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Retinol dehydrogenase 13 protects the mouse retina from acute light damage.

Wang H, Cui X, Gu Q, Chen Y, Zhou J, Kuang Y, Wang Z, Xu X - Mol. Vis. (2012)

Bottom Line: We found that there was no obvious difference in phenotype or function between Rdh13 knockout and wild-type mice.Increased expression levels of CytC, CytC-responsive apoptosis proteinase activating factor-1 (Apaf-1) and caspases 3, and other mitochondria apoptosis-related genes (nuclear factor-kappa B P65 [P65] and B-cell lymphoma 2-associated X protein [Bax]) were observed in Rdh13(-/-) mice.The mechanism may involve inhibition of the mitochondrial apoptosis pathway.

View Article: PubMed Central - PubMed

ABSTRACT

Purpose: To investigate whether retinol dehydrogenase 13 (RDH13) can protect the retina from acute light-induced damage.

Methods: We generated Rdh13 knockout mice using molecular biologic methods and assessed the associated morphological and functional changes under room-light conditions by hematoxylin-eosin (H&E), transmission electron microscopy (TEM), and scotopic electroretinography. Then, the light-damage model was established by exposure to diffuse white light (3,000 lx) for 48 h. Twenty-four h after light exposure, H&E was used for the histological evaluation. The thickness of the outer-plus-inner-segment and the outer nuclear layer was measured on sections parallel to the vertical meridian of the eye. An electroretinography test was performed to assess the functional change. Furthermore, the impairment of mitochondria was detected by TEM. Finally, the expression of cytochrome c (CytC) and other apoptosis-related proteins was detected by western blot.

Results: We found that there was no obvious difference in phenotype or function between Rdh13 knockout and wild-type mice. In Rdh13(-/-) mice subjected to intense light exposure, the photoreceptor outer-plus-inner-segment and outer nuclear layer were dramatically shorter, and the amplitudes of a- and b-waves under scotopic conditions were significantly attenuated. Distinctly swollen mitochondria with disrupted cristae were observed in the photoreceptor inner segments of Rdh13(-/-) mice. Increased expression levels of CytC, CytC-responsive apoptosis proteinase activating factor-1 (Apaf-1) and caspases 3, and other mitochondria apoptosis-related genes (nuclear factor-kappa B P65 [P65] and B-cell lymphoma 2-associated X protein [Bax]) were observed in Rdh13(-/-) mice.

Conclusions: Rdh13 can protect the retina against acute light-induced retinopathy. The mechanism may involve inhibition of the mitochondrial apoptosis pathway.

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Related in: MedlinePlus

Full-field electroretinogram responses and apoptosis detection in Rdh13+/+ and Rdh13−/− mice. A: The scotopic electroretinogram responses of Rdh13+/+ and Rdh13−/− mice at 10 months of age were recorded. B: The amplitudes of a- and b-waves for either genotype was plotted as the mean±SD (n=5, each group), *: p>0.05. C: The terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining showed that there was no obvious apoptosis in both Rdh13+/+ and Rdh13−/− mice at 10 months of age. D: Apoptosis genes expression in Rdh13+/+, Rdh13+/−, and Rdh13−/− mice was detected by western blots. There was no increased expression level of apoptosis genes in all genotypes, which was in accordance with the result of TUNEL.TNF-α, tumor necrosis factor alpha; Fas, TNF receptor superfamily member 6; Bax, B-cell lymphoma 2-associated X protein; P65, nuclear factor-kappa B P65; RPE, retinal pigment epithelia; OS, outer segments; IS, inner segments; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.
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f4: Full-field electroretinogram responses and apoptosis detection in Rdh13+/+ and Rdh13−/− mice. A: The scotopic electroretinogram responses of Rdh13+/+ and Rdh13−/− mice at 10 months of age were recorded. B: The amplitudes of a- and b-waves for either genotype was plotted as the mean±SD (n=5, each group), *: p>0.05. C: The terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining showed that there was no obvious apoptosis in both Rdh13+/+ and Rdh13−/− mice at 10 months of age. D: Apoptosis genes expression in Rdh13+/+, Rdh13+/−, and Rdh13−/− mice was detected by western blots. There was no increased expression level of apoptosis genes in all genotypes, which was in accordance with the result of TUNEL.TNF-α, tumor necrosis factor alpha; Fas, TNF receptor superfamily member 6; Bax, B-cell lymphoma 2-associated X protein; P65, nuclear factor-kappa B P65; RPE, retinal pigment epithelia; OS, outer segments; IS, inner segments; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.

Mentions: To evaluate whether visual function was impaired in Rdh13 KO mice, Ganzfield ERG was performed on 10-month-old Rdh13+/+ and Rdh13 −/− mice. The typical ERG records are displayed in Figure 4A. There was no significant difference in the amplitudes of the a- and b-waves of the scotopic ERG response between dark-adapted Rdh13+/+ and Rdh13−/− mice (a-wave amplitude: 181.75±19.44 versus 171.32±36.95 μv, p>0.05; b-wave amplitude: 864.45±74.18 versus 730.95±151.52 μv, p>0.05; Figure 4B). Thus, Rdh13 deletion did not have a significant effect on the rod-mediated light response. TUNEL detection showed that there was no obvious apoptosis in 10-month-old Rdh13−/− mice (Figure 4C), as determined by western blot, which showed no increased expression levels of caspases 3, P65, or other mitochondria apoptosis-related genes (Fas, TNF-α and Bax; Figure 4D).


Retinol dehydrogenase 13 protects the mouse retina from acute light damage.

Wang H, Cui X, Gu Q, Chen Y, Zhou J, Kuang Y, Wang Z, Xu X - Mol. Vis. (2012)

Full-field electroretinogram responses and apoptosis detection in Rdh13+/+ and Rdh13−/− mice. A: The scotopic electroretinogram responses of Rdh13+/+ and Rdh13−/− mice at 10 months of age were recorded. B: The amplitudes of a- and b-waves for either genotype was plotted as the mean±SD (n=5, each group), *: p>0.05. C: The terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining showed that there was no obvious apoptosis in both Rdh13+/+ and Rdh13−/− mice at 10 months of age. D: Apoptosis genes expression in Rdh13+/+, Rdh13+/−, and Rdh13−/− mice was detected by western blots. There was no increased expression level of apoptosis genes in all genotypes, which was in accordance with the result of TUNEL.TNF-α, tumor necrosis factor alpha; Fas, TNF receptor superfamily member 6; Bax, B-cell lymphoma 2-associated X protein; P65, nuclear factor-kappa B P65; RPE, retinal pigment epithelia; OS, outer segments; IS, inner segments; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3351414&req=5

f4: Full-field electroretinogram responses and apoptosis detection in Rdh13+/+ and Rdh13−/− mice. A: The scotopic electroretinogram responses of Rdh13+/+ and Rdh13−/− mice at 10 months of age were recorded. B: The amplitudes of a- and b-waves for either genotype was plotted as the mean±SD (n=5, each group), *: p>0.05. C: The terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining showed that there was no obvious apoptosis in both Rdh13+/+ and Rdh13−/− mice at 10 months of age. D: Apoptosis genes expression in Rdh13+/+, Rdh13+/−, and Rdh13−/− mice was detected by western blots. There was no increased expression level of apoptosis genes in all genotypes, which was in accordance with the result of TUNEL.TNF-α, tumor necrosis factor alpha; Fas, TNF receptor superfamily member 6; Bax, B-cell lymphoma 2-associated X protein; P65, nuclear factor-kappa B P65; RPE, retinal pigment epithelia; OS, outer segments; IS, inner segments; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.
Mentions: To evaluate whether visual function was impaired in Rdh13 KO mice, Ganzfield ERG was performed on 10-month-old Rdh13+/+ and Rdh13 −/− mice. The typical ERG records are displayed in Figure 4A. There was no significant difference in the amplitudes of the a- and b-waves of the scotopic ERG response between dark-adapted Rdh13+/+ and Rdh13−/− mice (a-wave amplitude: 181.75±19.44 versus 171.32±36.95 μv, p>0.05; b-wave amplitude: 864.45±74.18 versus 730.95±151.52 μv, p>0.05; Figure 4B). Thus, Rdh13 deletion did not have a significant effect on the rod-mediated light response. TUNEL detection showed that there was no obvious apoptosis in 10-month-old Rdh13−/− mice (Figure 4C), as determined by western blot, which showed no increased expression levels of caspases 3, P65, or other mitochondria apoptosis-related genes (Fas, TNF-α and Bax; Figure 4D).

Bottom Line: We found that there was no obvious difference in phenotype or function between Rdh13 knockout and wild-type mice.Increased expression levels of CytC, CytC-responsive apoptosis proteinase activating factor-1 (Apaf-1) and caspases 3, and other mitochondria apoptosis-related genes (nuclear factor-kappa B P65 [P65] and B-cell lymphoma 2-associated X protein [Bax]) were observed in Rdh13(-/-) mice.The mechanism may involve inhibition of the mitochondrial apoptosis pathway.

View Article: PubMed Central - PubMed

ABSTRACT

Purpose: To investigate whether retinol dehydrogenase 13 (RDH13) can protect the retina from acute light-induced damage.

Methods: We generated Rdh13 knockout mice using molecular biologic methods and assessed the associated morphological and functional changes under room-light conditions by hematoxylin-eosin (H&E), transmission electron microscopy (TEM), and scotopic electroretinography. Then, the light-damage model was established by exposure to diffuse white light (3,000 lx) for 48 h. Twenty-four h after light exposure, H&E was used for the histological evaluation. The thickness of the outer-plus-inner-segment and the outer nuclear layer was measured on sections parallel to the vertical meridian of the eye. An electroretinography test was performed to assess the functional change. Furthermore, the impairment of mitochondria was detected by TEM. Finally, the expression of cytochrome c (CytC) and other apoptosis-related proteins was detected by western blot.

Results: We found that there was no obvious difference in phenotype or function between Rdh13 knockout and wild-type mice. In Rdh13(-/-) mice subjected to intense light exposure, the photoreceptor outer-plus-inner-segment and outer nuclear layer were dramatically shorter, and the amplitudes of a- and b-waves under scotopic conditions were significantly attenuated. Distinctly swollen mitochondria with disrupted cristae were observed in the photoreceptor inner segments of Rdh13(-/-) mice. Increased expression levels of CytC, CytC-responsive apoptosis proteinase activating factor-1 (Apaf-1) and caspases 3, and other mitochondria apoptosis-related genes (nuclear factor-kappa B P65 [P65] and B-cell lymphoma 2-associated X protein [Bax]) were observed in Rdh13(-/-) mice.

Conclusions: Rdh13 can protect the retina against acute light-induced retinopathy. The mechanism may involve inhibition of the mitochondrial apoptosis pathway.

Show MeSH
Related in: MedlinePlus