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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

Light damage in Rdh13-/- mice. Light damage was induced in Rdh13+/+ and Rdh13−/− mice by 48 h exposure to diffuse white light (3,000 lx). Twenty-four h dark-adaption after light exposure, and morphological and functional assays were performed as described in Methods. A: Hematoxylin-eosin (H&E) staining showed that the outer-plus-inner-segment and outer nuclear layers of the retina from Rdh13−/− mice that were exposed to light obviously disintegrated. B: The thicknesses of the outer-plus-inner-segment and outer nuclear layers of all genotypes exposed to the light and the control group was valued. Values were mean±SD (n=5, each group). There were statistically significant differences in the thickness of the outer-plus-inner-segment and outer nuclear layer between the light exposed Rdh13−/− mice and the other three groups at any distance point. C: The scotopic electroretinogram response of Rdh13+/+ and Rdh13−/− mice, which were recorded in all groups. D: The amplitudes of a- and b-waves in all genotypes was plotted as the mean±SD (n=5, each goup); *: p<0.05. E: Mitochondria in photoreceptor inner segments of Rdh13+/+ and Rdh13−/− mice exposed to the light were detected by transmission electron microscopy. Distinctly swollen mitochondria with disrupted cristae were observed in Rdh13−/− mice (arrows). F and G: Cytochrome c (CytC) and apoptotic gene expression in all groups were analyzed by Western Blot, which revealed that levels of CytC, apoptosis proteinase activating factor-1 (Apaf-1), cleaved caspase 3, nuclear factor-kappa B P65 (p65) and B-cell lymphoma 2-associated X protein (Bax) were clearly increased in the cytoplasm of Rdh13−/− mice; ONH, optic nerve head; RPE, retinal pigment epithelia; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; UV, ultraviolet.
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f5: Light damage in Rdh13-/- mice. Light damage was induced in Rdh13+/+ and Rdh13−/− mice by 48 h exposure to diffuse white light (3,000 lx). Twenty-four h dark-adaption after light exposure, and morphological and functional assays were performed as described in Methods. A: Hematoxylin-eosin (H&E) staining showed that the outer-plus-inner-segment and outer nuclear layers of the retina from Rdh13−/− mice that were exposed to light obviously disintegrated. B: The thicknesses of the outer-plus-inner-segment and outer nuclear layers of all genotypes exposed to the light and the control group was valued. Values were mean±SD (n=5, each group). There were statistically significant differences in the thickness of the outer-plus-inner-segment and outer nuclear layer between the light exposed Rdh13−/− mice and the other three groups at any distance point. C: The scotopic electroretinogram response of Rdh13+/+ and Rdh13−/− mice, which were recorded in all groups. D: The amplitudes of a- and b-waves in all genotypes was plotted as the mean±SD (n=5, each goup); *: p<0.05. E: Mitochondria in photoreceptor inner segments of Rdh13+/+ and Rdh13−/− mice exposed to the light were detected by transmission electron microscopy. Distinctly swollen mitochondria with disrupted cristae were observed in Rdh13−/− mice (arrows). F and G: Cytochrome c (CytC) and apoptotic gene expression in all groups were analyzed by Western Blot, which revealed that levels of CytC, apoptosis proteinase activating factor-1 (Apaf-1), cleaved caspase 3, nuclear factor-kappa B P65 (p65) and B-cell lymphoma 2-associated X protein (Bax) were clearly increased in the cytoplasm of Rdh13−/− mice; ONH, optic nerve head; RPE, retinal pigment epithelia; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; UV, ultraviolet.

Mentions: When Rdh13−/− mice were exposed to 3,000 lx of white light for 48 h, the outer-plus-inner-segment and outer nuclear layers around the central area (500 μm from the optic nerve head), where the light of strongest intensity penetrates, disintegrated (Figure 5A). The thickness values of the outer-plus-inner-segment were reduced compared to those observed in the Rdh13+/+mice, especially the outer segment (100 μm superior: 6.57±1.02 versus 14.43±1.45 μm, p<0.05; 100 μm inferior: 7.64±1.36 versus 14.79±2.64 μm, p<0.05; 500 μm superior: 14.95±2.41 versus 27.27±1.14 μm, p<0.05; 500 μm inferior: 13.88±1.11 versus 25.70±3.16 μm, p<0.05) (Figure 5B). There was also a distinct difference in the thickness of the outer nuclear layer between the Rdh13−/− and Rdh13+/+mice (100 μm superior: 13.07±4.21 versus 26.62±2.84 μm, p<0.05; 100 μm inferior: 15.01±1.39 versus 24.68±1.70 μm, p<0.05; 500 μm superior: 26.84±2.78 versus 50.52±3.63 μm, p<0.05; 500 μm inferior: 23.86±2.47 versus 40.25±3.71 μm, p<0.05; Figure 5B). These findings were matched to data obtained by the full-field ERG, which revealed markedly decreased mean amplitudes of a- and b-waves under scotopic conditions in the Rdh13−/− mice compared with the Rdh13+/+mice (a-wave amplitude: 55.08±6.73 versus 89.38±3.86 μv, p<0.05; b-wave amplitude: 257.73±69.57 versus 389.30±68.25 μv, p<0.05; Figure 5C,D). TEM showed that the mitochondria in the photoreceptor inner segments of the Rdh13−/− mice were distinctly swollen and contained disrupted cristae. In contrast, the morphology of the mitochondria in the Rdh13+/+ mice was only minimally affected (Figure 5E). Western blot analysis revealed that levels of cytochrome C (CytC), apoptosis proteinase activating factor-1 (Apaf-1), caspase 3, p65, and Bax were clearly increased in the cytoplasm of Rdh13−/− mice (Figure 5F,G).


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)

Light damage in Rdh13-/- mice. Light damage was induced in Rdh13+/+ and Rdh13−/− mice by 48 h exposure to diffuse white light (3,000 lx). Twenty-four h dark-adaption after light exposure, and morphological and functional assays were performed as described in Methods. A: Hematoxylin-eosin (H&E) staining showed that the outer-plus-inner-segment and outer nuclear layers of the retina from Rdh13−/− mice that were exposed to light obviously disintegrated. B: The thicknesses of the outer-plus-inner-segment and outer nuclear layers of all genotypes exposed to the light and the control group was valued. Values were mean±SD (n=5, each group). There were statistically significant differences in the thickness of the outer-plus-inner-segment and outer nuclear layer between the light exposed Rdh13−/− mice and the other three groups at any distance point. C: The scotopic electroretinogram response of Rdh13+/+ and Rdh13−/− mice, which were recorded in all groups. D: The amplitudes of a- and b-waves in all genotypes was plotted as the mean±SD (n=5, each goup); *: p<0.05. E: Mitochondria in photoreceptor inner segments of Rdh13+/+ and Rdh13−/− mice exposed to the light were detected by transmission electron microscopy. Distinctly swollen mitochondria with disrupted cristae were observed in Rdh13−/− mice (arrows). F and G: Cytochrome c (CytC) and apoptotic gene expression in all groups were analyzed by Western Blot, which revealed that levels of CytC, apoptosis proteinase activating factor-1 (Apaf-1), cleaved caspase 3, nuclear factor-kappa B P65 (p65) and B-cell lymphoma 2-associated X protein (Bax) were clearly increased in the cytoplasm of Rdh13−/− mice; ONH, optic nerve head; RPE, retinal pigment epithelia; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; UV, ultraviolet.
© Copyright Policy - open-access
Related In: Results  -  Collection

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f5: Light damage in Rdh13-/- mice. Light damage was induced in Rdh13+/+ and Rdh13−/− mice by 48 h exposure to diffuse white light (3,000 lx). Twenty-four h dark-adaption after light exposure, and morphological and functional assays were performed as described in Methods. A: Hematoxylin-eosin (H&E) staining showed that the outer-plus-inner-segment and outer nuclear layers of the retina from Rdh13−/− mice that were exposed to light obviously disintegrated. B: The thicknesses of the outer-plus-inner-segment and outer nuclear layers of all genotypes exposed to the light and the control group was valued. Values were mean±SD (n=5, each group). There were statistically significant differences in the thickness of the outer-plus-inner-segment and outer nuclear layer between the light exposed Rdh13−/− mice and the other three groups at any distance point. C: The scotopic electroretinogram response of Rdh13+/+ and Rdh13−/− mice, which were recorded in all groups. D: The amplitudes of a- and b-waves in all genotypes was plotted as the mean±SD (n=5, each goup); *: p<0.05. E: Mitochondria in photoreceptor inner segments of Rdh13+/+ and Rdh13−/− mice exposed to the light were detected by transmission electron microscopy. Distinctly swollen mitochondria with disrupted cristae were observed in Rdh13−/− mice (arrows). F and G: Cytochrome c (CytC) and apoptotic gene expression in all groups were analyzed by Western Blot, which revealed that levels of CytC, apoptosis proteinase activating factor-1 (Apaf-1), cleaved caspase 3, nuclear factor-kappa B P65 (p65) and B-cell lymphoma 2-associated X protein (Bax) were clearly increased in the cytoplasm of Rdh13−/− mice; ONH, optic nerve head; RPE, retinal pigment epithelia; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; UV, ultraviolet.
Mentions: When Rdh13−/− mice were exposed to 3,000 lx of white light for 48 h, the outer-plus-inner-segment and outer nuclear layers around the central area (500 μm from the optic nerve head), where the light of strongest intensity penetrates, disintegrated (Figure 5A). The thickness values of the outer-plus-inner-segment were reduced compared to those observed in the Rdh13+/+mice, especially the outer segment (100 μm superior: 6.57±1.02 versus 14.43±1.45 μm, p<0.05; 100 μm inferior: 7.64±1.36 versus 14.79±2.64 μm, p<0.05; 500 μm superior: 14.95±2.41 versus 27.27±1.14 μm, p<0.05; 500 μm inferior: 13.88±1.11 versus 25.70±3.16 μm, p<0.05) (Figure 5B). There was also a distinct difference in the thickness of the outer nuclear layer between the Rdh13−/− and Rdh13+/+mice (100 μm superior: 13.07±4.21 versus 26.62±2.84 μm, p<0.05; 100 μm inferior: 15.01±1.39 versus 24.68±1.70 μm, p<0.05; 500 μm superior: 26.84±2.78 versus 50.52±3.63 μm, p<0.05; 500 μm inferior: 23.86±2.47 versus 40.25±3.71 μm, p<0.05; Figure 5B). These findings were matched to data obtained by the full-field ERG, which revealed markedly decreased mean amplitudes of a- and b-waves under scotopic conditions in the Rdh13−/− mice compared with the Rdh13+/+mice (a-wave amplitude: 55.08±6.73 versus 89.38±3.86 μv, p<0.05; b-wave amplitude: 257.73±69.57 versus 389.30±68.25 μv, p<0.05; Figure 5C,D). TEM showed that the mitochondria in the photoreceptor inner segments of the Rdh13−/− mice were distinctly swollen and contained disrupted cristae. In contrast, the morphology of the mitochondria in the Rdh13+/+ mice was only minimally affected (Figure 5E). Western blot analysis revealed that levels of cytochrome C (CytC), apoptosis proteinase activating factor-1 (Apaf-1), caspase 3, p65, and Bax were clearly increased in the cytoplasm of Rdh13−/− mice (Figure 5F,G).

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