Choroidal blood flow decreases with age: an MRI study.

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.

Decreased retinal-choroidal blood flow in retinitis pigmentosa as measured by MRI.

PURPOSE: To evaluate retinal and choroidal blood flow (BF) using high-resolution magnetic resonance imaging (MRI) as well as visual function measured by the electroretinogram (ERG) in patients with retinitis pigmentosa (RP). METHODS: MRI studies were performed in 6 RP patients (29-67 years) and 5 healthy volunteers (29-64 years) on a 3-Tesla scanner with a custom-made surface coil. Quantitative BF was measured using the pseudo-continuous arterial spin-labeling technique at 0.5 × 0.8 × 6.0 mm. Full-field ERGs of all patients were recorded. Amplitudes and implicit times of standard ERGs were analyzed. RESULTS: Basal BF in the posterior retinal-choroid was 142 ± 16 ml/100ml/min (or 1.14 ± 0.13 µl/mm(2)/min) in the control group and was 70 ±19 ml/100ml/min (or 0.56 ± 0.15 µl/mm(2)/min) in the RP group. Retinal-choroidal BF was significantly reduced by 52 ± 8 % in RP patients compared to controls (P<0.05). ERG a- and b-wave amplitudes of RP patients were reduced, and b-wave implicit times were delayed. There were statistically significant correlations between a-wave amplitude and BF value (r=0.9, P<0.05) but not between b-wave amplitude and BF value (r =0.7, P=0.2). CONCLUSIONS: This study demonstrates a novel non-invasive MRI approach to measure quantitative retinal and choroidal BF in RP patients. We found that retinal-choroidal BF was markedly reduced and significantly correlated with reduced amplitudes of the a-wave of the standard combined ERG.

A low cost color visual stimulator for fMRI.

This low cost visual stimulator was developed for use in small animal imaging. The stimulator uses a single tri-color LED for each eye and can output red, green, or blue light or any combination of the three. When all three LED colors are illuminated at the same time achromatic light is the output. The stimulator is almost entirely implemented in software with only minimal electronics. The LEDs are controlled via the parallel port of a desktop computer. Flicker frequency, wavelength, intensity and waveform shape are under software control. The LEDs are coupled to fiber optic cables which run into the MRI scanner room leaving the LEDs and the power source in the control room. Calibration with a radiometer shows the light output to be very linear from zero to full intensity. The stimulator was used in fMRI visual stimulation studies performed on Sprague Dawley rats with an 11.7Tesla magnet. As the stimulator is software driven, modifications to accommodate other protocols and extensions for new functionality can be readily incorporated. With this in mind, the visual stimulator circuit diagram and software including source code are available upon request.

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.

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.

A preliminary randomized, double-blind, placebo-controlled study of the safety and efficacy of ondansetron in the treatment of cocaine dependence.

Prior studies have demonstrated inefficacy among dopamine receptor antagonists for treating cocaine dependence. An alternative approach would be to investigate the ability of indirect inhibitors of cortico-mesolimbic dopamine release, such as the 5-HT(3) receptor antagonist ondansetron, to reduce cocaine's reinforcing effects. We hypothesized that ondansetron might be more efficacious than placebo at reducing cocaine intake and promoting abstinence in cocaine-dependent individuals. In a pilot randomized, double-blind, 10-week controlled trial, 63 treatment-seeking, cocaine-dependent men and women received ondansetron (0.25 mg, 1.0 mg, or 4.0 mg twice daily) or placebo. Up to three times per week, participants were assessed on several measures of cocaine use, including urine benzoylecgonine. Cognitive behavioral therapy was administered weekly. Ondansetron was well tolerated, causing no serious adverse events. The ondansetron 4.0 mg group had the lowest dropout rate among all treatment groups and a greater rate of improvement in percentage of participants with a cocaine-free week compared with the placebo group (p = 0.02), whereas the ondansetron 1.0 mg group had a lower rate of improvement in percentage of weekly mean non-use days than did placebo recipients (p = 0.04). These results suggest the possibility of a non-linear dose-response function, with evidence supporting efficacy for the 4.0 mg group.

Retinal function assessed by ERG before and after induction of ocular aspergillosis and treatment by the anti-fungal, micafungin, in rabbits.

This study was conducted to evaluate the effectiveness of a new antifungal drug, micafungin, and standard antifungal drugs against endophthalmitis induced in a rabbit by intravitreal injection of Aspergillus fumigatus, an important fungal pathogen. Effectiveness was evaluated by the preservation of b-wave amplitude at 72 h after injection of the fungus relative to the b-wave amplitude at baseline before any intravitreal injections. A 0.06 ml inoculum of 10(6) conidia of A. fumigatus was injected into the vitreous of the right eye of all rabbits; and, 12 h later, a 0.06 ml solution containing one of 3 antifungal drugs or saline was injected into the vitreous of both eyes. All three antifungal drugs produced significant b-wave preservation at 72 h in infected eyes compared to that in infected eyes receiving saline injections. There was no statistically significant difference between the effects of micafungin and amphotericin B in the right eyes with fungal endophthalmitis, and both produced significantly more preservation of b-wave amplitude than voriconazole. Amphotericin B, but neither micafungin nor voriconazole produced significant reduction of the b-wave amplitude in the left eyes.

Tonopen measurement of intraocular pressure in mice.

PURPOSE: To standardize a method of non-invasive measurement of intraocular pressure (IOP) in mice. METHODS: Cannulated-eye study: IOP was measured simultaneously with a Tonopen and by direct cannulation of the vitreous compartment while pressure was manipulated in steps between 10 and 45 mmHg by a saline reservoir via a second vitreal cannula (five mice, one rat). Non-cannulated-eye study: Tonopen and servo-null measurements were performed in independent groups (48 mice) to verify Tonopen measurements in non-cannulated-eyes. Topical brimonidine (0.15%) was used to decrease IOP. RESULTS: In the rat, there was a similar relationship between Tonopen readings and direct measurements via cannulation of the eye as previously reported. Although readings from mice eyes were higher in variability than those obtained from the rat, the measurements were reproducible and the correlation between the invasive and the non-invasive methods was good (r = 0.97). The IOP lowering effect of brimonidine was detected with Tonopen as well as servo-null measurements (p < 0.001) and the results with both techniques were similar. CONCLUSION: The Tonopen can be used for rapid and reproducible measurements of IOP in mice. The method is easy to apply and can provide a useful means for IOP measurement in mouse models of induced ocular hypertension, in knock-out and transgenic mice, or in pharmacological studies.

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 subject's 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 degrees 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.

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.

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.