Mao PDF
Biochemistry and Molecular Biology
Mitochondrial Oxidative Stress in the Retinal Pigment
Epithelium Leads to Localized Retinal Degeneration
Haoyu Mao,1 Soo Jung Seo,1 Manas R. Biswal,1 Hong Li,1 Mandy Conners,1 Arathi Nandyala,1
Kyle Jones,1 Yun-Zheng Le,2 and Alfred S. Lewin1
1
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, Florida, United States
Departments of Medicine, Endocrinology, and Cell Biology and Harold Hamm Diabetes Center, University of Oklahoma Health
Sciences Center, Oklahoma City, Oklahoma, United States
2
Correspondence: Alfred S. Lewin,
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Box
100266, Gainesville, FL 32610, USA; lewin@ufl.edu. Submitted: April 21, 2014
Accepted: June 12, 2014
Citation: Mao H, Seo SJ, Biswal MR, et al. Mitochondrial oxidative stress in the retinal pigment epithelium leads to localized retinal degeneration. Invest Ophthalmol Vis Sci.
2014;55:4613–4627. DOI:10.1167/ iovs.14-14633 PURPOSE. Oxidative stress in the RPE is widely accepted as a contributing factor to AMD. We have previously shown that ribozyme-mediated reduction in the antioxidant enzyme manganese superoxide dismutase (MnSOD) leads to some of the features of geographic atrophy in mice. To develop a mouse model independent of viral injection, we used a conditional knockout of the Sod2 gene in the RPE to elevate mitochondrial oxidative stress in that cell layer.
METHODS. Experimental mice in which exon 3 of Sod2 was flanked by loxP sites were also transgenic for PVMD2-rtTA and tetO-PhCMV cre, so that cre recombinase was expressed only in the RPE. Pups of this genotype (Sod2flox/floxVMD2cre) were induced to express cre recombinase by feeding doxycycline-laced chow to nursing dams. Controls included mice of this genotype not treated with doxycycline and doxycycline-treated Sod2flox/flox mice lacking the cre transgene. Expression of cre in the RPE was
Mitochondrial Oxidative Stress in the Retinal Pigment
Epithelium Leads to Localized Retinal Degeneration
Haoyu Mao,1 Soo Jung Seo,1 Manas R. Biswal,1 Hong Li,1 Mandy Conners,1 Arathi Nandyala,1
Kyle Jones,1 Yun-Zheng Le,2 and Alfred S. Lewin1
1
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, Florida, United States
Departments of Medicine, Endocrinology, and Cell Biology and Harold Hamm Diabetes Center, University of Oklahoma Health
Sciences Center, Oklahoma City, Oklahoma, United States
2
Correspondence: Alfred S. Lewin,
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Box
100266, Gainesville, FL 32610, USA; lewin@ufl.edu. Submitted: April 21, 2014
Accepted: June 12, 2014
Citation: Mao H, Seo SJ, Biswal MR, et al. Mitochondrial oxidative stress in the retinal pigment epithelium leads to localized retinal degeneration. Invest Ophthalmol Vis Sci.
2014;55:4613–4627. DOI:10.1167/ iovs.14-14633 PURPOSE. Oxidative stress in the RPE is widely accepted as a contributing factor to AMD. We have previously shown that ribozyme-mediated reduction in the antioxidant enzyme manganese superoxide dismutase (MnSOD) leads to some of the features of geographic atrophy in mice. To develop a mouse model independent of viral injection, we used a conditional knockout of the Sod2 gene in the RPE to elevate mitochondrial oxidative stress in that cell layer.
METHODS. Experimental mice in which exon 3 of Sod2 was flanked by loxP sites were also transgenic for PVMD2-rtTA and tetO-PhCMV cre, so that cre recombinase was expressed only in the RPE. Pups of this genotype (Sod2flox/floxVMD2cre) were induced to express cre recombinase by feeding doxycycline-laced chow to nursing dams. Controls included mice of this genotype not treated with doxycycline and doxycycline-treated Sod2flox/flox mice lacking the cre transgene. Expression of cre in the RPE was
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