Cystic macular oedema on spectral-domain optical coherence tomography in choroideremia patients without cystic changes on fundus examination.

PURPOSE: To determine the prevalence of cystic macular oedema (CME) in patients with choroideremia (CHM) by using spectral-domain optical coherence tomography (SD-OCT). METHODS: A total 16 patients affected with CHM were enrolled in the study. All patients underwent a complete eye examination. SD-OCT was performed using an OPKO spectral-domain OCT/SLO instrument. RESULTS: The average age of the study patients was 44.0 ± 16.0 years (range, 13-63 years). Out of the 16 patients with CHM, 10 patients (62.5%) showed a degree of CME on SD-OCT testing in at least one eye, and 8 patients (50%) showed CME in both eyes. CONCLUSIONS: Because of its notable prevalence, it would seem prudent to screen CHM patients by SD-OCT for the possible presence of CME and to identify those amenable to future treatment strategies for their macular oedema.

Clinical value, normative retinal sensitivity values, and intrasession repeatability using a combined spectral domain optical coherence tomography/scanning laser ophthalmoscope microperimeter.

PURPOSE: To establish normative values for macular light sensitivity and to determine the intrasession fluctuation of perimetric responses using the OPKO/OTI microperimeter. METHODS: A total of 32 visually normal subjects participated in the study. A standardized grid pattern was used for testing, which consisted of 28 points arranged concentrically in three circles that occupied an area of 11° (in diameter) within the central macula. Each subject participated in at least two tests. Parameters evaluated included: overall mean macular sensitivity for test 1 and 2, overall difference in mean macular sensitivity between tests, and the mean sensitivity for each circle. The relationship between sensitivity and age was also examined. RESULTS: The overall median sensitivity for test 1 was 16.8 decibels (dB) and for test 2 was 16.9 dB. The median sensitivities for test 1 and test 2 were not significantly different (P = 0.72). The mean intrasession sensitivity difference was 0.13 dB. The variability of the sensitivity difference between tests decreased as mean sensitivity increased. The sensitivity values averaged across the two tests for inner, middle, and outer circles ranged from 14.3 to 18.8 dB (median value of 16.9 dB), 13.8-18.3 dB (median value of 17.2 dB), and 11.3-18.3 dB (median value of 16.6 dB), respectively. Linear regression analysis showed a 0.5 dB sensitivity loss for each decade of life. CONCLUSION: We documented a narrow range of intrasession fluctuation using the OPKO/OTI microperimeter. The establishment of normative sensitivity values will facilitate monitoring the loss of macular visual function in patients with retinal disease.

Selective thinning of the perifoveal inner retina as an early sign of hydroxychloroquine retinal toxicity.

PURPOSE: To evaluate macular thickness profiles using spectral-domain optical coherence tomography (SDOCT) and image segmentation in patients with chronic exposure to hydroxychloroquine. METHODS: This study included eight patients with chronic exposure to hydroxychloroquine (group 1) and eight controls (group 2). Group 1 patients had no clinically evident retinal toxicity. All subjects underwent SDOCT imaging of the macula. An image segmentation technique was used to measure thickness of six retinal layers at 200 microm intervals. A mixed-effects model was used for multivariate analysis. RESULTS: By measuring total retinal thickness either at the central macular (2800 microm in diameter), the perifoveal region 1200-microm-width ring surrounding the central macula), or the overall macular area (5200 microm in diameter), there were no significant differences in the thickness between groups 1 and 2. On an image segmentation analysis, selective thinning of the inner plexiform+ganglion cell layers (P=0.021) was observed only in the perifoveal area of the patients in group 1 compared with that of group 2 by using the mixed-effects model analysis. CONCLUSION: Our study results suggest that chronic exposure to hydroxychloroquine is associated with thinning of the perifoveal inner retinal layers, especially in the ganglion cell and inner plexiform layers, even in the absence of functional or structural clinical changes involving the photoreceptor or retinal pigment epithelial cell layers. This may be a contributing factor as the reason most patients who have early detectable signs of drug toxicity present with paracentral or pericentral scotomas.

Effects of chronic exposure to hydroxychloroquine or chloroquine on inner retinal structures.

PURPOSE: To evaluate peripapillary retinal nerve fibre layer (RNFL) thickness and macular inner and outer retinal thickness using spectral domain optical coherence tomography (Sd-OCT) in patients with chronic exposure to hydroxychloroquine or chloroquine. METHODS: Subjects were divided into three groups: Group I, four patients with hydroxychloroquine or chloroquine toxicity with abnormal fundus; Group II, eight patients with chronic exposure without fundus changes; and Group III, eight visually normal controls. Peripapillary RNFL thinning for an individual quadrant was based on measurements of less than the 5th percentile from at least two out of four segments in the quadrant. Macular scans on Groups I and II were carried out to compare the thickness of the inner, outer, and full-thickness retina to that of Group III. RESULTS: The mean ages in Groups I, II, and III were 57.6+/-8.0, 54.9+/-11.0 and 53.7+/-10.5 years, respectively (P=0.83). Median (range) duration of exposure was 7.5 (5-12) years in Group I, and was 10 (6-35) years in Group II. Seven (88%) of eight eyes in Group I showed peripapillary RNFL thinning in at least one quadrant, whereas none of Groups II and III did so. Using macular scans, Group I showed significant thinning of the inner, outer, and full-thickness retina compared to Group III (P<0.001). Group II had significant thinning only of the inner retina compared to Group III (P<0.001). CONCLUSIONS: OCT is useful to detect peripapillary RNFL thinning in clinically evident retinopathy, and selective thinning of the macular inner retina can be detected in the absence of clinically apparent fundus changes.

Predicting the pathogenicity of RPE65 mutations.

To assist in distinguishing disease-causing mutations from nonpathogenic polymorphisms, we developed an objective algorithm to calculate an "estimate of pathogenic probability" (EPP) based on the prevalence of a specific variation, its segregation within families, and its predicted effects on protein structure. Eleven missense variations in the RPE65 gene were evaluated in patients with Leber congenital amaurosis (LCA) using the EPP algorithm. The accuracy of the EPP algorithm was evaluated using a cell-culture assay of RPE65-isomerase activity The variations were engineered into plasmids containing a human RPE65 cDNA and the retinoid isomerase activity of each variant was determined in cultured cells. The EPP algorithm predicted eight substitution mutations to be disease-causing variants. The isomerase catalytic activities of these RPE65 variants were all less than 6% of wild-type. In contrast, the EPP algorithm predicted the other three substitutions to be non-disease-causing, with isomerase activities of 68%, 127%, and 110% of wild-type, respectively. We observed complete concordance between the predicted pathogenicities of missense variations in the RPE65 gene and retinoid isomerase activities measured in a functional assay. These results suggest that the EPP algorithm may be useful to evaluate the pathogenicity of missense variations in other disease genes where functional assays are not available.

Retinal nerve fibre layer thickness analysis in X-linked retinoschisis using Fourier-domain OCT.

PURPOSE: To evaluate the presence of retinal nerve fibre layer (RNFL) defects in patients with X-linked retinoschisis (XLRS) using high-speed, high-resolution, Fourier domain OCT (FD-OCT). METHODS: Twenty-four patients with XLRS seen by the authors were enrolled in the study. All patients underwent a complete eye examination. FD-OCT was performed using Optovue technology. A quadrant of the RNFL was considered to be thinned if at least two of the four segments in the quadrant were reduced in thickness. RESULTS: The average age of the 24 patients in the study was 28.8+/-14.7 years. Thinning of the RNFL in one quadrant was seen in 10 patients (41.7%), and thinning in two or more quadrants was seen in 8 patients (33.3%). Thinning in the inferior quadrant was most commonly seen and was observed in 12 patients (50%), followed by the temporal quadrant in 8 patients (33.3%), nasal quadrant in 4 patients (16.7%), and the superior quadrant in 4 patients (16.7%). CONCLUSIONS: Among our 24 patients with XLRS, 15 patients (62.5%) showed a thinning of the RNFL in one or more quadrants in at least one eye and 9 patients (37.5%) in both eyes. High-speed, high-resolution FD-OCT may be useful to determine the presence of possible changes in RNFL thickness in patients with XLRS. Reductions in RNFL thickness in such patients could be relevant in their selection for future therapeutic trials.

The prevalence of cystoid macular oedema on optical coherence tomography in retinitis pigmentosa patients without cystic changes on fundus examination.

PURPOSE: To determine the prevalence of cystoid macular oedema (CME) by optical coherence tomography (OCT) in retinitis pigmentosa (RP) patients with no evidence of cystic macular lesions on fundus examination. METHODS: We included 63 RP patients with no evidence of cystic-appearing macular changes on fundus examination. All patients underwent a complete ocular examination including best-corrected visual acuity using an ETDRS (Early Treatment Diabetic Retinopathy Study) chart, intraocular pressure measurement, anterior segment examination, and a detailed fundus examination. On 50 of the 63 patients, Fourier-domain OCT was performed using the radial slicer protocol. An additional 13 of the 63 patients were scanned using the macular thickness protocol on a time-domain OCT unit. The diagnosis of CME was defined by the presence of hyporeflective lacunae with well-defined boundaries on at least two of the scans. RESULTS: The mean age of patients included in the study was 36 years (range 9-71 years). Out of the 63 patients examined, 20 showed CME in at least one eye (32%), whereas 11 patients showed CME in both eyes (18%). CONCLUSIONS: Our findings demonstrate that a substantial number of RP patients with CME, as determined by OCT, may not show cystic changes by direct ophthalmoscopy or contact lens biomicroscopy. Knowledge of the high frequency for CME in such patients can serve to identify those who may be amenable to current or future treatment strategies of their macular oedema and can potentially impact on future therapeutic trials where visual acuity is used as an outcome measure.

Prevalence of cystic macular lesions in patients with Usher II syndrome.

PURPOSE: To evaluate the prevalence of cystic macular lesions in patients with Usher II syndrome. METHODS: All Usher type II patients seen in the inherited eye disease clinic at the University of Illinois at Chicago between January 2002 and December 2007 were included (n=76). Each participating patient underwent a detailed clinical examination, including best-corrected visual acuity, slit-lamp biomicroscopy and dilated fundus examination. The presence of cystoid lesions was determined by optical coherence tomography (OCT), fundus fluorescein angiogram (FFA), fundus photographs and/or clinical examination. RESULTS: A cystic-appearing macular change was observed in at least one eye in 19 out of the 76 patients (25%), 13 on the basis of OCT, five using FFA (two solely with the use of FFA and three based on clinical notes and FFA findings) and one based solely on clinical notes. Of the 18 patients with CME, determined by OCT or FFA, five (27.8%) showed either a funduscopically normal-appearing macula (n=4) or an atrophic appearing macular change (n=1). CONCLUSIONS: One-fourth of our total cohort of Usher II patients had cystic macular lesions. Moreover, a funduscopically normal-appearing macula was observed in 22% (n=4) of our 18 patients with cystic-appearing macular lesions on OCT and/or FFA testing. On the basis of the reasonably high prevalence of cystic macular lesions in our cohort, it would seem prudent to evaluate Usher II patients for the presence of cystoid macular oedema.

The prevalence of cystoid macular oedema in retinitis pigmentosa patients determined by optical coherence tomography.

AIMS: To determine the prevalence of cystoid macular oedema (CMO) in retinitis pigmentosa (RP) patients of various genetic subtypes using optical coherence tomography (OCT). METHODS: We performed a complete ocular examination on 124 RP patients including best corrected visual acuity, intraocular pressure measurement, anterior segment and a detailed fundus exam. OCT images were then acquired using two different units. The presence of hypo-reflective lacunae was used to diagnose CMO. RESULTS: Of the 124 patients, 47 showed CMO in at least one eye (38%), while 34 showed CMO in both eyes (27%). The prevalence of CMO in at least one eye for autosomal dominant (AD) patients was 52%, for autosomal recessive (AR) 39%, isolated 39%, Usher II 35% and none in the X linked recessive (XL) group. Using a chi-square analysis, no statistical significant difference was found for the prevalence of "bilateral CMO" (p = 0.60) or "CMO in at least one eye" (p = 0.59) among the AD, AR, isolated and Usher II genetic subtypes. CONCLUSION: Because of its notable prevalence, it would seem prudent to screen RP patients by OCT for the possible presence of CMO, to identify those amenable to treatment and also for future treatment trials when monitoring visual acuity.

Attainment of educational levels in patients with Leber's congenital amaurosis.

PURPOSE: To assess the educational level attained by patients legally blind with Leber's congenital amaurosis (LCA). DESIGN: Cross-sectional assessment. INTERVENTION: None. MAIN OUTCOME MEASURE: Highest educational level attained by 55 patients with LCA. RESULTS: A total of 55 patients with LCA were included in the study. Of the 55, 54 finished high school. In addition, 36 patients (65%) completed a college education and received a bachelor's degree, and 5 additional patients (9%) were recently accepted to college, whereas 3 others (5%) were currently attending college classes. Further, 18 patients were either pursuing (n = 3) or had attained (n = 15) an educational level beyond a bachelor's degree. CONCLUSIONS: Compromised visual function does not preclude the successful attainment of an academic education in patients with LCA who are substantially visually impaired from birth. These data have clinically relevant implications for the parents of children with LCA and for the patients themselves in providing a tone of optimism for their potential of attaining competitive academic achievements.

Interocular amplitude differences of the full field electroretinogram in normal subjects.

AIMS: To determine the interocular amplitude response difference of the electroretinogram (ERG) in normal subjects. METHODS: 79 subjects, without retinal changes of clinical significance, underwent ERG testing. They included 63 men and 16 women, with a mean age of 44 (SD 12) years and range of 18-65 years. Isolated rod, scotopic maximal, dark adapted 30 Hz flicker, photopic single flash, and light adapted 30 Hz flicker responses were recorded in both eyes following the International Society for Clinical Electrophysiology of Vision (ISCEV) standard protocol. The interocular percentage differences of the ERG b-wave amplitudes were calculated and presented as percentiles (25th, 50th, 75th, 95th), means (SD), and medians. RESULTS: The median interocular percentage differences in the b-wave amplitudes for the above ERG stimulus responses were 10%, 8%, 10%, 11%, and 10%, respectively. The mean interocular percentage differences were 11%, 11%, 12%, 13%, and 14%. The 95th percentiles for the interocular percentage differences were 28%, 27%, 36%, 33%, and 35%, respectively. CONCLUSIONS: The interocular percentage differences in the ERG b-wave amplitudes for five different stimulus responses were similar in our cohort of individuals without clinically significant retinal changes and ranged from a median of 8-11% and a 95th percentile of 27-36%. Our findings should be useful for determining sample sizes in future therapeutic trials on retinal diseases with monocular therapeutic strategies and may also have application for the more accurate detection of asymmetric retinal disease.

Retrospective, longitudinal, and cross sectional study of visual acuity impairment in choroideraemia.

BACKGROUND/AIMS: Few studies have reported on the change in visual acuity (VA) in patients with choroideraemia. In order to determine the degree and rate of VA impairment associated with this disease, the central VA was analysed in a large group of patients with choroideraemia. METHODS: The authors completed a retrospective, cross sectional review of 115 patients with choroideraemia from three tertiary care centres. A longitudinal analysis was performed on 45 of these patients who met the inclusion criteria of at least three visits over a minimum period of 4.5 years. Multiple linear regression analysis was used to explore the 5 year rate of VA change while controlling for initial VA and initial age. Multiple logistic regression was also used to investigate VA impairment. RESULTS: In the cross sectional group (n = 115), 84% (87/103) of patients under the age of 60 had a VA of 20/40 or better while 33% (4/12) of patients 60 years of age or older had a VA of 20/200 or worse at their most recent visit. The majority of the patients (93%) in the longitudinal subgroup of 45 patients had a VA of 20/30 or better at their initial visit. The mean 5 year rate of VA change was 0.09 logMAR equivalent (approximately one line on the Lighthouse chart). CONCLUSION: In this cohort of patients with choroideraemia, there was typically a slow rate of VA loss and the prognosis for central VA retention was, as a group, favourable until the seventh decade.

Novel frameshift mutations in CRX associated with Leber congenital amaurosis.

Mutations in CRX, a photoreceptor-specific transcription factor, can cause Leber congenital amaurosis (LCA), cone-rod dystrophy (CORD), and retinitis pigmentosa (RP), all of which feature severe visual impairment. Upon screening 55 patients with Leber congenital amaurosis, 75 patients with cone-rod dystrophy, 13 with cone dystrophy, and 36 with recessive or isolate RP for changes in the CRX sequence, we found two patients with Leber congenital amaurosis who carried heterozygously one of two novel frameshift mutations. The first mutation, Tyr191(1-bp del), was a de novo change and the second change, Pro263(1-bp del) was inherited from the proband's affected father. Both mutations are predicted to encode mutant versions of CRX with altered carboxy termini. We also found a previously reported missense mutation, Arg41Gln, heterozygously in a 47-year-old patient with a form of RP. The missense change Val242Met was found in an isolate case of CORD and no controls; however, its pathogenicity remains uncertain because only limited segregation analysis was possible. A nonpathogenic missense change, Ala158Thr, was found to be a variant present at relatively high frequency among African-Americans.

CORD9 a new locus for arCRD: mapping to 8p11, estimation of frequency, evaluation of a candidate gene.

PURPOSE: To determine the locus of the mutant gene causing autosomal recessive cone-rod dystrophy (arCRD) in a consanguineous pedigree, to evaluate a candidate gene expressed in retina that maps to this locus, and to estimate the percentage of arCRD cases caused by mutations in this gene. METHODS: DNAs from family members were genotyped for markers covering the entire genome at an average spacing of approximately 9 centimorgans (cM). The data were input into a pedigree computer program to produce output files used to calculate lod scores. Significant linkage was revealed at 8cen, prompting the genotyping of a number of additional markers. Exons of a candidate gene were sequenced directly by standard fluorescent dideoxy methods. Haplotype analysis was performed with markers in this locus in 13 multiplex and 2 simplex CRD families in which neither parent had disease. RESULTS: Four-point linkage analysis gave a maximum lod score of approximately 7.6 at both D8S1769 and GATA101H09 in the large consanguineous family. Recombination events defined an interval of 8.7 cM between D8S1820 and D8S532 within which the gene must lie. This 8p11 locus (CORD9) is immediately distal to but distinct from the RP1 autosomal dominant RP (adRP) locus. Two islands of homozygosity were found in this locus: The alleles of 6 of 10 markers in one of the islands and 2 of 4 in the other were homozygous. The UniGene cluster Hs.8719 (UniGene System, provided by the National Center for Biotechnology Information and available at http://www.ncbi.nlm.nih.gov/UniGene), which tags a gene with significant homology to Dual Specificity Phosphatase 3, maps within the CORD9 interval and is highly expressed in the retina. To evaluate this gene as a potential disease candidate, intron-exon structure was determined, and exons were screened in the consanguineous family. No variants were found that could be related to disease. Haplotype analysis of 15 other families with CRD, using markers at CORD9, excluded this locus in 9 of 15. CONCLUSIONS: A new arCRD locus (CORD9) has been identified corresponding to a yet unidentified gene in the 8.7-cM interval D8S1820-D8S532. No mutations were found in one candidate gene in affected members of the primary study family. Haplotype analysis of a cohort of 13 multiplex and 2 simplex families with CRD ruled out the CORD9 gene in 9 of 15 of the families. To date, a total of 126 loci carrying gene mutations causing various forms of retinal degeneration have been mapped, and the mutant gene has been identified in 64 of them. However, only 2 loci for arCRD have been documented. This is the report of a third.

High-frequency attenuation of the cone ERG and ON-response deficits in X-linked retinoschisis.

PURPOSE: Individuals with X-linked retinoschisis (XLRS) show a comparatively greater reduction of the ON response than the OFF response of the electroretinogram (ERG) of the cone system. At high temporal frequencies, they also show a marked attenuation of the flicker ERG that has been attributed to an abnormal cone photoreceptor response. The purpose of this study was to determine whether the high-frequency response attenuation contributes to the abnormal ERG ON response in XLRS. METHODS: Light-adapted ERGs were recorded from three patients with XLRS and from three control subjects, by using rapid-on and rapid-off sawtooth flicker to emphasize ON and OFF responses, respectively, and by using low-pass sawtooth flicker, from which the high temporal frequencies had been removed to mimic the high-frequency attenuation in XLRS. RESULTS: For the control subjects, removing the high stimulus frequencies reduced the amplitude of the b-wave component of the ON response but had little effect on the amplitude of the d-wave component of the OFF response. In the patients with XLRS, the b-wave component of the ON response was already diminished using the full sawtooth stimulus, and removing the higher stimulus frequencies had no further effect. Patients' ERG responses to the 16-Hz stimulus fundamental alone were also abnormal, in that an initial response component normally present in the ERG was absent. CONCLUSIONS: The overall pattern of findings indicates that two factors contribute to the preferential ON-response deficit in XLRS: first, a high-frequency attenuation of the cone photoreceptor response that effectively produces a low-pass stimulus for the postreceptoral pathway and that affects the ON response more than the OFF response and, second, a relatively greater attenuation of the ON- than of the OFF-bipolar cell response that is evident in the aberrant response to the sawtooth fundamental.

Retinal findings in melanoma-associated retinopathy.

PURPOSE: To report unusual fundus findings in two cases of melanoma-associated retinopathy. METHODS: Observational case reports. The histories of two patients with melanoma-associated retinopathy were reviewed. Sera from both patients were examined for antibodies against retinal bipolar cells. Immunofluorescence was performed on cryostat sections of unfixed normal human retinas. Sera and IgG from both patients were tested against a known melanoma-associated retinopathy patient as well as a control subject. RESULTS: Our patients had metastatic, cutaneous melanoma and a clinical syndrome consistent with melanoma-associated retinopathy. Both patients had serum antibodies that were reactive against retinal bipolar cells. They had unusual fundus changes not previously described in association with melanoma-associated retinopathy. One patient also developed vitiligo. CONCLUSION: Patients with metastatic cutaneous melanoma and melanoma-associated retinopathy may present unusual fundus lesions that may be caused by autoimmunity which is part of the clinical picture of melanoma-associated retinopathy or metastatic melanoma.

An analysis of allelic variation in the ABCA4 gene.

PURPOSE: To assess the allelic variation of the ATP-binding transporter protein (ABCA4). METHODS: A combination of single-strand conformation polymorphism (SSCP) and automated DNA sequencing was used to systematically screen this gene for sequence variations in 374 unrelated probands with a clinical diagnosis of Stargardt disease, 182 patients with age-related macular degeneration (AMD), and 96 normal subjects. RESULTS: There was no significant difference in the proportion of any single variant or class of variant between the control and AMD groups. In contrast, truncating variants, amino acid substitutions, synonymous codon changes, and intronic variants were significantly enriched in patients with Stargardt disease when compared with their presence in subjects without Stargardt disease (Kruskal-Wallis P < 0.0001 for each variant group). Overall, there were 2480 instances of 213 different variants in the ABCA4 gene, including 589 instances of 97 amino acid substitutions, and 45 instances of 33 truncating variants. CONCLUSIONS: Of the 97 amino acid substitutions, 11 occurred at a frequency that made them unlikely to be high-penetrance recessive disease-causing variants (HPRDCV). After accounting for variants in cis, one or more changes that were compatible with HPRDCV were found on 35% of all Stargardt-associated alleles overall. The nucleotide diversity of the ABCA4 coding region, a collective measure of the number and prevalence of polymorphic sites in a region of DNA, was found to be 1.28, a value that is 9 to 400 times greater than that of two other macular disease genes that were examined in a similar fashion (VMD2 and EFEMP1).

Origin of deficits in the flicker electroretinogram of the cone system in X-linked retinoschisis as derived from response nonlinearities.

The aim of this study was to identify the origin of a high-frequency attenuation in the flicker electroretinogram (ERG) of patients with X-linked retinoschisis (XLRS) through an analysis of nonlinearities in the ERG response. The ERGs of six patients with XLRS and six age-similar control subjects were recorded in response to stimuli that consisted of pairs of sinusoids that had varying temporal frequencies and that differed by either 8 or 16 Hz. Compared with the control subjects, the patients with XLRS showed a significant reduction in the amplitude of the difference frequency to high-frequency stimuli that paralleled the high-frequency attenuation of their ERG response fundamental. This result indicates that a response attenuation at an initial linear filter, most likely photoreceptoral, was a major determinant of the reduced ERG amplitude of the XLRS patients at high temporal frequencies. Additional analyses of nonlinearities in the ERG responses provided evidence of a postreceptoral component to the flicker ERG deficits of the XLRS patients, as well.

Mutations in the CRB1 gene cause Leber congenital amaurosis.

OBJECTIVES: To test the hypothesis that mutations in the CRB1 gene cause Leber congenital amaurosis (LCA) and, if so, to describe the ocular phenotype of patients with LCA who harbor CRB1 sequence variations. PATIENTS: One hundred ninety probands with a clinical diagnosis of LCA were selected from a cohort of 233 probands ascertained in 5 different countries. The remaining 43 probands (18%) were excluded because they harbored sequence variations in previously identified LCA genes. METHODS: One hundred ninety unrelated individuals with LCA were screened for coding sequence mutations in the CRB1 gene with single-strand conformation polymorphism analysis followed by automated DNA sequencing. RESULTS: Twenty-one of the 190 probands (9% of the total cohort of 233) and 2 (1.4%) of 140 controls harbored amino acid-altering sequence variations in the CRB1 gene (P =.003). CONCLUSIONS: In our cohort of patients with LCA, coding sequence variations were observed in the CRB1 gene more frequently than in any of the other 5 known LCA-associated genes. Likely disease-causing sequence variations have now been identified in 64 (28%) of 233 subjects in this cohort. CLINICAL RELEVANCE: Molecular diagnosis can confirm and clarify the diagnosis in an increasing fraction of patients with LCA. As genotype data accumulate, clinical phenotypes associated with specific mutations may be established. This will facilitate the counseling of patients regarding their visual prognosis and the likelihood of associated systemic anomalies.

On-response deficit in the electroretinogram of the cone system in X-linked retinoschisis.

PURPOSE: The purpose of this study was to evaluate the hypothesis that the reduced b-wave to a-wave ratio of the brief-flash electroretinogram (ERG) of the cone system typically observed in X-linked retinoschisis (XLRS) represents a relatively greater deficit in the ON response (response to light onset) than the OFF response (response to light offset). A second purpose was to investigate the use of sawtooth flicker as a stimulus for eliciting ERG ON and OFF responses. METHODS: Light-adapted, full-field ERGs were recorded in six patients with XLRS and six age-similar control subjects in response to 8-Hz rapid-on and rapid-off sawtooth flicker to emphasize ON and OFF responses, respectively. ERG responses were analyzed in terms of the amplitudes and implicit times of the a-wave, b-wave, and d-wave components. RESULTS: There was no significant difference between the patients with XLRS and the control subjects for either the amplitude of the a-wave of the ON response or the amplitude of the d-wave of the OFF response. However, the amplitude of the b-wave of the ON response was reduced significantly in the patients with XLRS, resulting in a significantly reduced b-wave to d-wave ratio. The patients' implicit times were increased significantly for all waveform components. CONCLUSIONS: The reduced b-wave to d-wave ratio of the ERG of the cone system in these patients with XLRS is consistent with a relative dysfunction of the cone ON bipolar cell pathway in this disorder. The results show further that sawtooth flicker is a promising stimulus for eliciting well-defined ERG waveforms that can provide a quantitative assessment of the properties of ON and OFF responses in retinal disease.