Model Of Retinal Detachment Research Articles

A Novel Experimental Mouse Model of Retinal Detachment: Complete Functional and Histologic Recovery of the Retina

To establish an experimental mouse model of retinal detachment (RD) created by corneal puncture (CP). Mouse corneas were punctured with a 30.5-gauge beveled needle, and the anterior chamber was penetrated. Histologic and functional changes of the retina were examined by light microscopy and electroretinography (ERG). Certain retinal cellular responses were examined by immunofluorescence microscopy. Internucleosomal DNA fragmentation in the retina was determined by terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick-end labeling (TUNEL). RESULTS. CP caused transient leakage of aqueous humor along the needle shaft and immediate formation of multiple retinal blebs, which shrank and flattened within 24 hours. Bleb formation was associated with detachment of the neuroretina from the retinal pigment epithelium (RPE). After CP, the RPE cells underwent extensive transformation during retinal detachment/reattachment, but they resumed normal morphology on retinal reattachment around 10 to 13 days after CP. Relative to pre-CP ERG amplitudes, the punctured eyes showed decreases of 45% and 24% in scotopic and 7% and 12% in photopic b- and a-wave amplitudes, respectively, within 10 to 20 minutes after CP. The ERG amplitudes recovered fully by 12 hours after CP. No infiltrated cells were observed in the subretinal space, and no proliferating or TUNEL-positive cells were observed in the retina of the punctured eyes. Puncturing the mouse cornea can create transient RD, and the functional and histologic changes in the retina can subsequently recover. This experimental mouse model of RD mimics human traction and serous RD.

Effects of Supplemental Erythropoietin on Its Receptor Expression and Signal Transduction Pathways in Rat Model of Retinal Detachment

Purpose: The aim of this study was to investigate the effects of supplemental erythropoietin (EPO) on its receptor (EPOR) and signal transduction pathways in rat model of retinal detachment (RD).Methods: To investigate the effect of EPO on EPOR expression in RD rats 100, 200 or 400 ng EPO was injected into the vitreous cavity immediately after RD model was induced. Western blot and immunohistochemistry analyses were performed to measure EPOR expression. To investigate the effect of EPO on signal transduction pathways in RD rats single dose of 400 ng EPO was injected into the vitreous cavity immediately after RD model was induced. The total and phosphorylated levels of JAK2, Akt, ERK-1/2, STAT5 and NF-κB were assessed by western blot.Results: Western blot analysis showed that, compared with the normal control group, EPOR expression in the neurosensory retina was significantly increased in experimental RD groups (P < 0.05), but the differences were not significant between experimental RD groups (P > 0.05). Immunohistochemical examination indicated that EPOR staining on retinas became strongly positive 3 days after RD, with no significant difference in staining intensities between the treatment groups. Phosphorylated levels of JAK2, Akt, ERK-1/2, STAT5, and NF-κB were enhanced 3 days after RD, but only JAK2, Akt, and ERK-1/2 phosphorylation was further enhanced by 400 ng EPO treatment (P < 0.05).Conclusions: Supplementary EPO cannot affect EPOR expression in detached retina, but EPO may activate both PI-3K/Akt and MAPK/ERK-1/2 signal transduction pathways in RD model.

Safety and efficacy of intravitreal injection of recombinant erythropoietin for protection of photoreceptor cells in a rat model of retinal detachment

To elucidate the safety and efficacy of exogenous erythropoietin (EPO) for the protection of photoreceptor cells in a rat model of retinal detachment (RD). Recombinant rat EPO (400 ng) was injected into the vitreous cavity of normal rats to observe the eye manifestations. Retinal function was assessed by flash electroretinograms. Histopathological examination of retinal tissue was performed at 14 days and 2 months after injection, respectively. To investigate the inhibitory effect of EPO on photoreceptor cell apoptosis in RD rats, 100, 200, or 400 ng EPO was injected into the vitreous cavity immediately after RD model establishment. Apoptosis of photoreceptor cells was determined at 3 days after injection. Caspase-3 activation was measured by western blot analysis and immunofluorescence, respectively, and the level of Bcl-X(L) expression was analyzed by western blot. Intravitreal injection of EPO 400 ng into normal rats had no significant impact on retinal function, morphology, or structure. Apoptosis of retinal photoreceptor cells apparently increased after RD and was significantly reduced following EPO treatment. The thickness of the outer nuclear layer in the RD + 400 ng group was significantly thicker than that in other experimental RD groups both at 14 days and at 2 months after RD (P < 0.05). Western blot and immunofluorescence analyses showed decreased caspase-3 activation and increased Bcl-X(L) expression following EPO treatment. Intravitreal injection of EPO 400 ng is safe, and EPO may suppress caspase-3 activation and enhance Bcl-X(L) expression, resulting in inhibition of apoptosis and protection of photoreceptor cells.

Intraocular expression and release of high-mobility group box 1 protein in retinal detachment

High-mobility group box 1 (HMGB1) protein is a multifunctional protein, which is mainly present in the nucleus and is released extracellularly by dying cells and/or activated immune cells. Although extracellular HMGB1 is thought to be a typical danger signal of tissue damage and is implicated in diverse diseases, its relevance to ocular diseases is mostly unknown. To determine whether HMGB1 contributes to the pathogenesis of retinal detachment (RD), which involves photoreceptor degeneration, we investigated the expression and release of HMGB1 both in a retinal cell death induced by excessive oxidative stress in vitro and in a rat model of RD-induced photoreceptor degeneration in vivo. In addition, we assessed the vitreous concentrations of HMGB1 and monocyte chemoattractant protein 1 (MCP-1) in human eyes with RD. We also explored the chemotactic activity of recombinant HMGB1 in a human retinal pigment epithelial (RPE) cell line. The results show that the nuclear HMGB1 in the retinal cell is augmented by death stress and upregulation appears to be required for cell survival, whereas extracellular release of HMGB1 is evident not only in retinal cell death in vitro but also in the rat model of RD in vivo. Furthermore, the vitreous level of HMGB1 is significantly increased and is correlated with that of MCP-1 in human eyes with RD. Recombinant HMGB1 induced RPE cell migration through an extracellular signal-regulated kinase-dependent mechanism in vitro. Our findings suggest that HMGB1 is a crucial nuclear protein and is released as a danger signal of retinal tissue damage. Extracellular HMGB1 might be an important mediator in RD, potentially acting as a chemotactic factor for RPE cell migration that would lead to an ocular pathological wound-healing response.

Expression Profiling after Retinal Detachment and Reattachment: A Possible Role for Aquaporin-0

Retinal detachment (RD) is associated with acute visual loss caused by anatomic displacement of the photoreceptors and with chronic visual loss/disturbance caused by retinal remodeling and photoreceptor cell death, which may occur even after successful reattachment. The P2Y(2) receptor agonist INS37217 improves the rate of retinal reattachment in animal models of induced RD, and has been shown to also significantly enhance the rate of ERG recovery in a mouse model of RD. The identification of genes modulated by INS37217 may allow further drug discovery for treating RD and edema. To identify genes involved in RD and subsequent reattachment, a retinal microarray screen was performed using a mouse model of RD in the presence or absence of INS37217. Ninety-two genes were identified as differentially expressed across three time points, most of which were upregulated in the presence of this agonist. Furthermore, it was shown that RD alters the expression of aquaporin-0 (AQP-0), and this modulation is prevented by treatment with INS37217. The presence of AQP-0 in retinal bipolar cells was also demonstrated, whereas it was previously thought to be specific to the lens. Mice lacking functional alleles of AQP-0 had a phototransduction deficit as assessed by electroretinography; however, their photoreceptor structure was normal, indicative of a problem with signal transmission between neurons. This study establishes the genes involved in RD and reattachment, and also demonstrates for the first time a physiologically significant role for AQP-0 in retinal function.

Expression Pattern of Erythropoietin and Erythropoietin Receptor in Experimental Model of Retinal Detachment

Purpose: Erythropoietin (EPO), known for its role in erythroid cell differentiation, has been suggested to have a neuroprotective effect on retinal neurons. The aim of the current study was to investigate the expression pattern of EPO and erythropoietin receptor (EPOR) in the detached retina of a rat model of retinal detachment (RD). Methods: Forty-eight albino Sprague-Dawley rats were randomized into normal control group (n = 6) and RD model group (n = 42). The rat RD model was established by slow injection of 1.4% sodium hyaluronate into the upper half of the subretinal space, in which the upper half of the neurosensory retina was detached from the underlying retinal pigment epithelium (RPE). The expression of EPO and EPOR mRNA/protein was determined at 1, 3, 6, 12, 24, 48, and 72 hr after sodium hyaluronate injection by semiquantitative RT-PCR and Western blot analysis, respectively. Meanwhile, the distributions of EPO and EPOR were examined by immunohistochemistry. Results: EPO and EPOR mRNA levels increased gradually after sodium hyaluronate injection and peaked 48 hr later. EPO mRNA level became significantly higher than that in the normal control group at 12 hr after injection, and EPOR mRNA level became significantly higher at 6 hr (p < 0.05). Moreover, the protein levels of EPO and EPOR presented a similar tendency to the mRNA level and became significantly higher than those in normal control group at 3 hr after injection (p < 0.05). Immunohistochemical results showed that both EPO and EPOR were strongly expressed all over the neurosensory retina 48 hr after injection, but EPOR was not found in the rod outer segment. Only weak expression of EPO and EPOR was noticed in the retina of normal control group. Conclusions: The levels of EPO and EPOR in detached retina increase with the duration of retinal detachment and reach their peaks at 48 hr; most layers of detached neurosensory retina can express EPO and EPOR. It is indicated that the EPO/EPOR system might play an important role in protection of retinal neurons during RD; supplementation with exogenous EPO might promote the survival of retinal neurons.