Ron Ofri

Guidelines for clinical electroretinography in the dog

These procedures described for the dog ERG were approved at the 1st European Conference on Veterinary Visual Electrophysiology in Vienna, Austria, May 30, 2000. Dr. Narfström was Chair of the Committee for a Harmonized ERG Protocol, appointed by the European College of Veterinary Ophthalmology (ECVO), and Dr. Ofri was secretary. The other coauthors are committee members. Guidelines for ERG procedures in other animal species for clinical and laboratory studies are planned for in the future and the present guidelines are planned to be revised on a biannual basis. A brief report of the recommended procedures is available in the Conference Proceedings book.

T-cell-based vaccination for morphological and functional neuroprotection in a rat model of chronically elevated intraocular pressure

Acute or chronic glaucoma is often associated with an increase in intraocular pressure (IOP). In many patients, however, therapeutic pressure reduction does not halt disease progression. Neuroprotection has been proposed as a complementary therapeutic approach. We previously demonstrated effective T-cell-based neuroprotection in experimental animals vaccinated with the synthetic copolymer glatiramer acetate (copolymer-1, Cop-1), a weak agonist of self-antigens. This study was undertaken to test different routes and modes of vaccination with Cop-1 as treatment modalities for protection against retinal ganglion cell (RGC) death caused by chronic elevation of IOP in rats, and to determine whether anatomical neuroprotection is accompanied by functional neuroprotection. In a chronic model of unilaterally high IOP, Cop-1 vaccination, with or without an adjuvant, protected rats against IOP-induced loss of RGCs by eliciting a systemic T-cell-mediated response capable of cross-reacting with self-antigens residing in the eye. In rats deprived of T cells, Cop-1 (unlike treatment with α2-adrenoreceptor agonists) was not protective of RGCs, substantiating the contention that its beneficial effect is not conferred directly but is T-cell-mediated. Pattern electroretinography provided evidence of functional protection. Thus, vaccination with adjuvant-free Cop-1 can protect RGCs from the consequences of elevated IOP in rats. This protection is manifested both morphologically and functionally. These findings can be readily implemented for the development of a therapeutic vaccination to arrest the progression of glaucoma.

Cataract development in diabetic sand rats treated with α‐lipoic acid and its γ‐linolenic acid conjugate

Diabetes commonly leads to long‐term complications such as cataract. This study investigated the effects of α‐lipoic acid (LPA) and its γ‐linolenic acid (GLA) conjugate on cataract development in diabetic sand rats.

Gene Augmentation Therapy Restores Retinal Function and Visual Behavior in a Sheep Model of CNGA3 Achromatopsia

Achromatopsia is a hereditary form of day blindness caused by cone photoreceptor dysfunction. Affected patients suffer from congenital color blindness, photosensitivity, and low visual acuity. Mutations in the CNGA3 gene are a major cause of achromatopsia, and a sheep model of this disease was recently characterized by our group. Here, we report that unilateral subretinal delivery of an adeno-associated virus serotype 5 (AAV5) vector carrying either the mouse or the human intact CNGA3 gene under the control of the red/green opsin promoter results in long-term recovery of visual function in CNGA3-mutant sheep. Treated animals demonstrated shorter maze passage times and a reduced number of collisions with obstacles compared with their pretreatment status, with values close to those of unaffected sheep. This effect was abolished when the treated eye was patched. Electroretinography (ERG) showed marked improvement in cone function. Retinal expression of the transfected human and mouse CNGA3 genes at the mRNA level was shown by polymerase chain reaction (PCR), and cone-specific expression of CNGA3 protein was demonstrated by immunohistochemisrty. The rescue effect has so far been maintained for over 3 years in the first-treated animals, with no obvious ocular or systemic side effects. The results support future application of subretinal AAV5-mediated gene-augmentation therapy in CNGA3 achromatopsia patients.

Guidelines for clinical electroretinography in the dog: 2012 update

The full-field, flash electroretinogram (ERG) is now a widely used test of canine retinal function for the clinical diagnosis of hereditary retinal dystrophies and other causes of retinal degeneration, assessment of retinal function in patients with opaque media, ruling out of generalized retinal diseases in patients with sudden loss of vision and in ophthalmological research, as well as in pharmaceutical and toxicological screening for deleterious side effects of drugs and other chemical compounds. In 2002, the first guidelines for clinical ERGs in this species adopted by the European College of Veterinary Ophthalmologists were published. This work provides an update of these guidelines.

Clinical electrophysiology in veterinary ophthalmology - the past, present and future

The aim of this review is to introduce the reader to the world of clinical veterinary electroretinography. An important indication for ERG recordings in the dog is the early diagnosis of progressive retinal atrophy, an inherited form of photoreceptor degeneration, analogous to retinitis pigmentosa in humans. In most of the 20 canine breeds in which the disease has been studied electrophysiologically, changes in the ERG appear long before the appearance of clinical signs. This early diagnosis is a vital tool in efforts to eradicate the disease through preventive breeding. Pre-operative screening of canine cataract patients is another common indication for electroretinography in the dog. The ERG is also used to diagnose inherited and nutritional photoreceptor degenerations in the cat, and retinal disorders in a number of other animal species. The abundance of animal species (and breeds) seen by the veterinary ophthalmologist lends additional importance to the problem of a harmonized ERG recording protocol. The European College of Veterinary Ophthalmologists has set up a special committee to formulate guidelines for such a protocol. International meetings and wetlabs are also being organized as part of an effort to improve the quality of electrophysiological diagnosis that veterinary ophthalmologists provide their patients.

Reduced expression of Pax6 in lens and cornea of mutant mice leads to failure of chamber angle development and juvenile glaucoma

Heterozygous mutations in PAX6 are causative for aniridia, a condition that is frequently associated with juvenile glaucoma. Defects in morphogenesis of the iridocorneal angle, such as lack of trabecular meshwork differentiation, absence of Schlemms canal and blockage of the angle by iris tissue, have been described as likely causes for glaucoma, and comparable defects have been observed in heterozygous Pax6-deficient mice. Here, we employed Cre/loxP-mediated inactivation of a single Pax6 allele in either the lens/cornea or the distal optic cup to dissect in which tissues both alleles of Pax6 need to be expressed to control the development of the tissues in the iridocorneal angle. Somatic inactivation of one allele of Pax6 exclusively from epithelial cells of lens and cornea resulted in the disruption of trabecular meshwork and Schlemms canal development as well as in an adhesion between iris periphery and cornea in juvenile eyes, which resulted in the complete closure of the iridocorneal angle in the adult eye. Structural changes in the iridocorneal angle presumably caused a continuous increase in intraocular pressure leading to degenerative changes in optic nerve axons and to glaucoma. In contrast, the inactivation of a single Pax6 allele in the distal optic cup did not cause obvious changes in iridocorneal angle formation. We conclude that the defects in iridocorneal angle formation are caused by non-autonomous mechanisms due to Pax6 haploinsufficiency in lens or corneal epithelial cells. Pax6 probably controls the expression of signaling molecules in lens cells that regulate the morphogenetic processes during iridocorneal angle formation.

Intraocular pressure and tear production in five herbivorous wildlife species

The intraocular pressure and rate of tear production were measured in 18 addax antelopes (Addax nasomaculatus), four impalas (Aepyceros melampus), 11 wide-lipped rhinoceroses (Ceratotherium simum), 10 white-tailed wildebeests (Connochaetes gnou) and seven scimitar-horned oryxes (Oryx dammah). The animals were anaesthetised with an intramuscular injection of etorphine hydrochloride and acepromazine maleate, and the Schirmer tear test I was used to evaluate tear production, and applanation tonometry was used to evaluate the intraocular pressure. The mean (sd) rate of tear production ranged from 17.6 (3.1) mm/minute in the rhinoceros to 28.8 (8.3) mm/minute in the addax. The intraocular pressure ranged from 8.0 (1.2) mmHg in the impala to 32.1 (10.4) mmHg in the rhinoceros. The rate of tear production in the addax and the intraocular pressure in the rhinoceros appear to be the highest values of these variables to have been reported in any species.

Acute blindness associated with monoclonal gammopathy induced by Ehrlichia canis infection

Ehrlichia canis infection was diagnosed in a Labrador retriever presented with a primary complaint of acute blindness. Ocular signs on admission included bilateral hyphema, retinal haemorrhage and retinal detachment. Serum protein electrophoresis results revealed monoclonal gammopathy. This report discusses and suggests the pathogenesis of ocular bleeding in canine monocytic ehrlichiosis. Blood hyperviscosity, elevation in oncotic pressure, vasculitis, thrombocytopenia and platelet dysfunction are all proposed to be important factors in the pathogenesis of acute blindness in canine monocytic ehrlichiosis.

A novel day blindness in sheep: epidemiological, behavioural, electrophysiological and histopathological studies.

Four genetically related Improved Awassi sheep flocks had sporadic births of lambs with congenital visual impairments that differed from other known forms of sheep blindness. Pedigree analysis suggested an autosomal recessive mode of inheritance. Behavioural studies of 4-month old affected lambs showed that their day vision (but not night vision) was impaired. Electrophysiological results at this age demonstrated diminished function of cones but not rods. Histopathological and immunohistochemical evaluation of affected retinas from 5-month old lambs revealed both red-green and blue cones, suggesting that the behavioural day blindness and reduced cone electroretinograms reflect cone dysfunction rather than severe cone photoreceptor loss. Awassi day blindness may be a form of achromatopsia.