What Is Your Diagnosis?

In collaboration with the American College of Veterinary Radiology.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Endogenous Opioid Signaling in the Mouse Retina Modulates Pupillary Light Reflex.

Opioid peptides and their receptors are expressed in the mammalian retina; however, little is known about how they might affect visual processing. The melanopsin-expressing intrinsically photosensitive retinal ganglion cells (ipRGCs), which mediate important non-image-forming visual processes such as the pupillary light reflex (PLR), express ß-endorphin-preferring, µ-opioid receptors (MORs). The objective of the present study was to elucidate if opioids, endogenous or exogenous, modulate pupillary light reflex (PLR) via MORs expressed by ipRGCs. MOR-selective agonist [D-Ala2, MePhe4, Gly-ol5]-enkephalin (DAMGO) or antagonist D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP) was administered via intravitreal injection. PLR was recorded in response to light stimuli of various intensities. DAMGO eliminated PLR evoked by light with intensities below melanopsin activation threshold but not that evoked by bright blue irradiance that activated melanopsin signaling, although in the latter case, DAMGO markedly slowed pupil constriction. CTAP or genetic ablation of MORs in ipRGCs slightly enhanced dim-light-evoked PLR but not that evoked by a bright blue stimulus. Our results suggest that endogenous opioid signaling in the retina contributes to the regulation of PLR. The slowing of bright light-evoked PLR by DAMGO is consistent with the observation that systemically applied opioids accumulate in the vitreous and that patients receiving chronic opioid treatment have slow PLR.

Unfolding the diagnosis of subspectacular fluid opacity in a corn snake (Pantherophis guttatus).

OBJECTIVE: To present the results of clinical, surgical, and histopathologic procedures and how these were compared with the initial presumptive clinical diagnosis in a corn snake (Pantherophis guttatus) presenting with subspectacular fluid opacity; and to improve upon currently established surgical enucleation techniques in the snake. ANIMAL STUDIED: An 8-month-old corn snake was presented for enlarged globe OD. PROCEDURES: The following diagnostics were performed: systemic and ophthalmic examinations, complete blood count, cytology and culture of subspectacular fluid, and histopathology of enucleated globe and spectacle. Enucleation was performed in a routine fashion with the addition of a porcine small intestinal submucosa bioscaffold graft (SISplus™; Avalon Medical, Stillwater, MN), sutured over the orbit. RESULTS: Systemic examination revealed signs of maxillary stomatitis. Ophthalmic examination revealed semitransparent fluid in the subspectacular space. Complete blood count was unremarkable. Cytology of fluid obtained via subspectacular centesis was acellular, and culture grew Clostridium perfringens, which was consistent with the clinical suspicion of right maxillary stomatitis. Histopathology of the enucleated globe revealed spectaculitis, characterized by regional heterophilic inflammation, and no evidence of lymph dissection in the (peri)ocular tissues. The final diagnosis was a subspectacular abscess. Follow-up revealed that the SIS graft provided excellent healing and cosmesis of the surgical site. CONCLUSIONS: While there are reports of lymphatic fluid dissection between skin layers during ecdysis, which can result in an opaque spectacle, the fluid opacity in this case was attributed to a subspectacular abscess secondary to an ascending oral infection. Addition of biological wound dressing may contribute to positive post-enucleation outcome in the snake.

Iridocorneal angle assessment of companion rabbits using gonioscopy, spectral-domain optical coherence tomography (Optovue iVue® ), high-resolution ultrasound, and Pentacam® HR imaging.

OBJECTIVE: Iridocorneal angle (ICA) narrowing is a known risk factor for primary glaucoma in multiple species, but has not been described in companion rabbits. This study aimed to develop an ICA grading scheme for companion rabbits to enable early glaucoma predisposition diagnosis. ANIMALS STUDIED: Twenty healthy rabbits of varying breeds and ages. PROCEDURES: Rabbits received complete ophthalmic examinations, including gonioscopy, and imaging of the ICA using spectral-domain optical coherence tomography (SD-OCT), Scheimpflug imaging (Pentacam® HR), and high-resolution ultrasound (HRUS). Angle opening distance (AOD) and angle recess area (ARA) of the ICA were measured and assessed for agreement using a Bland-Altman analysis. A five-stage gonioscopy grading scheme was created, and Spearman-rank test assessed for correlation between gonioscopy grades and ICA measurements. Differences among age and sex were analyzed with a nonparametric ANOVA and Wilcoxon rank-sum test, respectively. RESULTS: Analysis revealed AOD medians of 0.28mm for SD-OCT [95% CI: 0.24-0.31], 0.20mm for Pentacam® HR [95% CI: 0.18-0.21], and 0.25mm for HRUS [95% CI: 0.22-0.28]. The median ARA was 0.14mm2 for SD-OCT [95% CI: 0.117-0.163], 0.09mm2 for Pentacam® HR [95% CI: 0.082-0.100], and 0.06mm2 for HRUS [95% CI: 0.046-0.054]. The association between gonioscopy grade and SD-OCT ARA was significant (P < 0.05), and there was a significant difference (P < 0.001) between imaging modalities for both ARA and AOD. CONCLUSIONS: Gonioscopy grade correlated well with SD-OCT ARA. Therefore, SD-OCT is recommended as a noncontact method for evaluating companion rabbit ICA. Each imaging device should not be used interchangeably for ICA evaluation.

µ-Opioid Receptor Activation Directly Modulates Intrinsically Photosensitive Retinal Ganglion Cells.

Intrinsically photosensitive retinal ganglion cells (ipRGCs) encode light intensity and trigger reflexive responses to changes in environmental illumination. In addition to functioning as photoreceptors, ipRGCs are post-synaptic neurons in the inner retina, and there is increasing evidence that their output can be influenced by retinal neuromodulators. Here we show that opioids can modulate light-evoked ipRGC signaling, and we demonstrate that the M1, M2 and M3 types of ipRGCs are immunoreactive for µ-opioid receptors (MORs) in both mouse and rat. In the rat retina, application of the MOR-selective agonist DAMGO attenuated light-evoked firing ipRGCs in a dose-dependent manner (IC50 < 40 nM), and this effect was reversed or prevented by co-application of the MOR-selective antagonists CTOP or CTAP. Recordings from solitary ipRGCs, enzymatically dissociated from retinas obtained from melanopsin-driven fluorescent reporter mice, confirmed that DAMGO exerts its effect directly through MORs expressed by ipRGCs. Reduced ipRGC excitability occurred via modulation of voltage-gated potassium and calcium currents. These findings suggest a potential new role for endogenous opioids in the mammalian retina and identify a novel site of action-MORs on ipRGCs-through which opioids might exert effects on reflexive responses to environmental light.

Surgically induced astigmatism in canines following sutured dorsonasal vs dorsotemporal clear corneal incisions.

OBJECTIVES: To investigate use of the Pentacam® HR for evaluation of surgically induced corneal astigmatism (SIA) in canines undergoing bilateral phacoemulsification and determine differences between dorsonasal and dorsotemporal clear corneal incisions. ANIMALS: Client-owned canines undergoing bilateral phacoemulsification. PROCEDURES: Patients received anterior segment imaging pre-operatively, immediately post-operatively, and 2-4 months post-operatively (follow-up). Total corneal refractive power was used to determine SIA. Surgically induced astigmatism was compared between right and left eyes, representing dorsotemporal and dorsonasal incisions, respectively. Repeated measures analyses were used between time points and paired t test compared SIA between eyes. RESULTS: Complete imaging series were obtained for seven patients. Follow-up imaging occurred at a median of 112 days (range 60-132 days) post-operatively. For repeated measures analyses, significant differences were found between pre- and immediate post-operative values (P < 0.01), and between immediate post-operative and follow-up values (P < 0.01). There was no significant difference between pre-operative and follow-up values. Surgically induced astigmatism was significantly different between right and left eyes, with values of 2.01 ± 1.24 D and 3.05 ± 1.58 D at 3 mm radius (P < 0.05), and 2.04 ± 1.18 D and 3.06 ± 1.27 D at 4 mm radius (P < 0.05) for dorsotemporal and dorsonasal incisions, respectively. CONCLUSIONS: Preliminary investigation revealed improvement of corneal SIA 2-4 months post-operatively, but development of significantly more SIA in dorsonasal vs dorsotemporal incisions. This prompts consideration of patient or microscope rotation to create a more dorsotemporal incision when possible.

Anterior segment parameters in normal dogs using the Pentacam® HR Scheimpflug system.

OBJECTIVE: To describe and compare normative anterior segment parameters between canine age groups using the Pentacam® HR Scheimpflug camera (Pentacam). ANIMALS STUDIED: Thirty-six sedated dogs (60 eyes) of varying ages and breeds were imaged with the Pentacam; only nondiseased anterior segments were included. PROCEDURES: Dogs were divided into three age groups: Group 1 (1-5 years), Group 2 (6-10 years), and Group 3 (11-15 years). Values assessed included central corneal thickness (CCT), anterior and posterior corneal elevation (ACE/PCE), anterior and posterior corneal curvature metrics, corneal volume (CV), anterior and posterior corneal astigmatism (AA/PA), anterior chamber depth (ACD), anterior chamber volume (ACV), and anterior chamber angle (ACA). Tukey-adjusted pairwise comparisons were performed. RESULTS: Overall CCT (mean ± SD) was 631.07 ± 59.91 µm. Central corneal thickness was 608.60 ± 48.63 µm for Group 1, 648.57 ± 51.06 µm for Group 2, and 635.37 ± 73.71 µm for Group 3. Anterior corneal elevation (ACE) measured 9.08 ± 0.58 mm, PCE measured 8.04 ± 0.50 mm, and CV was 58.13 ± 5.39 mm3 . Astigmatism values were 1.34 ± 0.94 D for AA and 0.46 ± 0.44 D for PA. Anterior chamber values were 3.76 ± 0.56 mm for ACD, 383.68 ± 66.24 mm3 for ACV, and 23.62 ± 29.33˚ for ACA. Significant differences were found between Groups 1 and 2 for CV (55.08 ± 4.08 mm3 and 60.32 ± 4.19 mm3 , respectively), (P = 0.02). CONCLUSIONS: Corneal volume significantly increased between Group 1 and Group 2. Central corneal thickness increased from Group 1 to Group 3, but was not significant with the current sample size. There were no other differences between age groups.

Canine central corneal thickness measurements via Pentacam-HR® , optical coherence tomography (Optovue iVue® ), and high-resolution ultrasound biomicroscopy.

OBJECTIVE: To compare central corneal thickness (CCT) values in canine eyes using Pentacam-HR® Scheimpflug imaging (Pentacam), Optovue® iVue spectral-domain optical coherence tomography (SD-OCT), and high-resolution ultrasound biomicroscopy (UBM) and generate normative canine Pentacam CCT values. ANIMALS STUDIED: Twenty-four client-owned dogs (37 eyes) with nondiseased cornea(s) presenting to the Colorado State University Veterinary Teaching Hospital. PROCEDURES: Corneal images were acquired via Pentacam, SD-OCT, and UBM in the listed order. Machine-calculated values of CCT from Pentacam and SD-OCT were compared to operator-measured values from UBM. Bland-Altman analysis was performed to evaluate agreement between instruments. RESULTS: Mean CCT ± SD measured by Pentacam was 629.73 ± 64.57 µm, by SD-OCT was 610.56 ± 57.48 µm, and by UBM was 689.77 ± 55.93 µm. On average, Pentacam CCT was 19.17 ± 32.90 µm (3%) thicker than SD-OCT and 65.12 ± 44.52 µm (10.3%) thinner than UBM. SD-OCT was on average 82.47 µm (13.5% ) thinner than UBM. The 95% limits of agreement were (-45.31, 83.65), (-152.38, 22.13), and (-126.674, -38.270) for Pentacam vs. SD-OCT, Pentacam vs. UBM, and SD-OCT vs. UBM respectively. All differences were statistically significant (P < 0.01). CONCLUSIONS: Considering there is an average of 7.5% normal diurnal variation in canine CCT, a 3.0% difference between Pentacam and SD-OCT values is likely not clinically relevant. However, Pentacam measurements were both statistically and clinically significantly different from UBM and SD-OCT measurements.

Comparison between Pentacam-HR and optical coherence tomographycentral corneal thickness measurements in healthy feline eyes.

OBJECTIVES: To measure the central corneal thickness (CCT) in healthy feline eyes with Scheimpflug imaging (Pentacam, Pentacam(®) -HR) and to compare these values with those obtained with spectral-domain optical coherence tomography (OCT, Optovue(®) iVue). ANIMALS STUDIED: Thirty one sedated Domestic Short-haired cats. PROCEDURES: Two repeated CCT measurements were obtained from both eyes using Pentacam measured at the pupil center and corneal apex (CCTpupil and CCTapex ) and using SD-OCT (CCTOCT ). Agreement between the imaging modalities for CCT and intradevice repeatability was evaluated with Bland-Altman analysis. Mixed modeling was used to test for a difference between methods. RESULTS: The calculated mean ± SD CCT was CCTOCT  = 584.93 ± 39.05 µm, CCTpupil  = 608.25 ± 47.26 µm, and CCTapex  = 606.41 ± 44.18 µm. There was a statistically significant difference between CCTOCT vs. CCTpupil (P < 0.0007) and CCTOCT vs. CCTapex (P < 0.0003) (n = 59 eyes). The 95% limits of agreement (LoA) for CCTOCT vs. CCTpupil was (-89.57 µm, 40.04 µm) and for CCTOCT vs. CCTapex was (-79.62 µm, 33.79 µm) (n = 59 eyes). 95% LoA between repeated CCT measurements by SD-OCT was (-10.23 µm, 9.32 µm) and by Pentacam was (-22.66 µm, 18.30 µm) at pupil and (-15.81 µm, 16.17 µm) at corneal apex (n = 11 eyes). CONCLUSIONS: SD-OCT and Pentacam provide excellent CCT measurement repeatability. Pentacamapex is our recommended clinical setting for use of the Pentacam. The level of agreement between SD-OCT and Pentacam for CCT is good.