Novel Methodology for Creating Macaque Retinas with Sortable Photoreceptors and Ganglion Cells

Purpose: The ability to generate macaque retinas with sortable cell populations would be of great benefit to both basic and translational studies of the primate retina. The purpose of our study was therefore to develop methods to achieve this goal by selectively labeling, in life, photoreceptors (PRs) and retinal ganglion cells (RGCs) with separate fluorescent markers. Methods: Labeling of macaque (Macaca fascicularis) PRs and RGCs was accomplished by subretinal delivery of AAV5-hGRK1-GFP, and retrograde transport of micro-ruby™ from the lateral geniculate nucleus, respectively. Retinas were anatomically separated into different regions. Dissociation conditions were optimized, and cells from each region underwent fluorescent activated cell sorting (FACS). Expression of retinal cell type- specific genes was assessed by quantitative real-time PCR to characterize isolated cell populations. Results: We show that macaque PRs and RGCs can be simultaneously labeled in-life and enriched populations isolated by FACS. Recovery from different retinal regions indicated efficient isolation/enrichment for PRs and RGCs, with the macula being particularly amendable to this technique. Conclusions: The methods and materials presented here allow for the identification of novel reagents designed to target retinal ganglion cells and/or photoreceptors in a species that is phylogenetically and anatomically similar to human. These techniques will enable screening of intravitreally- delivered AAV capsid libraries for variants with increased tropism for PRs and/or RGCs and the evaluation of vector tropism and/or cellular promoter activity of gene therapy vectors in a clinically relevant species

Evolving Techniques in the Treatment of Macular Detachment Caused by Optic

Fourteen patients with macular detachments caused by optic nerve pits were treated for progressive visual loss, cystoid macular changes , or atrophy of the macular retinal pigment epithelium. Photocoagulation of the temporal disc border alone was unsuccessful in two cases but promptly reat­ tached the macula of two patients immobilized after laser surgery. Vitrectomy and gas tamponade improved vision and flattened the macula of three patients over various periods. The detachment recurred in one patient. Prompt and sustained macular reattachment with improved vision was noted after photoco­ agulation, vitrectomy, and gas tamponade in eight patients, although four re­ quired second operations. The prompt reattachment and visual recovery noted in these eight patients surpasses the reported 25% rate of spontaneous resolu­ tion. (Key words : cystoid degeneration, gas, laser, macula, macular hole, optic nerve, optic nerve pit, photocoagulation, retinal detachment, retinal pigment epithelial atrophy, retinoschisis, tamponade.) Ophthalmology 95:889-896. 198

Evolving Techniques in the Treatment of Macular Detachment Caused by Optic Nerve Pits

Fourteen patients with macular detachments caused by optic nerve pits were treated for progressive visual loss, cystoid macular changes, or atrophy of the macular retinal pigment epithelium. Photocoagulation of the temporal disc border alone was unsuccessful in two cases but promptly reattached the macula of two patients immobilized after laser surgery. Vitrectomy and gas tamponade improved vision and flattened the macula of three patients over various periods. The detachment recurred in one patient. Prompt and sustained macular reattachment with improved vision was noted after photocoagulation, vitrectomy, and gas tamponade in eight patients, although four required second operations. The prompt reattachment and visual recovery noted in these eight patients surpasses the reported 25% rate of spontaneous resolution

Clinical and Epidemiologic Research Associations Between Abnormal Rod-Mediated Dark Adaptation and Health and Functioning in Older Adults With Normal Macular Health

PURPOSE. Delayed rod-mediated dark adaptation (DA) is characteristic of early age-related macular degeneration (AMD) and also can be observed in some older adults in normal macular health. We examine cross-sectional associations between rod-mediated DA and risk factors for AMD in older adults in normal macular health. METHODS. The sample consisted of adults aged ‡60 years old in normal macular health per grading of fundus photos using an established disease classification system. Rod-mediated DA was measured psychophysically following a photobleach using a computer-automated dark adaptometer with targets centered at 58 on the inferior vertical meridian. The speed of DA was characterized by the rod-intercept value, with abnormal DA defined as rod-intercept ‡ 12.3 minutes. We assessed several health and functional characteristics that the literature has suggested increase AMD risk (e.g., smoking, alcohol use, inflammatory markers, apolipoproteins, low luminance visual acuity, chronic medical conditions, body mass, family history). RESULTS. Among 381 participants (mean age, 68.5 years; SD, 5.5), 78% had normal and 22% had abnormal DA, with the prevalence of abnormal DA increasing with age. After ageadjustment, abnormal DA was associated with increased odds of elevated C-reactive protein (CRP), heavy use of or abstention from alcohol, high blood pressure, and drop in visual acuity under mesopic conditions. CONCLUSIONS. Despite having normal macular health according to accepted definitions of AMD presence, approximately one-quarter of older adults recruited from primary eye care clinics had abnormal DA, which was associated with known risk factors for AMD, including elevated CRP. Keywords: aging, dark adaptation, rod function, age-related macular degeneration S cotopic dysfunction is characteristic of early age-related macular degeneration (AMD) even though visual acuity remains relatively unimpaired. 10-12 Persons with early AMD tend to exhibit deficits in rod-mediated light sensitivity that are more severe than conemediated deficits measured in the same retinal areas

Multicenter trial of Cryotherapy for Retinopathy of Prematurity: Ophthalmological outcomes at 10 years

Objective: To evaluate outcomes at 10 years after randomization for eyes undergoing cryotherapy vs eyes serving as controls, for patients enrolled in the Multicenter Trial of Cryotherapy for Retinopathy of Prematurity (CRYO-ROP). Methods: The randomized cohort originally consisted of 291 preterm children with birth weights less than 1251 g who developed a defined threshold of ROP severity in one or both eyes. Patients with bilateral threshold ROP (n = 240) were randomly assigned to receive cryotherapy to one eye and no cryotherapy to the other eye. Those with ROP of less severity than threshold in the fellow eye ( asymmetric ; n = 51) were randomly assigned to cryotherapy or no cryotherapy in the eye with threshold ROP. Ten years later, a tester who was masked to treatment status of each eye measured distance and near visual acuity, with unfavorable outcome being 20/200 or worse. Patients also were evaluated by study-certified ophthalmologists who assessed ROP residua primarily in the posterior pole of the fundus, with unfavorable outcome being a posterior retinal fold or worse. Results: For the 247 children examined, both functional and structural primary outcomes showed fewer unfavorable outcomes in treated vs control eyes: 44.4% vs 62.1% (P\u3c.001) for distance visual acuity and 27.2% vs 47.9% (P\u3c.001) for fundus status. Near acuity results were similar to those for distance (42.5% vs 61.6%; P\u3c.001). Total retinal detachments had continued to occur in control eyes, increasing from 38.6% at 5 1/2 years to 41.4% at 10 years, while treated eyes remained stable (at 22.0%). A previously disturbing subgroup trend that more control eyes than treated eyes had visual acuity of 20/40 or better (in the 5 1/2-year report) was no longer present at 10 years; eyes that received cryotherapy were found at least as likely as control eyes to have 20/40 or better visual acuity. Conclusions: At 10 years, eyes that had received cryotherapy were much less likely than control eyes to be blind. A previous trend for a higher proportion of sighted control eyes than sighted treated eyes to show acuity in the normal range was not confirmed. The results show long-term value from cryotherapy in preserving visual acuity in eyes with threshold ROP