The role of the retinal pigment epithelial cells on the viability and renewal of photoreceptors has been well demonstrated in the Royal College of Surgeons (RCS) strain of rat. These rats are characterized by an inherited time-dependent degeneration of their photoreceptors. This degeneration is apparently due to the inability of the retinal pigment epithelial cells to adequately ingest fragments of photoreceptor membrane that are shed during the course of photoreceptor membrane renewal. The buildup of photoreceptor material in the interphotoreceptor space ultimately leads to the degeneration of photoreceptors in these animals. With regard to the pigment epithelial cells, neither the mechanism mediating the ingestion process in normal rats nor the nature of the defect of this process in RCS rats is understood.It is the goal of this proposed research to assay for the presence of phospholipase C in retinal pigment epithelial (RPE) cells and to determine possible modulators of the enzyme in an attempt to associate this with the process of phagocytosis. / Department of Biology

The Royal College of Surgeons (RCS) rat is an established animal model used to study human retinal dystrophies. This study investigated whether kidney dysfunction accompanies the eye abnormalities seen in this model. Overnight urine collection procedures were used to measure protein excretion in 2, 12, and 22 month old female pigmented RCS rats and control rats (RDY). Clearance experiments were performed in anesthetized rats to measure glomerular filtration rate (GFR) and renal plasma flow rate (RPF). There was an age-related increase in protein excretion in both RCS and RDY rats, but the protein excretion was significantly higher in the RCS rats at 2 and 22 months of age. Whole kidney GFR and RPF were significantly lower in the 22 month old RCS rats, when compared to age-matched RDY rats. These findings suggest that the RCS rat exhibits both kidney and eye abnormalities. / Department of Physiology and Health Science

Tez (Tıpta Uzmanlık) - Süleyman Demirel Üniversitesi, Tıp Fakültesi, Göz Hastalıkları Anabilim Dalı, 2002. / Kaynakça var.

Thesis (M.A.)--York University, 2005. Graduate Programme in Psychology. / Typescript. Includes bibliographical references (leaves 58-62). Also available on the Internet. MODE OF ACCESS via web browser by entering the following URL: http://gateway.proquest.com/openurl?url%5Fver=Z39.88-2004&res%5Fdat=xri:pqdiss &rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&rft_dat=xri:pqdiss:MR11906

In the mammalian retina, a small population of retinal ganglion cells are intrinsically photosensitive due to the expression of the photopigment melanopsin. These intrinsically photosensitive retinal ganglion cells (ipRGCs) integrate their own intrinsic light response with that of rod and cone photoreceptors to drive a variety of physiological and behavioural responses to light. Recently, however, a subset of these cells have been shown to project to the dorsal Lateral Geniculate Nucleus (dLGN) of the visual thalamus, where they directly contribute to visual perception. In the case of retinal degenerations (the most common being retinitis pigmentosa which affects up to 1:2000 people worldwide), the death of the rod and cone photoreceptors results in complete visual blindness with no available treatment. At least some ipRGCs survive retinal degeneration and can continue signalling light information to the dLGN, suggesting that these cells could support some form of visual perception. However, to-date little is known about this projection during retinal degeneration. Thus, characterising its anatomy and physiology is key to determining the quality of visual information that is conveyed to the dLGN during retinal degeneration and what prevents these cells supporting behaviourally relevant vision. A subset of ipRGCs target the dLGN and continue signalling light information even at advanced stages of retinal degeneration. However, it is unknown whether all ipRGC subtypes survive following the death of rod and cone photoreceptors, and whether they retain normal dendritic architecture following reorganisation of the remnant neural retina. We set out to answer these questions using the multi-colour labelling technique Brainbow. In doing so, we design and describe a unique methodology and toolset, based on Principal Component Analysis (PCA), to analyse 3-Dimensional (3D) multi-colour images. We then demonstrate its utility by identifying, isolating and reconstructing the 3D morphology of individual ipRGCs from a population of labelled cells in the degenerate retina and quantitatively characterise their dendritic architecture. The results indicate that all known ipRGC subtypes are resilient to the effects of outer photoreceptor degeneration. Melanopsin responses in the dLGN have been shown to support global brightness perception in mice with advanced retinal degeneration. However, to-date, it is unknown whether these cells can encode spatial information. Using in-vitro and in-vivo electrophysiological recordings from mice in advanced stages of retinal degeneration, we demonstrate for the first time that ipRGCs in the retina, and their target neurones in the dLGN, possess discrete spatial receptive fields. These receptive fields are large and lack a centre-surround organisation. The retinotopic organisation of these cells' projections would suggest they could support spatial vision. However the poor temporal resolution of the deafferented melanopsin response is the most significant limitation precluding melanopsin signalling from supporting behaviourally relevant vision under naturalistic viewing conditions. Considering these temporal limitations, we finally investigated if melanopsin could contribute to visual perception at earlier stages of degeneration which is more representative of clinical conditions in humans. Here, vision can rely on both the intrinsic melanopsin-driven light response and residual cone function. Using silent substitution in combination with in-vivo electrophysiological recordings from the dLGN of mice in early-stage degeneration, we identify a number of cone-driven responses which could support normal visual function. However, we were unable to detect a significant and robust contribution of melanopsin signalling to these residual light-responses using our silent substitution stimuli in both retinally degenerate and wildtype mice at these age. However, we did find a significant contribution to the Olivary Pretectal Nucleus (OPN) of visually intact mice at equivalent ages, and to the adult dLGN. Supported by anatomical data, this suggests that there is a specific temporal delay in the maturation of ipRGCs which project to the dLGN during development of the visual system.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 6 uppsatser.

This research investigated the influence of the normal aging process on truly photopic achromatic and chromatic retinal increment threshold over an extensive range of colored targets. It was found that for both achromatic and chromatic retinal thresholds there is a significant correlation between advancing age and the stimulus luminance intensity at threshold across the visible spectrum. This correlation is greater for the short wavelength range than for both the middle and the long wavelength ranges (r² (achromatic)= 0.43; r²(chromatic) = 0.49. In addition, a small exploratory study was completed which examined the role of prereceptoral changes in these age-related differences in color vision function. Results from this preliminary study (Experiment II) support the supposition that prereceptoral factors cannot account for all of the age-related losses that are seen in visual function, particularly in the middle and long wavelength ranges of the visible spectrum. / Arts, Faculty of / Psychology, Department of / Graduate

Vision provides a critical interface with the physical world. This work examines visual development and vision loss in mice to glean the influence of the retinal state on visual connections. I first assessed the impact of retinal activity on the eye-specific segregation of retinal afferents in the lateral geniculate nucleus (LGN) of young Gβ5 -/- mice. Gβ5 is the fifth member of the β subfamily of heterotrimeric G proteins. Gβ5 binds and stabilizes the R7 family of regulators of G-protein signaling (RGS), which accelerate Gi/o GTP hydrolysis. Gβ5 -/- mice, which lack R7RGS activity, have malformed synapses in the outer plexiform layer (OPL) and impaired OPL transmission. Altered spontaneous retinal activity in Gβ5-/- mice at P7, P12, P14, and P28 correlates with impaired eye-specific segregation of retinal afferents in the LGN at corresponding timepoints. However, Gβ5-/- mice exhibit a normal transition from cholinergic to glutamatergic drive that corresponds with a temporary recovery of refinement at P10. Thus the abnormal-normal-abnormal pattern of activity in the retina is coupled with abnormal-normal-abnormal segregation. This activity-segregation profile suggests activity may instruct early retinogeniculate development. nob mice, which also exhibit impaired OPL transmission, have aberrant retinal waves that align with loss of segregation. nobxGβ5-/- mice have similar levels of segregation as Gβ5-/- at P21, but activity only similar P14 nobxGβ5-/- and Gβ5-/- RGCs. This suggests that the critical period of eye-specific segregation closes shortly after P14 and that R7RGS activity is critically important to postnatal RGCs. Next, I investigated the aged visual system via the retinofugal projections of mice with retinal remodeling after photoreceptor degeneration (PD). ΔCT mice, with mild remodeling, and TG9N mice, with aggressive remodeling, retain gross anatomical and physiological connectivity in the presence of attenuated visual activity compounded by organic remodeling. However, the magnitude of pupillary light responses in PD mice was diminished. Reduced melanopsin signal in the retina, not downstream anomalies, explains this functional deficiency. These observations suggest that changes to eye-specific segregation are limited once projections are established, regardless of retinal activity or remodeling. These observations bode well for future retina-based treatments of vision loss.

The general aim of the thesis was to deduce whether selected naturally occurring flavonoids (genistein, epicatechin gallate (EC), epigallocatechin gallate (EGCG), baicalin) attenuate various secondary insults that may cause death of ganglion cells in primary open angle glaucoma (POAG). An ischemic insult to the rat retina significantly causes the inner retina to degenerate indexed by changes of various antigens, proteins and mRNAs located to amacrine and ganglion cells. These changes are blunted in animals treated with genistein as has been shown for ECGC. Studies conducted on cells (RGC-5 cells) in culture showed that hydrogen peroxide, L-buthionine sulfoximine (BSO)/glutamate and serum deprivation (mimicking oxidative stress), rotenone, sodium azide (affecting mitochondria function in specific ways) and light (where the mitochondria are generally affected) all generated reactive oxygen species and caused death of RGC-5 cells. EGCG was able to attenuate cell death caused by hydrogen peroxide, sodium azide and rotenone. Only EC was able to attenuate BSO/glutamate-induced cell death, in addition to cell death caused by hydrogen peroxide and rotenone. Genistein had no positive effect on cell death in experiments carried out on RGC-5 cells. Exposure of RGC-5 cells to flavonoids showed that EC and EGCG increased the mRNA expression of endogenous antioxidants such as HO-l (heme oxygenase 1) and Nrf-2 (nuclear erythroid factor-z-related factor 2). Light insult, rotenone and sodium azide activate the p38 (protein kinase 38) pathway, while only light and rotenone activate the JNK (c-Jun amino-terminal kinase) pathway. Serum deprivation affects mitochondrial apoptotic proteins causing an increase in the ratio of Bax/Bcl2 (Bax: Bcl-2-associated X protein; Bcl-2: B-cell lymphoma 2). An insult of light to RGC-5 cells, unlike that induced by sodium azide, is inhibited by necrostatin-I and causes an activation of AlF (apoptosis-inducing factor) with alpha-fodrin being unaffected. These studies suggest that ganglion cell death caused by insults as may occur in POAG involves various cellular signaling pathways. The selected flavonoids have diverse actions in increasing cellular defense mechanisms, and in negating the effects of ischemia and specific types of oxidative stress. The results argue for the possible use of flavonoids in the treatment of POAG to slow down ganglion cell death.

A neuroproteção é um dos tópicos mais relevantes aplicados à neurociência. As junções comunicantes (JC), formadas pelas conexinas (Cx) estão envolvidas na neurodegeneração após lesão. Estudos com animais KO apresentam resultados contraditórios sobre papel das JCs. O objetivo deste trabalho foi analisar o papel das Cxs a partir do trauma mecânico na retina, modelo que permite a visualização do foco, penumbra, e áreas adjacentes à lesão. Observamos regulação distinta das Cx36 e Cx43 durante a neurodegeneração. A Cx36 não se alterou e a Cx43 apresentou desorganização e aumento da imunorreatividade após 7 dias, concomitantemente com GFAP. Células amácrinas apoptóticas encontram-se acopladas a células vizinhas por Cx36. O papel funcional das JCs foi avaliado, utilizando bloqueadores, para verificar a viabilidade/morte de células. Carbenoxolone (CBX), reduziu o espalhamento da apoptose, após 4h, enquanto a quinina, teve o mesmo efeito após 1h. A distribuição de núcleos apoptóticos confirmou que a utilização de bloqueadores de JCs reduz a propagação da apoptose. A quinina, mas não o CBX, diminuiu a expressão de caspases iniciais e efetoras. O controle da permeabilidade de canais de JCs pode participar de estratégias de neuroproteção. / The neuroprotection stands out as one of the most pursued hot topics in applied neurosciences. The gap junctions (GJ), formed by connexin (Cx) are involved in neurodegeneration injury. Studies using KO animal models endowed apparently contradictory results in relation to the role of coupling in neuroprotection. The aim of this study was to analyze the role of Cx-mediated communication in focal lesion induced by mechanical trauma in the retina, a model that alow the visualization of the focus, penumbra and adjacent areas. We observed distinct regulation of Cx36 and Cx43 during neurodegeneration. The Cx36 did not change during the lesion progression and Cx43 showed disorganized pattern and upregulated after 7 days, the same as GFAP. Apoptotic amacrine cells are coupled with health neighborhood cells by Cx36. The functional role of GJ was evaluated using blockers to verify the viability/cell death. Carbenoxolone (CBX) reduced the spread of apoptosis after 4h while quinine had the same effect after 1h. The distribution of apoptotic nuclei confirmed that the use of GJ blockers reduced the propagation of apoptosis. Quinine, but not CBX, decreases initial and effector caspases expression. The control of GJ channels permeability can participate in neuroprotection strategies.