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.     

Aqueous humor myocilin protein levels in normal, genetic carriers, and glaucoma Beagles

Objective The gene (myocilin: MYOC) has been attributed to be involved in over 6% of inherited types of human glaucoma, the highest correlation for any gene to date. This study determines myocilin protein levels in the aqueous humor (AH) of normal laboratory quality, genetic carrier (offspring of normal laboratory quality and POAG Beagles), and primary open angle glaucoma (POAG) Beagles. Materials and methods Eighteen dogs were used and classified as either normal, carrier or having mild, moderate or advanced POAG. A 0.1‐mL sample of AH was drawn from the anterior chamber of each dog in the study and frozen on dry ice. A modified Coomassie stain and Western blot, using a polyclonal rabbit antihuman myocilin antibody (Santa Cruz Biotechnologies, Santa Cruz, CA), was run on each sample to compare the myocilin levels. A purified human trabecular meshwork excreted myocilin protein sample was used as a control (Alcon Research Laboratories, Fort Worth, TX) and its band/densitometry measurement was defined as one unit of myocilin for comparisons. Results Comparisons of AH myocilin levels differed among normal laboratory quality, genetic carrier, and POAG Beagles at different stages of the disease. In the normal laboratory, Beagles the AH myocilin measured 0.817 ± 0.075 units (mean ± SEM); in the carrier Beagles the AH myocilin was 3.117 ± 0.290 units. As POAG progressed, myocilin protein levels also increased to 6.097 ± 0.810, 8.844 ± 1.079, and 17.228 ± 1.198 units in the early, moderate, and advanced forms, respectively. Overall comparisons between normal, carrier and all POAG Beagles combined showed significant differences (P < 0.0010). Individual comparisons between normal and carrier eyes showed significant differences (P < 0.0193). Comparisons between normal and all POAG eyes also showed significant differences (P < 0.0426). Conclusion This study shows myocilin protein is present in normal Beagles, markedly increased in POAG Beagles, and mildly increased in genetic carrier Beagles. There is a strong correlation between amounts of AH myocilin protein and the presence and severity of POAG. The exact role of AH myocilin levels in the genesis of ocular hypertension remains unresolved, but myocilin may adversely affect AH outflow.     

Immunohistochemical analysis of lens epithelial-derived membranes following cataract extraction in the dog

The objective of the study was to characterize the morphologic and immunohistochemical features of lens epithelial-derived proliferative membranes from the anterior segment of canine globes. These features were correlated with those previously identified for diseases resulting from lens epithelial cell (LEC) proliferation including posterior capsular opacification, traumatic subcapsular cataract, and subcapsular plaques associated with hypermature cataracts. Sixteen canine globes were removed as a result of glaucoma or other complications following cataract extraction. Light microscopic and immunohistochemical analysis was performed on sections from formalin-fixed, paraffin-embedded globes. The tissues were stained with a variety of antibodies for cellular markers for LECs, growth factors or other cellular constituents relevant to cellular metaplasia and proliferation. The membranes were composed of monolayers or multilayers of spindle-shaped cells on the external surfaces of the anterior and posterior lens capsule, ciliary processes, iris leaflets, and iridocorneal angle, and they could be seen extending from an obvious monolayer of LEC within the capsular sac. Variably, scattered pigment cells, presumably of uveal origin, were concurrently present. Cellular components of the membranes stained positive for vimentin, transforming growth factor-beta, basic fibroblast growth factor, and smooth muscle actin. An amorphous eosinophilic extracellular matrix consisting predominately of collagen was associated with the membranes. Proliferative anterior segment membranes following cataract surgery were morphologically and immunohistochemically similar to cellular and matrix components of posterior capsular opacification and capsular plaques seen with hypermature cataracts, both of which result from metaplasia and proliferation of LEC. The presence of these LEC-derived membranes in association with secondary glaucoma suggests that exuberant proliferation of LEC outside the confines of the lens capsular sac may cause pathologic alterations in the eye following cataract surgery in the dog.     

Comparative Doppler imaging of the ophthalmic vasculature in normal Beagles and Beagles with inherited primary open-angle glaucoma

The objective of this study was to compare orbital and ocular vasculature velocity, measured by Doppler imaging, in normal Beagles and Beagles with inherited primary open-angle glaucoma. Eight normal Beagles and 13 Beagles with different stages of primary open-angle glaucoma were evaluated twice with a 2–4-week period between measurements. Doppler imaging was performed with the dogs anesthetized, and the Doppler transducer applied directly on the corneal surface. The majority of the orbital vasculature (external ethmoidal artery; internal ophthalmic artery and vein; and external ophthalmic artery and vein) and ocular blood vessels (anterior ciliary artery and veins; long posterior ciliary arteries; short posterior ciliary arteries; primary retinal arteries; and the vortex veins) were identified and Doppler blood velocity parameters were determined. The glaucomatous dogs demonstrated significant differences in the Doppler velocity parameters of several orbital vessels (external ethmoidal, external ophthalmic, and internal ophthalmic arteries), and several ocular vessels (anterior ciliary, short posterior ciliary, and long posterior ciliary arteries). These differences included decreased blood velocities, and increased pulsatility and resistive indexes. The Doppler blood flow velocities of the primary retinal arteries were unchanged between the normal and glaucomatous dogs. In the glaucomatous dogs, the Doppler imaging suggests increased vascular resistance downstream in both the orbital and ocular vasculature. These blood velocity parameter changes may be primary or secondary, and may offer therapeutic opportunities to increase perfusion, prolong the retina and optic nerve head function, and maintain vision in the canine glaucomas.     

Ultrastructural changes in laminar optic nerve capillaries of beagles with primary open-angle glaucoma

Ultrastructural examination of optic nerve capillaries in the canine lamina cribrosa revealed many spherical, membrane-bound, electron-dense inclusions that closely resembled Weibel-Palade bodies, in pericytes and endothelial cells of preglaucomatous, early, moderately, and advanced affected Beagles with hereditary primary open-angle glaucoma. This ultrastructural difference between the laminar capillary endothelial cells of normal and glaucomatous Beagles could represent a functional vascular disorder, because Weibel-Palade bodies are associated with microcirculatory abnormalities.     

Morphologic changes in the lamina cribrosa of beagles with primary open-angle glaucoma

Optic nerve axoplasmic flow is known to be impaired at the scleral lamina cribrosa in Beagles with hereditary primary open-angle glaucoma and is similar to the condition in human beings. Trypsin and detergent digestion to remove all neural, vascular, and glial cellular tissue revealed a well-developed scleral lamina cribrosa in the normal dog in this study. Beagles with primary open-angle glaucoma had signs of mechanical distortion of the anterior lamina cribrosa prior to the detection of ophthalmoscopic changes in the optic nerve head. This change in the supporting architecture of the optic nerve began early and increased in severity as the disease process progressed.     

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.     

Progressive changes in ophthalmic blood velocities in Beagles with primary open angle glaucoma

Objective To measure changes in the ocular and orbital blood flow velocities by color Doppler imaging (CDI) in beagles with primary open angle glaucoma as the disease progressed from early to advanced stages. Methods CDI measurements were performed periodically on 13 glaucomatous Beagles during the nontreated mild, moderate and advanced stages of POAG over the course of 4 years. CDI was performed with the dogs lightly anesthetized (butorphanol 0.1 mg/kg IV, acepromazine maleate 0.02 mg/kg IV, and atropine sulfate 0.05 mg/kg) while the CD transducer was placed directly on the cornea anesthetized with 0.5% tetracaine hydrochloride. Intraocular pressure (IOP) by pneumatonography or TonoPen XL, heart rate and mean arterial blood pressure were measured at the beginning, middle and end of each study. The ophthalmic vessels examined included: external ophthalmic arteries and veins, long and short posterior ciliary arteries, anterior ciliary arteries and veins, primary retinal arteries, and vortex veins. Recordings of each vessel included peak systolic velocity (PSV), end diastolic velocity (EDV) and time averaged velocity (TAV), and when possible the resistive index (RI) and pulsatility index (PI) were computed. Results CDI abnormalities were present before intraocular pressure exceeded the normal range. As the animals aged, and the glaucoma progressed with higher levels of IOP, significant changes occurred in nearly all vessels, and generally included a major increase in RI (P < 0.001) and an increase in the PI (P < 0.001). Mean arterial blood pressure (105 ± 18 mmHg) and heart rate (118 ± 33/min) remained reasonably constant. The IOP gradually increased as the disease progressed (early and normotensive: 19.4 ± 3.9 mmHg; moderate: 29.7 ± 2 mmHg; and advanced: 44.5 ± 6 mmHg). The ocular veins seemed most influenced early on in the disease. Late in the disease, ocular venous blood flow could not be consistently demonstrated. An increase in the PI of ocular veins occurred in the moderately and severely affected glaucomatous Beagles. As the IOP increased, there were trends of increasing resistive index and pulsatility index in most arteries, and periods of marked decreased velocities of the vortex and external ophthalmic veins in severe cases. Conclusion CDI measurements in Beagles with primary open angle glaucoma during the course of 4 years indicate easily measurable and repeatable progressive blood flow abnormalities before the elevation of IOP and, thereafter, with gradually increased levels of IOP.     

Transmission electron microscopy studies in an experimental model of canine atopic dermatitis

Impairment of skin barrier function has been hypothesized in canine atopic dermatitis (AD). In this prospective, controlled study, the ultrastructure of the upper epidermal layers was investigated using an experimental model of canine AD. Seven atopic Beagles sensitized to Dermatophagoides farinae and four healthy Beagles were used as controls. Both normal and atopic dogs were challenged with D. farinae for 3 days. Clinical signs were scored and skin biopsies were taken from the inguinal area before and 3 days after allergen exposure. Samples were processed to enhance lipid visibility and evaluated by Transmission Electron Microscopy. Emphasis was placed on evaluation of the lipid lamellae (LL), and lamellar bodies (LB) of the stratum corneum.
After allergen challenge, atopic Beagles developed severe pruritic dermatitis while no skin lesions were noted in the controls. Ultrastructurally, before allergen challenge, atopic Beagles displayed focally severe abnormalities in LL organization and wider intercellular spaces containing abnormal lipid material. In atopic Beagles, LBs were frequently found inside corneocytes while this finding was not observed in the controls. After allergen challenge, further increase of intercellular spaces was observed in the stratum corneum of atopic Beagles while no appreciable changes were observed in the normal dogs. Intercellular spaces in atopic Beagles were filled with abundant amounts of abnormal lipid material and highly disorganized LL. It is concluded that baseline differences in the ultrastructure of the skin exist between normal and experimentally sensitized atopic Beagles and that these changes are aggravated by allergen challenge and the resulting flare-up of dermatitis.     

Long-term complete chimerism and stable hematopoiesis in beagles after fetal liver hematopoietic stem cell transplantation

Karyotype analysis of bone marrow cells was performed from 6 Beagles that had received sex-mismatched fetal liver hematopoietic stem cell grafts 2.4 to 6.5 years earlier. Two dogs received dog leukocyte antigen-matched fetal liver cells and 4 received dog leukocyte antigen-mismatched cells. In each dog, all 25 evaluated metaphase spreads were of donor genotype. All dogs had normal hemograms and repopulation of bone marrow and no dog developed graft-vs-host disease.     

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.     

犬青光眼的研究进展

犬青光眼是犬眼科疾病中很严重的疾病,影响宠物的生活质量,给宠物和畜主带来极大的痛苦.文章对犬青光眼的发生、分类、临床症状、诊断及治疗等进行综述,为犬青光眼的进一步防治提供参考.     

Scanning electron microscopy of corrosion casts of the optic nerve microcirculation in dogs

Scanning electron microscopy of vascular corrosion casts of the optic nerve region in normal and glaucomatous Beagles demonstrated that the blood supply to the laminar optic nerve is derived from short posterior ciliary arteries, cilioretinal arteries, and longitudinal pial vessels. The short posterior ciliary arteries formed a ring of striated pillars around the scleral canal. The central retinal artery was not present in the dog. Differences between the casts in normal and glaucomatous dogs were not detected.     

Myofibroblasts in the accessory ligament (distal check ligament) and the deep digital flexor tendon of foals

OBJECTIVE: To demonstrate myofibroblasts in the accessory ligament of the deep digital flexor tendon (ie, distal check ligament) and deep digital flexor tendon of clinically normal foals. SAMPLE POPULATION: Tissue specimens from 25 foals that were necropsied for reasons unrelated to this study and unrelated to musculoskeletal disease. PROCEDURE: The distal check ligament and deep digital flexor tendon of both forelimbs were examined histologically. Myofibroblasts were identified by immunohistochemical staining specific for alpha-smooth muscle actin (alpha-SMA). RESULTS: Most of the cells in the distal check ligament and deep digital flexor tendon of all foals stained positive for alpha-SMA. CONCLUSION AND CLINICAL RELEVANCE: Myofibroblasts made up most of the cells in the distal check ligament and deep digital flexor tendon of clinically normal foals. These cells have contractile ability and therefore, may play a role in flexure contracture of these tendons. The ability of tetracycline to chelate calcium or decrease the expression of the contractile protein alpha-smooth muscle actin could inhibit the myofibroblasts' ability to contract, thus providing a rationale for tetracycline administration as a treatment of distal interphalangeal joint flexor deformity in foals.     

Th1/Th2 balance in canine peripheral blood lymphocytes--a flow cytometric study

The canine immune system undergoes continuous remodeling with advancing age. We measured the Th1/Th2 balance in peripheral blood lymphocytes (PBLs) obtained from 23 Beagles ranging in age from 0.5 to 11.8 years by flow cytometric analysis using intracellular cytokine staining. The percentage of CD4 cells producing interferon-gamma (Th1) increased with age. The percentage of CD4 cells producing interleukin-4 (Th2) increased but much less so. In conclusion, we demonstrated that the Th1/Th2 balance in canine peripheral blood could be measured by this flow cytometry technique and the Th1/Th2 balance inclined to dominance of the Th1 subpopulation in PBLs as the dog matured.     

Establishment and characterization of canine rhabdomyosarcoma cell line CMS-C

A novel canine tumor cell line designated as the CMS-C cell line was established from pleomorphic rhabdomyosarcoma (RMS) raised in the prostate gland of a 14-year-old intact male mixed-breed dog. CMS-C cells displayed the same immunohistochemical characteristics (positive for vimentin and desmin and negative for cytokeratin and smooth muscle actin) as the original tumor cells and express myoD1 and UCP3, known as striated muscle-specific molecules, as shown by RT-PCR assay. Therefore, the established CMS-C cell line appears to be of rhabdomyoblast cell origin. The CMS-C cell line established from pleomorphic RMS will be a useful tool for further studies about canine RMS.     

Isolation and measurement of carbonic anhydrase isoenzyme III in plasma, sera, and tissues of dogs

OBJECTIVE: To purify canine carbonic anhydrase isoenzyme III (CA-III) and determine plasma, serum, and tissue concentrations of CA-III in healthy dogs and dogs with experimentally induced muscle damage. ANIMALS: 121 healthy Beagles. PROCEDURE: Muscle was obtained from 2 Beagles after euthanasia, and CA-III was purified and characterized by use of column chromatography and electrophoresis, respectively. A CA-III-specific ELISA was developed to determine concentrations of CA-III in plasma of 116 dogs and tissues of 1 dog. Serum creatine kinase (CK) activity and CA-III concentration were also determined before and after induction of muscle damage by IM injection of 2 ml of 10% lidocaine to 2 dogs. RESULTS: Canine CA-III had a molecular weight of 28 kd and an isoelectric point of 8.2. Mean (+/- SD) concentration of CA-III in plasma of healthy dogs was 16.91 +/- 9.55 ng/ml. The highest tissue concentration of CA-III was detected in skeletal muscle. Serum concentration of CA-III increased and peaked within the first 2 to 3 hours after induction of muscle damage. The increase in CA-III concentration was more rapid than that of CK activity, and concentration reached its maximum and returned to baseline sooner than did CK activity. CONCLUSIONS AND CLINICAL RELEVANCE: The CA-III ELISA we developed was a sensitive method for determining CA-III concentrations in plasma, serum samples, and tissue specimens of dogs. Use of this ELISA requires only a small volume of serum and may enable the study of changes in CA isoenzyme concentrations associated with muscle disorders in dogs.     

Doppler imaging of the ophthalmic vasculature of the normal dog: blood velocity measurements and reproducibility

The purpose of this study was to assess the feasibility and reproducibility of color Doppler imaging (CDI) of the vasculature of the normal canine orbit and eye. Eight normal Beagles were evaluated by Doppler imaging. The goals of the study were to determine the location, spectral waveform morphology, specific blood velocity parameters, and reproducibility for the ophthalmic and orbital vessels most frequently identified in the normal dog. Vessels identified a majority of the time (> 50%) included: external ophthalmic artery, dorsal external ophthalmic vein, ventral external ophthalmic vein, internal ophthalmic artery, anterior ciliary artery and vein, short and long posterior ciliary arteries, primary retinal arteries, and vortex veins. Other vessels imaged less frequently included: external ethmoidal artery (50%), and primary retinal veins (25%). For each blood vessel the time averaged velocity, peak systolic velocity, minimum diastolic velocity, pulsatility index, and resistive index were determined. The ophthalmic and orbital vessels have unique spectral waveforms and velocities which serve as a basis for identification. Reproducibility of the most commonly imaged vessels of the canine eye and orbit with Doppler imaging was high (< 10% variation). Doppler imaging has the potential for determining noninvasively and consecutively the blood velocity parameters found in orbital and ocular diseases, including orbital inflammations and neoplasms; intraocular inflammations and neoplasms; vascular diseases including systemic vascular disease (hypertension), vasculopathies, and anemia; the glaucomas; and documentable follow-up after medical and/or surgical treatment of these diseases.     

Restricted dog leucocyte antigen (DLA) class II haplotypes and genotypes in Beagles

Beagles are commonly used in vaccine trials as part of the regulatory approval process. Genetic restriction within this breed and the impact this might have on vaccine responses are rarely considered. This study was designed to characterise diversity of dog leucocyte antigen (DLA) class II genes in a breeding colony of laboratory Beagles, whose offspring are used in vaccine studies. DLA haplotypes were determined by PCR and sequence-based typing from genomic DNA extracted from blood. Breeding colony Beagles had significantly different DLA haplotype frequencies in comparison with pet Beagles and both groups showed limited DLA diversity. Restricted DLA class II genetic variability within Beagles might result in selective antigen presentation and vaccine responses that are not necessarily representative of those seen in other dog breeds.     

A comparative histochemical and morphometric study of canine skeletal muscle.

The purpose of this study was to determine whether there were differences in skeletal muscle properties in the hindlimb muscles of different types of dogs. Muscle samples were obtained from the gracilis, sartorius cranial head, sartorius caudal head and tibialis anterior muscles of mixed-breed and hound-type dogs and Beagles. Fiber type, fiber size and capillary morphometry determinations of each muscle from each dog were made from sections stained for myofibrillar ATPase activity. Individual animals were bilaterally symmetric for all measured variables. Fiber type, fiber size and capillary geometry varied between dogs of a given type and muscles within a given dog. There were no differences between dog types for fiber type or fiber size; significant variation in log(muscle)/log(body) mass ratios between dog types was observed for all muscles. The results indicate that for a given muscle, significant variation can occur in skeletal muscle characteristics between different types of dogs and that these differences can be independent of differences in exercise history.