X. Zhou

Screening for Retinopathy of Prematurity in China: A Neonatal Units–Based Prospective Study

PURPOSE To analyze the incidence and severity of retinopathy of prematurity (ROP) in China, and to explore the workload implications of applying different criteria. METHODS A prospective, neonatal units-based study undertaken in two tertiary level hospitals in Shanghai, China, from January 1, 2010 to December 31, 2012. All infants with birth weight (BW) of 2000 g or less and/or gestational age (GA) of 34 weeks or less were screened for ROP. Retinopathy of prematurity was classified using the international classification, and was treated in accordance with the recommendations of the Early Treatment for Retinopathy of Prematurity Cooperative Group. RESULTS A total of 2825 (93.7%) of 3014 eligible infants were screened, and ROP was diagnosed in 503 infants (17.8%). One hundred ninety-one infants (6.8%) had type 1 or worse ROP and were treated with laser or vitrectomy. The mean GA of ROP patients was 29.9 ± 2.1 weeks and their mean BW was 1425 ± 266 g. Infants who needed treatment for ROP had a mean GA of 29.3 ± 2.1 weeks and mean BW of 1331 ± 330 g. Among these treated infants, 18 infants (9.4%) exceeded the United Kingdoms (UK) screening criteria, and 28 (14.7%) exceeded the criteria used in the United States (US). If narrower criteria, as in GA less than or equal to 33 weeks and/or BW less than or equal to 1750 g were adopted, almost 16.9% fewer infants would not have been examined, with no infant missing treatment. CONCLUSIONS Larger, older infants are at risk in China and screening criteria used in the US and UK may not be suitable for China. Further population-based studies are recommended to determine the necessity of modifying the current ROP screening protocol.

Alpha7 nicotinic acetylcholine receptor agonist promotes retinal ganglion cell function via modulating GABAergic presynaptic activity in a chronic glaucomatous model

Alpha-7 nicotinic acetylcholine receptor (α7-nAChR) agonists can prevent glutamate-induced excitotoxicity in cultured retinal ganglion cells (RGCs). However, the neuroprotective effects and the mechanism of action of PNU-282987, an α7-nAChR agonist, in a chronic in vivo rat glaucoma model are poorly understood. We found that elevated intraocular pressure (IOP) downregulated retinal α7-nAChR expression. Electroretinography revealed that the amplitude of the photopic negative response (PhNR) decreased in parallel with the loss of RGCs caused by elevated IOP. PNU-282987 enhanced RGC viability and function and decreased terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive signals in RGCs. Patch-clamp recordings revealed differences in the baseline frequencies and decay times of the miniature GABAergic inhibitory postsynaptic currents (mIPSCs) of RGCs between control and glaucomatous retinal slices. The results of western blotting and immunostaining showed that glutamic acid decarboxylase 65/67 and GABA deficits persisted in glaucomatous retinas and that these deficits were reversed by PNU-282987. Patch-clamp recordings also showed that PNU-282987 significantly increased the frequency and amplitude of the GABAergic mIPSCs of RGCs. The protective effects of PNU-292987 were blocked by intravitreal administration of selective GABAA receptor antagonists. The modulation of GABAergic synaptic transmission by PNU-282987 causes de-excitation of ganglion cell circuits and suppresses excitotoxic processes.

Quercetin Declines Apoptosis, Ameliorates Mitochondrial Function and Improves Retinal Ganglion Cell Survival and Function in In Vivo Model of Glaucoma in Rat and Retinal Ganglion Cell Culture In Vitro

Glaucoma is a progressive neuropathy characterized by the loss of retinal ganglion cells (RGCs). Strategies that delay or halt RGC loss have been recognized as potentially beneficial for rescuing vision in glaucoma patients. Quercetin (Qcn) is a natural and important dietary flavonoid compound, widely distributed in fruits and vegetables. Mounting evidence suggests that Qcn has numerous neuroprotective effects. However, whether Qcn exerts neuroprotective effects on RGC in glaucoma is poorly understood. In this study, we investigated the protective effect of Qcn against RGC damage in a rat chronic ocular hypertension (COHT) model in vivo and hypoxia-induced primary cultured RGC damage in vitro, and we further explored the underlying neuroprotective mechanisms. We found that Qcn not only improved RGC survival and function from a very early stage of COHT in vivo, it promoted the survival of hypoxia-treated primary cultured RGCs in vitro via ameliorating mitochondrial function and preventing mitochondria-mediated apoptosis. Our findings suggest that Qcn has direct protective effects on RGCs that are independent of lowering the intraocular pressure (IOP). Qcn may be a promising therapeutic agent for improving RGC survival and function in glaucomatous neurodegeneration.

Cannabinoids Regulate the Diameter of Pericyte-Containing Retinal Capillaries in Rats

Background/Aims: Cannabinoids are vasoactive substances that act as key regulators of arterial tone in the blood vessels supplying peripheral tissues and the central nervous system. We therefore investigated the effect of cannabinoids on retinal capillaries and pericytes. Methods: The effects of cannabinoids on capillary diameters were determined using an ex vivo whole-mount rat retinal model. Western blotting, quantitative PCR, and immunohistochemistry were performed to explore the underlying mechanism. Results: Endogenous cannabinoid 2-arachidonoylglycerol and anandamide and exogenous cannabinoid (R-(+)-WIN55212-2) dilated the noradrenaline-precontracted capillaries in a concentration-dependent manner (1 µM to 0.1 mM). The extent of vasorelaxation was positively correlated with changes in pericyte width. The effects of R-(+)-WIN55212-2 on vasorelaxation and pericyte width were inhibited by a cannabinoid receptor type-1 (CB1) antagonist, AM251 or rimonabant (SR141716A), the nitric oxide synthase inhibitor l-NAME, and the guanylate cyclase inhibitor ODQ. They were also abolished by the removal of the endothelium, but not by the cannabinoid receptor-2 antagonist SR144528, the endothelial cannabinoid receptor antagonist O-1918, or the cyclooxygenase inhibitor indomethacin. Conclusion: The exogenous cannabinoid R-(+)-WIN55212-2 promotes the vasorelaxation of pericyte-containing rat retinal capillaries. This effect of R-(+)-WIN55212-2 is dependent on CB1 and the nitric oxide–cyclic guanosine monophosphate pathway, and requires an intact endothelium.

Decreased d-Serine Levels Prevent Retinal Ganglion Cell Apoptosis in a Glaucomatous Animal Model

Purpose The purpose of this study was to determine d-Serine and d-Serine synthetase serine racemase (SR) expression and whether decreased d-Serine expression has protective effects on retinal ganglion cells (RGCs) in a glaucomatous animal model. Methods The rat chronic intraocular hypertension (COH) model was generated as a glaucomatous animal model by cauterizing three episcleral veins. Quantitative analysis of RGC survival was determined by the counting of retrograde FluoroGold-labeled RGCs. The level of d-Serine in the retinas and aqueous humor was determined by Ultra High Performance Liquid Chromatography coupled to triple-quadrupole Mass Spectrometry (UHPLC-MS/MS). Retinal expression of serine racemase (SR) protein was determined by immunohistochemistry and Western blot analysis. The TUNEL assay was used to detect cell apoptosis. Results The content of d-Serine increased significantly in the glaucomatous retina of the COH model 2 weeks after surgery compared with the control retina. d-Serine synthetase SR expression in the right glaucomatous eye increased slightly after surgery compared with that in the left control eye and remained at this high level for 6 weeks after surgery. SR-positive cells were located mainly in the ganglion cell layer (GCL) of the retina. d-Amino acid oxidase (DAAO) treatment significantly increased RGC survival in the glaucomatous eyes, and the TUNEL assay was used to confirm that DAAO reduced the number of TUNEL-positive cells in glaucomatous eyes. However, excess d-Serine could not exacerbate RGC loss in the COH model. Conclusions Increased d-Serine and SR expressions in the retina of the COH model were detected. DAAO treatment significantly increased RGC survival in the glaucomatous eyes. These results suggest that decreased d-Serine expression has protective effects on RGCs.

Differential Modulation of GABAA and NMDA Receptors by an α7-nicotinic Acetylcholine Receptor Agonist in Chronic Glaucoma

Presynaptic modulation of γ-aminobutyric acid (GABA) release by an alpha7 nicotinic acetylcholine receptor (α7-nAChR) agonist promotes retinal ganglion cell (RGC) survival and function, as suggested by a previous study on a chronic glaucomatous model from our laboratory. However, the role of excitatory and inhibitory amino acid receptors and their interaction with α7-nAChR in physiological and glaucomatous events remains unknown. In this study, we investigated GABAA and N-methyl-D-aspartate (NMDA) receptor activity in control and glaucomatous retinal slices and the regulation of amino acid receptor expression and function by α7-nAChR. Whole-cell patch-clamp recordings from RGCs revealed that the α7-nAChR specific agonist PNU-282987 enhanced the amplitude of currents elicited by GABA and reduced the amplitude of currents elicited by NMDA. The positive modulation of GABAA receptor and the negative modulation of NMDA receptor (NMDAR) by PNU-282987-evoked were prevented by pre-administration of the α7-nAChR antagonist methyllycaconitine (MLA). The frequency and the amplitude of glutamate receptor-mediated miniature glutamatergic excitatory postsynaptic currents (mEPSCs) were not significantly different between the control and glaucomatous RGCs. Additionally, PNU-282987-treated slices showed no alteration in the frequency or amplitude of mEPSCs relative to control RGCs. Moreover, we showed that expression of the α1 subunit of the GABAA receptor was downregulated and the expression of the NMDAR NR2B subunit was upregulated by intraocular pressure (IOP) elevation, and the changes of high IOP were blocked by PNU-282987. In conclusion, retina GABAA and NMDARs are modulated positively and negatively, respectively, by activation of α7-nAChR in in vivo chronic glaucomatous models.