Effects of short-term exposure to red or near-infrared light on axial length in young human subjects.

PURPOSE: To determine whether visible light is needed to elicit axial eye shortening by exposure to long wavelength light. METHODS: Incoherent narrow-band red (620 ± 10 nm) or near-infrared (NIR, 875 ± 30 nm) light was generated by an array of light-emitting diodes (LEDs) and projected monocularly in 17 myopic and 13 non-myopic subjects for 10 min. The fellow eye was occluded. Light sources were positioned 50 cm from the eye in a dark room. Axial length (AL) was measured before and after the exposure using low-coherence interferometry. RESULTS: Non-myopic subjects responded to red light with significant eye shortening, while NIR light induced minor axial elongation (-13.3 ± 17.3 µm vs. +6.5 ± 11.6 µm, respectively, p = 0.005). Only 41% of the myopic subjects responded to red light exposure with a decrease in AL and changes were therefore, on average, not significantly different from those observed with NIR light (+0.2 ± 12.1 µm vs. +1.1 ± 11.2 µm, respectively, p = 0.83). Interestingly, there was a significant correlation between refractive error and induced changes in AL after exposure to NIR light in myopic eyes (r(15) = -0.52, p = 0.03) and induced changes in AL after exposure to red light in non-myopic eyes (r(11) = 0.62, p = 0.02), with more induced axial elongation with increasing refractive error. CONCLUSIONS: Incoherent narrow-band red light at 620 nm induced axial shortening in 77% of non-myopic and 41% of myopic eyes. NIR light did not induce any significant changes in AL in either refractive group, suggesting that the beneficial effect of red laser light therapy on myopia progression requires visible stimulation and not simply thermal energy.

Imposed positive defocus changes choroidal blood flow in young human subjects.

PURPOSE: It has previously been found that imposing positive defocus changes axial length and choroidal thickness after only 30 min. In the present study, we investigated whether these changes may result from an altered choroidal blood flow. METHODS: Eighteen young adult subjects watched a movie from a large screen (65 in.) in a dark room at 2 m distance. A 15-min wash-out period was followed by 30 min of watching the movie with a monocular positive defocus (+ 2.5D). Changes in axial length and ocular blood flow were measured before and after the defocus, by using low-coherent interferometer (LS 900, Haag-Streit, Switzerland) and a laser speckle flowgraphy (LSFG) RetFlow unit (Nidek Co., LTD, Japan), respectively. Three regions were analyzed: (1) the macular area, where choroidal blood flow can be measured, (2) the optic nerve head (ONH), and (3) retinal vessel segments. RESULTS: Changes in choroidal blood flow were significantly and negatively correlated with changes in axial length that followed positive defocus in exposed eyes (R = - 0.67, p < 0.01). The absolute values of changes in choroidal blood flow in the defocused eyes were significantly larger than in the fellow control eyes (2.35 ± 2.16 AU vs. 1.37 ± 1.44 AU, respectively, p < 0.05). ONH and retinal blood flow were not associated with the induced changes in axial length. CONCLUSIONS: Positive defocus selectively alters choroidal, but not retinal or ONH blood flow in young human subjects after short-term visual exposure. The results suggest that blood flow modulation is involved in the mechanism of choroidal responses to optical defocus.

Studies on the interactions of retinal dopamine with choroidal thickness in the chicken.

PURPOSE: Recently, an increasing number of studies relied on the assumption that visually induced changes in choroidal thickness can serve as a proxy to predict future axial eye growth. The retinal signals controlling choroidal thickness are, however, not well defined. We have studied the potential roles of dopamine, released from the retina, in the choroidal response in the chicken. METHODS: Changes in retinal dopamine release and choroidal thickness changes were induced by intravitreal injections of either atropine (250 µg or 360 nMol), atropine combined with a dopamine antagonist, spiperone (500 µMol), or spiperone alone and were tracked by optical coherence tomography (OCT). To visually stimulate dopamine release, other chicks were exposed to flicker light of 1, 10, or 400 Hz (duty cycle 0.2) and choroidal thickness was tracked. In all experiments, dopamine and 3,4-Dihydroxyphenylacetic acid (DOPAC) were measured in vitreous, retina, and choroid by high-performance liquid chromatography with electrochemical detection (HLPC-ED). The distribution of the rate-limiting enzyme of dopamine synthesis, tyrosine hydroxylase (TH), neuronal nitric oxide synthase (nNOS), vascular endothelial growth factor (VEGF), and alpha2A adrenoreceptors (alpha2A-ADR) was studied in the choroid by immunofluorescence. RESULTS: The choroid thickened strongly in atropine-injected eyes, less so in atropine + spiperone-injected eyes and became thinner over the day in spiperone alone-, vehicle-, or non-injected eyes. Flickering light at 20 lx, both 1 and 10 Hz, prevented diurnal choroidal thinning, compared to 400 Hz, and stimulated retinal dopamine release. Correlation analysis showed that the higher retinal dopamine levels or release, the thicker became the choroid. TH-, nNOS-, VEGF-, and alpha2A adrenoreceptor-positive nerve fibers were localized in the choroid around lacunae and in the walls of blood vessels with colocalization of TH and nNOS, and TH and VEGF. CONCLUSIONS: Retinal DOPAC and dopamine levels were positively correlated with choroidal thickness. TH-positive nerve fibers in the choroid were closely associated with peptides known to play a role in myopia development. Findings are in line with the hypothesis that dopamine is related to retinal signals controlling choroidal thickness.

Impact of cone abundancy ratios and light spectra on emmetropization in chickens.

We had previously found that chicken eyes with normal visual experience grow larger when they have more L cones, relative to M cones. It is not known whether also S cone abundancies may affect eye size, whether cone abundancy ratios can also affect the amount of deprivation myopia that is induced by diffusers in front of the eyes, and whether broadband white light with added energy in the blue may reduce the development of deprivation myopia. Therefore, chickens were monocularly treated with diffusers and raised under three different light conditions with increasing amounts of energy in the blue but with matched total illuminance. L, M and S cones were counted in fresh retinal tissues after the experiments. It was found that adding energy in the blue did not significantly inhibit deprivation myopia, nor did it make uncovered eyes more hyperopic. However, more S cones, relative to L cones, were correlated with more hyperopic refractions in eyes with normal vision. M to L, L to S and M to S cone ratios were also correlated with the amount of induced deprivation myopia. Interestingly, in deprivation myopia, the correlations between cone abundancy ratios with refractive states had reverted signs: eyes with more S cones developed more myopia. Since cone abundancy ratios remained correlated in both eyes, no matter whether eyes had normal vision, were deprived or were exposed to different light spectra, they appear genetically determined. We conclude that, among other factors, inherited cone abundancy ratios determine both normal refractive development and deprivation myopia in the chicken while adding more blue light to a broadband light spectrum had no effect.

Positions of the horizontal and vertical centre of rotation in eyes with different refractive errors.

PURPOSE: To determine how the position of the centre of rotation of the eyeball is related to axial length and refractive error when horizontal and vertical eye movements are performed. METHODS: A custom-built eye tracker was used that determined the centre of rotation of the eye (COR) from lateral displacements of the pupil centre. Horizontal and vertical eye movements were studied in the right eyes, and each measurement performed five times in 59 subjects (32 females) with an average age of 36.6 ± 9.1 years. Spherical equivalent refractive errors ranged from -9.7 to +6.8 D with an average error of -1.5 ± 2.9 D. Axial lengths were measured with the ZEISS IOL Master 500. RESULTS: The mean horizontal centre of rotation (COR) of the right eye for a saccade from 0° to ±11.9° was 15.3 ± 1.5 mm behind the corneal apex, while the average vertical COR for the same angle of eccentricity was 12.5 ± 1.4 mm, indicating that the horizontal COR was 2.8 ± 1.7 mm behind the vertical COR. In right eyes, horizontal COR was significantly correlated with axial length (r = 0.28, p = 0.02) but not with the spherical equivalent refractive error (r = 0.39, p = 0.90). Similarly, vertical COR was significantly correlated with axial length (r = 0.25, p = 0.03) but not with the spherical equivalent refractive error (r = 0.17, p = 0.90). CONCLUSIONS: While it might be expected that the COR is dependent on axial length, the correlation was not strong. Interestingly, the location of the COR was substantially different for horizontal and vertical eye movements which may relate to the flatter curvature of the eyeball in the vertical meridian, compared to the horizontal, as described in previous studies.

Mice Lacking Gpr179 with Complete Congenital Stationary Night Blindness Are a Good Model for Myopia.

Mutations in GPR179 are one of the most common causes of autosomal recessive complete congenital stationary night blindness (cCSNB). This retinal disease is characterized in patients by impaired dim and night vision, associated with other ocular symptoms, including high myopia. cCSNB is caused by a complete loss of signal transmission from photoreceptors to ON-bipolar cells. In this study, we hypothesized that the lack of Gpr179 and the subsequent impaired ON-pathway could lead to myopic features in a mouse model of cCSNB. Using ultra performance liquid chromatography, we show that adult Gpr179-/- mice have a significant decrease in both retinal dopamine and 3,4-dihydroxyphenylacetic acid, compared to Gpr179+/+ mice. This alteration of the dopaminergic system is thought to be correlated with an increased susceptibility to lens-induced myopia but does not affect the natural refractive development. Altogether, our data added a novel myopia model, which could be used to identify therapeutic interventions.

Functional integration of eye tissues and refractive eye development: Mechanisms and pathways.

Refractive eye development is a tightly coordinated developmental process. The general layout of the eye and its various components are established during embryonic development, which involves a complex cross-tissue signaling. The eye then undergoes a refinement process during the postnatal emmetropization process, which relies heavily on the integration of environmental and genetic factors and is controlled by an elaborate genetic network. This genetic network encodes a multilayered signaling cascade, which converts visual stimuli into molecular signals that guide the postnatal growth of the eye. The signaling cascade underlying refractive eye development spans across all ocular tissues and comprises multiple signaling pathways. Notably, tissue-tissue interaction plays a key role in both embryonic eye development and postnatal eye emmetropization. Recent advances in eye biometry, physiological optics and systems genetics of refractive error have significantly advanced our understanding of the biological processes involved in refractive eye development and provided a framework for the development of new treatment options for myopia. In this review, we summarize the recent data on the mechanisms and signaling pathways underlying refractive eye development and discuss new evidence suggesting a wide-spread signal integration across different tissues and ocular components involved in visually guided eye growth.

A retrospective analysis of the therapeutic effects of 0.01% atropine on axial length growth in children in a real-life clinical setting.

BACKGROUND: Several randomized controlled studies have demonstrated the beneficial effects of 0.01% atropine eye drops on myopia progression in children. However, treatment effects may be different in a routine clinical setting. We performed a retrospective analysis of our clinical data from children to investigate the effect of 0.01% atropine eye drops on myopia progression in a routine clinical setting. METHODS: Atropine-treated children were asked to instill one drop of 0.01% atropine in each eye every evening at 5 days a week. Myopic children who did not undergo atropine treatment served as controls. Objective refraction and ocular biometry of 80 atropine-treated and 103 untreated children at initial visit and 1 year later were retrospectively analyzed. RESULTS: Myopic refractions in the treated and untreated children at initial visit ranged from -0.625 to -15.25 D (-4.21 ± 2.90 D) and from -0.125 to -9.375 D (-2.92 ± 1.77 D), respectively. Ages at initial visit ranged from 3.2 to 15.5 years (10.1 ± 2.7 years) in the treated and from 3.4 to 15.5 years (11.2 ± 3.0 years) in untreated children. Two-factor ANOVA for age and atropine effects on axial length growth confirmed that axial length growth rates declined with age (p<0.0001) and revealed a significant inhibitory effect of atropine on axial length growth (p<0.0015). The atropine effect on axial length growth averaged to 0.08 mm (28%) inhibition per year. Effects on refraction were not statistically significant. CONCLUSION: The observed atropine effects were not very distinctive: Statistical analysis confirmed that atropine reduced axial length growth, but to an extent of minor clinical relevance. It was also shown that beneficial effects of 0.01% atropine may not be obvious in each single case, which should be communicated with parents and resident ophthalmologists.

Effects of Single and Repeated Intravitreal Applications of Atropine on Choroidal Thickness in Alert Chickens.

INTRODUCTION: Atropine, a muscarinic antagonist, is known since the 19th century to inhibit myopia development in children. One of its effects is that it stimulates choroidal thickening. Thicker choroids, in turn, have been linked to myopia inhibition. We used the atropine-stimulated choroidal response in the chicken to learn more about the time courses and amplitudes of the effects of atropine, as well as whether repeated applications lead to accumulation or desensitization. METHODS: Intravitreal injections containing 250 µg atropine sulfate were performed in 1 eye around 10:00 in the morning, the fellow eye received vehicle. Chickens with bilateral vehicle injections served as controls. Choroidal thickness was measured over the day for every 2-3 h in alert animals, using spectral domain optical coherence tomography, with 3-5 independent measurements in each eye. Three experiments were done - (1) single injection and time course measured over 1 day, (2) single injection and time course measured over 4 days, and (3) daily injections and time course measured over 4 days for measuring the effects of atropine on vitreal, retinal, and choroidal dopamine, and 3,4-dihydroxyphenylacetic acid levels by using high-performance liquid chromatography with electrochemical detection. RESULTS: Atropine induced an increase in choroidal thickness by about 60 percent, with a peak amplitude after about 2 h. The effect persisted only for a few hours and had nearly disappeared by evening. Initially, similar amounts of choroidal thickening were observed in vehicle-injected fellow eyes but recovery to baseline was faster. When atropine was injected daily for 4 days, choroids thickened every day with similar amplitudes and time courses, with no signs of either accumulation or desensitization effects. Interestingly, while dopamine release from the retina was stimulated by atropine and followed approximately, the time course of choroidal thickening, its tissue concentration dropped in the choroid. CONCLUSIONS: Even at relatively high intravitreal doses, effects of atropine on choroidal thickness remained transient, similar to its effects on retinal dopamine. With repeated application every day, the diurnal patterns of choroidal thickening could be reproduced for 4 days with similar amplitudes and time courses. The transient nature of the effects of atropine on the choroid may be relevant for application protocols of atropine against myopia.

Interactions of cone abundancies, opsin expression, and environmental lighting with emmetropization in chickens.

We had previously found that M to L cone abundancy ratios in the chicken retina are correlated with vitreous chamber depth and refractive state in chickens eyes, when they have normal visual exposure but not when they develop deprivation myopia. The finding suggests an interaction between cone abundancies and emmetropization. In the current study, we analyzed how stable this correlation was against changes in environmental variables and strain differences. We found that the correlation was preserved in two chicken strains, as long as they were raised in the laboratory facilities and not in the animal facilities of the institute. To determine the reasons for this difference, spectral and temporal lighting parameters were better adjusted in both places, whereas temperature, humidity, food, diurnal lighting cycles and illuminance were already matched. It was also verified that both strains of chickens had the same cone opsin amino acid sequences. The correlation between M to L cone abundancy and ocular biometry is highly susceptible to changes in environmental variables. Yet undetermined differences in lighting parameters were the most likely reasons. Other striking findings were that green cone opsin mRNA expression was downregulated when deprivation myopia developed. Similarly, red opsin mRNA was downregulated when chicks wore red spectacles, which made them more hyperopic. In summary, our experiments show that photoreceptor abundancies, opsin expression, and the responses to deprivation, and therefore emmetropization, are surprisingly dependent on subtle differences in lighting parameters.

Studies on retinal mechanisms possibly related to myopia inhibition by atropine in the chicken.

PURPOSE: While low-dose atropine eye drops are currently widely used to inhibit myopia development in children, the underlying mechanisms are poorly understood. Therefore, we studied possible retinal mechanisms and receptors that are potentially involved in myopia inhibition by atropine. METHODS: A total of 250 µg atropine were intravitreally injected into one eye of 19 chickens, while the fellow eyes received saline and served as controls. After 1 h, 1.5 h, 2 h, 3 h, and 4 h, eyes were prepared for vitreal dopamine (DA) measurements, using high-pressure liquid chromatography with electrochemical detection. Twenty-four animals were kept either in bright light (8500 lx) or standard light (500 lx) after atropine injection for 1.5 h before DA was measured. In 10 chickens, the α2A-adrenoreceptor (α2A-ADR) agonists brimonidine and clonidine were intravitreally injected into one eye, the fellow eye served as control, and vitreal DA content was measured after 1.5 h. In 6 chickens, immunohistochemical analyses were performed 1.5 h after atropine injection. RESULTS: Vitreal DA levels increased after a single intravitreal atropine injection, with a peak difference between both eyes after 1.97 h. DA was also enhanced in fellow eyes, suggesting a systemic action of intravitreally administered atropine. Bright light and atropine (which both inhibit myopia) had additive effects on DA release. Quantitative immunolabelling showed that atropine heavily stimulated retinal activity markers ZENK and c-Fos in cells of the inner nuclear layer. Since atropine was recently found to also bind to α2A-ADRs at doses where it can inhibit myopia, their retinal localization was studied. In amacrine cells, α2A-ADRs were colocalized with tyrosine hydroxylase (TH), glucagon, and nitric oxide synthase, peptides known to play a role in myopia development in chickens. Intravitreal atropine injection reduced the number of neurons that were double-labelled for TH and α2A-ADR. α2A-ADR agonists clonidine and brimonidine (which were also found by other authors to inhibit myopia) severely reduced vitreal DA content in both injected and fellow eyes, compared to eyes of untreated chicks. CONCLUSIONS: Merging our results with published data, it can be concluded that both muscarinic and α2A-adrenergic receptors are expressed on dopaminergic neurons and both atropine and α2A-ADR antagonists stimulate DA release whereas α2A-ADR agonists strongly suppress its release. Stimulation of DA by atropine was enhanced by bright light. Results are in line with the hypothesis that inhibition of deprivation myopia is correlated with DA stimulation, as long as no toxicity is involved.

Effects of 7-Methylxanthine on Deprivation Myopia and Retinal Dopamine Release in Chickens.

INTRODUCTION: Intake of 7-methylxanthine (7-MX), an adenosine receptor (AR) antagonist, has been shown to inhibit school myopia in children and deprivation myopia in rhesus monkeys, but the underlying mechanisms are not known. Also retinal dopamine seems to be involved in the control of eye growth, and in the brain, ARs and dopamine receptors interact widely by heteromerization. We have studied whether 7-MX can inhibit deprivation myopia also in chickens and whether inhibition may involve the retinal dopamine system. METHODS: 7-MX was applied by either tube-feeding (100 µg/g body weight, twice a day) or intravitreal injection (12.5 µg, every other day). Forty-eight 2-week-old chicks wore unilateral diffusers and were randomly assigned to either the tube-feeding group (involving 7-MX, vehicle [xanthan gum], or no feeding, for 13 days) or the intravitreal injection group (involving 7-MX, vehicle, or DMSO, for 8 days). Refractions (REs), ocular biometry (AL, VCD), and scleral and choroidal thickness (ChT) were measured before and after treatment. Dopamine and dihydroxyphenylacetic acid (DOPAC) content were determined in retina and vitreous by HPLC at the end of the experiments. RESULTS: No matter how 7-MX was applied, it did not inhibit deprivation myopia in chicks. No significant differences were observed in RE, VCD, AL, and scleral fibrous layer thickness. Feeding 7-MX produced more choroidal thinning in the open contralateral eye compared to control eyes in the vehicle-fed group (-40 ± 14 vs. -1 ± 7 µm, unpaired t test, p < 0.05). DOPAC and dopamine concentration in vitreous and DOPAC concentration in retina did not change with 7-MX. Vitreal dopamine content was significantly decreased in deprived eyes in the groups fed with the vehicle xanthan gum (paired t test, p < 0.01) but not in 7-MX-treated eyes, perhaps indicating a small effect of 7-MX on dopamine. CONCLUSIONS: In our study, 7-MX had no effect on DM in chicks and only minor effects on ChT and retinal dopamine. It remains unclear whether 7-MX inhibits myopia through a retinal mechanism or whether it acts directly on choroid and sclera. In the latter case, the finding that myopia is suppressed in mammals but not birds might be explained by differences in scleral structure.

Demyelination and shrinkage of axons in the retinal nerve fiber layer in chickens developing deprivation myopia.

Placing diffusers in front of the eyes induces deprivation myopia in a variety of animal models. As a result of the low pass filtering of the retinal images, less spatial information is available to the retina which should reduce neural activity. Since it has been found that myelination of axons in the central nervous system is modulated by neuronal activity, we have studied whether ganglion cell axons may shrink in response to the restricted visual input. Young chickens were treated for 5 h or 7 days with frosted diffusers to induce deprivation myopia. Nerve fiber layer thickness was measured in vivo, using B-scan OCT. Refractive states were tracked by IR photoretinoscopy, and UV fundus reflectivity by a custom-built device which flashed an LED centered in the camera aperture and recorded pupil brightness after refractive errors were corrected by trial lenses. Moreover, structure and histology of the retinal nerve fibers layer (RNFL) were analyzed ex vivo using transmission electron microscopy and immunohistochemistry. Since chicks have both non-myelinated and myelinated fibers in their RNFL, the thickness of myelin sheaths (G ratio) was measured, as well as the percentage of myelinated axons and the diameters of unmyelinated axons. Short-term deprivation caused an increase in UV fundus reflectivity already after 5 h (measured as pixel grey levels in the pupil: 28 ±â€¯5 vs. 36 ±â€¯10, p < 0.05) and thinning of the myelin sheaths (higher G ratio), compared to untreated control eyes (0.74 ±â€¯0.01 vs. 0.79 ±â€¯0.03, p < 0.05). Neither axon diameters (0.81 ±â€¯0.05 µm vs. 0.82 ±â€¯0.15 µm) nor thickness of the RNFL had changed after only 5 h (42.9 ±â€¯1.3 µm vs. 42.3 ±â€¯2.5 µm). However, after 7 days of diffuser wear, axons had become thinner (0.56 ±â€¯0.14 µm vs. 0.78 ±â€¯0.09 µm vs, p < 0.05), which could explain the thinning of the RNFL (36.3 ±â€¯2.7 µm vs. 42.1 ±â€¯2.4 µm, p < 0.01). Furthermore, myopic eyes had 38% less myelinated axons than untreated eyes as determined by immunohistochemical labelling against myelin basic protein (immunopositive areas in the central retina 1406 ±â€¯341 µm2 vs. 2185 ±â€¯290 µm2 in controls, p < 0.001). Myelin sheaths in the remaining axons remained unchanged (G ratio 0.76 ±â€¯0.02 vs. 0.76 ±â€¯0.03). Our study shows that deprivation myopia is associated with a significant loss of myelinated axons and shrinkage of the axon diameters of certain fibers in the RNFL. Early changes were already detected after 5 h and were accompanied by an increased fundus reflectivity in UV light. These parameters could therefore serve as the biomarkers for myopia development, at least in the chicken.

Prolonged nearwork affects the ciliary muscle morphology.

There is evidence for a possible link between myopia development and near vision. We investigated the effect of prolonged nearwork on ciliary muscle (CM) morphology and accommodation in 18 myopic and 17 emmetropic subjects (age 19 to 25). The CM was imaged during far (0.25 D) and near vision (4 D) using optical coherence tomography (OCT), and accommodation to a step pulse (0.25 D - 4 D - 0.25 D, 15 s each) was assessed by eccentric infrared photorefraction before and after a 30-min reading task at 25 cm. OCT images were analyzed using a custom-developed semi-automatic segmentation algorithm to determine CM thickness (CMT) profiles and selective CMT readings. Accommodation was assessed using a non-linear model. On average, the CM got thinner after nearwork, predominantly at 0.0-1.4 mm posterior to the scleral spur in emmetropes, and at 1.0-1.9 mm in myopes. Selective CMT readings confirmed a significant thinning after nearwork (univariate ANOVA F1,66 = 26.313, p < 0.001), without any influence of the subjects' refractive state (F1,66 = 1.887, p = 0.174) or the target distance (F1,66 = 0.014, p = 0.907). The mean accommodation response for targets at infinity was significantly increased after nearwork (F1,32 = 7.775, p = 0.009), with a larger myopic shift in myopes (F1,32 = 11.310, p = 0.002). No change in velocity of accommodation was found. Sharing properties of striated muscles, the CM was expected to increase its thickness, but the opposite was found. Previous studies suggesting sustained nearwork to result in a CM spasm cannot be confirmed by the data presented here. Further research exploring the possible impact of sympathetic innervation is necessary as it is activated during intense nearwork.

M to L cone ratios determine eye sizes and baseline refractions in chickens.

Following a hypothesis raised by M. and J. Neitz, Seattle, we have tested whether the abundance and the ratio of Long wavelength-sensitive (L) to Middle wavelength-sensitive (M) cones may affect eye size and development of myopia in the chicken. Fourteen chickens were treated with frosted plastic diffusers in front of one eye on day 10 post-hatching for a period of 7 days to induce deprivation myopia. Ocular dimensions were measured by A-scan ultrasonography at the beginning and at the end of the treatment and development of refractive state was tracked using infrared photorefraction. At the end of the treatment period, L and M cone densities and ratios were analyzed in retinal flat mounts of both myopic and control eyes, using the red and yellow oil droplets as markers. Because large numbers of cones were counted (>10000), software was written in Visual C++ for automated cone detection and density analysis. (1) On average, 9.7 ± 1.7D of deprivation myopia was induced in 7 days (range from 6.8D to 13.7D) with an average increase in axial length by 0.65 ± 0.20 mm (range 0.42 mm-1.00 mm), (2) the increase in vitreous chamber depth was correlated with the increase in myopic refractive error, (3) average central M cone densities were 10,498 cells/mm2, and L cone densities 9574 cells/mm2. In the periphery, M cone densities were 6343 cells/mm2 and L cones 5735 cells/mm2 (4) M to L cone ratios were highly correlated in both eyes of each animal (p < 0.01 in all cases), (5) the most striking finding was that ratios of M to L cones were significantly correlated with vitreous chamber depths and refractive states in the control eyes with normal vision, both in the central and peripheral retinas (p < 0.05 to p < 0.01), (6) M to L cone ratios did however not predict the amount of deprivation myopia that could be induced. M and L cone ratios are most likely genetically determined in each animal. The more L cones, the deeper the vitreous chambers and the more myopic were the refractions in eyes. M to L cone ratios may determine the set point of emmetropization and thereby ultimately the probability of becoming myopic. Deprivation myopia was not determined by M to L cone ratios.

Short-term effects of low-concentration atropine eye drops on pupil size and accommodation in young adult subjects.

PURPOSE: A single eye drop containing 0.01% atropine every evening has previously been found to inhibit myopia progression in young adults. We have tested the short-term effects of very low-dose atropine eye drops on pupil sizes and accommodation in young adult subjects. METHODS: Fourteen eyes of young adult subjects participated in the clinical observation. A single eye drop was applied with concentrations of either 0.01%, 0.005%, or 0.001% in the evening. Baseline parameters were measured before atropine application. Changes of pupil sizes, under photopic and mesopic conditions, as well as accommodation amplitudes were observed over the next day and analyzed by paired the Wilcoxon signed-rank test. RESULTS: The pupil was significantly dilated 12 h after instillation of 0.01% atropine eye drops, both under photopic (3.3 ± 0.5 mm vs. 4.9 ± 0.9 mm) and mesopic (4.8 ± 0.7 mm vs. 6.1 ± 0.7 mm) conditions. Pupil sizes recovered over the day but were still significantly larger in the evening, compared to the baseline parameters measured on the day before (3.9 ± 0.5 mm vs. 5.3 ± 0.6 mm). The subjective near point of accommodation was reduced from 8.0 ± 2.4 to 6.6 ± 2.8 dpt in the morning and to 7.0 ± 2.9 dpt in the evening. At 0.005%, the pattern of results remained still similar, although the magnitude of the effects was generally smaller. At 0.001%, pupil sizes were still weakly significantly larger in the morning. CONCLUSIONS: At a dose of 0.01%, clinically significant short-term effects were detected on pupil size and accommodation for at least 24 h. At the lowest dose of 0.001%, only tiny effects on pupil size were detectable.

Changes in dopamine and ZENK during suppression of myopia in chicks by intense illuminance.

High ambient illuminances have been found to slow the development of deprivation myopia in several animal models. Almost complete inhibition of myopia was observed in chickens when intermittent episodes of high illuminance were alternated with standard office illuminance (50% duty cycle, alternate periods of 1 min 15,000 lux and 1 min 500 lux, continued for 10 h per day), or when illuminances were increased to 40,000 lux. Since the mechanisms by which bright light suppresses myopia are poorly understood, we have studied the roles of two well-established signaling molecules in myopia, dopamine and ZENK, in the chicken. In line with previous studies, we found that retinal dopamine release (as reflected by vitreal DOPAC content) was severely reduced during development of deprivation myopia. We found that illuminance of 15,000 lux, provided by quartz-halogen lamps, partially rescued the drop in retinal dopamine release. The finding is in line with the assumption that dopamine is involved in the light-induced inhibition of myopia. No differences in vitreal DOPAC were found when bright light was provided continuously or with 1:1 min alternating exposure with 500 lux. As previously described by others, wearing diffusers suppressed the expression of ZENK protein in glucagonergic amacrine cells (GACs) but neither continuous nor 1:1 min alternating bright to normal light could rescue the suppression of ZENK in GACs. While it is well known that light increases global retinal ZENK mRNA and protein levels, the changes of ZENK protein induced specifically in GACs by diffuser wear appear independent of light levels.

Myopia-What is Old and What is New?

The recent "boom of myopia," described predominantly for East Asia, is assumed to result from increasingly demanding education programs that include extensive near work (and perhaps also extensive use of computers) and little exposure to bright light as found outdoors. Already in 1892, Hermann Cohn stated that the prevalence of myopia is related to the educational level which is related to the economic status of a country. It is not much appreciated that the rates of myopia were already high among school children in central Europe in the middle of the 19th century, as described by Hermann Cohn. From extensive research in recent times, three major approaches have emerged to interfere with myopia progression in children: (1) promoting exposure to bright light and enforce outdoor activity, (2) adapting/improving optical corrections and visual behavior to generate inhibitory signals for eye growth in the retina, and (3) applying atropine eye drops at low doses. However, Hermann Cohn had already proposed that low luminances during school work promote myopia development and requested that lighting in the classrooms needs to be at least "10 meter candles" (equivalent to an illuminance of 10 lux). Different from today, he explained the link between low light and myopia by shorter reading distances that he observed at low luminances of the reading surface (<<1 cd/m). He suggested that short reading distances should be avoided in children and described several devices to control them. He further suggested that reading duration should be limited and urged myopes to choose professions that do not involve extensive near work. He also studied the effects of atropine against myopia but concluded that the side effects make it less useful than simply "3-4 weeks without reading." In summary, a number of his findings were re-discovered today, but they are now much better supported by data, and their interpretations have changed, at least in some aspects.