Thomas T. Norton

Regulation of the mechanical properties of tree shrew sclera by the visual environment

Experiments in several species have shown that the axial elongation rate of the developing eye can be increased or decreased by manipulating the visual environment, indicating that a visually guided emmetropization mechanism controls the enlargement of the vertebrate eye during postnatal development. Previous studies in tree shrews (Tupaia glis belangeri) suggest that regulation of the mechanical properties of the sclera may be an important part of the mechanism that controls the axial elongation rate in this mammal. To learn whether the mechanical properties of the sclera change when the axial elongation rate is increased or decreased under visual control, uniaxial mechanical tests were performed on 3-mm wide strips of tree shrew sclera. The creep rate was measured under 1, 3, and 5 g of tension, maintained for 30 min at each level. The modulus of elasticity was calculated from the elastic extension that occurred when the force was increased from 0 to 1 g, 1 to 3 g, and 3 to 5 g. Both were measured in the sclera of both eyes from animals exposed to four experimental conditions: (1) Normal development, at intervals from the day of natural eyelid opening (day 1 of visual experience [VE]) to greater than 5 years of age; (2) Monocular form deprivation (MD), for varying lengths of time; (3) Recovery from MD; (4) Monocular -5 D lens treatment. The creep rate was low in normal animals (1-2% elongation/h), did not change significantly between day 1 and day 75 of VE, and was not significantly different between the two eyes. Four days of MD produced a 200-300% increase in creep rate in the sclera from deprived eyes. Creep rate remained similarly elevated after 11 and 21 days of MD. After 2 days of recovery, which followed 11 days of MD, the creep rate of sclera from the recovering eyes was below normal levels. In animals that wore a monocular -5 D lens for up to 21 days, creep rate increased, and then decreased, in concert with the increase, and decrease, in axial elongation rate as the eyes compensated for the lens. The modulus of elasticity of the sclera was not significantly affected by any manipulation. The temporal correspondence between changes in axial elongation rate and changes in creep rate support the hypothesis that regulation of the time-dependent mechanical properties of fibrous mammalian sclera plays a role in controlling axial elongation rate during both normal emmetropization and the development of refractive errors.

Reduced extracellular matrix in mammalian sclera with induced myopia

The purpose of this study was to learn whether visual form deprivation, which produces myopia in the deprived eye, alters the scleral extracellular matrix in tree shrew, a mammal closely related to primates. Axial myopia was induced in 10 tree shrews by monocular deprivation imposed with a translucent diffuser. The other eye in each animal was an untreated control. After 21 days of deprivation the refractive state and axial component dimensions were measured and the eyes were assayed for levels of DNA, hydroxyproline, and sulfated glycosaminoglycans (GAGs) in samples of the sclera and the cornea. In comparison to the open control eye, the deprived eyes became myopic and elongated. In the sclera, DNA levels were not significantly changed from the control eye. Sulfated GAG levels were significantly lower in the deprived eyes, as compared to the control eyes, at the posterior pole (-15.6%), at the nasal equatorial region (-18.1%), and in the rest of the sclera (-11.6%). The hydroxyproline level was significantly lower only at the posterior pole (-11.8%). Levels of sulfated GAGs were significantly reduced relative to DNA and relative to hydroxyproline in the total sclera. No significant changes were found in the cornea. The lower level of sulfated GAGs throughout the sclera of the deprived eyes, as compared with the control eyes, suggests that the deprived sclera contained less proteoglycan, or that the proteoglycans were less glycosylated or less sulfated. In contrast, the regional reduction of hydroxyproline suggests that collagen accumulation was specifically reduced only at the posterior pole of deprived eyes. These results suggest that form deprivation slows or reverses the normal process of extracellular matrix accumulation in the sclera of this mammal. This may allow the sclera to be more distensible, permitting the vitreous chamber elongation and resultant myopia.

The development of experimental myopia and ocular component dimensions in monocularly lid-sutured tree shrews (Tupaia belangeri)

Tree shrews were monocularly deprived (MD) from the day of eye opening for periods of 15, 30, 45, 60 and 75 days. The initial structural change after 15 days of MD was a flattening of the corneal curvature in the deprived eye causing a hyperopic increase in refraction, relative to the fellow control eye. A relative myopia was first observed after 30 days of deprivation and increased as the length of MD increased. Animals monocularly deprived for 75 days consistently showed high degrees of myopia (greater -10 D). An increase in vitreous chamber depth was found after 30 days of deprivation and continued to increase, relative to the control eye, throughout the developmental period under investigation. There was a strong correlation (r = 0.84) between increase in vitreous chamber depth and the amount of experimentally-induced myopia. Anterior chamber depth was shallower in the deprived eyes of all animals. The crystalline lens was also consistently thinner in the deprived eye. Based on optical modeling, the observed myopia was consistent with the changes in ocular component dimensions. The susceptible period for experimental myopia begins about 15 days after eye opening.

Normal development of refractive state and ocular component dimensions in the tree shrew (Tupaia belangeri)

The normal development of refractive state, ocular components and simple visually-guided behaviors was examined in maternally-reared tree shrews. Six groups consisting of 5 animals each were anesthetized and examined after 0, 15, 30, 45, 60 and 75 days of normal binocular visual exposure. Measures in the 75-day group provided values for an improved schematic eye of the tree shrew. Cycloplegic refraction showed a marked hyperopia (+25 D) at eye opening which decreased rapidly during the first 15 days of visual exposure and stabilized near the value (+5 D) expected in an eye of this axial length (approx. 7.8 mm). Corneal radius increased slightly during development. Anterior segment depth, measured by A-scan ultrasonography, seemed to complete most of its development at an earlier age (15-30 days of visual exposure) than did other ocular parameters. Lens thickness increased steadily throughout development. Vitreous chamber depth increased rapidly until 15 days of visual exposure, and then decreased because the lens thickness increased more rapidly than axial length. Crude orienting to, and following of, large objects developed shortly after eye opening (median age at onset, 5 and 6 days, respectively). Triggered visual placing responses developed at about the same time that the refractive state completed the rapid drop from highly hyperopic values. The slowed rate of ocular development after 15 days of visual exposure may be related to increased retinal activity that is permitted by neural maturation and by the presence of a relatively well-focussed retinal image. The increased activity may influence the final dimensions of the eye to coordinate the axial length with the focal length of the eye.

Effect of interrupted lens wear on compensation for a minus lens in tree shrews.

BACKGROUND When a young animal wears a monocular minus (concave) lens that shifts the focal plane away from the cornea, the vitreous chamber elongates over a period of days, shifting the retinal location to compensate for the altered focal plane. We examined the effect of removing the lens for a portion of each day on the amount of compensation in tree shrews. METHODS Starting 24 days after natural eye opening, juvenile tree shrews wore a goggle frame that held a -5 D lens in front of one eye, with an open frame around the fellow control eye. The goggle was removed for 0, 0.5, 1, 2, or 7 h each day (N = 5, 5, 5, 5, and 3 animals per group, respectively), starting 0.5 h after the start of each 14 h light-on period. After 21 days of treatment, measures were made of the cycloplegic refractive state (streak retinoscopy) and the ocular component dimensions (A-scan ultrasound). Normal animals that experienced 14 h each day with no lens (N = 3) were also examined. RESULTS The treated eyes of the 0 h group developed full refractive compensation for the lens (treated eye - control eye, mean +/- SEM = -5.8+/-1.1 D) and had increased vitreous chamber depth (0.13+/-0.02 mm) and axial length (0.12+/-0.02 mm) relative to the untreated control eye. The groups in which the lens was removed for 0.5 and 1 h each day showed partial compensation for the -5 D lens, both in refractive state (-4.2+/-0.4 D; -2.9+/-1.6 D) and in vitreous chamber depth (0.12+/-0.02 mm; 0.09+/-0.02 mm). The 2, 7, and 14 h (normal) groups showed no significant refractive or axial compensation. In the 0.5 and 1 h groups, A-scan ultrasound showed a thinning of the region between the front of the retina and back of the sclera. CONCLUSIONS The eyes of tree shrews can tolerate altered monocular visual stimulation produced by a minus lens worn for 12 h of a 14-h light cycle without developing an induced myopia. However, when the lens is worn more than 12 of 14 h each day, compensation appears to increase linearly with decreased lens-off time. If the eyes of human children respond similarly to defocus from near work or other sources, it would seem that the defocus must be present almost all the time to induce myopia. If defocus contributes to human myopia through a compensation mechanism, then an increase in the amount of time that focused images are present should reduce myopic progression.

W-like response properties of interlaminar zone cells in the lateral geniculate nucleus of a primate (Galago crassicaudatus)

Recent anatomical studies have suggested that the cells located in the interlaminar zones (ILZs) of the primate dorsal lateral geniculate nucleus (LGN) relay visual information from the retina to the striate cortex in a manner similar to that of W-cells in the LGN of cat. In the present study, we examined this idea directly by recording the response properties of single cells localized to the ILZs in the prosimian primate, Galago crassicaudatus. The properties of the cells in the ILZs were found to be physiologically distinct from the X-like and Y-like properties of the parvocellular and magnocellular LGN layers. Moreover, the small cells located in the interlaminar zones were physiologically similar to the W-like cells found in the specialized small-celled koniocellular layers in these primates. As is the case with the koniocellular layer cells, the ILZ cells exhibited a broad range of properties which, as a group, were distinguished by the following characteristics: the ILZ cells had long latencies to stimulation of the optic chiasm (mean, 3.95 ms) and to antidromic stimulation from striate cortex (mean, 3.31 ms) and had relatively large receptive-field centers (mean, 1.79 degrees). They also had low maintained discharge rates (5.5 spikes/s), relatively long response latencies to light (mean onset, 82 ms; peak, 112 ms) and low peak firing rates (59 spikes/s). Few (25%) had standard receptive-field organization (ON-center, OFF-surround, or vice versa). Only 29% responded well to sine-wave gratings and all were influenced by non-visual (auditory and tactile) stimuli.(ABSTRACT TRUNCATED AT 250 WORDS)

The susceptible period for deprivation-induced myopia in tree shrew

To examine the susceptible period for deprivation-induced myopia, six groups of tree shrew pups (Tupaia glis belangeri) were monocularly deprived for 12 days with an opaque occluder starting 7, 15, 21, 33, 48, or 63 days after natural eyelid opening. Compared to the untreated fellow control eye, significant myopia and vitreous chamber elongation were produced by the deprivation in all six groups. The effect was greater in the middle three groups in comparison with the youngest and the two oldest groups and the amount of induced myopia and axial elongation was not proportional to the normal rate of axial growth. The peak period of susceptibility was between approximately 15 and 45 days after eye opening during the juvenile, slow-elongation phase of ocular development when the eye is within 7% of its adult axial length. Significant myopia and axial elongation were also induced in adult animals by 70 days of monocular deprivation. To examine recovery from deprivation-induced myopia, the occluder was removed at the end of the 12 day deprivation period. After an additional 48 days of binocular visual experience, no significant myopia was present in the previously deprived eyes in any experimental group. During the recovery period, the elongation rate of the previously deprived eyes was reduced in comparison with the control eyes while normal corneal flattening and lens development continued, thus reducing the myopia. No difference in corneal curvature, relative to the untreated control eyes, was found after deprivation or after the recovery period. Data are presented which suggests that changes in the thickness of the choroid may occur in this mammal during deprivation and recovery that are in the same direction, but of smaller magnitude, than those reported in the chicken. The results of this study provide evidence that visually guided emmetropization occurs in this mammalian species during a period of ocular development analogous to the juvenile period in humans.

Center/surround relationships of magnocellular, parvocellular, and koniocellular relay cells in primate lateral geniculate nucleus

As in other primates, the lateral geniculate nucleus (LGN) of the prosimian primate, bush baby (Galago crassicaudatus), contains three morphologically and physiologically distinct cell classes [magnocellular (M), parvocellular (P), and koniocellular (K)] (Norton & Casagrande, 1982; Casagrande & Norton, 1991). The present study examined quantitatively the center/surround relationships of cells in all three classes. Estimates of receptive-field center size (Rc) and sensitivity (Kc) and of surround size (Rs) and sensitivity (Ks) were obtained from 47 LGN relay cells by fitting a difference of Gaussians function to contrast-sensitivity data. For M and P cells, center size (Rc) increases with eccentricity but is about two times larger for M than for P cells at a given eccentricity. Surround size (Rs) increases with eccentricity for P but not for M or K cells. The center sensitivity (Kc) is inversely related to center size (Rc) and surround sensitivity (Ks) is inversely related to surround size (Rs) for cells in all classes, a result consistent with the sensitivity regulation that is produced by light adaptation. High spatial-frequency cutoff (acuity) is inversely related to center size (Rc). However, the peak contrast sensitivity is relatively independent of Rc. The ratio of the integrated strength (volume) of the surround to the volume of the center remains relatively constant (median, 0.87) across all three cell classes. This ratio is an excellent predictor of a cells rolloff in contrast sensitivity at low spatial frequencies: cells with a low surround/center ratio have less low-frequency rolloff. Although M, P, and K cells generally display similar center/surround relationships, differences in center size and the other parameters between the classes distinguish most M, P, and K cells. These findings demonstrate that both similarities and differences in the visual-response properties of primate LGN cells in these three parallel afferent pathways can be explained by basic center/surround relationships.

Lid-suture myopia in tree shrews with retinal ganglion cell blockade.

To determine whether central communication of retinal signals is necessary for the development of an experimentally induced myopia, tree shrews were exposed to monocular deprivation (MD) while the action potentials of retinal cells in the deprived eye were blocked with intravitreally injected tetrodotoxin (TTX-MD animals). TTX injections (0.6 microgram in 3 microL) and MD began about 15 days after eye opening, at the start of the susceptible period for the development of lid-suture myopia. Six injections were given, one every second day to produce 12 days of MD and TTX-blockade. Control TTX animals (TTX-open) received TTX in one eye, but not MD, on the same injection schedule and were always found to be behaviorally unresponsive to visual stimuli through the injected eye indicating that TTX blocked central communication of action potentials. Other control animals received intravitreally injected saline in either an open eye (saline-open), or an MD eye (saline-MD). A sham-injected group (sham-inj-MD) received MD and all anesthetic and surgical manipulations except for penetration of the sclera. In all groups, one eye in each animal was an untreated control. Two effects were found. All MD groups, including the TTX-MD animals, developed a significant vitreous chamber elongation in the deprived eye, indicating that an experimental myopia developed despite ganglion cell blockade. Thus, retinal mechanisms in tree shrew can detect the presence of a degraded visual image and produce an experimental myopia that does not depend on the receipt of visual messages by central neural structures. In addition, eyes in which the sclera was punctured had smaller vitreous chamber depths than comparable uninjected eyes, indicating that puncturing the sclera reduced the normal elongation. These data suggest that forces within the eye normally contribute to its expansion and may be resisted by the choroid and/or the sclera.

Reduction in choroidal blood flow occurs in chicks wearing goggles that induce eye growth toward myopia

Goggles that degrade the retinal image produce axial enlargement of the ocular globe and large myopic refractive errors. Many authors have assumed that visual image degradation itself leads to myopia. Hodos and co-authors have shown, however, that goggled eyes in chicks are considerably warmer than normal. Such temperature changes may either underlie or be a consequence of alterations in choroidal blood flow (CBF). Since alterations in CBF could affect eye growth, we explored the effect of monocular goggling on CBF in chicks. Plastic goggles were glued over one eye in four-day old chicks and the goggles were left in place for 12 or 14 days. Fourteen days after the goggling, CBF was measured using laser Doppler velocimetry. Three groups of chicks were studied: 1) chicks with goggles for 14 days; 2) chicks with goggles for 12 days followed by no goggles for the two days; 3) age matched non-goggled chicks. A -scan ultrasonography confirmed that the visual deprivation produced vitreous chamber elongation in the goggled eye and that the degree of elongation for the goggled eye was the same for the two goggled groups. The results were: 1) blood flow in non-goggled chicks was similar in both eyes; 2) blood flow was significantly reduced in the goggled eye in chicks wearing goggles for 14 days- 37% of control; and 3) blood flow was still significantly reduced in the goggled eye in chicks whose goggles were removed two days before measurement- 51% of control. These results show that CBF is reduced by goggles that result in myopic eye growth.(ABSTRACT TRUNCATED AT 250 WORDS)