IOVS
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

(Investigative Ophthalmology and Visual Science. 2006;47:1177-1184.)
© 2006 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.05-0965
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (13)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Chen, Y.
Right arrow Articles by Ma, J.-x.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chen, Y.
Right arrow Articles by Ma, J.-x.

RPE65 Gene Delivery Restores Isomerohydrolase Activity and Prevents Early Cone Loss in Rpe65–/– Mice

Ying Chen, Gennadiy Moiseyev, Yusuke Takahashi, and Jian-xing Ma

From the Department of Cell Biology, Department of Medicine, The University of Oklahoma, Health Sciences Center, Oklahoma City, Oklahoma.

PURPOSE. Recent in vitro evidence has shown that RPE65 is the isomerohydrolase that converts all-trans retinyl ester to 11-cis retinal, the chromophore for visual pigments in vertebrates. Homozygous RPE65 knockout (Rpe65–/–) mice lack 11-cis retinoids and have early cone degeneration. The purpose of this study is to determine whether RPE65 gene delivery restores the isomerohydrolase activity and normal profile of endogenous retinoids in Rpe65–/– mice.

METHODS. Adenovirus-expressing RPE65 (Ad-RPE65) was injected into the subretinal space of Rpe65–/– mice. The expression of RPE65 was determined by immunohistochemistry and Western blot analysis. The isomerohydrolase activity was measured in vitro in eyecup homogenates. Endogenous retinoid profile in the eyecups was analyzed by high-performance liquid chromatography (HPLC). Photoreceptor-specific gene expression was determined with real-time RT-PCR. Cone degeneration was determined by cone-specific staining and counting cones in flatmounted retina.

RESULTS. High levels of RPE65 expression from the Ad-RPE65 injection generated robust isomerohydrolase activity in the eyecup of Rpe65–/– mice, at levels comparable to those in wild-type (wt) mice. Consequently, the RPE65 gene delivery resulted in substantial amounts of 11-cis retinal in Rpe65–/– mice. The RPE65 gene delivery prevented the downregulation of cone-specific genes, including both cone opsins and cone tranducin {alpha} subunit in Rpe65–/– mice. Moreover, the Ad-RPE65 injection also prevented massive cone degeneration at early ages of Rpe65–/– mice.

CONCLUSIONS. RPE65 gene delivery generates isomerohydrolase activity and restores retinoid profile in Rpe65–/– mice. Regeneration of 11-cis retinal is essential for survival of cone photoreceptors.




This article has been cited by other articles:


Home page
 Proc. Natl. Acad. Sci. USAHome page
A. V. Cideciyan, T. S. Aleman, S. L. Boye, S. B. Schwartz, S. Kaushal, A. J. Roman, J.-j. Pang, A. Sumaroka, E. A. M. Windsor, J. M. Wilson, et al.
Human gene therapy for RPE65 isomerase deficiency activates the retinoid cycle of vision but with slow rod kinetics
PNAS, September 30, 2008; 105(39): 15112 - 15117.
[Abstract] [Full Text] [PDF]

Home page
 IOVSHome page
J. Fan, B. Rohrer, J. M. Frederick, W. Baehr, and R. K. Crouch
Rpe65-/- and Lrat-/- Mice: Comparable Models of Leber Congenital Amaurosis
Invest. Ophthalmol. Vis. Sci., June 1, 2008; 49(6): 2384 - 2389.
[Abstract] [Full Text] [PDF]

Home page
 Hum Mol GenetHome page
M. Samardzija, J. von Lintig, N. Tanimoto, V. Oberhauser, M. Thiersch, C. E. Reme, M. Seeliger, C. Grimm, and A. Wenzel
R91W mutation in Rpe65 leads to milder early-onset retinal dystrophy due to the generation of low levels of 11-cis-retinal
Hum. Mol. Genet., January 15, 2008; 17(2): 281 - 292.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y. Chen, Y. Hu, K. Lu, J. G. Flannery, and J.-x. Ma
Very Low Density Lipoprotein Receptor, a Negative Regulator of the wnt Signaling Pathway and Choroidal Neovascularization
J. Biol. Chem., November 23, 2007; 282(47): 34420 - 34428.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. G. Jacobson, T. S. Aleman, A. V. Cideciyan, E. Heon, M. Golczak, W. A. Beltran, A. Sumaroka, S. B. Schwartz, A. J. Roman, E. A. M. Windsor, et al.
Human cone photoreceptor dependence on RPE65 isomerase
PNAS, September 18, 2007; 104(38): 15123 - 15128.
[Abstract] [Full Text] [PDF]

Home page
 IOVSHome page
A. Wenzel, J. von Lintig, V. Oberhauser, N. Tanimoto, C. Grimm, and M. W. Seeliger
RPE65 Is Essential for the Function of Cone Photoreceptors in NRL-Deficient Mice
Invest. Ophthalmol. Vis. Sci., February 1, 2007; 48(2): 534 - 542.
[Abstract] [Full Text] [PDF]

Home page
 IOVSHome page
Y. Takahashi, G. Moiseyev, Y. Chen, and J.-x. Ma
The Roles of Three Palmitoylation Sites of RPE65 in Its Membrane Association and Isomerohydrolase Activity
Invest. Ophthalmol. Vis. Sci., December 1, 2006; 47(12): 5191 - 5196.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2006 by the Association for Research in Vision and Ophthalmology