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Membrane frizzled-related protein is necessary for the normal development and maintenance of photoreceptor outer segments

Published online by Cambridge University Press:  01 September 2008

JUNGYEON WON ,
RICHARD S. SMITH ,
NEAL S. PEACHEY ,
JIANG WU ,
WANDA L. HICKS ,
JÜRGEN K. NAGGERT  and
PATSY M. NISHINA
JUNGYEON WON
Affiliation:
The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine
RICHARD S. SMITH
Affiliation:
The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine
NEAL S. PEACHEY
Affiliation:
Research Service, Cleveland VA Medical Center, Cleveland, Ohio Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, Ohio
JIANG WU
Affiliation:
Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, Ohio
WANDA L. HICKS
Affiliation:
The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine
JÜRGEN K. NAGGERT
Affiliation:
The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine
PATSY M. NISHINA*
Affiliation:
The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine
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Abstract

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Type
Erratum
Information
Visual Neuroscience , Volume 25 , Issue 5-6 , September 2008 , pp. 713 - 716
Copyright
Copyright © Cambridge University Press 2008

(This article appeared in Volume 25, Number 4, 2008, pp. 563–574).

Due to an error in the Production office, incorrect files for figures 2, 3, and 10 were used in the production of the original published version of this article. The Publisher apologizes for this error, and reproduces the correct figure files herein.

Fig. 2. Localization of MFRP in mouse retina (A–H). Retinal sections obtained from P7 (A, B), P14 (C, D), P21 (E, F), and P28 (G, H, L) and stained with anti-hMFRP (red). Nuclear counterstaining was done with DAPI (blue). Inserts show higher magnification images. Higher magnification image of control (L) was obtained using confocal microscopy to show the MFRP localization in the RPE. The arrows indicate the RPE nuclei. Western analysis using anti-mMFRP (I) and anti-hMFRP (J). Eyes for Western analysis were obtained at 2 months. Anti-actinin 4 was used as a loading control. (K) Western analysis of Myc-MFRPrd6 overexpressed in COS-7 cells. Cells were harvested 48 h after transfection. Lysate from Myc-MFRPrd6 transfected cells and nontransfected control (NT) were separated on 10% SDS-PAGE gel; transferred to nitrocellulose membrane; and stained with anti-c-Myc, anti-mMFRP, and anti-hMFRP. RPE, retinal pigment epithelium; ONL, outer nuclear layer; INL, inner nuclear layer; GC, ganglion cell layer. Bars = 10 μm.

Fig. 3. Localization of C1QTNF5 in mouse retina (A–H). Retinal sections obtained from P7 (A, B), P14 (C, D), P21 (E, F), and P28 (G, H) and stained with anti-C1QTNF5 (red). Nuclear counterstaining was done with DAPI (blue). Inserts show higher magnification images. (I) Western blot analysis with anti-C1QTNF5 was performed as described in Fig. 2. RPE, retinal pigment epithelium; ONL, outer nuclear layer; INL, inner nuclear layer; GC, ganglion cell layer. Bar=10 μm.

Fig. 10. Mfrprd6/rd6 mice have phagocytic defects (A–D). The retinal sections in panels C and D were obtained 1 h after light stimulation while the sections in panels A and B were obtained at the same time without the light stimulation from 2-month control (A, C) and Mfrprd6/rd6 (B, D) mice. Retinal sections were stained with anti-rhodopsin (RHO; red) to determine phagocytic uptake of rod OS in RPE, and anti-ezrin (EZRIN; green) was used to distinguish the RPE–photoreceptor border. The boxed area is enlarged in the insert. In control mice, rhodopsin uptake (arrows) was increased with light stimulation (C) compared to no light stimulation (A). Quantification of rhodopsin-positive material in the RPE (E). Western analysis of whole-eye lysates from 2-month control and Mfrprd6/rd6 mice and immunoprecipitated products of MERTK (F). Phospo-MERTK is shown by immunoblot with anti-phospho-Tyrosine (pY) antibody following immunoprecipitation with anti-MERTK (Mer). To quantify the band intensity, the membrane was reblotted with anti-MERTK. The immunoblot of 10% of lysate input was performed with primary antibody as indicated. RPE, retinal pigment epithelium; ELM, external limiting membrane; ONL, outer nuclear layer; OS, outer segment. Bar = 10 μm.

Figures 2, 3, and 10 appear on the next three pages.

Figure 0

Fig. 2. Localization of MFRP in mouse retina (A–H). Retinal sections obtained from P7 (A, B), P14 (C, D), P21 (E, F), and P28 (G, H, L) and stained with anti-hMFRP (red). Nuclear counterstaining was done with DAPI (blue). Inserts show higher magnification images. Higher magnification image of control (L) was obtained using confocal microscopy to show the MFRP localization in the RPE. The arrows indicate the RPE nuclei. Western analysis using anti-mMFRP (I) and anti-hMFRP (J). Eyes for Western analysis were obtained at 2 months. Anti-actinin 4 was used as a loading control. (K) Western analysis of Myc-MFRPrd6 overexpressed in COS-7 cells. Cells were harvested 48 h after transfection. Lysate from Myc-MFRPrd6 transfected cells and nontransfected control (NT) were separated on 10% SDS-PAGE gel; transferred to nitrocellulose membrane; and stained with anti-c-Myc, anti-mMFRP, and anti-hMFRP. RPE, retinal pigment epithelium; ONL, outer nuclear layer; INL, inner nuclear layer; GC, ganglion cell layer. Bars = 10 μm.

Figure 1

Fig. 3. Localization of C1QTNF5 in mouse retina (A–H). Retinal sections obtained from P7 (A, B), P14 (C, D), P21 (E, F), and P28 (G, H) and stained with anti-C1QTNF5 (red). Nuclear counterstaining was done with DAPI (blue). Inserts show higher magnification images. (I) Western blot analysis with anti-C1QTNF5 was performed as described in Fig. 2. RPE, retinal pigment epithelium; ONL, outer nuclear layer; INL, inner nuclear layer; GC, ganglion cell layer. Bar=10 μm.

Figure 2

Fig. 10. Mfrprd6/rd6 mice have phagocytic defects (A–D). The retinal sections in panels C and D were obtained 1 h after light stimulation while the sections in panels A and B were obtained at the same time without the light stimulation from 2-month control (A, C) and Mfrprd6/rd6 (B, D) mice. Retinal sections were stained with anti-rhodopsin (RHO; red) to determine phagocytic uptake of rod OS in RPE, and anti-ezrin (EZRIN; green) was used to distinguish the RPE–photoreceptor border. The boxed area is enlarged in the insert. In control mice, rhodopsin uptake (arrows) was increased with light stimulation (C) compared to no light stimulation (A). Quantification of rhodopsin-positive material in the RPE (E). Western analysis of whole-eye lysates from 2-month control and Mfrprd6/rd6 mice and immunoprecipitated products of MERTK (F). Phospo-MERTK is shown by immunoblot with anti-phospho-Tyrosine (pY) antibody following immunoprecipitation with anti-MERTK (Mer). To quantify the band intensity, the membrane was reblotted with anti-MERTK. The immunoblot of 10% of lysate input was performed with primary antibody as indicated. RPE, retinal pigment epithelium; ELM, external limiting membrane; ONL, outer nuclear layer; OS, outer segment. Bar = 10 μm.