Aging changes in the retina of male albino rat: a histological, ultrastructural and immunohistochemical study
Abstract
Background: Degenerative changes caused by aging may affect the eye, especially the retina. Such changes occur as a part of normal physiological process and may be irreversible. The aim of the study was to demonstrate the influence of aging on the morphology of the retina to provide a basis to explain the pathogenesis of age-associated decline in visual acuity, scotopic and photopic sensitivity.
Materials and methods: Forty male albino rats were used and divided into four age groups (group I: age of cortical maturity, group II: middle-aged, group III: aged group and group IV: senile group). The rats were sacrificed, the eye balls were enucleated. Intra-vitreal injections of formalin for haematoxylin and eosin and immunohistochemical sections, glutaraldehyde for toluidine blue semithin and E/M ultra-thin sections were performed. Measurements and quantitative histomorphometric estimation of the layers of the retina were done.
Results: Light microscopic examination revealed age-dependent attenuation of photoreceptor striations. Aged and senile groups presented pyknotic, widely- -spaced nuclei of the outer nuclear layer. The inner nuclear layer was thinned out to 2 or 3 cellular rows. Retinal capillaries showed progressive dilatation and congestion. Statistical analysis proved significant thinning of the retina with variable degrees of thinning of the constituting layers. Decreased arborisation with age was confirmed with quantification of synaptophysin-immunostained sections. Glial fibrillary acidic protein immunostaining revealed the picture of reactive gliosis. On the ultrastructural level, the retinal pigment epithelium exhibited major alterations with aging. Numerous phagosomes, lipofuscin and melanolipofusin granules appeared within the cells, together with exaggerated basal infoldings. The pho- toreceptor nuclei became degenerated and the perinuclear space was widened.
Conclusions: Rat retinae clearly undergo age-related morphological changes. Such changes provide a cellular base for explanation of decreased vision in humans with aging other than reflection errors. Effect of aging was not only qualitative, but also quantitative.
Keywords: retinaageingultrastructureimmunohistochemicalhistology
References
- Aggarwal P, Nag TC, Wadhwa S. Age-related decrease in rod bipolar cell density of the human retina: an immunohistochemical study. J Biosci. 2007; 32(2): 293–298.
- Bancroft JD, Gamble M. Immunohistochemical technique. In: Theory and practice of histological techniques, 6th ed. Churchill Livingstone, London 2008: 433–472.
- Bianchi E, Ripandelli G, Taurone S, et al. Age and diabetes related changes of the retinal capillaries: An ultrastructural and immunohistochemical study. Int J Immunopathol Pharmacol. 2016; 29(1): 40–53.
- Bonilha VL. Age and disease-related structural changes in the retinal pigment epithelium. Clin Ophthalmol. 2008; 2(2): 413–424.
- Boulton ME. Studying melanin and lipofuscin in RPE cell culture models. Exp Eye Res. 2014; 126: 61–67.
- Boya P, Esteban-Martínez L, Serrano-Puebla A, et al. Autophagy in the eye: Development, degeneration, and aging. Prog Retin Eye Res. 2016; 55: 206–245.
- Calkins DJ. Age-related changes in the visual pathways: blame it on the axon. Invest Ophthalmol Vis Sci. 2013; 54(14): ORSF37–ORSF41.
- Cattoretti G, Pileri S, Parravicini C, et al. Antigen unmasking on formalin-fixed, paraffin-embedded tissue sections. J Pathol. 1993; 171(2): 83–98.
- Chernoivanenko IS, Matveeva EA, Gelfand VI, et al. Mitochondrial membrane potential is regulated by vimentin intermediate filaments. FASEB J. 2015; 29(3): 820–827.
- Chirco KR, Sohn EH, Stone EM, et al. Structural and molecular changes in the aging choroid: implications for age-related macular degeneration. Eye. 2017; 31(1): 10–25.
- Dan C, Jian-Bin T, Hui W, et al. Synaptophysin expression in rat retina following acute high intraocular pressure. Acta Histochem Cytochem. 2008; 41(6): 173–178.
- de Pablo Y, Nilsson M, Pekna M, et al. Intermediate filaments are important for astrocyte response to oxidative stress induced by oxygen-glucose deprivation and reperfusion. Histochem Cell Biol. 2013; 140(1): 81–91.
- Donato AJ, Black AD, Jablonski KL, et al. Aging is associated with greater nuclear NF kappa B, reduced I kappa B alpha, and increased expression of proinflammatory cytokines in vascular endothelial cells of healthy humans. Aging Cell. 2008; 7(6): 805–812.
- Dorfman AL, Cuenca N, Pinilla I, et al. Immunohistochemical evidence of synaptic retraction, cytoarchitectural remodeling, and cell death in the inner retina of the rat model of oygen-induced retinopathy (OIR). Invest Ophthalmol Vis Sci. 2011; 52(3): 1693–1708.
- Feng L, Sun Z, Han H, et al. No age-related cell loss in three retinal nuclear layers of the Long-Evans rat. Vis Neurosci. 2007; 24(6): 799–803.
- Fernández-Sánchez L, de Sevilla Müller LP, Brecha NC, et al. Loss of outer retinal neurons and circuitry alterations in the DBA/2J mouse. Invest Ophthalmol Vis Sci. 2014; 55(9): 6059–6072.
- Hippert C, Graca AB, Barber AC, et al. Müller glia activation in response to inherited retinal degeneration is highly varied and disease-specific. PLoS One. 2015; 10(3): e0120415.
- Joly S, Pernet V, Samardzija M, et al. Pax6-positive Müller glia cells express cell cycle markers but do not proliferate after photoreceptor injury in the mouse retina. Glia. 2011; 59(7): 1033–1046.
- Karthaus M, Falkenstein M. Functional Changes and Driving Performance in Older Drivers: Assessment and Interventions. Geriatrics. 2016; 1(2): 12.
- Keeling E, Lotery AJ, Tumbarello DA, et al. Impaired cargo clearance in the retinal pigment epithelium (RPE) underlies irreversible blinding diseases. Cells. 2018; 7(2).
- Klaassen I, Van Noorden CJF, Schlingemann RO. Molecular basis of the inner blood-retinal barrier and its breakdown in diabetic macular edema and other pathological conditions. Prog Retin Eye Res. 2013; 34: 19–48.
- Litts KM, Messinger JD, Freund KB, et al. Inner segment remodeling and mitochondrial translocation in cone photoreceptors in age-related macular degeneration with outer retinal tubulation. Invest Ophthalmol Vis Sci. 2015; 56(4): 2243–2253.
- Luke MPS, LeVatte TL, O'Reilly AM, et al. Effect of NCAM on aged-related deterioration in vision. Neurobiol Aging. 2016; 41: 93–106.
- Ma W, Coon S, Zhao L, et al. A2E accumulation influences retinal microglial activation and complement regulation. Neurobiol Aging. 2013; 34(3): 943–960.
- Martinez-De Luna RI, Ku RY, Aruck AM, et al. Müller glia reactivity follows retinal injury despite the absence of the glial fibrillary acidic protein gene in Xenopus. Dev Biol. 2017; 426(2): 219–235.
- Nadal-Nicolás FM, Sobrado-Calvo P, Jiménez-López M, et al. Long-Term Effect of Optic Nerve Axotomy on the Retinal Ganglion Cell Layer. Invest Ophthalmol Vis Sci. 2015; 56(10): 6095–6112.
- Nadal-Nicolás FM, Vidal-Sanz M, Agudo-Barriuso M. The aging rat retina: from function to anatomy. Neurobiol Aging. 2018; 61: 146–168.
- Nag T, Wadhwa S. Ultrastructure of the human retina in aging and various pathological states. Micron. 2012; 43(7): 759–781.
- Nieves-Moreno M, Martínez-de-la-Casa JM, Morales-Fernández L, et al. Impacts of age and sex on retinal layer thicknesses measured by spectral domain optical coherence tomography with Spectralis. PLoS One. 2018; 13(3): e0194169.
- Rodríguez-Muela N, Koga H, García-Ledo L, et al. Balance between autophagic pathways preserves retinal homeostasis. Aging Cell. 2013; 12(3): 478–488.
- Samuel MA, Zhang Y, Meister M, et al. Age-related alterations in neurons of the mouse retina. J Neurosci. 2011; 31(44): 16033–16044.
- Sanes JR, Zipursky SL. Design principles of insect and vertebrate visual systems. Neuron. 2010; 66(1): 15–36.
- Sengupta P. The laboratory rat: relating its age with human's. Int J Prev Med. 2013; 4(6): 624–630.
- Szabadfi K, Estrada C, Fernandez-Villalba E, et al. Retinal aging in the diurnal Chilean rodent (Octodon degus): histological, ultrastructural and neurochemical alterations of the vertical information processing pathway. Front Cell Neurosc. 2015; 9.
- Warburton S, Davis WE, Southwick K, et al. Proteomic and phototoxic characterization of melanolipofuscin: correlation to disease and model for its origin. Mol Vis. 2007; 13: 318–329.
- Yao J, Jia L, Shelby SJ, et al. Circadian and noncircadian modulation of autophagy in photoreceptors and retinal pigment epithelium. Invest Ophthalmol Vis Sci. 2014; 55(5): 3237–3246.
- Yassa HD. Age-related changes in the optic nerve of Sprague-Dawley rats: an ultrastructural and immunohistochemical study. Acta Histochem. 2014; 116(6): 1085–1095.
- Zhao JJ, Ouyang H, Luo J, et al. Induction of retinal progenitors and neurons from mammalian Müller glia under defined conditions. J Biol Chem. 2014; 289(17): 11945–11951.
