Joseph M. Harrison

Dysfunctional Light-Evoked Regulation of cAMP in Photoreceptors and Abnormal Retinal Adaptation in Mice Lacking Dopamine D4 Receptors

Dopamine is a retinal neuromodulator that has been implicated in many aspects of retinal physiology. Photoreceptor cells express dopamine D4 receptors that regulate cAMP metabolism. To assess the effects of dopamine on photoreceptor physiology, we examined the morphology, electrophysiology, and regulation of cAMP metabolism in mice with targeted disruption of the dopamine D4 receptor gene. Photoreceptor morphology and outer segment disc shedding after light onset were normal in D4 knock-out (D4KO) mice. Quinpirole, a dopamine D2/D3/D4 receptor agonist, decreased cAMP synthesis in retinas of wild-type (WT) mice but not in retinas of D4KO mice. In WT retinas, the photoreceptors of which were functionally isolated by incubation in the presence of exogenous glutamate, light also suppressed cAMP synthesis. Despite the similar inhibition of cAMP synthesis, the effect of light is directly on the photoreceptors and independent of dopamine modulation, because it was unaffected by application of the D4 receptor antagonist l-745,870. Nevertheless, compared with WT retinas, basal cAMP formation was reduced in the photoreceptors of D4KO retinas, and light had no additional inhibitory effect. The results suggest that dopamine, via D4 receptors, normally modulates the cascade that couples light responses to adenylyl cyclase activity in photoreceptor cells, and the absence of this modulation results in dysfunction of the cascade. Dark-adapted electroretinogram (ERG) responses were normal in D4KO mice. However, ERG b-wave responses were greatly suppressed during both light adaptation and early stages of dark adaptation. Thus, the absence of D4 receptors affects adaptation, altering transmission of light responses from photoreceptors to inner retinal neurons. These findings indicate that dopamine D4 receptors normally play a major role in regulating photoreceptor cAMP metabolism and adaptive retinal responses to changing environmental illumination.

Flash visual evoked potentials in the hypomyelinated mutant mouse shiverer

Myelin basic protein (MBP) is an essential component of central nervous system (CNS) myelin, as demonstrated by shiverer mutant mice that have deletions of most of the Mbp structural gene. These mutants do not produce detectable MBP protein, and their CNS is hypomyelinated. Although the function of the visual pathway is presumed to be adversely affected by hypomyelination of the optic nerve, it has never been studied. We compared flash visual evoked potentials (FVEPs) of shiverer homozygotes with those of their wild-type littermates in order to characterize any dysfunction. There was a statistically significant delay in the implicit times of a negative component peaking at 85 ms and a large positive component peaking at 170 ms in the FVEPs of the shiverer mice. The amplitudes of the two components did not differ significantly in the shiverers and wild-type controls. Barring a retinal pathology, which cannot be excluded by these data, the delayed FVEP of the shiverer can likely be attributed to effects of hypomyelination of the optic nerve, optic tract and visual radiations on conduction time in the visual pathway and subsequent further post-synaptic delays.

Dorzolamide Hydrochloride and Visual Function in Normal Eyes

PURPOSE To determine by a pilot study whether standard treatment with the topical carbonic anhydrase inhibitor dorzolamide hydrochloride influences visual function under normal breathing conditions, during carbon dioxide inhalation, or during hyperventilation, and to establish criteria for future larger-scale studies. METHODS We enrolled 12 normal subjects into this randomized double-masked placebo-controlled crossover study. Each subject was treated with either dorzolamide 2% or placebo, three times daily, for 4 days. After a 2-week washout period, the alternative topical agent was used under identical testing conditions. On day 2 of each treatment phase, contrast sensitivities to sinusoidal gratings of 1 and 4 cycles per degree (cpd) were assessed. On day 4, mean deviation values from full-threshold 10-2 visual fields were obtained. Three sets of each visual function test were obtained before each treatment phase, and in sequence on each testing day, during normal breathing (baseline), inhalation of carbon dioxide-enriched air, and hyperventilation while intraocular pressure was monitored. RESULTS Contrast sensitivity at 4 cpd decreased significantly (P < .01) during carbon dioxide supplementation with placebo but showed no significant change with dorzolamide. The decrease in contrast sensitivity accompanying hyperventilation was attenuated (by nearly 50% at 1 cpd) during dorzolamide treatment. Dorzolamide treatment was associated with higher perimetry mean deviation values under each treatment condition and was statistically significant (P < .05) at baseline. CONCLUSIONS Dorzolamide appears to enhance contrast sensitivity in normal subjects during physiologic hypercapnia and hypocapnia at 4 and 1 cpd, respectively. Also, under normal breathing conditions, dorzolamide therapy increases perimetric light sensitivity.

Lamina-Specific Functional MRI of Retinal and Choroidal Responses to Visual Stimuli

PURPOSE To demonstrate lamina-specific functional magnetic resonance imaging (MRI) of retinal and choroidal responses to visual stimulation of graded luminance, wavelength, and frequency. MATERIALS AND METHODS High-resolution (60 × 60 μm) MRI was achieved using the blood-pool contrast agent, monocrystalline iron oxide nanoparticles (MION) and a high-magnetic-field (11.7 T) scanner to image functional changes in the normal rat retina associated with various visual stimulations. MION functional MRI measured stimulus-evoked blood-volume (BV) changes. Graded luminance, wavelength, and frequency were investigated. Stimulus-evoked fMRI signal changes from the retinal and choroidal vascular layers were analyzed. RESULTS MRI revealed two distinct laminar signals that corresponded to the retinal and choroidal vascular layers bounding the retina and were separated by the avascular layer in between. The baseline outer layer BV index was 2-4 times greater than the inner layer BV, consistent with higher choroidal vascular density. During visual stimulation, BV responses to flickering light of different luminance, frequency, and wavelength in the inner layer were greater than those in the outer layer. The inner layer responses were dependent on luminance, frequency, and wavelength, whereas the outer layer responses were not, suggesting differential neurovascular coupling between the two vasculatures. CONCLUSIONS This is the first report of simultaneous resolution of layer-specific functional responses of the retinal and choroid vascular layers to visual stimulation in the retina. This imaging approach could have applications in early detection and longitudinal monitoring of retinal diseases where retinal and choroidal hemodynamics may be differentially perturbed at various stages of the diseases.

Voluntary Alteration of Pattern Visual Evoked Responses

Pattern and flash visual evoked responses (VERs) were recorded from a large group of ophthalmologically normal subjects during two conditions: in one they were instructed to attend to the stimulus and in the other they were instructed to ignore the stimulus but maintain their gaze on the stimulus. Pattern VERs were recorded from 42 subjects. Fixation was constantly monitored during both attend and ignore conditions and no changes of fixation were noted at any time. The amplitude of the major positive wave of the pattern VERs produced by both 50- and 25-min checks was reduced significantly during the ignore condition. The implicit time of this positive wave did not differ significantly during the two conditions. The pattern VERs of eight subjects were extinguished and the VERs of another three subjects were unrecognizable during the ignore condition. Flash VERs produced by 10 flashes per second were recorded from 38 of the 42 subjects. There were no significant differences between the amplitudes recorded during the attend and ignore conditions.

Forward light scatter at one month after photorefractive keratectomy.

BACKGROUND Although it is known that backward light scatter increases transiently following most excimer laser photorefractive keratectomies (PRKs), it is not clear that there is a significant increase in forward light scatter, which is of primary concern for the patient. The object of this study was to determine if there is a significant change of forward light scatter at 1 month after (PRK) with an ablation zone diameter of 6 mm. METHODS Overlapping subsets of 24 normal myopic eyes were tested before (on the day of surgery) and 1 month after PRK, using three instruments: a Stray Light Meter (16 eyes); a Computerized Stray Light Meter (14 eyes); and a mesopic Increment Threshold-Glare Paradigm (six eyes). Differences between the two eyes before PRK were compared with the differences between the same eye before and after PRK, using repeated measured analysis of variance. In addition, increment threshold data obtained from 22 eyes after PRK were compared with those of 60 controls of the same age range and distribution by a t test. RESULTS None of the statistical comparisons approached significance at the alpha = 0.05 level. Changes in light scatter as small as a factor of 1.95 (Stray Light Meter) and 1.55 (Increment Threshold-Glare Paradigm) could be detected as significant with a high power (0.8). Changes larger than a factor of 21 could be detected with a power of 0.8 for the Computerized Stray Light Meter. CONCLUSIONS In these data, there is no support for the hypothesis that forward light scatter increases significantly 1 month after PRK with an ablation zone of 6 mm. Any increases in forward light scatter are unlikely to be greater than a factor of 1.5 to 2 under daytime or nighttime illumination conditions.

Lamina-Specific Anatomic Magnetic Resonance Imaging of the Human Retina

PURPOSE Magnetic resonance imaging (MRI) of the human retina faces two major challenges: eye movement and hardware limitation that could preclude human retinal MRI with adequate spatiotemporal resolution. This study investigated eye-fixation stability and high-resolution anatomic MRI of the human retina on a 3-Tesla (T) MRI scanner. Comparison was made with optical coherence tomography (OCT) on the same subjects. METHODS Eye-fixation stability of protocols used in MRI was evaluated on four normal volunteers using an eye tracker. High-resolution MRI (100 × 200 × 2000 μm) protocol was developed on a 3-T scanner. Subjects were instructed to maintain stable eye fixation on a target with cued blinks every 8 seconds during MRI. OCT imaging of the retina was performed. Retinal layer thicknesses measured with MRI and OCT were analyzed for matching regions of the same eyes close to the optic nerve head. RESULTS The temporal SDs of the horizontal and vertical displacements were 78 ± 51 and 130 ± 51 μm (±SD, n = 4), respectively. MRI detected three layers within the human retina, consistent with MRI findings in rodent, feline, and baboon retinas. The hyperintense layer 1 closest to the vitreous likely consisted of nerve fiber, ganglion cell, and inner nuclear layer; the hypointense layer 2, the outer nuclear layer and the inner and outer segments; and the hyperintense layer 3, the choroid. The MRI retina/choroid thickness was 711 ± 37 μm, 19% (P < 0.05) thicker than OCT thickness (579 ± 34 μm). CONCLUSIONS This study reports high-resolution MRI of lamina-specific structures in the human retina. These initial results are encouraging. Further improvement in spatiotemporal resolution is warranted.

Choroidal Blood Flow Decreases with Age: An MRI Study

Abstract Purpose: To verify that a visual fixation protocol with cued eye blinks achieves sufficient stability for magnetic resonance imaging (MRI) blood-flow measurements and to determine if choroidal blood flow (ChBF) changes with age in humans. Methods: The visual fixation stability achievable during an MRI scan was measured in five normal subjects using an eye-tracking camera outside the MRI scanner. Subjects were instructed to blink immediately after recorded MRI sound cues but to otherwise maintain stable visual fixation on a small target. Using this fixation protocol, ChBF was measured with MRI using a 3 Tesla clinical scanner in 17 normal subjects (24–68 years old). Arterial and intraocular pressures (IOP) were measured to calculate perfusion pressure in the same subjects. Results: The mean temporal fluctuations (standard deviation) of the horizontal and vertical displacements were 29 ± 9 μm and 38 ± 11 μm within individual fixation periods, and 50 ± 34 μm and 48 ± 19 μm across different fixation periods. The absolute displacements were 67 ± 31 μm and 81 ± 26 μm. ChBF was negatively correlated with age (R = −0.7, p = 0.003), declining 2.7 ml/100 ml/min per year. There were no significant correlations between ChBF versus perfusion pressure, arterial pressure, or IOP. There were also no significant correlations between age versus perfusion pressure, arterial pressure, or IOP. Multiple regression analysis indicated that age was the only measured independent variable that was significantly correlated with ChBF (p = 0.03). Conclusions: The visual fixation protocol with cued eye blinks was effective in achieving sufficient stability for MRI measurements. ChBF had a significant negative correlation with age.

Contrast sensitivity and disability glare in the middle years

Spatial contrast sensitivity and disability glare were measured in a large sample (n = 90, 30 per decade) of middle-aged subjects, aged 21-50 years, who had clear media and were ophthalmologically normal. We found no significant differences in the contrast sensitivities as a function of age in the middle years for (1) gratings generated on a monitor; (2) interference gratings generated in the retinal plane; (3) gratings in the presence of glare; and (4) mesopic increment thresholds with and without glare. The large sample size provides sufficient statistical power (0.8) for one to conclude that contrast sensitivity, optical quality, and foveal neural sensitivity are unlikely to vary more than 0.1 log unit between the ages of 21 and 50 in ophthalmologically normal subjects with clear media.

Modification of the Heidelberg retinal flowmeter to record pattern and flicker induced blood flow changes

Purpose: We tested a prototype stimulator interfaced with a commercially available scanning laser ophthalmoscope designed to measure retinal capillary perfusion (Heidelberg Retina Flowmeter (HRF)). The add-on stimulator optically superimposed the image of a monitor display on to the subjects retina coaxially with the imaging optics of the HRF. The purpose of the study was to determine if flicker and pattern stimulation presented in this manner could evoke changes in retinal perfusion that could be measured by the HRF. Methods: The prototype stimulator projected 55° visual angle circular fields of homogeneous flicker, alternating checkerboard, and multi-focal m-sequence hexagonal patterns on the retina of 10 human subjects during acquisition of images by the HRF. Results: Images were successfully acquired and processed. HRF perfusion values during flicker and pattern stimulation were not significantly different from control values. Conclusions: Results of the present study and a previously published study showing a flicker-induced increase in the HRF perfusion values are contradictory. Retinal perfusion measured by the HRF were not affected by flicker and pattern stimulation delivered through the prototype device. These data are not consistent with a large flicker or pattern induced increase in retinal perfusion. The instrumental modification appears promising. However, the raster scan stimulation technique or some other aspect of stimulation or image acquisition may account for the different results in the present study and previous studies in our laboratory and in the laboratories of other investigators.