Limited bandwidth short-wavelength light produces slowly-developing myopia in tree shrews similar to human juvenile-onset myopia.

During postnatal development, an emmetropization feedback mechanism uses visual cues to modulate the axial growth of eyes so that, with maturation, images of distant objects are in focus on the retina. If the visual cues indicate that the eye has become too long, it generates STOP signals that slow eye elongation. Myopia is a failure of this process where the eye becomes too long. The existing animal models of myopia have been essential in understanding the mechanics of emmetropization but use visual cues that lead to rapidly progressing myopia and don't match the stimuli that lead to human myopia. Form deprivation removes esssentially all spatial contrast. Minus lens wear accurately guides axial elongation to restore sharp focus: technically it is not a model of myopia! In contrast, childhood myopia involves a slow drift into myopia, even with the presence of clear images. We hypothesize that, in the modern visual environment, STOP signals are present but often are not quite strong enough to prevent myopic progression. Using tree shrews, small diurnal mammals closely related to primates, we have developed an animal model that we propose better represents this situation. We used limited bandwidth light to provide limited chromatic cues for emmetropization that are not quite enough to produce fully effective STOP signaling, resulting in a slow drift into myopia as seen in children. We hypothesize that this animal model of myopia may prove useful in evaluating anti-myopia therapies where form deprivation and minus lens wear would be too powerful.

Chromatically simulated myopic blur counteracts a myopiagenic environment.

There is a world-wide epidemic of myopia (nearsightedness), produced largely by human-made environmental visual cues that disrupt the emmetropization feedback mechanism that normally uses defocus cues to produce and maintain eyes in good focus. Previous studies have shown that the wavelength of light affects this process and that myopic defocus can slow the progression of myopia in children. We first asked if continuous exposure to a small cage with restricted viewing distance would produce an environmentally-induced myopia in tree shrews, small diurnal mammals closely related to primates. A group (n = 7) spent 11 days in a small cage with restricted viewing distance; one wall was a video display covered with Maltese crosses that included low-to-high spatial frequencies in the range visible to tree shrews. This group developed myopia (-1.2 ± 0.4 [stderr] D) that was significant relative to a colony group of seven animals (+1.0 ± 0.2 D) raised in mesh cages allowing more distant viewing. We then asked if chromatically-simulated myopic defocus, produced by blurring just the blue channel of the video display, would counteract this environmentally-induced myopia in a group of eight tree shrews. This group instead became significantly hyperopic (+4.0 ± 0.4 D) due to slowed axial elongation. These results demonstrate the high potency of chromatic cues in refractive regulation and may provide the basis for an anti-myopia treatment in humans.

DeBruyn and Casagrande manuscripts on tree shrew retinal ganglion cells as a basis for cross-species retina research.

The purpose of this brief communication is to make publicly available three unpublished manuscripts on the organization of retinal ganglion cells in the tree shrew. The manuscripts were authored in 1986 by Dr. Edward DeBruyn, a PhD student in the laboratory of the late Dr. Vivien Casagrande at Vanderbilt University. As diurnal animals closely related to primates, tree shrews are ideally suited for comparative analyses of visual structures including the retina. We hope that providing this basic information in a citable form inspires other groups to pursue further characterization of the tree shrew retina using modern techniques.

Amber light treatment produces hyperopia in tree shrews.

PURPOSE: Exposure to narrow-band red light, which stimulates only the long-wavelength sensitive (LWS) cones, slows axial eye growth and produces hyperopia in tree shrews and macaque monkeys. We asked whether exposure to amber light, which also stimulates only the LWS cones but with a greater effective illuminance than red light, has a similar hyperopia-inducing effect in tree shrews. METHODS: Starting at 24 ± 1 days of visual experience, 15 tree shrews (dichromatic mammals closely related to primates) received light treatment through amber filters (BPI 500/550 dyed acrylic) either atop the cage (Filter group, n = 8, 300-400 human lux) or fitted into goggles in front of both eyes (Goggle group, n = 7). Non-cycloplegic refractive error and axial ocular dimensions were measured daily. Treatment groups were compared with age-matched animals (Colony group, n = 7) raised in standard colony fluorescent lighting (100-300 lux). RESULTS: At the start of treatment, mean refractive errors were well-matched across the three groups (p = 0.35). During treatment, the Filter group became progressively more hyperopic with age (p < 0.001). By contrast, the Goggle and Colony groups showed continued normal emmetropization. When the treatment ended, the Filter group exhibited significantly greater hyperopia (mean [SE] = 3.5 [0.6] D) compared with the Goggle (0.2 [0.8] D, p = 0.01) and Colony groups (1.0 [0.2] D, p = 0.01). However, the refractive error in the Goggle group was not different from that in the Colony group (p = 0.35). Changes in the vitreous chamber were consistent with the refractive error changes. CONCLUSIONS: Exposure to ambient amber light produced substantial hyperopia in the Filter group but had no effect on refractive error in the Goggle group. The lack of effect in the Goggle group could be due to the simultaneous activation of the short-wavelength sensitive (SWS) and LWS cones caused by the scattering of the broad-band light from the periphery of the goggles.

How chromatic cues can guide human eye growth to achieve good focus.

The postnatal growing eye uses visual cues to actively control its own axial elongation to achieve and maintain sharp focus, a process termed emmetropization. The primary visual cue may be the difference in image sharpness as sensed by the arrays of short- and long-wavelength sensitive cone photoreceptors caused by longitudinal chromatic aberration: Shorter wavelengths focus in front of longer wavelengths. However, the sparse distribution of short-wavelength sensitive cones across the retina suggests that they do not have sufficient spatial sampling resolution for this task. Here, we show that the spacing of the short-wavelength sensitive cones in humans is sufficient for them, in conjunction with the longer wavelength cones, to use chromatic signals to detect defocus and guide emmetropization. We hypothesize that the retinal spacing of the short-wavelength sensitive cones in many mammalian species is an evolutionarily ancient adaption that allows the efficient use of chromatic cues in emmetropization.

Tree shrews do not maintain emmetropia in initially-focused narrow-band cyan light.

We asked if emmetropia, achieved in broadband colony lighting, is maintained in narrow-band cyan light that is well focused in the emmetropic eye, but does not allow for guidance from longitudinal chromatic aberrations (LCA) and offers minimal perceptual color cues. In addition, we examined the response to a -5 D lens in this lighting. Seven tree shrews from different litters were initially housed in broad-spectrum colony lighting. At 24 ± 1 days after eye opening (Days of Visual Experience, DVE) they were housed for 11 days in ambient narrow-band cyan light (peak wavelength 505 ± 17 nm) selected because it is in focus in an emmetropic eye. Perceptually, monochromatic light at 505 nm cannot be distinguished from white by tree shrews. While in cyan light, each animal wore a monocular -5 D lens (Cyan -5 D eyes). The fellow eye was the Cyan no-lens eye. Daily awake non-cycloplegic measures were taken with an autorefractor (refractive state) and with optical low-coherence optical interferometry (axial component dimensions). These measures were compared with the values of animals raised in standard colony fluorescent lighting: an untreated group (n = 7), groups with monocular form deprivation (n = 7) or monocular -5 D lens treatment (n = 5), or that experienced 10 days in total darkness (n = 5). Refractive state at the onset of cyan light treatment was low hyperopia, (mean ± SEM) 1.4 ± 0.4 diopters. During treatment, the Cyan no-lens eyes became myopic (-2.9 ± 0.3 D) whereas colony lighting animals remained slightly hyperopic (1.0 ± 0.2 D). Initially, refractions of the Cyan -5 D eyes paralleled the Cyan no-lens eyes. After six days, they gradually became more myopic than the Cyan no-lens eyes; at the end of treatment, the refractions were -5.4 ± 0.3 D, a difference of -2.5 D from the Cyan no-lens eyes. When returned to colony lighting at 35 ± 1 DVE, the no-lens eye refractions rapidly recovered towards emmetropia but, as expected, the refraction of the -5 D eyes remained near -5 D. Vitreous chamber depth in both eyes was consistent with the refractive changes. In narrow-band cyan lighting the emmetropization mechanism did not maintain emmetropia even though the light initially was well focused. We suggest that, as the eyes diverged from emmetropia, there were insufficient LCA cues for the emmetropization mechanism to utilize the developing myopic refractive error in order to guide the eyes back to emmetropia. However, the increased myopia in the Cyan -5 D eyes in the narrow-band light indicates that the emmetropization mechanism nonetheless detected the presence of the lens-induced refractive error and responded with increased axial elongation that partly compensated for the negative-power lens. These data support the conclusion that the emmetropization mechanism cannot maintain emmetropia in narrow-band lighting. The additional myopia produced in eyes with the -5 D lens shows that the emmetropization mechanism responds to multiple defocus-related cues, even under conditions where it is unable to use them to maintain emmetropia.

An opponent dual-detector spectral drive model of emmetropization.

In post-natal developing eyes a feedback mechanism uses optical cues to regulate axial growth so as to achieve good focus, a process termed emmetropization. However, the optical cues that the feedback mechanism uses have remained unclear. Here we present evidence that a primary visual cue may be the detection of different image statistics by the short-wavelength sensitive (SWS) and long-wavelength sensitive (LWS) cone photoreceptors, caused by longitudinal chromatic aberration (LCA). We use as a model system the northern tree shrew Tupaia belangeri, diurnal cone-dominated dichromatic mammals closely related to primates. We present an optical model in which the SWS and LWS photoreceptors each represent an image at different levels of defocus. The model posits that an imbalance between SWS and LWS image statistics directs eye growth towards the point at which these image statistics are in balance. Under spectrally broadband ("white") lighting, the focus of the eye is driven to a target point approximately in the middle of the visible spectrum, which is emmetropia. Calculations suggest that the SWS cone array, despite the sparse number of SWS cones, can plausibly detect the wavelength-dependent differences in defocus and guide refractive development. The model is consistent with the effects of various narrow-band illuminants on emmetropization in tree shrews. Simulations suggest that common artificial light spectra do not interfere with emmetropization. Simulations also suggest that multi-spectral multi-focal lenses, where the different optical zones of a multifocal lens have different spectral filtering properties, could be an anti-myopia intervention.

Gene expression signatures in tree shrew sclera during recovery from minus-lens wear and during plus-lens wear.

Purpose: In juvenile tree shrews that have developed minus lens-induced myopia, if lens treatment is discontinued, refractive recovery (REC) occurs. However, in age-matched juvenile animals, plus-lens wear (PLW) produces little refractive change, although the visual stimulus (myopia) is similar (an "IGNORE" response). Because the sclera controls axial elongation and refractive error, we examined gene expression in the sclera produced by PLW and compared it with the gene expression signature produced by REC to learn whether these similar refractive conditions produce similar, or differing, scleral responses. Methods: Eight groups of tree shrews (n = 7 per group) were examined. Four groups wore a monocular -5 D lens for 11 days until 35 days of visual experience (DVE). Lens wear was then discontinued, and the animals recovered for 0 h (REC-0), 2 h (REC-2h), 1 day (REC-1d), or 4 days (REC-4d). Starting at 35 DVE, three groups wore a monocular +5 D lens for 2 h (PLW-2h), 1 day (PLW-1d), or 4 days (PLW-4d). A normal group (PLW-0) was examined at 38 DVE to provide baseline measures. Using quantitative real-time PCR (qPCR), we examined scleral mRNA levels in recovering, plus-lens treated, and untreated control eyes for 55 candidate genes whose protein products included signaling molecules, metallopeptidases (MPs) and their inhibitors (tissue inhibitors of metallopeptidases [TIMPs]), and extracellular matrix proteins. Results: No refractive recovery was measured in the REC-2h group. The scleral mRNA expression pattern for recovering versus untreated control eyes after 2 h of recovery was similar to that found for the group (REC-0) that had no recovery time. Many genes in both groups still had downregulated expression in the treated eyes versus the control eyes. The REC-1d group showed little refractive recovery (0.1 ± 0.1 D, mean ± standard error of the mean [SEM]), and the mRNA expression pattern was similar to that of the REC-2h group, but had fewer statistically significantly downregulated genes in the recovering eyes. The REC-4d group recovered refractively by 2.6 ± 0.4 D, and displayed a "STOP" gene expression signature of mostly upregulated mRNA expression in the recovering eyes compared with the untreated control eyes. The PLW-0 (normal) group and the PLW-2h group showed no statistically significant differential gene expression. The PLW-1d group showed a small hyperopic shift (0.1 ± 0.2 D). Two genes were differentially expressed: NPR3 was upregulated in the plus lens-wearing eyes, and IGF1 was downregulated. The PLW-4d group showed a similar hyperopic shift (0.3 ± 0.4 D), confirming that the plus lens-induced 5 D of myopia produced little refractive change. In the sclera, there was an IGNORE pattern of general differential upregulation of genes in the treated eyes (22 upregulated, one downregulated) that was distinct from the STOP signature found in recovery. Ten genes were upregulated in the REC-4d group and the PLW-4d group. However, ten other genes were differentially expressed in recovery, but not in plus-lens wear, while 12 genes were differentially expressed in plus-lens wear but not in recovery. Conclusions: One day of recovery is not long enough for the emmetropization mechanism to produce significant gene expression changes in the sclera or refractive recovery. After 4 days, recovery and plus-lens wear produced altered scleral gene expression, but the patterns ("signatures") differed as to which genes showed altered expression, and whether the gene expression was up- or downregulated. Thus, myopia produced altered scleral mRNA expression in recovery and plus-lens wear, confirming that signals initiated by the retina reached the sclera, but the sclera in the elongated recovering eye responded differently from a normal sclera. This might have occurred because the recovering-eye sclera had remodeled during minus-lens compensation, making the sclera respond differently to the signals initiated by the retina. However, the myopia-produced retinal signals in plus lens-wearing animals also may have differed from those in the recovering eyes by the time the signals passed through the RPE and choroid to reach the sclera.

Noninvasive imaging of the tree shrew eye: Wavefront analysis and retinal imaging with correlative histology.

Tree shrews are small mammals with excellent vision and are closely related to primates. They have been used extensively as a model for studying refractive development, myopia, and central visual processing and are becoming an important model for vision research. Their cone dominant retina (∼95% cones) provides a potential avenue to create new damage/disease models of human macular pathology and to monitor progression or treatment response. To continue the development of the tree shrew as an animal model, we provide here the first measurements of higher order aberrations along with adaptive optics scanning light ophthalmoscopy (AOSLO) images of the photoreceptor mosaic in the tree shrew retina. To compare intra-animal in vivo and ex vivo cone density measurements, the AOSLO images were matched to whole-mount immunofluorescence microscopy. Analysis of the tree shrew wavefront indicated that the optics are well-matched to the sampling of the cone mosaic and is consistent with the suggestion that juvenile tree shrews are nearly emmetropic (slightly hyperopic). Compared with in vivo measurements, consistently higher cone density was measured ex vivo, likely due to tissue shrinkage during histological processing. Tree shrews also possess massive mitochondria ("megamitochondria") in their cone inner segments, providing a natural model to assess how mitochondrial size affects in vivo retinal imagery. Intra-animal in vivo and ex vivo axial distance measurements were made in the outer retina with optical coherence tomography (OCT) and transmission electron microscopy (TEM), respectively, to determine the origin of sub-cellular cone reflectivity seen on OCT. These results demonstrate that these megamitochondria create an additional hyper-reflective outer retinal reflective band in OCT images. The ability to use noninvasive retinal imaging in tree shrews supports development of this species as a model of cone disorders.

Matching the LenStar optical biometer to A-Scan ultrasonography for use in small animal eyes with application to tree shrews.

We describe an analysis strategy to obtain ultrasonography-matched axial dimensions of small animal eyes using the LenStar biometer. The LenStar optical low-coherence reflectometer is an attractive device for animal research due to its high precision, non-invasiveness, and the ability to measure the axial dimensions of cornea, anterior chamber, lens, vitreous chamber, and axial length. However, this optical biometer was designed for clinical applications in human eyes and its internal analysis provides inaccurate values when used on small eyes due to species-dependent differences in refractive indices and relative axial dimensions. The LenStar uses a near infrared light source to measure optical path lengths (OPLs) that are converted by the LenStar's EyeSuite software into geometrical lengths (GLs) based on the refractive indices and axial dimensions of the human eye. We present a strategy that extracts the OPLs, determines refractive indices specific for the small animal eye of interest and then calculates corrected GLs. The refractive indices are obtained by matching the LenStar values to ultrasonography values in the same eyes. As compared to ultrasounography, we found that the internal calculations of the LenStar underestimate the axial dimensions of all ocular compartments of the tree shrew eye: anterior segment depth by 6.17±4.50%, lens thickness by 1.37±3.06%, vitreous chamber depth by 29.23±2.35%, and axial length by 10.62±1.75%. Using tree shrew-specific refractive indices, the axial dimensions closely matched those measured by ultrasonography for each compartment. Our analysis strategy can be easily translated to other species by obtaining a similar paired data set using ultrasonography and LenStar, and applying our step by step procedures.

Juvenile Tree Shrews Do Not Maintain Emmetropia in Narrow-band Blue Light.

SIGNIFICANCE: In spectrally broad-band light, an emmetropization mechanism in post-natal eyes uses visual cues to modulate the growth of the eye to achieve and maintain near emmetropia. When we restricted available wavelengths to narrow-band blue light, juvenile tree shrews (diurnal dichromatic mammals closely related to primates) developed substantial refractive errors, suggesting that feedback from defocus-related changes in the relative activation of long- and short-wavelength-sensitive cones is essential to maintain emmetropia. PURPOSE: The purpose of this study was to examine the effects of narrow-band ambient blue light on refractive state in juvenile tree shrews that had completed initial emmetropization (decrease from hyperopia toward emmetropia). METHODS: Animals were raised in fluorescent colony lighting until they began blue-light treatment at 24 days of visual experience, at which age they had achieved age-normal low hyperopia (mean ± SEM refractive error, 1.2 ± 0.5 diopters). Arrays of light-emitting diodes placed atop the cage produced wavelengths of 457 (five animals) or 464 nm (five animals), flickered in a pseudo-random pattern (temporally broad band). A third group of five animals was exposed to steady 464-nm blue light. Illuminance on the floor of the cage was 300 to 500 human lux. Noncycloplegic autorefractor measures were made daily for a minimum of 11 days and up to 32 days. Seven age-matched animals were raised in colony light. RESULTS: The refractive state of all blue-treated animals moved outside the 95% confidence limits of the colony-light animals' refractions. Most refractions first moved toward hyperopia. Then the refractive state decreased monotonically and, in some animals, passed through emmetropia, becoming myopic. CONCLUSIONS: From the tree shrew cone absorbance spectra, the narrow-band blue light stimulated both long-wavelength-sensitive and short-wavelength-sensitive cones, but the relative activation would not change with the refractive state. This removed feedback from longitudinal chromatic aberration that may be essential to maintain emmetropia.

Axial Elongation in Myopic Children and its Association With Myopia Progression in the Correction of Myopia Evaluation Trial.

OBJECTIVES: Describe axial elongation using 14-year longitudinal data in a large, ethnically diverse group of myopic children, estimate age and axial length (AL) at stabilization, and evaluate associations between the progression and stabilization of AL and myopia. METHODS: Axial length was measured by A-scan ultrasonography annually. Axial length data were fit with individual polynomial functions and curve-based parameters (AL at stabilization and age at stabilization when annual rate of axial elongation ≤0.06 mm) were estimated. For myopia progression, noncycloplegic spherical equivalent refractions were fit with Gompertz functions. RESULTS: Four hundred thirty-one participants, with AL and myopia data fit successfully, were classified into four cohorts: Younger (n=30); Older (n=334); AL Stabilized at Baseline (n=19); and AL Not Stabilized (n=48). At AL stabilization, for participants in the Younger and Older Cohorts, mean (SD) age and AL were 16.3 (2.4) years and 25.2 (0.9) mm, respectively. No associations were found between age at AL stabilization and ethnicity, sex, or number of myopic parents. At stabilization, sex and number of myopic parents (both P<0.003), but not ethnicity, were significantly associated with AL. Axial length and myopia progression curves were highly correlated overall (all r>0.77, P<0.0001). However, unlike AL, the amount of myopia did not differ significantly between males and females. CONCLUSIONS: In most of the participants, AL increased rapidly at younger ages and then slowed and stabilized. The close association between growth and stabilization of AL and myopia is consistent with the suggestion that axial elongation is the primary ocular component in myopia progression and stabilization.

The hyperopic effect of narrow-band long-wavelength light in tree shrews increases non-linearly with duration.

During postnatal refractive development, an emmetropization mechanism uses refractive error to modulate the growth rate of the eye. Hyperopia (image focused behind the retina) produces what has been described as "GO" signaling that increases growth. Myopia (image focused in front of the retina) produces "STOP" signaling that slows growth. The interaction between GO and STOP conditions is non-linear; brief daily exposure to STOP counteracts long periods of GO. In young tree shrews, long-wavelength (red) light, presented 14 h per day, also appears to produce STOP signals. We asked if red light also shows temporal non-linearity; does brief exposure slow the normal decrease in hyperopia in infant animals? At 11 days after eye opening (DVE), infant tree shrews (n = 5/group) began 13 days of daily treatment (red LEDs, 624 ±â€¯10 or 636 ±â€¯10 nm half peak intensity bandwidth) at durations of 0 h (normal animals, n = 7) or 1, 2, 4, or 7 h. Following each daily red period, colony lighting resumed. A 14 h red group had no colony lights. Refractive state was measured daily; ocular component dimensions at the end of the 13-day red-light period. Even 1 h of red light exposure produced some hyperopia. The average hyperopic shift from normal rose exponentially with duration (time constant 2.5 h). Vitreous chamber depth decreased non-linearly with duration (time constant, 3.3 h). After red treatment was discontinued, refractions in colony lighting recovered toward normal; the initial rate was linearly related to the amount of hyperopia. The red light may produce STOP signaling similar to myopic refractive error.

Altered gene expression in tree shrew retina and retinal pigment epithelium produced by short periods of minus-lens wear.

Hyperopic refractive error is detected by retinal neurons, which generate GO signals through a direct emmetropization signaling cascade: retinal pigment epithelium (RPE) into choroid and then into sclera, thereby increasing axial elongation. To examine signaling early in this cascade, we measured gene expression in the retina and RPE after short exposure to hyperopia produced by minus-lens wear. Gene expression in each tissue was compared with gene expression in combined retina + RPE. Starting 24 days after normal eye opening, three groups of juvenile tree shrews (n = 7 each) wore a monocular -5 D lens. The untreated fellow eye served as a control. The "6h" group wore the lens for 6 h; the "24h" group wore the lens for 24 h; each group provided separate retina and RPE tissues. Group "24hC" wore the lens for 24 h and provided combined retina + RPE tissue. Quantitative PCR was used to measure the relative differences (treated eye vs. control eye) in mRNA levels for 66 candidate genes. In the retina after 6 h, mRNA levels for seven genes were significantly regulated: EGR1 and FOS (early intermediate genes) were down-regulated in the treated eyes. Genes with secreted protein products, BMP2 and CTGF, were down-regulated, whilst FGF10, IL18, and SST were up-regulated. After 24 h the pattern changed; only one of the seven genes still showed differential expression; BMP2 was still down-regulated. Two new genes with secreted protein products, IGF2 and VIP, were up-regulated. In the RPE, consistent with its role in receiving, processing, and transmitting GO signaling, differential expression was found for genes whose protein products are at the cell surface, intracellular, in the nucleus, and are secreted. After 6 h, mRNA levels for 17 genes were down-regulated in the treated eyes, whilst four genes (GJA1, IGF2R, LRP2, and IL18) were up-regulated. After 24 h the pattern was similar; mRNA levels for 14 of the same genes were still down-regulated; only LRP2 remained up-regulated. mRNA levels for six genes no longer showed differential expression, whilst nine genes, not differentially expressed at 6 h, now showed differential expression. In the combined retina + RPE after 24 h, mRNA levels for only seven genes were differentially regulated despite the differential expression of many genes in the RPE. Four genes showed the same expression in combined tissue as in retina alone, including up-regulation of VIP despite significant VIP down-regulation in RPE. Thus, hyperopia-induced GO signaling, as measured by differential gene expression, differs in the retina and the RPE. Retinal gene expression changed between 6 h and 24 h of treatment, suggesting evolution of the retinal response. Gene expression in the RPE was similar at both time points, suggesting sustained signaling. The combined retina + RPE does not accurately represent gene expression in either retina or, especially, RPE. When gene expression signatures were compared with those in choroid and sclera, GO signaling, as encoded by differential gene expression, differs in each compartment of the direct emmetropization signaling cascade.

Long-wavelength (red) light produces hyperopia in juvenile and adolescent tree shrews.

In infant tree shrews, exposure to narrow-band long-wavelength (red) light, that stimulates long-wavelength sensitive cones almost exclusively, slows axial elongation and produces hyperopia. We asked if red light produces hyperopia in juvenile and adolescent animals, ages when plus lenses are ineffective. Animals were raised in fluorescent colony lighting (100-300 lux) until they began 13days of red-light treatment at 11 (n=5, "infant"), 35 (n=5, "juvenile") or 95 (n=5, "adolescent") days of visual experience (DVE). LEDs provided 527-749 lux on the cage floor. To control for the higher red illuminance, a fluorescent control group (n=5) of juvenile (35 DVE) animals was exposed to ∼975 lux. Refractions were measured daily; ocular component dimensions at the start and end of treatment and end of recovery in colony lighting. These groups were compared with normals (n=7). In red light, the refractive state of both juvenile and adolescent animals became significantly (P<0.05) hyperopic: juvenile 3.9±1.0 diopters (D, mean±SEM) vs. normal 0.8±0.1D; adolescent 1.6±0.2D vs. normal 0.4±0.1D. The fluorescent control group refractions (0.6±0.3D) were normal. In red-treated juveniles the vitreous chamber was significantly smaller than normal (P<0.05): juvenile 2.67±0.03mmvs. normal 2.75±0.02mm. The choroid was also significantly thicker: juvenile 77±4µmvs. normal 57±3µm (P<0.05). Although plus lenses do not restrain eye growth in juvenile tree shrews, the red light-induced slowed growth and hyperopia in juvenile and adolescent tree shrews demonstrates that the emmetropization mechanism is still capable of restraining eye growth at these ages.

Optic Nerve Tilt, Crescent, Ovality, and Torsion in a Multi-Ethnic Cohort of Young Adults With and Without Myopia.

Purpose: The purpose of this article is to evaluate optic nerve head (ONH) characteristics in an ethnically diverse cohort of young U.S. adults. Methods: In this study, 409 myopes and 206 nonmyopes (median age 22 years) completed measures including biometry and spectral domain optical coherence tomography from enface (ovality and torsion) and cross-sectional (tilt and crescent width) scans. Associated factors were evaluated using multivariable models. Results: In myopic versus nonmyopic right eyes, median tilt (6.0° vs. 2.4°; P < 0.0001) and frequency of crescents (49% vs. 10%; P < 0.0001) were higher in myopes. Right eyes with crescents had higher median tilts (8.8° [myopic], 9.0° [nonmyopic]) than those without crescent (2.5° [myopic], 2.1° [nonmyopic]), irrespective of refractive group (both P < 0.0001). Torsion was similar between groups, with a slight difference in ovality (0.89 vs. 0.91; P < 0.03). Data in the left eyes were similar, and modeling was done only for the right myopic eyes. Multivariable models showed that an increased tilt was associated with ethnicity (P < 0.001), the presence of crescent (P < 0.001), and smaller ONH diameter (P < 0.0031), with interactions between ethnicity and crescent (P = 0.002). Specifically, ONH tilt was significantly higher in Asian eyes without crescent (P < 0.0001 for all comparisons), and crescent width was associated with increased tilt in non-Asian eyes (P < 0.02). Crescent width was associated with ethnicity (greatest in Asians) and disc tilt. Interactions were observed between tilt and ethnicity, whereby tilt had a greater effect on crescent width in non-Asian eyes, and crescent width was associated with increased tilt in non-Asian eyes. Conclusions: The data clarify the influence of ethnicity and myopia on ONH characteristics in young adults and may inform future studies of biomechanical properties or of retinal pathology of the myopic eye.

Intravitreally-administered dopamine D2-like (and D4), but not D1-like, receptor agonists reduce form-deprivation myopia in tree shrews.

We examined the effect of intravitreal injections of D1-like and D2-like dopamine receptor agonists and antagonists and D4 receptor drugs on form-deprivation myopia (FDM) in tree shrews, mammals closely related to primates. In eleven groups (n = 7 per group), we measured the amount of FDM produced by monocular form deprivation (FD) over an 11-day treatment period. The untreated fellow eye served as a control. Animals also received daily 5 µL intravitreal injections in the FD eye. The reference group received 0.85% NaCl vehicle. Four groups received a higher, or lower, dose of a D1-like receptor agonist (SKF38393) or antagonist (SCH23390). Four groups received a higher, or lower, dose of a D2-like receptor agonist (quinpirole) or antagonist (spiperone). Two groups received the D4 receptor agonist (PD168077) or antagonist (PD168568). Refractions were measured daily; axial component dimensions were measured on day 1 (before treatment) and day 12. We found that in groups receiving the D1-like receptor agonist or antagonist, the development of FDM and altered ocular component dimensions did not differ from the NaCl group. Groups receiving the D2-like receptor agonist or antagonist at the higher dose developed significantly less FDM and had shorter vitreous chambers than the NaCl group. The D4 receptor agonist, but not the antagonist, was nearly as effective as the D2-like agonist in reducing FDM. Thus, using intravitreally-administered agents, we did not find evidence supporting a role for the D1-like receptor pathway in reducing FDM in tree shrews. The reduction of FDM by the dopamine D2-like agonist supported a role for the D2-like receptor pathway in the control of FDM. The reduction of FDM by the D4 receptor agonist, but not the D4 antagonist, suggests an important role for activation of the dopamine D4 receptor in the control of axial elongation and refractive development.

The wavelength composition and temporal modulation of ambient lighting strongly affect refractive development in young tree shrews.

Shortly after birth, the eyes of most animals (including humans) are hyperopic because the short axial length places the retina in front of the focal plane. During postnatal development, an emmetropization mechanism uses cues related to refractive error to modulate the growth of the eye, moving the retina toward the focal plane. One possible cue may be longitudinal chromatic aberration (LCA), to signal if eyes are getting too long (long [red] wavelengths in better focus than short [blue]) or too short (short wavelengths in better focus). It could be difficult for the short-wavelength sensitive (SWS, "blue") cones, which are scarce and widely spaced across the retina, to detect and signal defocus of short wavelengths. We hypothesized that the SWS cone retinal pathway could instead utilize temporal (flicker) information. We thus tested if exposure solely to long-wavelength light would cause developing eyes to slow their axial growth and remain refractively hyperopic, and if flickering short-wavelength light would cause eyes to accelerate their axial growth and become myopic. Four groups of infant northern tree shrews (Tupaia glis belangeri, dichromatic mammals closely related to primates) began 13 days of wavelength treatment starting at 11 days of visual experience (DVE). Ambient lighting was provided by an array of either long-wavelength (red, 626 ± 10 nm) or short-wavelength (blue, 464 ± 10 nm) light-emitting diodes placed atop the cage. The lights were either steady, or flickering in a pseudo-random step pattern. The approximate mean illuminance (in human lux) on the cage floor was red (steady, 527 lux; flickering, 329 lux), and blue (steady, 601 lux; flickering, 252 lux). Refractive state and ocular component dimensions were measured and compared with a group of age-matched normal animals (n = 15 for refraction (first and last days); 7 for ocular components) raised in broad spectrum white fluorescent colony lighting (100-300 lux). During the 13 day period, the refraction of the normal animals decreased from (mean ± SEM) 5.8 ± 0.7 diopters (D) to 1.5 ± 0.2 D as their vitreous chamber depth increased from 2.77 ± 0.01 mm to 2.80 ± 0.03 mm. Animals exposed to red light (both steady and flickering) remained hyperopic throughout the treatment period so that the eyes at the end of wavelength treatment were significantly hyperopic (7.0 ± 0.7 D, steady; 4.7 ± 0.8 D, flickering) compared with the normal animals (p < 0.01). The vitreous chamber of the steady red group (2.65 ± 0.03 mm) was significantly shorter than normal (p < 0.01). On average, steady blue light had little effect; the refractions paralleled the normal refractive decrease. In contrast, animals housed in flickering blue light increased the rate of refractive decrease so that the eyes became significantly myopic (-2.9 ± 1.3 D) compared with the normal eyes and had longer vitreous chambers (2.93 ± 0.04 mm). Upon return to colony lighting, refractions in all groups gradually returned toward emmetropia. These data are consistent both with the hypothesis that LCA can be an important visual cue for postnatal refractive development, and that short-wavelength temporal flicker provides an important cue for assessing and signaling defocus.

A sloped piecemeal Gaussian model for characterising foveal pit shape.

PURPOSE: High-quality optical coherence tomography (OCT) macular scans make it possible to distinguish a range of normal and diseased states by characterising foveal pit shape. Existing mathematical models lack the flexibility to capture all known pit variations and thus characterise the pit with limited accuracy. This study aimed to develop a new model that provides a more robust characterisation of individual foveal pit variations. METHODS: A Sloped Piecemeal Gaussian (SPG) model, consisting of a linear combination of a tilted line and a piecemeal Gaussian function (two halves of a Gaussian connected by a separate straight line), was developed to fit retinal thickness data with the flexibility to characterise different degrees of pit asymmetry and pit bottom flatness. It fitted the raw pit data between the two rims of the fovea to improve accuracy. The model was tested on 3488 macular scans from both eyes of 581 young adults (376 myopes and 206 non-myopes, mean (S.D.) age 21.9 (1.4) years). Estimates for retinal thickness, wall height and slope, pit depth and width were derived from the best-fitting model curve. Ten variations of Gaussian and Difference of Gaussian models were fitted to the same scans and compared with the SPG model for goodness of fit (by Root mean square error, RMSE), model complexity (by the Bayesian Information Criteria) and model fidelity. RESULTS: The SPG model produced excellent goodness of fit (mean RMSE = 4.25 and 3.89 µm; 95% CI: 4.20, 4.30 and 3.86, 3.93 for fitting horizontal and vertical profiles respectively). The SPG model showed pit asymmetry, with average nasal walls 17.6 (11.6) µm higher and 0.96 (0.61)° steeper than temporal walls and average superior walls 7.0 (12.2) µm higher and 0.41 (0.65)° steeper than the inferior walls. The SPG model also revealed a continuum of human foveal shapes, from round bottoms to extended flat bottoms (up to 563 µm). 49.1% of foveal profiles were best fitted with a flat bottom >30 µm wide. Compared with the other tested models, the SPG was the preferred model overall based on the Bayesian Information Criteria. CONCLUSIONS: The SPG is a new parsimonious mathematical model that improves upon other models by accounting for wall asymmetry and flat pit bottoms, providing an excellent fit and more faithful characterisation of typical foveal pit shapes and their known variations. This new model may be helpful in distinguishing normal foveal shape variations by refractive status as well by other characteristics such as sex, ethnicity and age.