Biometry and UBM measurements for the cohort of healthy Eurasier dogs (cohort 2).

<p>Data are expressed as means ± standard deviation (SD) and <i>n</i> is the number of eyes examined in each group whereas <i>N</i> is the number of dogs in each group. We did not include all the eyes, because some of the images could not be exploited.</p><p>AGL: axial globe length; ACL: anterior chamber length; CCW: ciliary cleft width; CCL: ciliary cleft length; DSAC: distance from Schwalbe's line to the anterior capsule of the lens.</p><p><b>^*</b><i>p</i><0.05 versus the male group; °°°p<0.001 versus females; <b>^##</b><i>p</i><0.01 versus the adult males and <b>^¶¶</b><i>p</i><0.01 versus the young male group (Kruskal-Wallis ANOVA followed by a Dunn's post-hoc test).</p><p>Biometry and UBM measurements for the cohort of healthy Eurasier dogs (cohort 2).</p

Determinations of biomarkers of oxidative stress in the plasma of Eurasier dogs.

<p>(a) Plasma glutathione peroxidase (GP) activity in all EDs (black bar), compared with a cohort of randomized control dogs (white bar). (b) Measurement of plasma taurine concentrations in EDs (black bar), compared with control dogs (white bar). (c-d) Determination of the concentrations of the taurine's precursors, methionine and cysteine, in plasma from EDs (black bar) and control dogs (white bar). (e): Measurement of plasma GP activity in the various subgroups of the ED cohort. (f): Measurement of plasma taurine concentration in the various subgroups of EDs. (g) Correlation between GP activity and plasma taurine concentration in the adult male subgroup of EDs, demonstrating a significant association of low levels of GP activity with a low plasma taurine concentration (<i>p</i><0.05; Spearman's correlation test). This correlation was shown to be linear (r²>0.52). ***<i>p</i><0.001 versus the control dogs (Mann Whitney test for a, b,c and Kruskal-Wallis ANOVA followed by the Dunn's post-hoc test for d, e, f).</p

Biochemical markers of inflammation determined in plasma from the cohort of healthy Eurasier dogs (cohort 2).

<p>Data are expressed as means ± standard deviation (SD) for each group. <i>N</i> is the number of dogs in each group.</p><p>CRP: C-reactive protein.</p><p>Biochemical markers of inflammation determined in plasma from the cohort of healthy Eurasier dogs (cohort 2).</p

Correlation between intraocular pressure (IOP) and axial globe length biometric parameters in the various subgroups of the ED cohort.

<p>(a-c) Correlation between IOP and axial globe length (AGL) in all EDs from the cohort: increasing IOP was significantly associated with decreasing AGL (<i>p</i> = 0.05, Spearman's test; a). Similar significant correlations were also found for adult EDs (<i>p</i> = 0.03; b) and for female adult EDs (<i>p</i> = 0.05; c). The linear regression analysis (hatched lines) demonstrated the non-linear status of the correlations, because <i>r</i>²<0.5 in all cases (a-c). (d-f) Correlation between IOP and distance between Schwalbe's line and the anterior capsule of the lens (DSAC): increasing IOP was significantly associated with increasing DSAC in male EDs (<i>p</i> = 0.03; e) and adult female EDs (<i>p</i> = 0.05; f), whereas no significant correlation between these parameters was found when all EDs were considered together (<i>p</i> = 0.17; d). Linear regression analysis (hatched lines) indicated that the significant correlations observed were not linear (<i>r</i>²<0.52 in all cases; d-f).</p

Morphological appearance of the pectinate ligament in the cohort of healthy Eurasier dogs (cohort 2).

<p>The PL was observed by gonioscopy. In cases of pectinate ligament abnormalities, quantification was performed by assigning a percentage of the 360° after systematically viewing the entire circumference of the ICA, and a grade was attributed according to the percentage of the ICA affected (AFEP/MHOC). Grade 0 corresponds to an absence of goniodysplasia (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111873#pone-0111873-g001" target="_blank">figure 1a</a>), grade 1 corresponds to mild goniodysplasia (stage 1 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111873#pone-0111873-g001" target="_blank">figure 1b</a>), grade 2 corresponds to moderate goniodysplasia (stage 2 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111873#pone-0111873-g001" target="_blank">figure 1c</a>), and grade 3 corresponds to severe goniodysplasia (stage 3 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111873#pone-0111873-g001" target="_blank">figure 1d</a>). PLA grades are provided separately for the two eyes because PLA did not progress uniformly in the two eyes.</p><p>Data are expressed as the number of eyes affected by the various grades of goniodysplasia for each group of EDs. <i>N</i> is the number of dogs in each group.</p><p>Morphological appearance of the pectinate ligament in the cohort of healthy Eurasier dogs (cohort 2).</p

Biometry of the dog eyeball.

<p>Representative image from a normal dog obtained by B-mode ultrasound: a standard 10 MHz transducer was placed directly on the cornea after the instillation of topical anesthetic into the eye. A vertical axial scan was performed, making it possible to measure axial globe length (AGL) — the length from the corneal epithelial surface (CES) to the inner surface of the retina (ISR). The normal lens is translucent and only the posterior lens capsule is visible, as a highly reflective concave line. The anterior chamber and the vitreous are clear and anechoic and appear as homogeneous black areas. The scale bar represents 1 mm.</p

Ocular abnormalities and tonometry in the cohort of healthy Eurasier dogs (cohort 2).

<p>Abnormalities were observed on clinical examinations other than gonioscopy examination. Data represent the numbers for each category of abnormalities observed in each subgroup of EDs and the total numbers of abnormalities (with a percentage) are presented in the last line.</p><p>Tonometric data are expressed as means ± standard deviation (SD).</p><p><i>n</i> refers to the number of eyes examined in each group whereas <i>N</i> is the number of dogs in each group.</p><p>IOP: intraocular pressure.</p><p>Ocular abnormalities and tonometry in the cohort of healthy Eurasier dogs (cohort 2).</p

Iridocorneal angle (ICA) in dogs: images from ICA sections versus ultrasound biomicroscopy.

<p>(a-b) Normal histological sections of the ICA (stained with hematoxilin/eosin) are shown on the left, with the corresponding UBM images on the right. ICA structures in (a) are compared with those obtained by UBM (b). The pectinate ligament corresponds to the anterior delineation of the ciliary cleft. The trabecular meshwork, which fills the ciliary cleft (CC), is clearly visible. (c-d) The width of the entrance of the ciliary cleft (CCW) was measured by determining the distance between the corneoscleral limbus and the iris root as follows. A line outlining the pectinate ligament was traced. Another line, outlining the inner wall of the sclera in the anterior portion of the CC, was then traced. The CCW corresponds to a line perpendicular to the inner wall of the sclera and crossing the pectinate ligament halfway along its length. (e-f) The length of the CC (CCL) was determined as the distance from the pectinate ligament (or the most anterior visible portion of the uveal trabecular meshwork) to the anterior portion of the ciliary body as follows. A line was traced outlining the pectinate ligament. The CCL corresponds to a line parallel to the long axis of the CC, crossing the pectinate ligament halfway along its length at the anterior and reaching the ciliary muscle at the posterior. (g-h) Determination of the distance between Schwalbe's line (the peripheral termination of Descemet's membrane, i.e. the borderline between the cornea and sclera) and the anterior capsule of the lens (DSAC). The scale bar corresponds to 1 mm for all images. The ICA sections were obtained from the same dog as the UBM images.</p

Pectinate ligament abnormalities (PLA) evaluated by gonioscopic examination in Eurasier dogs.

<p>(a) Normal pectinate ligament (PL) visible when rarefaction of the initial fibrilar sheet was almost complete, by 2 to 4 weeks after birth, leaving strands of intertwining collagen, progressively encased by attenuate trabecular cells, confluent with the anterior surface of the iris (stage 0). (b-d) The entire ICA and opening of the CC were systematically examined for the presence of a PLA, defined as abnormally broad and thickened pectinate ligament fibers (stage 1) (<b>b</b>) or solid sheets (stage 2) (<b>c</b>) of PL tissue, with or without “holes” (stage 3) (<b>d</b>). (e) Percentage of PLA (including grade 1, 2 and 3; black bars) with respect to normal pectinate ligament (white bars) in each category of the ED cohort. *<i>p</i><0.05 and ***<i>p</i><0.001 indicate significant differences in the prevalence of PLA between the indicated groups (χ² test).</p

Taurine prevents the RGC death in NMDA-treated retinal explants.

<p>A) Digitalized reconstruction of a whole flat-mounted retinal explant immunolabeled with the POU4F1 (Brn3a) antibody. B–E) Representative enlarged fields from flat-mounted retinal explants acquired with the automated platform, showing POU4F1-immunopositive RGCs in a control untreated condition (Control; B) after 100 µM NMDA application (NMDA; C), after co-application of NMDA with taurine (1 mM; NMDA+Taurine; D) or after co-application of NMDA with MK801 (100 µM; NMDA+MK801; E) for 4 days. F) Quantification of RGC densities from whole flat-mounted retinal explants using the automated counting platform in Control group (n = 33, white bar); NMDA group: (n = 31, hatched bar); NMDA+Taurine group (n = 23, black bar) and Taurine group (n = 6, grey bar). G) Quantification of POU4F1-immunopositive RGC densities from whole flat-mounted explants under the control group (white bar), or the NMDA group (hatched bar), the NMDA plus MK 801 group (black bar), or finally the MK-801 group (grey bar) (n = 6 for each group). Data are expressed as means ± s.e.m. from n independent experiments. ***p<0.001, *p<0.05; One-way ANOVA followed by a Bonferroni post-hoc test. The scale bar represents 100 µm in panels (B–E).</p