Localization of smooth muscle actin in the iridocorneal angle of normal and spontaneous glaucomatous beagle dogs

Purpose To date, our knowledge of the canine trabecular meshwork (TM) with regard to contractility is incomplete. It is important to understand the potential contractile capability within the TM and possible changes associated with spontaneous hypertensive glaucoma. To that end we have examined the presence of actin, including smooth muscle (SM) actin, in the normal and glaucomatous canine iridocorneal angle (ICA) morphologically and immunohistochemically. Methods Sections from the ICAs of 12 Beagles with inherited glaucoma (3 months to 6 years old) and age‐matched normal Beagles were treated with target retrieval, protein and power blocked and sequentially incubated with the primary antibody (rat anticanine SM actin) and the secondary antibody (rabbit antirat immunoglobulin), followed by peroxidase labeled streptavidin and incubation with substrate‐chromogen solution (AEC). Smooth muscle fibers that lined an artery within canine heart tissue were used as positive controls. Separate specimens were prepared for ultrastructual observation. Results Ultrastructurally, cells within the inner, posterior region of the corneoscleral TM and outer, posterior region of the uveal TM contained many microfilaments, 6 nm in diameter (i.e. actin). Immunohistochemistry demonstrated that cells within these regions possessed SM actin, having been greatest posteriorly, but extended anteriorly to a lesser extent. In the preglaucomatous affected dog the localization pattern for SM actin was identical to that seen in the normal dogs. With the progression of the disease the pattern disappeared. Conclusions The interior presence of myofibroblastic cells within the canine ICA suggests that these cells and the smooth muscle cells of the ciliary body along the same plane of orientation function to facilitate the removal of aqueous humor and are likely to be influenced by vascular mediators. The contractile apparatus for the ICA in the dog with inherited glaucoma appeared identical to that of the normal dog prior to expression of the disease, but weakened as the disease progressed.     

Relationship of the iridocorneal angle, as measured using ultrasound biomicroscopy, with post-operative increases in intraocular pressure post-phacoemulsification in dogs

Objective  To investigate the relationship of the iridocorneal angle as it appears on ultrasound biomicroscopy (UBM) to increases in IOP post-phacoemulsification in the canine eye.
Animals studied  47 eyes of 28 dogs of various age, sex, and breed.
Methods  The ciliary process and limbus were used as a reference points to measure the angle opening distance (AOD), which was set by multiplying 0.45 by the distance between the ciliary process and limbus (when measured from the ciliary process). Pressure measurements were taken at five set points: Before, immediately post-, one day post-, 1 week post-, and 1 month post-phacoemulsification.
Results  A weak relationship between the AOD and the IOP one day post-phacoemulsification (Intercept: 2.83, Slope: –1.24, P -value: 0.0155) was observed. No relationship immediately post-operative (Intercept: 3.45, Slope: –1.34, P -value: 0.0651), 1 week post-phacoemulsification (Intercept: 2.31, Slope: –0.01, P -value: 0.9829), 1 month post-phacoemulsification (Intercept: 1.85, Slope: –0.49, P -value: 0.1533) was observed. The pre-operative measurements made with UBM were: Distance from limbus to ciliary process (DLCP): (Minimum: 1.152, Maximum: 2.992, Mean: 1.91, SD: 0.468); AOD (Minimum: 0.104, Maximum: 0.764, Mean: 0.40, SD: 0.172).
Conclusions  The relationship between AOD as measured by UBM pre-operatively is weakly associated with IOP pressure elevations at day one post-phacoemulsification. Further study is required prospectively to establish the importance of this relationship. Initial measurements of the canine iridocorneal angle were created, suggesting a method to be used in the future to establish true canine normal measurements.     

Evaluation of the distance between Schwalbe’s line and the anterior lens capsule as a parameter for the correction of ultrasound biomicroscopic values of the canine iridocorneal angle

Objective To determine whether 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 lens capsule (SLD) is an applicable parameter for correcting raw ultrasound biomicroscopic values of the canine iridocorneal angle (ICA) and to establish a comparative system for the ICA values in different canine breeds with varied body sizes/weights. Animal studied The dogs were divided into four groups based on body weight (BW): <4 kg, 4–8 kg, 8–20 kg, and >20 kg, and 180 normotensive eyes were studied. Procedures The ICA microstructure was examined by ultrasound biomicroscopy (UBM) using 40‐MHz probes in dogs with or without anesthesia/sedation. Linear regression analysis and correlation coefficients were evaluated between SLD or SLD² and UBM measurements; subsequently, noncorrected and SLD‐corrected UBM values were statistically assessed. Results Significant linear correlations were detected between SLD and the ciliary cleft width, the minimum distance between the angle recess and the scleral venous plexus, and the scleral thickness. Positive linear correlations were also demonstrated between SLD² and the ciliary cleft area as well as the scleral venous plexus area. Raw UBM measurements were corrected by using a ratio with SLD on the distance or SLD² on the area. Although noncorrected UBM measurements increased with canine BW, SLD‐corrected UBM values remained similar with no significant statistical differences in any of the dogs. Conclusions Correction with SLD would be clinically useful for comparing UBM measurements of the ICA in dogs with different body sizes/weights.     

Schwalbe line's cell in the normal and glaucomatous dog

Objective The canine iridocorneal angle contains an operculum, which is similar to that in nonhuman primates and consists of a peripheral extension of the inner cornea that overlies the anterior‐most portion of the corneoscleral trabecular meshwork. This region contains cells, the Schwalbe line’s (SL) cells, that have been found to have secretory and epithelial characteristics. This region of the iridocorneal angle represents the nonfiltering portion and becomes altered early during spontaneous glaucoma in the Beagle. The present study describes the SL cell for the first time in the dog and changes associated with canine primary open angle glaucoma. Procedures The iridocorneal angles from 18 Beagles with inherited glaucoma (3 months–8 years old) and 17 normal, age‐matched Beagles were placed in 10% buffered formalin for light microscopic evaluation, or 2.5% glutaraldehyde for ultrastructural evaluation. Using at least three fields from each region of the iridocorneal angle (opercular, corneoscleral, and uveal) at ×1000 magnification, trabecular cell nuclei were counted. Results The operculum in the canine iridocorneal angle consisted of the peripheral extension of the corneal endothelium and underlying anterior‐most corneoscleral meshwork, having no direct contact with the angular aqueous plexus. The SL cells associated with operculum‐retained epithelial morphology (polyhedral in shape with rER, Golgi apparatus, and secretory vesicles) in both normal, pre‐and early glaucomatous dogs. In animals with moderate and advanced stages the SL cells often became less epithelial and secretory in appearance. The number of SL cells in normal dogs declined by approximately one‐third by the end of their first year with gradual loss thereafter. In the glaucoma group the decline was more substantial and continuous through the first three years. Conclusions The SL cell is morphologically a distinct cell type within the canine iridocorneal angle that is specifically associated with the nonfiltering portion of the corneoscleral trabecular meshwork. Changes in the SL cells of the glaucomatous dog occurred with regard to age and progression of the disease.     

Comparison of three rebound tonometers in normal and glaucomatous dogs

Objective

The objectives of the study were to compare intraocular pressure (IOP) readings across a wide range and obtained via three rebound tonometers in ADAMTS10-mutant Beagle-derived dogs with different stages of open-angle glaucoma (OAG) and normal control dogs and to investigate the effect of central corneal thickness (CCT).

Animals Studied

Measurements were performed on 99 eyes from 50 Beagle-derived dogs with variable genetics—16 non-glaucomatous and 34 with ADAMTS10-OAG. Seventeen OAG eyes were measured twice—with and without the use of IOP-lowering medications.

Procedures

IOP was measured in each eye using three tonometers with their “dog” setting—ICare® Tonovet (TV), ICare® Tonovet Plus® (TVP), and the novel Reichert® Tono-Vera® Vet (TVA)—in randomized order. CCT was measured with the Accutome® PachPen. Statistical analyses included one-way ANOVA, Tukey pairwise comparisons, and regression analyses of tonometer readings and pairwise IOP-CCT Pearson correlations (MiniTab®).

Results

A total of 116 IOP measurements were taken with each of the three tonometers. When comparing readings over a range of ~7–77 mmHg, mean IOPs from the TV were significantly lower compared with TVP (−4.6 mmHg, p < .001) and TVA (−3.7 mmHg, p = .001). We found no significant differences between TVA and TVP measurements (p = .695). There was a moderate positive correlation between CCT and IOP for TVA (r = 0.53, p < .001), TVP (r = 0.48, p < .001), and TV (r = 0.47, p < .001).

Conclusions

Our data demonstrate strong agreement between TVP and TVA, suggesting that the TVA may similarly reflect true IOP values in canines. CCT influenced IOP measurements of all three tonometers.     

Structure and presumptive function of the iridocorneal angle of the West Indian manatee (Trichechus manatus), short-finned pilot whale (Globicephala macrorhynchus), hippopotamus (Hippopotamus amphibius), and African elephant (Loxodonta africana)

The iridocorneal angles of prepared eyes from the West Indian manatee, short-finned pilot whale, hippopotamus and African elephant were examined and compared using light microscopy. The manatee and pilot whale demonstrated capacity for a large amount of aqueous outflow, probably as part of a system compensating for lack of ciliary musculature, and possibly also related to environmental changes associated with life at varying depths. The elephant angle displayed many characteristics of large herbivores, but was found to have relatively low capacity for aqueous outflow via both primary and secondary routes. The hippopotamus shared characteristics with both land- and water-dwelling mammals; uveoscleral aqueous outflow may be substantial as in the marine mammals, but the angular aqueous plexus was less extensive and a robust pectinate ligament was present. The angles varied greatly in size and composition among the four species, and most structures were found to be uniquely suited to the habitat of each animal.     

Abdominal ultrasonographic findings at diagnosis of osteosarcoma in dogs and association with treatment outcome

The purpose of this study was to describe abdominal ultrasonographic findings present at diagnosis of osteosarcoma (OSA) in dogs and to investigate for associations with treatment outcome. Medical records from 118 dogs diagnosed with OSA that had abdominal ultrasonography performed as part of their initial evaluation were reviewed. Fifty‐seven percent had ultrasonographic abnormalities identified. The organ with the highest frequency of ultrasonographic changes was the spleen. While most sonographic changes were considered to be either benign or of unknown clinical consequences, metastases were identified in three dogs (2.5%), two of which (1.7%) did not have other evidence of metastasis. Dogs with any ultrasonographic abnormality were less likely to receive definitive therapy (P = 0.005) and exhibited shorter survival, although the latter observation was not statistically significant (P = 0.071). However, the identification of lesions in either the liver (P = 0.021) or the kidney (P = 0.003) was statistically associated with shorter survival.     

Ultrasound biomicroscopy of the iridocorneal angle of the eye before and after phacoemulsification and intraocular lens implantation in dogs

OBJECTIVE: To compare the iridocorneal angle (ICA) and angle opening distance (AOD) in dogs with cataractous and noncataractous lenses; evaluate cataractous eyes ultrasono-graphically for association of postoperative ocular hypertension (POH) with the ICA, AOD, and postoperative echogenic anterior chamber debris; and evaluate intraobserver reliability associated with ICA and AOD measurements. ANIMALS: 56 dogs with 102 cataracts, and 23 clinically normal dogs. PROCEDURES: Ultrasound biomicroscopy was performed on 102 eyes of 56 dogs before and after cataract surgery and on 46 nondilated and dilated eyes of 23 clinically normal dogs. Cataract stage, ICA, AOD, and association with POH were assessed. RESULTS: Cataract stage and ICA or AOD were not significantly associated; however, ICA and AOD typically decreased with increasing cataract maturity. Before and after pupillary dilation, AODs were significantly smaller in cataractous eyes than in noncataractous eyes. Before surgery, ICA and AOD in eyes without pupillary dilation were significantly associated with POH. At > 13 degrees , odds of developing POH increased by 11% for each degree increase in the ICA. Postoperative anterior chamber debris was not associated with POH. Coefficient of variation for repeated measurements was 10% for the ICA and 9.5% for the AOD, suggesting good intraobserver reliability. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, dogs with larger ICA and AOD measurements before surgery were at greater risk of developing POH. This information may be useful for future studies to determine whether preventative treatment for POH administered prior to surgery may be beneficial.     

Pectinate ligament dysplasia and narrowing of the iridocorneal angle associated with glaucoma in the English Springer Spaniel

Objective The aim of the study was to evaluate if tonometry and gonioscopy could serve as predictors of later glaucoma development in the English Springer Spaniel, a breed prone to developing primary glaucoma. Animals studied Gonioscopy was performed on 279 English Springer Spaniel dogs, 119 males and 160 females, with clinically normal eyes. In addition, 14 dogs, five males and nine females were examined for glaucoma. Results A positive relation between pectinate ligament dysplasia (PLD) and narrowing of the relative width of the ciliary cleft (RWOCC) and also between PLD, RWOCC and age was demonstrated. The prevalence of PLD was 25.5%, which is higher than in other breeds reported. A positive association was demonstrated between PLD and glaucoma, as well as between narrowing of the RWOCC and glaucoma. Glaucoma was not observed in dogs with normal appearance of the iridocorneal angle. English Springer Spaniels related to dogs with glaucoma show more narrowing of the RWOCC and more PLD than unrelated dogs. Conclusion This study demonstrates the positive association between PLD and glaucoma, between narrowing of the iridocorneal angle and glaucoma and the effect of age on the iridocorneal angle. Mating of dogs with normal iridocorneal angles appears to reduce the presence and degree of abnormal appearance of the iridocorneal angle in the offspring. However, breeding only dogs with normal iridocorneal angles without consideration of their relationship to dogs with glaucoma is not a guarantee for preventing glaucoma in the offspring.     

Ultrasound is a poor predictor of early or overt liver or spleen metastasis in dogs with high‐risk mast cell tumours

Conflicting evidence exists regarding the importance of routine abdominal ultrasound (US) with hepatic and splenic fine needle aspiration (FNA) cytology during staging of canine mast cell tumours (MCT). The objective of this study was to correlate ultrasonographic and cytologic findings in dogs with strictly defined high‐risk MCTs and to determine the influence on outcome. Our hypothesis was that US poorly predicts visceral metastasis in high‐risk MCTs and that early metastasis is associated with improved outcome when compared to overt metastasis. US of liver and spleen correlated to cytologic results, categorized as no metastasis, early metastasis or overt metastasis. Of 82 dogs prospectively enrolled, 18% had early visceral metastasis and 7% had overt metastasis on cytology; 67% with visceral metastasis had regional LN metastasis. US was a poor predictor of metastasis with sensitivity, specificity, positive predictive value and negative predictive value for the spleen of 67%, 68%, 21% and 94%, respectively and for the liver of 29%, 93%, 56% and 82%, respectively. Median time to progression (TTP) for dogs with no metastasis, early metastasis and overt metastasis was not reached, 305 and 69 days, respectively (P < .001). Median survival time (MST) for the 3 groups were not reached, 322 and 81 days, respectively (P < .001). High Patnaik or Kiupel grade, early metastasis, overt metastasis and adequate local control were significantly associated with outcome. Early visceral metastasis was associated with poorer outcome compared to dogs without metastasis, however, a subset of dogs experienced long‐term control.