Extracellular Vesicles Isolated from Equine Adipose-Derived Stromal Stem Cells (ASCs) Mitigate Tunicamycin-Induced ER Stress in Equine Corneal Stromal Stem Cells (CSSCs).

Corneal ulcers, characterized by severe inflammation of the cornea, can lead to serious, debilitating complications and may be vision-threatening for horses. In this study, we aimed to investigate the role of endoplasmic reticulum (ER) stress in corneal stem progenitor cell (CSSC) dysfunction and explore the potential of equine adipose-derived stromal stem cell (ASC)-derived extracellular vesicles (EVs) to improve corneal wound healing. We showed that CSSCs expressed high levels of CD44, CD45, and CD90 surface markers, indicating their stemness. Supplementation of the ER-stress-inducer tunicamycin to CSSCs resulted in reduced proliferative and migratory potential, accumulation of endoplasmic reticulum (ER)-stressed cells in the G0/G1 phase of the cell cycle, increased expression of proinflammatory genes, induced oxidative stress and sustained ER stress, and unfolded protein response (UPR). Importantly, treatment with EVs increased the proliferative activity and number of cells in the G2/Mitosis phase, enhanced migratory ability, suppressed the overexpression of proinflammatory cytokines, and upregulated the anti-inflammatory miRNA-146a-5p, compared to control and/or ER-stressed cells. Additionally, EVs lowered the expression of ER-stress master regulators and effectors (PERK, IRE1, ATF6, and XBP1), increased the number of mitochondria, and reduced the expression of Fis-1 and Parkin, thereby promoting metabolic homeostasis and protecting against apoptosis in equine CSSCs. Our findings demonstrate that MSCs-derived EVs represent an innovative and promising therapeutic strategy for the transfer of bioactive mediators which regulate various cellular and molecular signaling pathways.

Schirmer tear test-1 with open or closed eyelids: An evaluation in brachycephalic and nonbrachycephalic dogs.

PURPOSE: Assess aqueous tear production when measured with the dogs' eyelids open or closed. METHODS: Thirty healthy dogs (15 Shih Tzus, 15 Labrador retrievers) were recruited. With the order of testing randomized for each dog, two sessions (separated by 30 min) of STT-1 testing were performed with the dogs' eyelids closed or open. Schirmer strip wetness (every 10 s for 60 s) and number of time(s) the strip dislodged during testing were recorded in each eye. Preferred STT-1 method was surveyed via a global Listserv of the veterinary ophthalmology community. RESULTS: STT-1 values were significantly higher in closed versus open eyes in Shih Tzus (18.6 ± 2.7 mm/min vs. 16.3 ± 2.5 mm/min; p = .002) and Labrador retrievers (21.6 ± 2.9 mm/min vs. 17.8 ± 3.2 mm/min, p < .001), findings that were also significant at times <60 s for either breed (p ≤ .004). Schirmer strips dislodged from six dogs with open eyelids and no dogs with closed eyelids. Maximal STT-1 difference with closed versus open eyelids was 13 mm/min in Labrador retrievers and 7 mm/min in Shih Tzus. Survey results from 275 veterinarians showed STT-1 performed with "closed eyelids" (38.5%), "open eyelids" (26.9%), or "never paid attention, sometimes closed, sometimes open" (34.6%). CONCLUSIONS: Eyelids status (closed or open) during STT-1 testing had a significant impact on aqueous tear secretion in brachycephalic and nonbrachycephalic dogs, highlighting the importance of consistency when repeating STT-1 in a canine patient. STT-1 differences are likely due to sustained reflex tearing throughout the test duration when the dogs' eyelids are closed.

The effect of systemic acetazolamide administration on intraocular pressure in healthy horses-A preliminary study.

OBJECTIVES: In equine glaucoma, topical treatment with carbonic anhydrase inhibitors (CAIs) is recommended. Oral acetazolamide, a systemic CAI, is used in horses with hyperkalemic periodic paralysis. Information regarding its effect on equine intraocular pressure (IOP) is scarce. The aim of the study was to determine the effect of oral acetazolamide treatment on IOP in horses, in a case-control study. ANIMALS: Ten healthy horses. PROCEDURES: Horses were treated with oral acetazolamide (4.4 mg/kg) BID for 1 week. Serum acetazolamide concentrations were determined by liquid chromatography/tandem mass spectrometry, and IOP were measured before treatment, daily during treatment, and at 48 and 72 h after treatment. RESULTS: Acetazolamide serum levels reached steady state at 72 h after the first oral dose. In a mixed effect model logistic regression, there was a significant decrease in IOP on the third treatment day, of 2.4 mmHg (p = .012) and 2.7 mmHg (p = .006) in the left (OS) and right eye (OD), respectively. On the seventh day, there was a decrease in 2.5 mmHg (p = .008) and 2.7 mmHg (p = .007) OS and OD, respectively. A significant increase occurred 48 h following treatment discontinuation (3.6 mmHg, p < .001 and 3.5 mmHg, p < .001 OS and OD, respectively). The area under the concentration versus time curve (AUC(0-10h)) was 1.1 ± 0.5 µg/mL*h, mean residence time 6.7 ± 4.3 h, peak plasma concentration (Cmax) 0.4 ± 0.4 µg/mL and time to reach Cmax 1.8 h. There was a significant increase in serum concentrations 1, 2, 48, 72, and 156 h following the first drug administration (p < .05). CONCLUSIONS: Further studies are required to determine whether acetazolamide is a potential treatment for equine glaucoma.

Retrospective evaluation of pre-surgical electroretinography results in a mixed-breed canine population presented for cataract removal surgery.

OBJECTIVE: Electroretinography (ERG) is used prior to cataract removal surgery to assess retinal function. We aimed to replicate and improve upon previous studies by performing a full ECVO protocol and by examining the retina post-surgery in all patients. ANIMALS STUDIED: One hundred twenty-seven eyes from 67 dogs were included in the study. PROCEDURES: A full ECVO protocol electroretinography, which includes extensive rod and cone analysis, was performed on all dogs presenting for cataract surgery. RESULTS: Our main findings were that amplitudes, but not implicit times of rod responses decreased with advanced cataracts. Amplitudes of the single flash rod and rod flicker responses were significantly lower in eyes with mature cataracts, and the former also decreased in hypermature cataracts. Cone flicker amplitude responses were also significantly lower in eyes with mature and hypermature cataracts. However, mixed single flash rod-cone and cone responses, with the exception of the mixed rod-cone a-wave amplitude in eyes with hypermature cataracts, were unaffected by cataract stage. The b-wave amplitude of the scotopic, mixed rod-cone, and photopic cone responses were affected by age and decreased by an average of 2.9, 7.5, and 1.5 µV/year, retrospectively (p < 0.01). CONCLUSIONS: Lower ERG amplitudes in canine cataract patients may result from aging or the presence of advanced cataracts and may not indicate the presence of retinal disease.

The 100 most-cited articles published in the Veterinary Ophthalmology journal between 1998 and 2022: A bibliometric study.

PURPOSE: The aim of this study was to bibliometrically analyze the 100 most-cited articles published in the Veterinary Ophthalmology (VO) journal. METHODS: Web of Science was searched for citations of VO articles published in 1998-2022. Tissue and species studied, and first and last author domicile and affiliation were recorded for the 100 most-cited articles and descriptively analyzed. RESULTS: Altogether, the 100 most-cited VO articles have cited a total of 5483 times. Most commonly, these were devoted to the cornea (23%), multiple tissues (19%), and glaucoma (16%). Studies on dogs (36%), horses (17%), and multiple species (15%) were most often cited. Most first/last authors were from the USA (n = 113), Brazil (n = 13), and France and Germany (n = 7 each), and most frequently affiliated with the University of Florida (n = 36), University of Wisconsin-Madison (n = 15), and Animal Health Trust, North Carolina State University, and Ohio State University (n = 6 each). KN Gelatt (n = 9), DE Brooks (n = 6), and FJ Ollivier and EO MacKay (n = 5 each) were the most frequent first or last authors. The greatest number of citations was for articles with KN Gelatt (n = 555), FJ Ollivier (n = 411), and DE Brooks (n = 372) as first or last authors. "The comparative morphology of the tapetum lucidum" by FJ Ollivier et al. (2002) is the most frequently cited article in VO history (n = 178). CONCLUSIONS: This study provides insight into the impact of publishing in VO and a more comprehensive understanding of trends and the most influential contributions to VO.

Accuracy of data in abstracts of veterinary ophthalmology research articles published in peer-reviewed journals.

OBJECTIVE: Assess the accuracy of abstracts in published veterinary ophthalmology articles. PROCEDURES: Abstracts and contents of 204 original research articles in veterinary ophthalmology published in seven peer-reviewed journals between 2016-2020 were reviewed. Abstracts were considered inconsistent if they contained data that were either missing from or inconsistent with corresponding data in the article's body. Each abstract was graded between 0 (inaccurate) to 3 (accurate), and each inconsistency was subjectively classified as minor or major. The influence of selected variables was assessed: journal, impact factor, year of publication, number of words in abstract, study type (prospective/retrospective), and characteristics of the corresponding author [institution (academia/private practice), country of domicile (native/non-native English), number of publications]. RESULTS: Most abstracts were accurate, with 1%, 4%, 9% and 86% receiving a score of 0, 1, 2 and 3, respectively. When detected, most inconsistencies were considered minor (77%). Although not statistically significant (p ≥ .130), the proportion of articles with a perfect score (=3) was higher in prospective (88%) vs. retrospective (81%) studies, academia (88%) vs. private practice (78%), and studies from corresponding authors domiciled in English (89%) vs. non-English (83%) speaking countries. A significant but very weak (r = -0.15 to -0.19; p ≤ .034) negative correlation was found between accuracy score and number of words, as well as 1-year and 5-year impact factors. CONCLUSIONS: Although relatively uncommon, data in abstracts that are inconsistent or missing from the article's body do occur in veterinary ophthalmology articles, and could adversely influence a reader's interpretation of study findings.

A retinoscopic survey of donkeys and goats.

PURPOSE: Assess the refractive states of donkeys and goats. METHODS: Forty-two donkeys and 28 goats were enrolled. The mean ± SD ages were 7.68 ± 7.33 years for donkeys and 4.26 ± 2.33 years for goats. Seven donkeys and one goat were <6 months old. Retinoscopy was performed in alert animals, following cycloplegia in goats but not in donkeys. Normality was determined using the Kolmogorov-Smirnov test. The two primary meridians and two eyes were compared using Pearson's correlation and paired Student's t-tests. The association between refractive states and age was examined using one-way ANOVA in donkeys and a paired Student's t-test in goats. One-sample t-tests were conducted to assess if the refractive error distributions were significantly different from "0". RESULTS: The mean ± SD spherical equivalent (SE) refractive errors of the right and left donkey eyes were -0.80 ± 1.03 D and -0.35 ± 0.95 D, respectively. The majority (86%) of the donkeys had an astigmatic refraction and eight (19%) had anisometropia. The mean SE refractive errors of the right and left goat eyes were -0.15 ± 1.1 D and -0.18 ± 1.2 D, respectively. The majority (54%) of the goat eyes had an astigmatic refraction and five (18%) had anisometropia. The right and left eye SE refractive errors were positively correlated in both species (both p = .9). Age was not correlated with refractive error in both donkeys (p = .09) and goats (p = .6). CONCLUSIONS: Both goats and donkeys are emmetropic.

Effect of anesthetic induction with propofol, alfaxalone or ketamine on intraocular pressure in cats: a randomized masked clinical investigation.

OBJECTIVE: To compare the effect of propofol, alfaxalone and ketamine on intraocular pressure (IOP) in cats. STUDY DESIGN: Prospective, masked, randomized clinical trial. ANIMALS: A total of 43 ophthalmologically normal cats scheduled to undergo general anesthesia for various procedures. METHODS: Following baseline IOP measurements using applanation tonometry, anesthesia was induced with propofol (n = 15), alfaxalone (n = 14) or ketamine (n = 14) administered intravenously to effect. Then, midazolam (0.3 mg kg-1) was administered intravenously and endotracheal intubation was performed without application of topical anesthesia. The IOP was measured following each intervention. Data was analyzed using one-way anova and repeated-measures mixed design with post hoc analysis. A p-value <0.05 was considered significant. RESULTS: Mean ± standard error IOP at baseline was not different among groups (propofol, 18 ± 0.6; alfaxalone, 18 ± 0.7; ketamine, 17 ± 0.5 mmHg). Following induction of anesthesia, IOP increased significantly compared with baseline in the propofol (20 ± 0.7 mmHg), but not in the alfaxalone (19 ± 0.8 mmHg) or ketamine (16 ± 0.7 mmHg) groups. Midazolam administration resulted in significant decrease from the previous measurement in the alfaxalone group (16 ± 0.7 mmHg), but not in the propofol group (19 ± 0.7 mmHg) or the ketamine (16 ± 0.8 mmHg) group. A further decrease was measured after intubation in the alfaxalone group (15 ± 0.9 mmHg). CONCLUSIONS AND CLINICAL RELEVANCE: Propofol should be used with caution in cats predisposed to perforation or glaucoma, as any increase in IOP should be avoided.

Histological and molecular characterization of glaucoma model induced by one or two injections of microbeads to the anterior chamber of mice.

PURPOSE: To characterize glaucoma-induced damage following injections of plastic microbeads into the anterior chamber of mice. METHODS: Mice were divided into three groups: a single plastic microbeads injection (n = 21); two consecutive plastic microbead injections to the right eye at 1-week intervals, 4 of which with two consecutive saline injections in the left eye (n = 15); and an additional control group of two consecutive saline injections at 1-week intervals (n = 6). Intraocular pressure (IOP) was measured weekly. Retinal thickness, ganglion cells (RGCs) and axonal loss, inflammatory and gliosis reactions were measured at week four. Molecular analysis using qRT-PCR in the microbeads injection groups focused on expression levels of inflammation and glaucoma-related genes. RESULTS: Mean IOP following single injection at 4 weeks was significantly elevated compared to baseline in injected eyes (14.5 ± 3.3 mmHg vs. 11.1 ± 2.5 mmHg, respectively, p = 0.003) and not in fellow eyes (13.2 ± 2.9 mmHg vs. 12.2 ± 2.9, respectively, NS). Six (35.3%) bead-injected eyes had IOP ≥ 17 mmHg compared with 2 (11.8%) saline-injected control eyes. Retinal thickness in injected and fellow eyes was 193.7 ± 15.5 µm and 223.9 ± 15.5 µm, respectively (p = 0.03). RGC loss in injected and fellow eyes was 16.0 ± 0.5 and 17.6 ± 0.7 cells per 200 µm, respectively (p = 0.005). Retinal gliosis, axonal loss and inflammatory cell infiltration to the bead-injected eyes were noted. Molecular analysis following double injection showed STAT3 expression decreased in the glaucoma-induced optic nerves (0.69 ± 0.3 vs. 1.16 ± 0.3, p = 0.04), but increased in the glaucoma-induced retinae (p = 0.05) versus saline; retinal IL-1ß decreased significantly (0.04 ± 0.04 vs. 0.36 ± 0.2, p = 0.02). TNF-α, NFkB and SOD-1 expression did not change. CONCLUSION: One/two injections of microbeads elevated IOP, with measurable neuronal damage. An inflammatory response was detected in the injured retina and optic nerve. The therapeutic significance of these findings should be explored.

Treatment with chloroquine is retinotoxic in captive African penguins (Speniscus demersus). Attenuation and recovery of electroretinographic responses.

PURPOSE: To evaluate the effect of prophylactic anti-malarial chloroquine treatment, and its cessation, on electroretinographic (ERG) responses of captive African penguins. METHODS: A brief ERG protocol ("QuickRetCheck") was recorded under mesopic conditions with manual restraint and no sedation or pupil dilation. Birds were recorded on two separate occasions, first while being treated with a daily chloroquine dose of 10 mg/kg for 12 days (n = 15, treatment group) and second after 4 months without chloroquine treatment (n = 6, off-treatment group). Three birds were recorded on both occasions. Three other birds from the flock that died were studied histopathologically. RESULTS: Scotopic responses were unmeasurable in either recording and therefore were not analyzed. Mean a- and b-wave amplitudes of the mixed rod-cone responses to standard (3 cd·s/m2 ) and high (10 cd·s/m2 ) intensity flashes were higher in the off-treatment group. No difference in implicit times was observed. Sex, age, and number of previous chloroquine treatments did not affect ERG responses. Histopathology revealed Plasmodium spp.in the lungs, liver, and brain, but not in the eyes, of the necropsied birds, and there were no signs of retinitis or retinopathy. CONCLUSIONS: Daily chloroquine treatment was associated with attenuated ERG responses in penguins, which improve following cessation of treatment. Further work is warranted to establish a chloroquine dose that is efficacious yet has minimal adverse effects. Our results suggest that ERG responses of captive penguins undergoing ERG for any indication (such as prior to cataract surgery), must be evaluated in light of the birds' anti-malaria treatment status.

Does the order of intraocular pressure measurement affect tonometry results?

PURPOSE: To determine whether the order of intraocular pressure (IOP) measurement affects readings, regardless of which eye is measured first. METHODS: Intraocular pressure was measured in 31 and 41 dogs using applanation and rebound tonometry, respectively. Initially, IOP was measured in the first (randomly chosen) eye (reading A), followed by measurement in the fellow eye (reading B), and a third (repeated) measurement in the first eye (reading C). After 15 minutes, measurements were repeated in reverse order (readings D - F). RESULTS: Applanation tonometry revealed significant differences between readings A & B (15.6 ± 2.3 and 14.8 ± 2.7 mm Hg, respectively, p = .02), A & C (15.6 ± 2.3 and 14.5 ± 2.4 mm Hg, respectively, p = .002), D & E (14.5 ± 2.3 and 13.7 ± 2.1 mm Hg, respectively, p = .02), D & F (14.5 ± 2.3 and 13.9 ± 1.9 mm Hg, respectively, p = .05), and A & E (15.6 ± 2.3 and 13.7 ± 2.1 mm Hg, respectively, p = .001). Rebound tonometry yielded similar results, with additional differences between B & C (19.1 ± 3.0 and 18.2 ± 2.4 mm Hg, respectively, p = .002) and E & F (18.7 ± 3.3 and 18.2 ± 3.3 mm Hg, respectively, p = .02). CONCLUSIONS: Intraocular pressure measured in the first eye, whether right or left, is higher than in the fellow eye. Repeated tonometry in the same visit could result in a significant IOP decrease, though the magnitude may not be clinically appreciable.

Is it necessary to wait several minutes between applications of different topical ophthalmic solutions? A preliminary study with tropicamide eye drops in healthy dogs.

OBJECTIVE: To evaluate the efficacy of topical tropicamide when placed at different time intervals before or after a saline drop. ANIMALS STUDIED: Eight healthy Labrador and golden retriever dogs. PROCEDURES: The effect of 1% tropicamide on pupillary diameter (PD) was measured over 240 min when administered alone (control) and then 1 and 5 min prior to, or following, application of a saline drop, with 1-week washout between each of the five trials. Data were analyzed using repeated-measures ANOVA and Tukey post hoc test. RESULTS: Only 6/110 pairwise comparisons among the 5 trials were statistically significant (p ≤ .035), with post-hoc analysis showing no significant differences (p ≥ .14) between the overall means of all trials. In all five trials, maximal PD was reached 30 min after tropicamide application and maintained until 210 min for 180 min (p = .0005). CONCLUSIONS: Our results suggest that waiting 1 min between applications of different ophthalmic solutions may be sufficient for maximal drug effect. Care should be taken when extrapolating these results to other species and different ophthalmic formulations.

Atypical free-floating iridociliary pigmented epithelial cysts and secondary glaucoma in a caiman (Caiman latirostris).

We describe a case of chronic ocular trauma that resulted in fixed and free-floating, pigmented epithelial iridociliary cysts, inflammation, and secondary glaucoma in a caiman (Caiman latirostris). A 20- to 25-year-old male caiman was presented with phthisis bulbi in the right eye, and congested episcleral vessels, corneal leukoma, disorganized anterior chamber, multifocal anterior synechia, and elevated intraocular pressure in the left eye. Ocular ultrasound of the left eye revealed round structures dispersed in the anterior and posterior chambers and vitreous cavity. Bilateral enucleation was performed, and gross pathology of the left eye revealed multiple pigmented cysts attached to the iris and posterior corneal surface causing marked distortion of the anterior uvea, and free-floating in the vitreous cavity. Histopathology demonstrated heavily pigmented cystic structures of iridociliary epithelium origin carpeting the anterior segment surfaces and causing obstruction of the iridocorneal angles, leading to secondary glaucoma. To the authors' knowledge, this is the first report of iridociliary cysts in wildlife species.

Atypical ocular Chelonoid herpesvirus manifestations in a captive Loggerhead turtle (Caretta caretta).

A captive loggerhead turtle (Caretta caretta) of unknown sex, 3 years of age, presented with bilateral mucoid secretions, severe chemosis, conjunctival hyperemia, and globe retraction. The animal was evaluated ophthalmologically and systemically, and hematological, microbiological, and conjunctival cytological and biopsy samples were collected for complementary diagnosis. The histopathological examination showed amphophilic intranuclear inclusions associated with severe inflammatory infiltrate. The diagnosis of Chelonid alphaherpesvirus 5 (ChAHV 5) was confirmed with end point PCR. Following systemic treatment with L-lysine, acyclovir and vitamin A, the ocular signs resolved. No amphophilic intranuclear inclusions were seen in a follow-up biopsy 5 months later, and there has been no recurrence of clinical ophthalmic signs during a 4-year follow-up. It is suggested that ChAHV 5 be considered as a differential diagnosis in captive marine turtles that present for conjunctival disease other than fibropapillomatosis.

Ocular anatomy and correlation with histopathologic findings in two common octopuses (Octopus vulgaris) and one giant Pacific octopus (Enteroctopus dofleini) diagnosed with inflammatory phakitis and retinitis.

PURPOSE: Review octopus ocular anatomy and describe the histopathologic findings in three octopuses diagnosed with phakitis and retinitis. ANIMALS: Two common octopuses (Octopus vulgaris) and one giant Pacific octopus (Enteroctopus dofleini) with a history of ophthalmic disease. METHODS: A literature search was performed for the ocular anatomy section. Both eyes from all three octopuses, and two control eyes, were submitted for histopathologic evaluation. Hematoxylin and eosin stain was used for standard histopathologic evaluation; GMS stain was used to screen for fungi, gram stain for bacteria; and Fite's acid fast stain for acid fast bacteria. RESULTS: Anatomically, the anterior chamber of the octopus has direct contact with ambient water due to an opening in the dorsal aspect of a pseudocornea. The octopus lens is divided into anterior and posterior segments. The anterior half is exposed to the environment through the opening into the anterior chamber. Neither part of the lens has a lens capsule. The retina is everted, unlike the inverted vertebrate retina, and consists of just two layers. Histopathology revealed inflammatory phakitis and retinitis of varying severity in all six eyes of the study animals. No intraocular infectious organisms were recognized but one common octopus eye had clusters of coccidian parasites, identified as Aggregata sp., in extraocular tissues and blood vessels. CONCLUSION: We describe inflammatory phakitis and retinitis in two species of octopuses. The underlying cause for the severe intraocular response may be direct intraocular infection, water quality, an ocular manifestation of a systemic disease, or natural senescence.

Baseline retinal OCT measurements in normal female beagles: The effects of eccentricity, meridian, and age on retinal layer thickness.

OBJECTIVE: Our aim was to generate baseline optical coherence tomography (OCT) measurements of retinal thickness in female Beagles and to determine how these are affected by meridian, eccentricity, and age. METHODS: Twenty-three female Beagles, including six puppies (<6 months old), six mature (1.8-8.2 years old), and 11 elderly dogs (>11 years old) were studied. Both retinas of each dog were scanned (in 4 principal meridians) using the Heidelberg Spectralis following ophthalmic examination, refraction, and sedation. In each eye and each meridian, total retinal, outer retinal, and nerve fiber layer (NFL) thickness were measured from the disc rim up to 6 mm peripherally. RESULTS: The canine retina is thickest dorsally and thinnest ventrally. Total retinal, outer retinal, and NFL thickness decrease progressively and significantly as a function of eccentricity. The greatest eccentricity-dependent thinning occurs dorsally. This thinning is due mostly to NFL tapering, while the eccentricity-dependent change in outer retinal thickness is more moderate, especially in the lateral meridian, possibly due to the presence of the visual streak. The retina is thickest in puppies, but there were no significant differences between mature and elderly dogs. CONCLUSIONS: Our results provide normative values for total, outer, and inner retinal thickness in female dogs and may facilitate OCT use in the diagnosis of canine glaucoma and inherited retinopathies.

Prognostic indicators and outcome in dogs undergoing temporary tarsorrhaphy following traumatic proptosis.

PURPOSE: Retrospectively evaluate prognostic indicators and outcome in dogs undergoing temporary tarsorrhaphy following traumatic proptosis. METHODS: Medical records (2004-2017) were reviewed for signalment, cause and duration of proptosis, and clinical findings on admission. The operating faculty member, post-operative medications, and ophthalmic findings at last recheck were recorded. Owners of dogs with blinded eyes were surveyed regarding final outcome and satisfaction. Data were analyzed using Fisher's exact, Likelihood ratio, and Mann-Whitney tests. RESULTS: At last recheck, 17.5 ± 7.3 (mean ± SD) days post-operatively, vision was present in 12/43 eyes (28%). Vision at last recheck was not correlated with breed, cause or duration of proptosis, or post-operative medications (P > .05), but was correlated with presence of direct and indirect pupillary light reflexes (PLRs) on admission (P = .001 and .02, respectively), and with assessment and surgery performed by veterinary ophthalmologists rather than surgery or emergency personnel (P = .015). Mean satisfaction scores (graded 0-10) of 22 owners contacted 59.6 ± 48.2 months after last recheck were 7.8 ± 2.6 (n = 6), 5.7 ± 4.9 (n = 3), and 8.8 ± 1.3 (n = 13) for dogs with phthisical eyes, eyes requiring long-term medications, and blind but otherwise normal eyes, respectively (P = .284). CONCLUSIONS: Contrary to previous studies, breed, and cause and duration of proptosis, did not significantly affect outcome. Testing of direct and indirect PLR is a simple and significant prognostic indicator. Patients assessed and operated by a veterinary ophthalmologist have better prognosis.

Sip1 regulates the generation of the inner nuclear layer retinal cell lineages in mammals.

The transcription factor Sip1 (Zeb2) plays multiple roles during CNS development from early acquisition of neural fate to cortical neurogenesis and gliogenesis. In humans, SIP1 (ZEB2) haploinsufficiency leads to Mowat-Wilson syndrome, a complex congenital anomaly including intellectual disability, epilepsy and Hirschsprung disease. Here we uncover the role of Sip1 in retinogenesis. Somatic deletion of Sip1 from mouse retinal progenitors primarily affects the generation of inner nuclear layer cell types, resulting in complete loss of horizontal cells and reduced numbers of amacrine and bipolar cells, while the number of Muller glia is increased. Molecular analysis places Sip1 downstream of the eye field transcription factor Pax6 and upstream of Ptf1a in the gene network required for generating the horizontal and amacrine lineages. Intriguingly, characterization of differentiation dynamics reveals that Sip1 has a role in promoting the timely differentiation of retinal interneurons, assuring generation of the proper number of the diverse neuronal and glial cell subtypes that constitute the functional retina in mammals.

Contrary to popular belief, chinchillas do not have a pure rod retina.

PURPOSE: To establish whether there is cone contribution to retinal function and structure in chinchillas (Chinchilla lanigera), in view of the prevailing notion that this species possesses a pure rod retina. METHODS: Photopic electroretinography (ERG) responses to high-intensity flashes (10 and 25 cd*s/m2 ) were recorded unilaterally in six pigmented chinchillas following 10 minutes of light adaptation (30 cd/m2 ). Retinas of two animals were studied histologically, and immunohistochemistry (IHC) was conducted to detect the presence of short and medium/long wavelength cone photoreceptors. RESULTS: ERG recordings revealed photopic responses, albeit of low amplitudes. Histopathology demonstrated presumptive cone inner segments in the photoreceptor layer. Presence of cone photoreceptors was confirmed by IHC. Cone density was higher in the central retina, and red/green cones outnumbered blue cones. CONCLUSIONS: Our results provide convincing evidence for the presence of functioning cone photoreceptors in the chinchilla retina, disproving the established belief that the species has a pure rod retina.

Can an in vivo imaging system be used to determine localization and biodistribution of AAV5-mediated gene expression following subretinal and intravitreal delivery in mice?

Recombinant adeno associated viruses (AAV) are the most commonly used vectors in animal model studies of gene therapy for retinal diseases. The ability of a vector to localize and remain in the target tissue, and in this manner to avoid off-target effects beyond the site of delivery, is critical to the efficacy and safety of the treatment. The in vivo imaging system (IVIS) is a non-invasive imaging tool used for detection and quantification of bioluminescence activity in rodents. Our aim was to investigate whether IVIS can detect localization and biodistribution of AAV5 vector in mice following subretinal (SR) and intravitreal (IVT) injections. AAV5 carrying firefly luciferase DNA under control of the ubiquitous cytomegalovirus (CMV) promoter was injected unilaterally IVT or SR (in the central or peripheral retina) of forty-one mice. Luciferase activity was tracked for up to 60 weeks in the longest surviving animals, using repeated (up to 12 times) IVIS bioluminescence imaging. Luciferase presence was also confirmed immunohistochemically (IHC) and by PCR in representative animals. In the SR group, IVIS readings demonstrated luciferase activity in all (32/32) eyes, and luciferase presence was confirmed by IHC (4/4 eyes) and PCR (12/12 eyes). In the IVT group, IVIS readings demonstrated luciferase activity in 7/9 eyes, and luciferase presence was confirmed by PCR in 5/5 eyes and by IHC (2/2 eyes). In two SR-injected animals (one each from the central and peripheral injection sites), PCR detected luciferase presence in the ipsilateral optic nerves, a finding that was not detected by IVIS or IHC. Our results show that when evaluating SR delivery, IVIS has a sensitivity and specificity of 100% compared with the gold standard PCR. When evaluating IVT delivery, IVIS has a sensitivity of 78% and specificity of 100%. These finding confirm the ability of IVIS to detect in-vivo localized expression of AAV following SR delivery in the retina up to 60 weeks post-treatment, using repeated imaging for longitudinal evaluation, without fading of the biological signal, thereby replacing the need for post mortem processing in order to confirm vector expression. However, IVIS is probably not sensitive enough, compared with genome detection, to demonstrate biodistribution to the optic nerve, as it could not detect luciferase activity in ipsilateral optic nerves following SR delivery in mice.