DRP1 inhibition rescues retinal ganglion cells and their axons by preserving mitochondrial integrity in a mouse model of glaucoma.

Glaucoma is the leading cause of irreversible blindness and is characterized by slow and progressive degeneration of the optic nerve head axons and retinal ganglion cell (RGC), leading to loss of visual function. Although oxidative stress and/or alteration of mitochondrial (mt) dynamics induced by elevated intraocular pressure (IOP) are associated with this neurodegenerative disease, the mechanisms that regulate mt dysfunction-mediated glaucomatous neurodegeneration are poorly understood. Using a mouse model of glaucoma, DBA/2J (D2), which spontaneously develops elevated IOP, as well as an in vitro RGC culture system, we show here that oxidative stress, as evidenced by increasing superoxide dismutase 2 (SOD2) and mt transcription factor A (Tfam) protein expression, triggers mt fission and loss by increasing dynamin-related protein 1 (DRP1) in the retina of glaucomatous D2 mice as well as in cultured RGCs exposed to elevated hydrostatic pressure in vitro. DRP1 inhibition by overexpressing DRP1 K38A mutant blocks mt fission and triggers a subsequent reduction of oxidative stress, as evidenced by decreasing SOD2 and Tfam protein expression. DRP1 inhibition promotes RGC survival by increasing phosphorylation of Bad at serine 112 in the retina and preserves RGC axons by maintaining mt integrity in the glial lamina of glaucomatous D2 mice. These findings demonstrate an important vicious cycle involved in glaucomatous neurodegeneration that starts with elevated IOP producing oxidative stress; the oxidative stress then leads to mt fission and a specific form of mt dysfunction that generates further oxidative stress, thus perpetuating the cycle. Our findings suggest that DRP1 is a potential therapeutic target for ameliorating oxidative stress-mediated mt fission and dysfunction in RGC and its axons during glaucomatous neurodegeneration. Thus, DRP1 inhibition may provide a new therapeutic strategy for protecting both RGCs and their axons in glaucoma and other optic neuropathies.

Inhibition of cyclophilin D by cyclosporin A promotes retinal ganglion cell survival by preventing mitochondrial alteration in ischemic injury.

Cyclosporin A (CsA) inhibits the opening of the mitochondrial permeability transition pore (MPTP) by interacting with cyclophilin D (CypD) and ameliorates neuronal cell death in the central nervous system against ischemic injury. However, the molecular mechanisms underlying CypD/MPTP opening-mediated cell death in ischemic retinal injury induced by acute intraocular pressure (IOP) elevation remain unknown. We observed the first direct evidence that acute IOP elevation significantly upregulated CypD protein expression in ischemic retina at 12 h. However, CsA prevented the upregulation of CypD protein expression and promoted retinal ganglion cell (RGC) survival against ischemic injury. Moreover, CsA blocked apoptotic cell death by decreasing cleaved caspase-3 protein expression in ischemic retina. Of interest, although the expression level of Bcl-xL protein did not show a significant change in ischemic retina treated with vehicle or CsA at 12 h, ischemic damage induced the reduction of Bcl-xL immunoreactivity in RGCs. More importantly, CsA preserved Bcl-xL immunoreactivity in RGCs of ischemic retina. In parallel, acute IOP elevation significantly increased phosphorylated Bad (pBad) at Ser112 protein expression in ischemic retina at 12 h. However, CsA significantly preserved pBad protein expression in ischemic retina. Finally, acute IOP elevation significantly increased mitochondrial transcription factor A (Tfam) protein expression in ischemic retina at 12 h. However, CsA significantly preserved Tfam protein expression in ischemic retina. Studies on mitochondrial DNA (mtDNA) content in ischemic retina showed that there were no statistically significant differences in mtDNA content among control and ischemic groups treated with vehicle or CsA. Therefore, these results provide evidence that the activation of CypD-mediated MPTP opening is associated with the apoptotic pathway and the mitochondrial alteration in RGC death of ischemic retinal injury. On the basis of these observations, our findings suggest that CsA-mediated CypD inhibition may provide a promising therapeutic potential for protecting RGCs against ischemic injury-mediated mitochondrial dysfunction.

Vascular tone pathway polymorphisms in relation to primary open-angle glaucoma.

AIMS: Vascular perfusion may be impaired in primary open-angle glaucoma (POAG); thus, we evaluated a panel of markers in vascular tone-regulating genes in relation to POAG. METHODS: We used Illumina 660W-Quad array genotype data and pooled P-values from 3108 POAG cases and 3430 controls from the combined National Eye Institute Glaucoma Human Genetics Collaboration consortium and Glaucoma Genes and Environment studies. Using information from previous literature and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, we compiled single-nucleotide polymorphisms (SNPs) in 186 vascular tone-regulating genes. We used the 'Pathway Analysis by Randomization Incorporating Structure' analysis software, which performed 1000 permutations to compare the overall pathway and selected genes with comparable randomly generated pathways and genes in their association with POAG. RESULTS: The vascular tone pathway was not associated with POAG overall or POAG subtypes, defined by the type of visual field loss (early paracentral loss (n=224 cases) or only peripheral loss (n=993 cases)) (permuted P≥0.20). In gene-based analyses, eight were associated with POAG overall at permuted P<0.001: PRKAA1, CAV1, ITPR3, EDNRB, GNB2, DNM2, HFE, and MYL9. Notably, six of these eight (the first six listed) code for factors involved in the endothelial nitric oxide synthase activity, and three of these six (CAV1, ITPR3, and EDNRB) were also associated with early paracentral loss at P<0.001, whereas none of the six genes reached P<0.001 for peripheral loss only. DISCUSSION: Although the assembled vascular tone SNP set was not associated with POAG, genes that code for local factors involved in setting vascular tone were associated with POAG.

Inhibition of oxidative stress by coenzyme Q10 increases mitochondrial mass and improves bioenergetic function in optic nerve head astrocytes.

Oxidative stress contributes to dysfunction of glial cells in the optic nerve head (ONH). However, the biological basis of the precise functional role of mitochondria in this dysfunction is not fully understood. Coenzyme Q10 (CoQ10), an essential cofactor of the electron transport chain and a potent antioxidant, acts by scavenging reactive oxygen species (ROS) for protecting neuronal cells against oxidative stress in many neurodegenerative diseases. Here, we tested whether hydrogen peroxide (100 µM H2O2)-induced oxidative stress alters the mitochondrial network, oxidative phosphorylation (OXPHOS) complex (Cx) expression and bioenergetics, as well as whether CoQ10 can ameliorate oxidative stress-mediated alterations in mitochondria of the ONH astrocytes in vitro. Oxidative stress triggered the activation of ONH astrocytes and the upregulation of superoxide dismutase 2 (SOD2) and heme oxygenase-1 (HO-1) protein expression in the ONH astrocytes. In contrast, CoQ10 not only prevented activation of ONH astrocytes but also significantly decreased SOD2 and HO-1 protein expression in the ONH astrocytes against oxidative stress. Further, CoQ10 prevented a significant loss of mitochondrial mass by increasing mitochondrial number and volume density and by preserving mitochondrial cristae structure, as well as promoted mitofilin and peroxisome-proliferator-activated receptor-γ coactivator-1 protein expression in the ONH astrocyte, suggesting an induction of mitochondrial biogenesis. Finally, oxidative stress triggered the upregulation of OXPHOS Cx protein expression, as well as reduction of cellular adeonsine triphosphate (ATP) production and increase of ROS generation in the ONH astocytes. However, CoQ10 preserved OXPHOS protein expression and cellular ATP production, as well as decreased ROS generation in the ONH astrocytes. On the basis of these observations, we suggest that oxidative stress-mediated mitochondrial dysfunction or alteration may be an important pathophysiological mechanism in the dysfunction of ONH astrocytes. CoQ10 may provide new therapeutic potentials and strategies for protecting ONH astrocytes against oxidative stress-mediated mitochondrial dysfunction or alteration in glaucoma and other optic neuropathies.

A new vicious cycle involving glutamate excitotoxicity, oxidative stress and mitochondrial dynamics.

Glutamate excitotoxicity leads to fragmented mitochondria in neurodegenerative diseases, mediated by nitric oxide and S-nitrosylation of dynamin-related protein 1, a mitochondrial outer membrane fission protein. Optic atrophy gene 1 (OPA1) is an inner membrane protein important for mitochondrial fusion. Autosomal dominant optic atrophy (ADOA), caused by mutations in OPA1, is a neurodegenerative disease affecting mainly retinal ganglion cells (RGCs). Here, we showed that OPA1 deficiency in an ADOA model influences N-methyl-D-aspartate (NMDA) receptor expression, which is involved in glutamate excitotoxicity and oxidative stress. Opa1(enu/+) mice show a slow progressive loss of RGCs, activation of astroglia and microglia, and pronounced mitochondrial fission in optic nerve heads as found by electron tomography. Expression of NMDA receptors (NR1, 2A, and 2B) in the retina of Opa1(enu/+) mice was significantly increased as determined by western blot and immunohistochemistry. Superoxide dismutase 2 (SOD2) expression was significantly decreased, the apoptotic pathway was activated as Bax was increased, and phosphorylated Bad and BcL-xL were decreased. Our results conclusively demonstrate that not only glutamate excitotoxicity and/or oxidative stress alters mitochondrial fission/fusion, but that an imbalance in mitochondrial fission/fusion in turn leads to NMDA receptor upregulation and oxidative stress. Therefore, we propose a new vicious cycle involved in neurodegeneration that includes glutamate excitotoxicity, oxidative stress, and mitochondrial dynamics.

Anterior chamber angle imaging with optical coherence tomography.

The technology of optical coherence tomography (OCT) has evolved rapidly from time-domain to spectral-domain and swept-source OCT over the recent years. OCT has become an important tool for assessment of the anterior chamber angle and detection of angle closure. Improvement in image resolution and scan speed of OCT has facilitated a more detailed and comprehensive analysis of the anterior chamber angle. It is now possible to examine Schwalbe's line and Schlemm's canal along with the scleral spur. High-speed imaging allows evaluation of the angle in 360°. With three-dimensional reconstruction, visualization of the iris profiles and the angle configurations is enhanced. This article summarizes the development and application of OCT for anterior chamber angle measurement, detection of angle closure, and investigation of the pathophysiology of primary angle closure.

Assessment of rates of structural change in glaucoma using imaging technologies.

PURPOSE: To review the ability of current imaging technologies to provide estimates of rates of structural change in glaucoma patients. PATIENTS AND METHODS: Review of literature. RESULTS: Imaging technologies, such as confocal scanning laser ophthalmoscopy (CSLO), scanning laser polarimetry (SLP), and optical coherence tomography (OCT), provide quantifiable and reproducible measurements of the optic disc and parapapillary retinal nerve fibre layer (RNFL). Rates of change as quantified by the rim area (RA) (for CSLO) and RNFL thickness (for SLP and OCT) are related to glaucoma progression as detected by conventional methods (eg, visual fields and optic disc photography). Evidence shows that rates of RNFL and RA loss are significantly faster in progressing compared with non-progressing glaucoma patients. CONCLUSION: Measurements of rates of optic disc and RNFL change are becoming increasingly precise and individualized. Currently available imaging technologies have the ability to detect and quantify progression in glaucoma, and their measurements may be suitable end points in glaucoma clinical trials.

Pattern electroretinogram association with spectral domain-OCT structural measurements in glaucoma.

PURPOSE: To describe the association between pattern electroretinogram (PERG) amplitude and spectral domain-optical coherence tomography (SD-OCT) macular thickness, retinal nerve fibre layer (RNFL) thickness and optic disc topography measurements. SUBJECTS AND METHODS: Both eyes (n = 132) of 66 glaucoma patients (mean age = 67.9 years) enrolled in the University of California, San Diego, CA, USA, Diagnostic Innovations in Glaucoma Study (DIGS) were included. Eyes were tested with PERG (Glaid PERGLA, Lace Elettronica, Pisa, Italy), RTVue SD-OCT (Optovue Inc., Fremont, CA, USA) GCC, and NHM4 protocols on the same day. Of the 66 enrolled patients, 43 had glaucoma defined by repeated abnormal standard automated perimetry (SAP) results in at least one eye and 23 were glaucoma suspects defined by a glaucomatous-appearing optic disc by physicians' examination in at least one eye and normal SAP results in both eyes. Associations (R(2)) were determined between PERG amplitude (µV) and SD-OCT macular ganglion cell complex (GCC) thickness (µm), macular thickness (µm), macular outer retinal thickness (macular thickness minus GCC thickness) (µm), RNFL thickness (µm), neuroretinal rim area (mm(2)), and rim volume (mm(3)). RESULTS: PERG amplitude was significantly associated with GCC thickness (R(2) = 0.179, P < 0.001), RNFL thickness (R(2) = 0.174, P < 0.001), and macular thickness (R(2) = 0.095, P<0.001). R(2) associations with other parameters were not significant (all P > 0.624). Significant associations remained for GCC and average RNFL thickness when age and intraocular pressure at the time of testing were included in multivariate models (both P ≤ 0.030). CONCLUSIONS: PERG amplitude is significantly (but weakly) associated with macular GCC thickness, RNFL thickness, and macular thickness. The lack of association between PERG amplitude and macular outer retinal thickness supports previous results, possibly suggesting that that the PERG is driven primarily by retinal ganglion cell (inner retinal) responses.

Agreement between Heidelberg Retina Tomograph-I and -II in detecting glaucomatous changes using topographic change analysis.

PURPOSE: To evaluate the suitability of including both Heidelberg Retina Tomograph-I (HRT-I) and HRT-II examinations in the same longitudinal series for HRT topographic change analysis (TCA) and to evaluate parabolic error correction (PEC) to improve the agreement between HRT-I and HRT-II examinations. METHODS: A total of 66 eyes from the University of California San Diego Diagnostic Innovations in Glaucoma Study with baseline HRT-I and HRT-II examinations obtained on the same day and ≥ 3 HRT-II follow-up examinations were included. Two TCA analyses, HRT-I examination at baseline (HRT-I-mixed series) and HRT-II examination at baseline (HRT-II-only series) were compared. Agreement between the HRT-I-mixed and HRT-II-only series were estimated using Bland-Altman plots. Agreement was assessed: (1) using the current HRT software settings (PEC applied only to HRT-II-only series), and (2) modified HRT settings (PEC also applied to HRT-I-mixed series). RESULTS: With current HRT software settings, the HRT-I-mixed series significantly overestimated change locations (ie, red pixels) compared with the HRT-II-only series as indicated by statistically significant proportional biases in the Bland-Altman analysis. By applying PEC to HRT-I-mixed series there were no statistically significant biases in the TCA parameter estimates compared with the HRT-II-only series. CONCLUSION: In some eyes, HRT-I and HRT-II baseline examinations are not interchangeable in TCA analysis without parabolic error correction. HRT-I-mixed series detected more changes characteristic of glaucoma when there were only minimal changes in the HRT-II-only series. Our results suggest that in the majority of cases, with PEC, HRT-I examinations may be included in a longitudinal series containing HRT-II examinations.

Comparisons of anterior segment biometry between Chinese and Caucasians using anterior segment optical coherence tomography.

PURPOSE: To compare anterior segment parameters between eyes of Chinese and Caucasians using anterior segment optical coherence tomography and to evaluate the association between these parameters and anterior chamber angle width between the two ethnic groups. METHODS: 60 Chinese and 60 Caucasians, 30 with open angles and 30 with narrow angles (defined as Shaffer grade < or =2 in > or =3 quadrants during dark room gonioscopy) in each group, were consecutively enrolled. One eye of each subject was randomly selected for imaging in a completely darkened room. Measurements, including anterior chamber depth (ACD), scleral spur-to-scleral spur distance (anterior chamber width (ACW)), anterior chamber angle width, iris convexity and iris thickness, were compared between the groups. The associations between angle opening distance and biometric measurements were evaluated with univariate and multivariate regression analyses. RESULTS: There were no differences in age, axial length, anterior chamber angle measurements, pupil diameter and iris convexity between Chinese and Caucasians in both open-angle and narrow-angle groups. However, the ACD and ACW were smaller and the iris was thicker in Chinese. In the multivariate analysis, the ACD was the most influential biometric parameter for angle opening distance in both Chinese and Caucasians. After adjusting the effects of axial length, age and sex, ACD and ACW were significantly smaller in Chinese. CONCLUSIONS: Chinese eyes had smaller ACD, smaller ACW and greater iris thickness than Caucasians. ACD was the most influential parameter in determining the angle width in both ethnic groups.

Predictors of atypical birefringence pattern in scanning laser polarimetry.

AIM: To evaluate predictors of atypical birefringence patterns (ABP) observed in scanning laser polarimetry. METHODS: A total of 179 eyes from 82 normal subjects and 97 glaucoma patients were included. The retinal nerve fibre layer in each eye was imaged sequentially with GDx variable corneal compensation (VCC) and GDx enhanced corneal compensation (ECC) (Carl Zeiss Meditec, Dublin, California). The associations between the typical scan score (TSS) and age, axial length, spherical error, parapapillary atrophy (PPA) and visual-field mean deviation (MD) were evaluated with univariate and multivariate regression analyses. RESULTS: 23.5% (42/179) and 5.0% (9/179) of subjects had ABP (TSS<80) with GDx VCC and GDx ECC, respectively. For both GDx VCC and ECC, the TSS was significantly correlated with age, axial length, spherical error and PPA, but not with visual-field MD. After adjusting the effect of covariates, the axial length/spherical error and PPA were significantly associated with GDx VCC TSS, whereas the axial length/spherical error was the only predictor for GDx ECC TSS. Myopic eyes were more likely to develop ABP in both GDx VCC and ECC. CONCLUSIONS: Axial length or spherical error is a significant predictor for ABP with both GDx VCC and GDx ECC. Caution should be exercised in interpreting the results of scanning laser polarimetry in eyes with a long axial length or myopia.

Agreement between spectral-domain and time-domain OCT for measuring RNFL thickness.

BACKGROUND/AIMS: To evaluate spectral-domain (SD) optical coherence tomography (OCT) reproducibility and assess the agreement between SD-OCT and Time-Domain (TD) OCT retinal nerve fibre layer (RNFL) measurements. METHODS: Three Cirrus-SD-OCT scans and one Stratus-TD-OCT scan were obtained from Diagnostic Innovations in Glaucoma Study (DIGS) healthy participants and glaucoma patients on the same day. Repeatability was evaluated using Sw (within-subject standard deviation), CV (coefficient of variation) and ICC (intraclass correlation coefficient). Agreement was assessed using correlation and Bland-Altman plots. RESULTS: 16 healthy participants (32 eyes) and 39 patients (78 eyes) were included. SD-OCT reproducibility was excellent in both groups. The CV and ICC for Average RNFL thickness were 1.5% and 0.96, respectively, in healthy eyes and 1.6% and 0.98, respectively, in patient eyes. Correlations between RNFL parameters were strong, particularly for average RNFL thickness (R(2) = 0.92 in patient eyes). Bland-Altman plots showed good agreement between instruments, with better agreement for average RNFL thickness than for sectoral RNFL parameters (for example, at 90 microm average RNFL thickness, 95% limits of agreement were -13.1 to 0.9 for healthy eyes and -16.2 to -0.3 microm for patient eyes). CONCLUSIONS: SD-OCT measurements were highly repeatable in healthy and patient eyes. Although the agreement between instruments was good, TD-OCT provided thicker RNFL measurements than SD-OCT. Measurements with these instruments should not be considered interchangeable.

Longitudinal profile of retinal ganglion cell damage assessed with blue-light confocal scanning laser ophthalmoscopy after ischaemic reperfusion injury.

AIM: To longitudinally investigate retinal ganglion cell (RGC) expression of Thy-1, a cell-surface glycoprotein specifically expressed in RGCs, with a blue-light confocal scanning laser ophthalmoscope, following retinal ischaemia induced by acute elevation of intraocular pressure. METHODS: A blue-light confocal scanning laser ophthalmoscope (bCSLO, 460 nm excitation and 490 nm detection) was used to image Thy1-cyan fluorescent protein (CFP) mice before and weekly for 4 weeks after transiently elevating the intraocular pressure to 115 mm Hg for 45 min (n = 4) or 90 min (n = 5) to induce ischaemic injury. Corresponding retinal areas before and after the intraocular pressure (IOP) elevation, during the period of ischaemic reperfusion, were compared, and the fluorescent spots (Thy-1 expressing RGCs) were counted. The longitudinal profile of CFP-expressing RGCs was modelled with a linear regression equation. The spatial distribution of RGC damage was analysed in the superior, nasal, inferior and temporal quadrants of the retina. RESULTS: No significant change was found at 4 weeks after 45 min of IOP elevation (n = 4, p = 0.465). The average RGC densities before and 4 weeks after IOP elevation were 1660 (SD 242) cells/mm2 and 1624 (209) cells/mm2, respectively. However, significant loss of CFP-expressing RGCs was detected at 1 week following 90 min of IOP elevation (n = 5, p<0.001). After this initial RGC loss, no significant change was detected subsequently. The proportion of RGC fluorescence remaining was variable and ranged from 14.5% to 79.5% at 4 weeks after the IOP elevation. The average RGC densities before and 4 weeks after IOP elevation were 1443 (162) cells/mm2 and 680 (385) cells/mm2, respectively. Diffuse loss of fluorescent RGCs was observed in the spatial distribution analysis. CONCLUSIONS: The longitudinal profile of Thy-1 expressing RGC fluorescence loss after ischaemic injury is non-progressive and unrelated to the duration of reperfusion.

Effects of scan circle displacement in optical coherence tomography retinal nerve fibre layer thickness measurement: a RNFL modelling study.

OBJECTIVE: To study the effect of optical coherence tomography (OCT) scan circle displacement on retinal nerve fibre layer (RNFL) measurement errors using cubic spline models. METHODS: Forty-nine normal subjects were included in the analysis. In one randomly selected eye in each subject, RNFL thickness around the optic disc was measured by taking 16 circular scans of different sizes (scan radius ranged from 1 to 2.5 mm). The RNFL profile in each eye was constructed with a mathematical model using a smoothing spline approximation. Scan circle (diameter 3.4 mm) RNFL measurements (total average, superior, nasal, inferior, and temporal RNFL thicknesses) obtained from eight directions (superior, superonasal, nasal, inferonasal, inferior, inferotemporal, temporal, and superotemporal) displaced at different distances (0.1, 0.2, 0.3, 0.4, 0.5, 0.6, and 0.7 mm) from the disc centre were then computed by a computer program and compared to the 'reference standard' where the scan circle is centred at the optic disc. RNFL measurement error was calculated as the absolute of (RNFL thickness(displaced) - RNFL thickness(reference standard)). RESULTS: The respective mean average, superior, nasal, inferior, and temporal RNFL measurement errors were 2.3+/-2.0, 4.9+/-4.5, 4.1+/-3.8, 6.2+/-7.6, and 3.8+/-3.5 microm upon 0.1 mm scan circle displacement, and 12.1+/-11.4, 27.8+/-18.4, 21.7+/-18.6, 34.8+/-22.9, and 15.2+/-10.7 microm upon 0.7 mm scan circle displacement. Significant differences of average and quadrant RNFL thicknesses were evident between centred and displaced scan circle measurements (all with P<0.001). RNFL measurement error increased in a monotonic fashion with increasing distance away from the disc and the change was direction-dependent. CONCLUSIONS: RNFL measurement error varies with the direction and distance of scan displacement. The superior and the inferior RNFL measurements are most vulnerable to scan displacement errors, whereas the average RNFL thickness is the least susceptible. Obtaining a well-centred scan is essential for reliable RNFL measurement in OCT.

Quantitative assessment of lens opacities with anterior segment optical coherence tomography.

AIM: To evaluate the reliability of lens density measurement with anterior segment optical coherence tomography (OCT) and its association with the Lens Opacity Classification System Version III (LOCS III) grading. METHODS: Fifty-five eyes from 55 age-related cataract patients were included. One eye from each subject was selected at random for lens evaluation. After dilation, lens photographs were taken with a slit lamp and graded against the LOCS III standardised condition. Anterior segment OCT imaging was performed on the same eyes with a high-resolution scan. The association between the anterior segment OCT nucleus density measurement and LOCS III nuclear opalescence (NO) and nuclear colour (NC) scores was evaluated with the Spearman correlation coefficient. Anterior segment OCT measurement precision, coefficient of variation (CVw), and intraclass correlation coefficient (ICC) were calculated. RESULTS: The mean NO and NC scores were 3.39 (SD 1.10) and 3.37 (SD 1.27), respectively. Significant correlations were found between anterior segment OCT nuclear density measurements and the LOCS III NO and NC scores (r = 0.77 and 0.60, respectively, both with p<0.001). The precision, CVw and ICC of anterior segment OCT measurement were 2.05 units, 4.55% and 0.98, respectively. CONCLUSION: Anterior segment OCT nucleus density measurement is reliable and correlates with the LOCS III NO and NC scores.

[Evaluation of focal arteriolar narrowing of retinal arterioles in glaucoma]. / Fokale Verengungen retinaler Arteriolen beim Glaukom.

PURPOSE: Vascular risk factors seem to play a role in the pathogenesis of glaucoma. This study was performed to compare the prevalence of focal arteriolar narrowing in glaucoma patients and normals. MATERIAL AND METHODS: Stereoscopic optic disc photographs of 40 normal subjects and 14 primary open-angle glaucoma (POAG) patients were reviewed independently in a masked fashion by two graders. Focal arteriolar narrowing within one disc diameter from the rim edge was evaluated based on two different methods: narrowing present if (1) the arteriole was wider distal to the narrowing and (2) if the arteriole was wider both distal and proximal to the narrowing. RESULTS: With both methods, focal arteriolar narrowing was significantly higher in glaucoma versus normal eyes. Focal arteriolar narrowing was observed with definition 1 in 35.0% of normals and 71.4% of POAG patients and with definition 2 in 12.5% of normals and 42.9% of POAG patients. CONCLUSIONS: Focal arteriolar narrowing is more frequent in glaucoma patients than in normals. Independent of the method for assessing arteriolar narrowing, however, the diagnostic value of arteriolar narrowing seems limited due to the high incidence in normals.

Detection of prostaglandin EP(1), EP(2), and FP receptor subtypes in human sclera.

PURPOSE: To examine the expression of five prostaglandin (PG) receptors, EP(1), EP(2), EP(3), EP(4), and FP and their corresponding mRNA transcripts in human sclera and cultured human scleral fibroblasts (HSFs). METHODS: Primary cultures of HSFs were established from donor eyes. Also, sclera from human donor eyes was snap frozen and sectioned. Immunocytochemistry was performed on HSFs and tissue sections with subtype-specific antibodies to the EP(1), EP(2), EP(3), EP(4), and FP receptors. The presence of mRNA for the receptor subtypes was examined from total RNA obtained from human sclera and confirmed with restriction digest analysis. RESULTS: Positive EP(1) and FP receptor immunoreactivity was observed in fibroblasts within the sections from human sclera. In primary cultures of HSFs, EP(1) and FP labeling was observed over the entire cell surface. EP(2) immunoreactivity within HSFs was mostly present in the juxtanuclear region. RT-PCR analysis of total RNA isolated from human sclera and HSFs confirmed the presence of EP(1), EP(2), and FP receptor subtypes. The identity of the polymerase chain reaction products was confirmed by restriction enzyme analysis. No mRNA or immunoreactivity above basal levels was detected for the EP(3) and EP(4) prostanoid receptor subtypes in tissue sections or primary cultures. CONCLUSIONS: The EP(1), EP(2), and FP receptor subtypes are present in HSFs, suggesting that these cells may respond to endogenous PGs and their structural analogues through interaction with these receptor subtypes.

Atrophy of relay neurons in magno- and parvocellular layers in the lateral geniculate nucleus in experimental glaucoma.

PURPOSE: To determine whether in glaucoma there is atrophy of relay neurons in magnocellular and/or parvocellular lateral geniculate nucleus (LGN) layers projecting to the visual cortex and to compare the degree of neuronal atrophy in magnocellular layers with that in parvocellular layers. METHODS: Seven cynomolgus monkeys with unilateral experimentally induced glaucoma and five control monkeys were studied. The left LGN neurons in magnocellular layer 1 and parvocellular layers 4 and 6, connected to the right glaucomatous eye were examined. Immunocytochemistry with antibody to parvalbumin was used to specifically label relay neurons connecting to the visual cortex. Neuronal cell body cross-sectional area was estimated using unbiased point-counting methodology. Experimental and control groups were compared using t-tests. Analysis of covariance (ANCOVA) tests were used to compare the percentage of decrease in mean neuronal area between layers 1, 4, and 6, as a function of percentage of optic nerve fiber loss or mean IOP. There was significant correlation between percentage of optic nerve fiber loss and mean IOP. RESULTS: The mean cross-sectional area of relay neurons in magnocellular layer 1 and parvocellular layers 4 and 6 were significantly decreased in glaucoma compared with controls by 28%, 37%, and 45%, respectively. Neuronal area decreased in a linear fashion, with increasing optic nerve fiber loss or increasing mean IOP for layers 1, 4, and 6. The percentage of neuronal shrinkage in each of parvocellular layers 4 and 6, as a function of optic nerve fiber loss (P = 0.05; P = 0.001, respectively) or mean IOP (P = 0.046; P = 0.0008, respectively), was greater than that seen in magnocellular layer 1. CONCLUSIONS: Relay neurons in the LGN, which project to the visual cortex, undergo significant shrinkage in glaucoma, and neurons in parvocellular layers undergo significantly more shrinkage than neurons in magnocellular layers.

Using optical imaging summary data to detect glaucoma.

PURPOSE: To evaluate the sensitivity and specificity for discriminating between early to moderate glaucomatous and normal eyes using summary data reports from the Heidelberg Retina Tomograph (HRT), the GDx Nerve Fiber Analyzer (GDx), and the Optical Coherence Tomograph (OCT). DESIGN: Comparative cross-sectional study PARTICIPANTS: One eye each of 50 normal subjects and 39 glaucoma patients with early to moderate visual field damage (mean deviation, -5.04 +/- 3.32 dB; range, -0.85 to -13.2 dB). METHODS: Three experienced graders masked to patient identity and diagnosis evaluated each summary data report from the HRT, GDx, and OCT independently. MAIN OUTCOME MEASURES: Each summary report was classified as either normal or glaucomatous. Sensitivity and specificity are reported for each grader, and agreement between graders is reported. RESULTS: For the HRT, sensitivity and specificity ranged from 64% to 75% and 68% to 80%, respectively. Agreement (kappa +/- standard error [SE]) between observers one and two, two and three, and one and three was 0.73 +/- 0.07, 0.77 +/- 0.07, and 0.67 +/- 0.08, respectively. For the GDx, sensitivity and specificity ranged from 72% to 82% and 56% to 82%, respectively. Agreement (kappa +/- SE) between observers one and two, two and three, and one and three was 0.66 +/- 0.08, 0.66 +/- 0.08, and 0.50 +/- 0.09, respectively. For the OCT, sensitivity and specificity ranged from 76% to 79% and 68% to 81%, respectively. Agreement (kappa +/- SE) between observers one and two, two and three, and one and three was 0.73 +/- 0.07, 0.58 +/- 0.08, and 0.51 +/- 0.09, respectively. CONCLUSIONS: When used alone, HRT, GDx, and OCT summary data reports can differentiate between normal and glaucomatous eyes with mild to moderate visual field loss. However, none of the instruments provided sensitivity and specificity that justify summary data reports being used as a screening tool for early to moderate glaucoma.