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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

Localization, retinal histology, and thickness measurements for the Rdh13 knockout mice. A: Immunofluorescence localization of Rdh13 (green) was shown in 3-month-old Rdh13+/+ and Rdh13−/− retina paraffin sections. B: western blot analysis of RDH13 protein in wild-type (WT) and homozygous mice revealed that there was no expression of Rdh13 in the retinas of Rdh13 knockout mice. C: Semi-thin sections of WT and homozygous mice retinas revealed no major differences in retinal histology at 3 and 10 months of age. D: Transmission electron microscopy (TEM) of the photoreceptor outer and inner segments and the outer nuclear layer in Rdh13+/+ and Rdh13−/− mice at 10 months of age revealed no apparent abnormalities. E: The outer-plus-inner-segment and outer nuclear layer thickness for Rdh13−/− and WT mice at the ages of 3 months and 10 months was valued. Values were mean±SD (n=5, each group). There were no statistically significant differences between the two genotypes at any distance point. ONH, optic nerve head; 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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f3: Localization, retinal histology, and thickness measurements for the Rdh13 knockout mice. A: Immunofluorescence localization of Rdh13 (green) was shown in 3-month-old Rdh13+/+ and Rdh13−/− retina paraffin sections. B: western blot analysis of RDH13 protein in wild-type (WT) and homozygous mice revealed that there was no expression of Rdh13 in the retinas of Rdh13 knockout mice. C: Semi-thin sections of WT and homozygous mice retinas revealed no major differences in retinal histology at 3 and 10 months of age. D: Transmission electron microscopy (TEM) of the photoreceptor outer and inner segments and the outer nuclear layer in Rdh13+/+ and Rdh13−/− mice at 10 months of age revealed no apparent abnormalities. E: The outer-plus-inner-segment and outer nuclear layer thickness for Rdh13−/− and WT mice at the ages of 3 months and 10 months was valued. Values were mean±SD (n=5, each group). There were no statistically significant differences between the two genotypes at any distance point. ONH, optic nerve head; 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: For Rdh13 localization in mouse eyes, immunofluorescence analysis showed that Rdh13 was expressed in the photoreceptor inner segment layer, with no apparent expression in the RPE, photoreceptor outer segments, or outer nuclear layer in WT mice (Figure 3A). Rdh13 expression was abolished in the retinas of Rdh13 KO mice, as determined by immunofluorescence (Figure 3A) and western blot (Figure 3B). Light micrographs of retina sections from Rdh13 KO and WT mice revealed no major differences in retinal histology at 3 and 10 months of age (Figure 3C, at 3 months and 10 months), with normal lamination and numbers of cells. Similarly, TEM at 10 months of age revealed no apparent abnormalities in retinal architecture (Figure 3D). There was no evidence of any abnormality in Bruch’s membrane, increased pigmented body and lipofuscin accumulation in retinal pigment epithelium cells, or photoreceptor debris. The thickness of the outer-plus-inner-segment and outer nuclear layer of the retina was similar at 1,400 µm to that of the optic nerve head for both genotypes at 3 months and 10 months of age (Figure 3E).


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)

Localization, retinal histology, and thickness measurements for the Rdh13 knockout mice. A: Immunofluorescence localization of Rdh13 (green) was shown in 3-month-old Rdh13+/+ and Rdh13−/− retina paraffin sections. B: western blot analysis of RDH13 protein in wild-type (WT) and homozygous mice revealed that there was no expression of Rdh13 in the retinas of Rdh13 knockout mice. C: Semi-thin sections of WT and homozygous mice retinas revealed no major differences in retinal histology at 3 and 10 months of age. D: Transmission electron microscopy (TEM) of the photoreceptor outer and inner segments and the outer nuclear layer in Rdh13+/+ and Rdh13−/− mice at 10 months of age revealed no apparent abnormalities. E: The outer-plus-inner-segment and outer nuclear layer thickness for Rdh13−/− and WT mice at the ages of 3 months and 10 months was valued. Values were mean±SD (n=5, each group). There were no statistically significant differences between the two genotypes at any distance point. ONH, optic nerve head; 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

License
Show All Figures
getmorefigures.php?uid=PMC3351414&req=5

f3: Localization, retinal histology, and thickness measurements for the Rdh13 knockout mice. A: Immunofluorescence localization of Rdh13 (green) was shown in 3-month-old Rdh13+/+ and Rdh13−/− retina paraffin sections. B: western blot analysis of RDH13 protein in wild-type (WT) and homozygous mice revealed that there was no expression of Rdh13 in the retinas of Rdh13 knockout mice. C: Semi-thin sections of WT and homozygous mice retinas revealed no major differences in retinal histology at 3 and 10 months of age. D: Transmission electron microscopy (TEM) of the photoreceptor outer and inner segments and the outer nuclear layer in Rdh13+/+ and Rdh13−/− mice at 10 months of age revealed no apparent abnormalities. E: The outer-plus-inner-segment and outer nuclear layer thickness for Rdh13−/− and WT mice at the ages of 3 months and 10 months was valued. Values were mean±SD (n=5, each group). There were no statistically significant differences between the two genotypes at any distance point. ONH, optic nerve head; 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: For Rdh13 localization in mouse eyes, immunofluorescence analysis showed that Rdh13 was expressed in the photoreceptor inner segment layer, with no apparent expression in the RPE, photoreceptor outer segments, or outer nuclear layer in WT mice (Figure 3A). Rdh13 expression was abolished in the retinas of Rdh13 KO mice, as determined by immunofluorescence (Figure 3A) and western blot (Figure 3B). Light micrographs of retina sections from Rdh13 KO and WT mice revealed no major differences in retinal histology at 3 and 10 months of age (Figure 3C, at 3 months and 10 months), with normal lamination and numbers of cells. Similarly, TEM at 10 months of age revealed no apparent abnormalities in retinal architecture (Figure 3D). There was no evidence of any abnormality in Bruch’s membrane, increased pigmented body and lipofuscin accumulation in retinal pigment epithelium cells, or photoreceptor debris. The thickness of the outer-plus-inner-segment and outer nuclear layer of the retina was similar at 1,400 µm to that of the optic nerve head for both genotypes at 3 months and 10 months of age (Figure 3E).

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