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.     

Ultrasound biomicroscopic findings of the iridocorneal angle in live healthy and glaucomatous dogs

By using ultrasound biomicroscopy (UBM), the cross-sectional structures of the entire iridocorneal angle (ICA) which are unable to assess with gonioscopic examination were evaluated objectively and quantitatively in live healthy and glaucomatous dogs. The ICAs of normotensive eyes in healthy dogs with normal open angle (NOR), a predisposition to primary closed angle glaucoma (PCAG) (PREDIS) and suffering from unilateral PCAG (UNI), as well as the ICAs of hypertensive eyes with acute and chronic PCAG (ACG and CRG), were assessed. The opening of the ciliary cleft in PREDIS was smaller than that in NOR. In UNI, the opening and area of the ciliary cleft were significantly decreased compared with those of NOR and PREDIS. ACG had widespread structural abnormalities including marked decrease in the ciliary cleft and scleral venous plexus, and a thinner sclera than those in normotensive eyes, whereas the ICA collapsed in CRG with the thinnest sclera. Medical therapy-responsive glaucomatous cases had wider ciliary cleft and scleral venous plexus than unresponsive ones. These findings suggest that the ciliary cleft and scleral venous plexus of the ICA are key structures contributing to not only the pathophysiology of canine glaucoma but also the responsiveness to medical therapy in glaucomatous eyes, and cross-sectional entire structures of the ICA should be evaluated quantitatively with UBM when diagnosing and managing canine glaucoma.     

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.     

Anterior lens capsule disruption and suspected malignant glaucoma in a dog

A Boston Terrier puppy presented with a full-thickness peripheral corneal defect, iris prolapse and anterior lens capsule tear in the left eye (OS). Phacofragmentation and primary repair of the corneal laceration was performed. At surgery, subluxation of the lens was also apparent. One day postoperative, there was severe corneal edema, diffuse hyphema, an intraocular pressure (IOP) of 65 mmHg and a small amount of vitreous that protruded from the corneal incision OS. Malignant glaucoma or pupillary block glaucoma were suspected. Intravenous mannitol was administered preoperatively and had no effect. An anterior vitrectomy was performed on the vitreous within the anterior chamber and pupil. One day postoperative the IOP was 16 mmHg in the right eye (OD) and 20 mmHg OS. Postoperative iridocyclitis was managed medically, and additional elevations in IOP were not recorded. Resolution of the elevated IOP following anterior vitrectomy was supportive of pupillary block or malignant glaucoma. Vision returned 3 weeks after the initial surgery. Two years after the initial injury, the eye is visual and comfortable with infrequent topical anti-inflammatory therapy.     

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.     

Enhanced expression of cyclooxygenase-2 in glaucomatous dog eyes

Objective Cyclooxygenase‐2 (COX‐2)‐derived prostaglandins (PGs) are shown to play important pathophysiologic roles in various disease states. Recently, the effectiveness of topical PGs in reducing intraocular pressure (IOP) has stimulated further interest in the physiologic function of COX‐2 and PGs in normal and glaucomatous eyes. Therefore, we investigated the cell‐type distribution and expression of COX‐2 in normal and glaucomatous dog eyes. Procedures Using isoform‐specific antibodies, we immunohistochemically evaluated COX‐2 expression in formalin‐fixed and paraffin‐embedded normal (n = 5) and glaucomatous (n = 17) dog eyes. Results In the normal eyes, only minimal COX‐2 immunoreactivity was observed in the ciliary epithelium. In the glaucomatous eyes, COX‐2 expression was further observed in the cornea and corneoscleral limbus. In the cornea, moderate to strong COX‐2 expression was observed in all corneal layers (epithelium, stromal cells and endothelium), with the greatest expression present in the epithelial layer. In the corneoscleral limbus area, COX‐2 immunoreactivity was noted in the stromal cells of sclera, trabecular meshwork and endothelial cells of the angular aqueous plexus. Conclusions Increased expression of COX‐2 in dog glaucomatous eyes suggests that COX‐2‐derived PGs may have a potential role in the pathogenesis of canine glaucoma.     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.     

Corneal squamous cell carcinoma in a dog: a case report

Purpose:  To report a case of primary corneal squamous cell carcinoma (SCC) in an English Bulldog. In addition, immunohistochemistry of the corneal tissue mass was performed using a panel of antibodies. A prominent feature of the present case was the clinical history of chronic keratitis due to eyelid abnormalities.
Results:  No papillomavirus antigen was detected in section of normal or neoplastic corneal tissue. The corneal epithelial cells were positive for pancytokeratins AE1/AE3 and MNF116, and E-cadherin. The neoplastic cells in close proximity to the normal epithelial lining were positive for both pancytokeratins and E-cadherin with gradual loss of staining toward the center of the neoplastic mass. Rare neoplastic cells demonstrated positive staining for caspase 3 and a large number was strongly positive for GADD45 and p53.
Conclusion and discussion:  The observed loss of the various cytokeratins, the strong p53 expression, and low numbers of caspase 3 positive cells were suggestive that a p53 mutation may have caused this primary corneal SCC. Over-expression of the tumor-suppressor gene p53 is likely to be a consequence of ultraviolet radiation exposure. Two factors, however, may have played a role in the formation of this primary corneal SCC: chronic irritation of the corneal surface (microtrauma) and exposure to UV radiation.     

Validation of the TonoVet® rebound tonometer in normal and glaucomatous cats

Objective  To validate intraocular pressure (IOP) readings obtained in cats with the TonoVet^® tonometer. Animals studied  IOP readings obtained with the TonoVet^® were compared to IOP readings determined by manometry and by the Tono‐Pen XL^? in 1 normal cat and two glaucomatous cats. TonoVet^® and Tono‐Pen XL^? readings were also compared in a further six normal and nine glaucomatous cats. Procedures  The anterior chambers of both eyes of three anesthetized cats were cannulated and IOP was varied manometrically, first increasing from 5 to 70 mmHg in 5 mmHg increments, then decreasing from 70 to 10 mmHg in 10 mmHg decrements. At each point, two observers obtained three readings each from both eyes, with both the TonoVet^® and Tono‐Pen XL^?. IOP was measured weekly for 8 weeks with both tonometers in six normal and nine glaucomatous unsedated cats. Data were analyzed by linear regression. Comparisons between tonometers and observers were made by paired student t‐test. Results  The TonoVet^® was significantly more accurate than the Tono‐Pen XL^? (P = 0.001), correlating much more strongly with manometric IOP. In the clinical setting, the Tono‐Pen XL^? underestimated IOP when compared with the TonoVet^®. Conclusions  Both the TonoVet^® and Tono‐Pen XL^? provide reproducible IOP measurements in cats; however, the TonoVet^® provides readings much closer to the true IOP than the Tono‐Pen XL^?. The TonoVet^® is superior in accuracy to the Tono‐Pen XL^? for the detection of ocular hypertension and/or glaucoma in cats in a clinical setting.     

Pectinate ligament dysplasia (PLD) and glaucoma in Flat Coated Retrievers. I. Objectives, technique and results of a PLD survey

Pectinate ligament dysplasia (PLD) in dogs is a congenital ocular abnormality affecting the iridocorneal angle, the presence of which may be associated with adult-onset primary glaucoma. This paper describes the objectives, methods and results of a survey of PLD in Flat Coated Retrievers in the UK, including the technique used for gonioscopic assessment of the degree of PLD. In a random sample of 389 Flat Coated Retrievers, 34.7% were found to have a degree of PLD. In a 'target' sample of 48 Flat Coated Retrievers (close relatives of dogs from the random sample with high degrees of PLD, or dogs presented with primary glaucoma), 83.3% were found to have a degree of PLD. The percentage of animals with a degree of PLD was only 6.0% in a 'control' sample of 100 dogs from other breeds. The relationship between PLD and canine glaucoma is discussed.     

Mast cell numbers in normal and glaucomatous canine eyes

Numbers of mast cells in the cornea, sclera, choroid, ciliary body, iris, and retina of sections of globes from 35 clinically normal dogs and 34 dogs with secondary glaucoma was determined. Fixed globes were trimmed along a vertical midsagittal plane and embedded in paraffin. Tissue sections, approximately 6 microns thick, were stained with toluidine blue for identification of mast cells. In normal globes, most of the mast cells were observed in the anterior portion of the uvea, and fewer mast cells were seen in the choroid and sclera. Mast cells were not observed in the retina and were seldom observed in the cornea of dogs with or without glaucoma. In sections of glaucomatous globes, mast cells were distributed evenly in the uvea and sclera, and fewer mast cells were present than in normal globes, regardless of the cause of glaucoma.     

Effects of oral isosorbide and glycerol on intraocular pressure,serum osmolality,and blood glucose in normal dogs

Objective To evaluate and compare the effects of oral isosorbide and glycerol on intraocular pressure (IOP), serum osmolality (SOSM), and blood glucose (BG) in normal dogs. Methods Ten normal dogs were administered an oral dose of either isosorbide (1.5 g/kg), glycerol (1.5 g/kg) or control (water, 2 mL/kg) in a double blind protocol. Prior to dosing, baseline IOP, SOSM, and BG were measured in all dogs. IOP was subsequently evaluated every 30 min for 6 h post‐dosing. BG and SOSM were reassessed at times 1, 2, 4, and 6 h post‐dosing. After 1‐week washout periods, every dog was subject to each of the three treatments. The dogs were held NPO for 4 h after dosing. Results The maximal decrease in IOP was 17% by 1 h and 13.5% by 30 min after glycerol and isosorbide administration, respectively. However, the overall changes in IOP were not significant when compared to the controls. SOSM increased above baseline after dosing with glycerol but decreased after isosorbide, which difference was significant at 1, 2, and 4 h post‐administration. BG significantly increased after administration of glycerol relative to the control but was not significantly affected by isosorbide. Conclusions Neither glycerol nor isosorbide significantly affected IOP when compared to the control. However, glycerol induced significant elevations in both BG relative to the control and SOSM relative to isosorbide.     

Pectinate ligament dysplasia and glaucoma in Flat Coated Retrievers. II. Assessment of prevalence and heritability

Pectinate ligament dysplasia (PLD) in dogs is a congenital ocular abnormality affecting the iridocorneal angle, the presence of which may be associated with adult-onset primary glaucoma. This study demonstrates a significant association between PLD and glaucoma in Flat Coated Retrievers, and also a higher prevalence of PLD in Flat Coated Retrievers compared with other breeds. It is shown that the association between PLD and glaucoma was robust and not due to a small increase in PLD with age. There was also a significant association between PLD in offspring and parents and, using the offspring–parents regression, the heritability of PLD was estimated as approximately 0.7. As glaucoma is significantly related to PLD, glaucoma may also be heritable. Thus, glaucoma can be controlled by gonioscopic examination and elimination of animals with a high degree of PLD from a breeding program. Breeding from only those animals with a moderate or low PLD value or score should limit the expected prevalence of glaucoma in the resulting population to < 0.2%. The preceding study describes in detail the PLD survey, gonioscopic technique and some initial results obtained prior to this epidemiological investigation.