry In Situ Hybridization Molecular Sequence Data newt Paired Box Transcription Factors - metabolism PAX6 Transcription Factor Pigment Epithelium of Eye - cytology Pigment Epithelium of Eye - physiology Polymerase Chain Reaction regeneration Regeneration - physiology Repressor Proteins - metabolism retina Retina - cytology Retina - physiology retinal pigment epithelium RPE65 Salamandridae Stem Cells - metabolism transdifferentiation
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Summary:	Adult newts can regenerate their entire retina through transdifferentiation of the retinal pigment epithelium (RPE). The objective of this study was to redescribe the retina regeneration process by means of modern biological techniques. We report two different antibodies (RPE‐No.112 and MAB5428) that recognize the newt homolog of RPE65, which is involved in the visual cycle and exclusively label the RPE cell‐layer in the adult newt eye. We analyzed the process of retinal regeneration by immunohistochemistry and immunoblotting and propose that this process should be divided into nine stages. We found that the RPE65 protein is present in the RPE‐derived new retinal rudiment at 14 days postoperative (po) and in the regenerating retinas at the 3–4 cell stage (19 days po). These observations suggest that certain characteristics of RPE cells overlap with those of retinal stem/progenitor cells during the period of transdifferentiation. However, RPE65 protein was not detected in either retinal stem/progenitor cells in the ciliary marginal zone (CMZ) of adult eyes or in neuroepithelium present during retina development, where it was first detected in differentiated RPE. Moreover, the gene expression of RPE65 was drastically downregulated in the early phase of transdifferentiation (by 10 days po), while those of Connexin43 and Pax‐6, both expressed in regenerating retinas, were differently upregulated. These observations suggest that the RPE65 protein in the RPE‐derived retinal rudiment may represent the remainder after protein degradation or discharge rather than newly synthesized protein. J. Comp. Neurol. 40:391–407, 2006. © 2006 Wiley‐Liss, Inc.
Bibliography:	Science and Technology Agency of Japan University of Tsukuba Ministry of Education, Science, and Culture of Japan istex:79BE5FA0CA12041AF0519A2D99116259F756B03C ark:/67375/WNG-SQ3JKK7H-C ArticleID:CNE20880 Japan Society for the Promotion of Science The first three authors contributed equally to this work. ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0021-9967 1096-9861
DOI:	10.1002/cne.20880