Download Poster Session

ARVO 2015 Annual Meeting Abstracts
275 Myopia
Monday, May 04, 2015 3:45 PM–5:30 PM
Exhibit Hall Poster Session
Program #/Board # Range: 2148–2182/B0001–B0035
Organizing Section: Anatomy and Pathology/Oncology
Contributing Section(s): Clinical/Epidemiologic Research
Program Number: 2148 Poster Board Number: B0001
Presentation Time: 3:45 PM–5:30 PM
The effect of combination of white and monochromatic light on
eye growth of normal chicks
Rachel Ka-man Chun1, Danyang Wang1, 2, King Kit Li1, Thomas
C Lam1, Quan Liu2, Chi Ho To1, 2. 1Laboratory of Experimental
Optometry, Centre for Myopia Research, School of Optometry, The
Hong Kong Polytechnic University, Hong Kong, Hong Kong; 2State
Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center,
Sun Yat-sen University, Guangzhou, China.
Purpose: To examine the effect of combination of white and
monochromatic light on eye growth of normal chicks
Methods: White Leghorn chicks aged 4 days (n = 8 in each group,
three groups in total) were raised in a cabinet of 80 x 80 x 125 cm
under three different lighting. They were white (585 nm), white
(585 nm) and red (630 nm), white (585 nm) and blue (450 nm).
The average luminances in these three environments were around
260 lux. The ratio of those light combination was 50:50. The ocular
parameters and refractive errors were measured before and after
14 days of exposure. The ocular parameters were measured by
high frequency A-scan ultrasonography while the refractive errors
were examined by streak retinoscopy. Percentage changes in the
ocular parameters and refractive errors relative to the baseline
were calculated and compared among different groups by two-way
ANOVA and Fisher’s least significant difference (LSD) post hoc test.
Results: After 14 days of exposure, the percentage increase in
anterior chamber depth (ACD), lens thickness, vitreous chamber
depth (VCD), retinal thickness and axial length were found to be
significantly greater in chicks raised under white and red light when
compared with those under white light only (p < 0.001). Besides,
more myopic shift was shown in the chicks under white and red light
(p < 0.05). However, the choroid did not demonstrate the thinning
as expected but significant thickening was found (mean choroidal
thickness ± SEM; white vs. white and red; 264.3 ± 4.43 mm vs. 280.8
± 5.18 mm).
Chicks raised in white and blue light had a smaller increase in ACD,
lens thickness, VCD and axial length when compared with chicks
under white light only (p< 0.001). Significant thinning of the retina
was found (percentage change of the retinal thickness ± SEM; white
vs. white and blue; -6.1 ± 1.2% vs -11.6 ± 0.7%, p = 0.001). Changes
in refractive errors and choroidal thickness were comparable to the
chicks under white light only.
Conclusions: Combination of the white and monochromatic light
provided two peak wavelengths across the spectrum. It significantly
affected the eye growth of normal chicks. Greater elongation of
eyeball was found in the chicks under white and red light than those
under white light only whereas ocular growth was slower in the
chicks under white and blue light. Monochromatic light seems to play
a role in modulating the ocular growth in chicks.
Commercial Relationships: Rachel Ka-man Chun, None;
Danyang Wang, None; King Kit Li, None; Thomas C Lam, None;
Quan Liu, None; Chi Ho To, None
Support: This work was supported by research grants GU986,
GU839, GUA32 and GYK89 from The Hong Kong Polytechnic
University and the Henry G. Leong Endowed Professorship in
Elderly Vision Health.
Program Number: 2149 Poster Board Number: B0002
Presentation Time: 3:45 PM–5:30 PM
Effects of 430nm monochromatic light on defocus-induced
myopia in guinea pigs
Yi-Feng Qian1, rui liu2, jinhui dai2. 1Department of Ophthalmology,
First Affiliated Hospital of Soochow University, Suzhou, China;
2
Department of Opthalmology, Eye & ENT Hospital of Fudan
University, Shanghai, China.
Purpose: To investigate the effects of 430nm monochromatic light
on defocus-induced myopia in guinea pigs.
Methods: Eighteen 2-week-old pigmented guinea pigs were
randomly assigned to two groups based on the mode of illumination:
short-wavelength light (SL) for 8 weeks and broad-band white light
(BL) for 8 weeks. All animals of the two groups were worn -5D
lenses on right eye. Biometric and refractive measurements were then
performed every 2 weeks. The illuminative parameters of all groups
were identical and the light quantum number was 3×10-4μmolcm-2s-1.
Results: After the beginning of the experiment, the right eyes of the
two groups decreased in refraction. At the end of the experiment,
relative myopia of the right eye was about 2.72D in the SL and about
3.03D in the BL when compared with the fellow eye. But a relative
hyperopia, about 1.2D, was induced in the SL compared with the BL
group in the end. From 4 to 8 week, there was significant difference
in radius of corneal curvature between the two eyes of the SL group.
But, there was no significant difference in corneal curvature between
the two eyes of the BL group. At the end of the experiment, there was
significant difference in radius of corneal curvature between the right
eyes of the two groups. The difference was not significant in vitreous
length of right eye between the two groups from beginning to the
end of experiment. There was no significant difference in vitreous
length between the two eyes of the SL group in the end. But finally,
significant difference existed in vitreous length between the two eyes
of the BL group. There were no significant inter-group or intra-group
differences in length of anterior segment and Lens thickness.
Conclusions: 430nm monochromatic light could interfere with
the development of defocus-induced myopia in guinea pigs. The
effect of the monochromatic light may be achieved by influencing
the developments of vitreous chamber and corneal curvature. The
recognition of defocus under the monochromatic light may be
achieved by the function from only one type of cone.
Commercial Relationships: Yi-Feng Qian, None; rui liu, None;
jinhui dai, None
Support: This work was supported by Grants 81400429,
81100689,81271040 from the National Natural Science Foundation
of China
Program Number: 2150 Poster Board Number: B0003
Presentation Time: 3:45 PM–5:30 PM
Antagonistic effects of atropine and timolol on the color and
luminance emmetropization mechanisms
Laura A. Goldberg, Frances J. Rucker. New England College of
Optometry, Boston, MA.
Purpose: The role of the autonomic nervous system in the color
and luminance emmetropization mechanisms is unknown. This
study analyzed the response to the non-selective, parasympathetic
antagonist, atropine, and the sympatholytic, beta-adrenergic
antagonist, timolol, in chicks subjected to illumination conditions
that selectively stimulate the color and luminance emmetropization
mechanisms.
Methods: Chicks were binocularly exposed eight hours each day,
for four days, to one of three illumination conditions: 2Hz sinusoidal
luminance flicker (LUM), 2Hz sinusoidal color flicker (B/Y), or
steady light. Mean illuminance was 680 lux. Eyes received daily
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
injections of either 20μl atropine (18nmol) (N=8), 2 drops of 0.5%
timolol (N=8), 20μl phosphate-buffered saline (N=8), or no injection
(N=8). Measurements of the axial dimensions of ocular components
and refraction were performed using A-scan ultrasonography,
photorefraction and a Hardinger Refractometer. In each illumination
condition, the saline effect was subtracted from the drug effect [Drug
(ΔX-ΔN) – Saline (ΔX-ΔN)].
Results: LUM flicker demonstrated opposite effects on eye growth
and refraction with atropine and timolol treatment. Atropine caused
a reduction in eye growth (-0.08 ± 0.02 mm, p=0.01) and a reduction
in vitreous chamber depth (-0.10 ± 0.02 mm, p=0.004), evoking a
hyperopic shift in refraction (3.40 ± 1.77 D), despite an antagonistic
increase in lens thickness (0.14 ± 0.05 mm, p=0.004). In contrast,
timolol elicited a myopic shift in refraction (-4.07 ± 0.92 D, p=0.001),
due to an increase in eye length (0.045 ± 0.030 mm). Color flicker
induced choroidal compensation for eye growth, preventing refractive
shifts with atropine and timolol. With atropine, hyperopia was not
observed, because a reduction in eye length (-0.05 ± 0.02 mm,
p=0.01) was compensated for by choroidal thinning (-0.05 ± 0.02
mm, p=0.03). With timolol, myopia did not occur because a reduction
in eye length (-0.05 ± 0.018 mm, p=0.02) was also compensated for
by choroidal thinning (-0.052 ± 0.015 mm, p=0.01).
Conclusions: The opposing growth and refractive effects of atropine
and timolol with luminance flicker, and the compensatory choroidal
compensation with color flicker, suggest a precise balancing
mechanism between the parasympathetic and sympathetic nervous
system, and the visual environment, in achieving emmetropization.
Commercial Relationships: Laura A. Goldberg, None; Frances J.
Rucker, None
Support: NEI T35 Student Research Fellowship and Beta Sigma
Kappa Student Research Grant
Program Number: 2151 Poster Board Number: B0004
Presentation Time: 3:45 PM–5:30 PM
Antagonistic Effect of Ciliary and Superior Cervical Ganglion
Sections on the Color and Luminance Emmetropization
Mechanisms
Frances J. Rucker1, Falk Schroedl2. 1Biomedical Science, New
England Coll of Optometry, Boston, MA; 2Paracelsus Medical
University, Salzburg, Austria.
Purpose: Longitudinal chromatic aberration produces changes in
retinal color and luminance contrast that guides emmetropization.
An eye exposed to luminance contrast becomes hyperopic, like an
atropine treated eye, an eye exposed to color contrast becomes more
myopic. This experiment investigates the role of the autonomic
nervous system in the control of emmetropization.
Methods: One to two week old, white leghorn chicks, underwent
unilateral lesion of the ciliary ganglion (CGX; N=16) or superior
cervical ganglion (SCGX; N=16). Animals were allowed to recover
for one week, and were then placed in cages illuminated with
sinusoidally modulated light (2 Hz: 80% contrast) that changed in
luminance (LUM) contrast or COLOR (red to green) contrast (mean
illumination 680 lux). Animals were kept in these illumination
conditions for three days (9am-5pm), and otherwise in the dark.
Changes in ocular components after the recovery period, and after
exposure to the illuminants, were measured with OCT (Lenstar) and
refraction with a Hartinger Refractometer. Changes in the lesioned
eye were compared with the unlesioned fellow eye.
Results: After recovery: CGX produced an eye with relative
hyperopia (2.01 ± 0.63D; p=0.006) and thinning of the anterior
chamber (25 ± 11 mm; p=0.037). SCGX produced an enlarged eye
(114 ± 26 mm; p<0.001) with longer vitreous chamber depth (154 ±
22 mm; p<0.001). With subsequent exposure to flicker: With CGX,
LUM prevented significant eye growth (47 ± 30 mm) but the choroid
thinned slightly (-17 ± 17 mm; p = 0.03) increasing vitreous depth (79
± 17 mm; p < 0.01) without refractive shift (-1.1 ± 1.45 D; p = 0.44).
The lens thickened (36 ± 8 mm; p = 0.002) and anterior chamber
thinned (-41 ± 12 mm; p = 0.009). COLOR increased eye growth (101
± 34 mm; p<0.05) but the choroid thickened (41 ± 19 mm), preventing
significant vitreal (59 ± 33 mm) and refractive change (0.38 ± 1.0 D).
With SCGX, LUM prevented vitreal growth (-13 ± 19 mm), and the
choroid thickened slightly (16 ± 6 mm; p = 0.03), without refractive
shift (0.3 ± 0.6 D). The lens thinned (32 ± 9 mm). COLOR increased
vitreal growth (40 ± 14 mm; p=0.02) without refractive shift (0.8 ±
1.0 D).
Conclusions: The results indicate common neural pathways for CGX
and LUM flicker, slowing growth and mostly affecting the anterior
eye, and for SCGX and color flicker, increasing growth and affecting
the posterior eye.
Commercial Relationships: Frances J. Rucker, None; Falk
Schroedl, None
Support: Research Promotion Fund of the Paracelsus University
S13/05/007-SCH: F. Rucker and F. Schroedl. Neural pathways for
Emmetropization
Program Number: 2152 Poster Board Number: B0005
Presentation Time: 3:45 PM–5:30 PM
Scotopic and Photopic Lighting Prevents Lens-induced Myopia
in Mice
Erica Landis1, Han na Park2, Megan Prunty2, 3, Curran Sidhu2,
P M. Iuvone2, 4, Machelle T. Pardue2, 3. 1Neuroscience, Emory
University, Atlanta, GA; 2Ophthalmology, Emory University,
Atlanta, GA; 3Rehab Center for Excellence, Atlanta VA, Atlanta, GA;
4
Pharmacology, Emory University, Atlanta, GA.
Purpose: The goal of this study was to determine the effects of
different ambient illumination levels on dopaminergic signaling in
the retina and on the susceptibility of the mouse eye to lens-induced
myopia. Previous studies have shown photopic lighting to protect
against myopia in both controlled animal studies and correlational
human population studies. Photopic lighting was tested as well as
scotopic lighting since rods may be needed for emmetropization
(Park et al IOVS 2014).
Methods: Male C57BL/6J mice were exposed to photopic (15,000
lux, n=38), mesopic (50 lux, n=36), or scotopic (0.005 lux, n=38)
lighting during the light phase of a 12:12 hr light cycle, starting at
postnatal day 23 (P23). At P28, half the mice received head-mounted
monocular lens defocus (-10D). Retinas were enucleated at P36 and
analyzed for dopamine and DOPAC levels via HPLC. At each time
point, the refractive error, corneal curvature, and ocular parameters
of the mice were measured. The DOPAC/dopamine ratios were
calculated as a measure of dopamine turnover.
Results: After two weeks of exposure to photopic, mesopic, or
scotopic light there was no myopic shift (OD minus OS) in the
refractive development of the control animals. Mice with lens
defocus under mesopic light had significantly larger myopic shifts
(normalized to P28, -4.741±0.608; p<0.005) by P34 compared to
mice exposed to photopic (-2.604±0.544) or scotopic (-1.807±0.608).
Additionally, in a subset of mice, the difference in DOPAC/dopamine
ratio between the lens defocused and opposite eyes was significantly
increased in mice exposed to scotopic light levels (0.022±0.007);
decreased in mice exposed to mesopic light levels (-0.020±0.003;
p<0.001); and showed no significant change in mice exposed to
photopic light (0.002 ±0.003). No significant differences were found
in corneal curvatures or axial lengths.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Conclusions: While photopic and scotopic light levels were
protective against lens induced myopia with increases in dopamine
turnover, mesopic light levels increased the development of lensinduced myopia with a decrease in dopamine turnover. This implies
that high and low intensities of light may prevent myopia, while
intermediate intensities, similar to indoor lighting, promote myopia.
Commercial Relationships: Erica Landis, None; Han na Park,
None; Megan Prunty, None; Curran Sidhu, None; P M. Iuvone,
None; Machelle T. Pardue, None
Support: NEI Grant EY016435, NEI Grant EY004864, NEI Core
Grant P30EY006360, Research to Prevent Blindness
Program Number: 2153 Poster Board Number: B0006
Presentation Time: 3:45 PM–5:30 PM
Retinal-specific Dopamine Knock-out Mice are Myopic
Michael A. Bergen3, 4, Han na Park1, Ranjay Chakraborty1, 4, Erica
G. Landis5, 4, Curran Sidhu1, P M. Iuvone1, 2, Machelle T. Pardue4, 1.
1
Ophthalmology, Emory University School of Medicine, Atlanta, GA;
2
Pharmacology, Emory University School of Medicine, Atlanta, GA;
3
Biology, Emory University, Atlanta, GA; 4Rehab R&D Center of
Excellence, Atlanta VA Medical Center, Decatur, GA; 5Neuroscience,
Emory University, Atlanta, GA.
Purpose: Dopamine has been implicated as a stop signal for
refractive eye growth based on pharmacological studies in chickens,
mammals, and primates. More recently, dopamine receptor knockout mice have been used to elucidate dopaminergic mechanisms of
refractive development (Huang et al. IOVS 2014). In this study, a
Cre-mediated, retinal-specific tyrosine hydroxylase knockout (KO)
mouse was studied to determine the effect of eliminating retinal
dopamine on refractive development and susceptibility to form
deprivation (FD) myopia.
Methods: KO mice were on a C57BL/6J background and were
homozygous for both the Chx10 Cre-recombinase and floxed tyrosine
hydroxylase alleles. Mice were randomly assigned to two groups, one
undergoing normal refractive development and the other undergoing
FD. Refractive development of KO mice and age-matched C57BL/6J
wild-type (WT) mice was measured every 2 weeks from post-natal
day 28 (P28) to P112. Under the FD paradigm, mice received a headmounted diffuser goggle at P28 over their right eye (OD) and were
measured weekly until P77. Measurements of refractive error, corneal
curvature, and ocular biometrics were obtained using an automated
photorefractor, a keratometer, and a spectral-domain optical
coherence tomography system, respectively.
Results: During normal refractive development, KO mice were
significantly more myopic (at P70, KO 2.74 ± 2.18 D, n=19; WT 7.08
± 1.47 D, n=9; p < 0.01) and had significantly shorter axial lengths (at
P56, KO 3.18 ± 0.01 mm; WT 3.23 ± 0.04 mm; p < 0.05) than their
WT counterparts. KO mice also had significantly steeper corneas than
WT mice (at P56, KO 1.42 ± 0.03 mm; WT 1.44 ± .03 mm; p < 0.05).
Both WT and KO form-deprived mice showed similar magnitudes of
myopic shift (difference of right and left eyes) (at P42, KO -2.29 ±
3.59 D, n=8; WT -3.85 ± 1.35 D; n=7).
Conclusions: Our results support the hypothesis that dopamine is
a stop signal for refractive eye growth. KO mice showed greater
variability in FD myopic shifts than WT mice, which may indicate
varying levels of retinal dopamine depletion in this model. Future
work will correlate dopamine and DOPAC levels with refractive error
and ocular parameters to comprehensively examine how dopamine
concentration affects refractive development
Commercial Relationships: Michael A. Bergen, None; Han na
Park, None; Ranjay Chakraborty, None; Erica G. Landis, None;
Curran Sidhu, None; P M. Iuvone, None; Machelle T. Pardue,
None
Support: NEI Grant EY016435, NIH Core grant P30EY006360,
Research to Prevent Blindness Grant
Program Number: 2154 Poster Board Number: B0007
Presentation Time: 3:45 PM–5:30 PM
Apomorphine attenuates form-deprivation myopia (FDM) by a
dopamine D2R-independent mechanism
Xiangtian Zhou, Furong Huang, Jiangfan Chen, Jia Qu. School
of Ophthalmology and Optometry, Wenzhou Medical College,
Wenzhou, China.
Purpose: The dopamine agonist apomorphine (APO) can profoundly
attenuate form-deprivation myopia (FDM) development in animals.
Here, we determine whether APO acts at dopamine D2 receptors
(D2R) to exert its effect on myopia development using D2R knockout
(KO) mice.
Methods: Wild-type (WT) littermates and D2R KO were subjected
to FDM at postnatal days 28-56. Both groups were intraperitoneally
injected daily with either APO (5 mg/kg/day) or vehicle for 4 weeks
(starting from postnatal day 28). Their body weight, refraction,
corneal radius of curvature and ocular axial components were
measured at the end of 4-week treatment.
Results: Consistent with our recent report, D2R KO attenuated
FDM development compared to WT littermates. As expected, APO
treatment attenuated myopia development compared with vehicle
treatment in WT mice. Importantly, APO treatment in D2R KO mice
further attenuated myopia development compared with the vehicle
treatment in D2R KO mice. In parallel with refractory changes, D2R
KO alone or APO alone also attenuated FDM-induced elongation
of vitreous chamber depth and axial length compared to their
corresponding controls. Moreover, combined treatment of D2R KO
an APO treatment attenuated FDM-induced elongation of vitreous
chamber depth and axial length compared to the D2R KO treated
with vehicle.
Conclusions: The inhibition of APO on FDM development was still
effective in absence of D2Rs, suggesting that APO attenuates myopia
development by a D2R-independent mechanism.
Commercial Relationships: Xiangtian Zhou, None; Furong
Huang, None; Jiangfan Chen, None; Jia Qu, None
Support: 973 project:2001CB504602
Program Number: 2155 Poster Board Number: B0008
Presentation Time: 3:45 PM–5:30 PM
Effectiveness of Low-dose Atropine on the Marmoset Eye
Alexandra Benavente-Perez1, Ann Nour1, Eric R. Ritchey2, David
Troilo1. 1Biological Sciences, SUNY College of Optometry, New
York, NY; 2Johnson & Johnson Vision Care, Inc, Jacksonville, FL.
Purpose: Low dose atropine (0.5%, 0.1% and 0.01%) has been
shown to reduce myopia progression in humans by up to 24.5%
(0.01%) to 40% (0.5%). The purpose of this study was to develop
reliable measures to evaluate the effects of low doses of atropine on
marmoset eyes for studies of myopia control.
Methods: Nine age-matched young adult marmosets were divided
into three treatment groups of low-dose atropine (0.1%, 0.01% or
0.005%). Subjects received one drop of atropine in one eye. The
contralateral eye served as control. Pupil diameter (PD) under fixed
background illumination (254lux) and pupillary light reflex (PLR)
to ophthalmoscope illumination were monitored at baseline and
after atropine every 5mins for the first 30mins, every hour for the
first 9hrs, and daily for 7 days. The accommodative response to 1D
of imposed hyperopic defocus was measured with a Shin-Nippon
autorefractor in one marmoset from each group for 5 days following
the atropine drop.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Results: The interocular PD difference, normalized to baseline,
reached maximum at 4hrs (0.1%), 3hrs (0.01%) and 2hrs (0.005%)
after instillation (+1.96±0.26mm,+1.46±0.28mm and +1.45±0.49mm
respectively). PDs returned to baseline levels 6 (0.1%) or 7 days
(0.01% and 0.005%) after instillation. The PLR was blocked 15 to
25mins after 0.1% instillation and recovered after 2 days, but was
always present for the 0.01% and 0.005% dosages. Accommodation
was reduced 3hrs after instillation of 0.1% and recovered after 4
days (baseline accommodation: 1.11±0.56D; 3hrs post: 0.05±1.08D,
p=0.03; 4days post: 1.45±1.20D, p>0.05). Lower doses of atropine
did not block the accommodative response (p>0.05).
Conclusions: A single drop of atropine had measurable effects on
pupil function lasting several days and showed a dose response. Only
the highest dose tested affected accommodation. While the effects of
extended treatment have yet to be examined, these results will help
guide further investigation of the effects of low-dose atropine for
myopia control using the marmoset model.
Commercial Relationships: Alexandra Benavente-Perez,
2Johnson & Johnson Vision Care, Inc (F); Ann Nour, Johnson &
Johnson Vision Care, Inc (F); Eric R. Ritchey, Johnson & Johnson
Vision Care, Inc (E); David Troilo, Johnson & Johnson Vision Care,
Inc (C), Johnson & Johnson Vision Care, Inc (F)
Support: Johnson & Johnson Vision Care, Inc
Program Number: 2156 Poster Board Number: B0009
Presentation Time: 3:45 PM–5:30 PM
The control effects of Fenofibrate on the axial length elongation
in lens-induced myopia chicken model
Panfeng Wang1, 2, Thomas C Lam2, Chi Ho To2. 1Zhongshan
Ophthalmic Center, Sun Yat-sen University, Guangzhou, China;
2
Laboratory of Experimental Optometry,Centre for Myopia Research,
School of Optometry, The Hong Kong Polytechnic University, Hong
Kong, Hong Kong.
Purpose: Apolipoprotein A1 (ApoA1), the major component of high
density lipoprotein, was suggested to be down-regulated in the retina
of lens-induced myopia (LIM) animal model. The growth of axial
length was reduced in LIM chicken possibly through an up-regulation
of the ApoA1. Fenofibrate, a peroxisome proliferator-activated
receptor α (PPARα) agonist, is the first line therapy to regulate lipid
metabolism by increasing the synthesis of ApoA1. The current study
investigated the efficacy of fenofibrate on the growth of eye axial in
lens-induced myopia chicken model.
Methods: At 4-day old, male chicken was divided into different
groups after gender determination by PCR method. In the LIM group,
negative powered lenses (-10D) were worn on the right eyes and the
left eyes were kept untreated as controls. In the treatment groups,
20uM, 100uM, 200uM fenofibrate delivered at an injection volume
of 10ul, were injected into the bottom of the vitreous chamber of
the right eye, and vehicle solutions were injected into the left eyes
as control on day 5. Then, lenses (-10D) were worn on both eyes
of treated chicken. An extra group of chicken without lens received
200uM fenofibrate intravitreal injection at the right eyes, and the left
eyes were kept untreated as normal control. A high-frequency A-scan
ultrasound system was used to measure the ocular parameters of all
the chicken before the treatment and on day 8. Chicken with body
weight growth abnormality or vitreous hemorrhage were ruled out.
The changes of ocular parameters among different treated eyes were
statistically analyzed by t-test.
Results: Fenofibrate dose dependently suppressed the growth of
ocular axial length, and statistical difference was recorded at the
concentration of 200uM. On day 8, LIM chicken (n=15) treated
by fenofibrate (200uM) had shorter axial length than LIM chicken
(n=13) by 32.7% (P= 5.15078E-07). The fenofibrate showed no
effect on the growth of ocular axial length of normal eyes. There was
no different between fenofibrate treated eyes and LIM eyes at the
choroid recovery changes after the negative lenses were taken off on
day 8.
Conclusions: 200uM of Fenofibrate is protective against LIM
development. The results not only demonstrate the therapeutic effects
of fenofibrate on myopia, but they also support the possible role of
PPARα-dependent mechanism in the development of myopia.
Commercial Relationships: Panfeng Wang, None; Thomas C
Lam, None; Chi Ho To, None
Support: XJ2012061
Program Number: 2157 Poster Board Number: B0010
Presentation Time: 3:45 PM–5:30 PM
Effect of oral administration of nicotinic acid on ocular growth of
lens-induced myopic chicks
Hu XIAO, Panfeng Wang, King Kit Li, Rachel Ka-man Chun,
Thomas C Lam, Chi Ho To. School of Optometry, The Hong Kong
Polytechnic University, Hong Kong, Hong Kong.
Purpose: To explore the effect of oral administration of nicotinic
acid on eye growth of normal and myopic chicks. The nicotinic acid
is the normal drug used in human to raising the HDL in blood. The
Apolipoprotein A1 is the potential in protect myopic eye growth in
previous studies.
Methods: White Leghorn chicks aged at 4 days (n = 48 in total) were
randomly allocated into 4 groups. Chicks in group A and B were
orally administered a single dose of nicotinic acid daily (150mg/
ml, 1ml per chick) while the chicks in group C and D were received
saline orally as control (1ml per chick). The oral administration last
for 11 consecutive days. After 7 days of oral administration, -10D
lenses were attached to both eyes of the chicks in group A and C
while the chicks in group B and D worn plano lenses for 4 days. The
refractive errors and ocular dimension components were examined
using streak retinoscopy and high resolution A-scan ultrasonography
before and after 11 days of the oral administration respectively. The
changes of refractive errors and vitreous chamber depth between 1
days and 11 days of oral administration were gained. T-test was used
to analysis the difference.
Results: After 4 days of lens wear, chicks with -10D lenses became
significantly more myopic than the chicks with plano lenses (A and
B group: P= 0.0274; C and D group: P= 0.0013; t-test). In the groups
with -10D lenses (group A and C), the changes in vitreous chamber
depth (VCD) in chicks treated with nicotinic acid (mean ± SEM;
0.568 ± 0.146mm) were significantly less than that of the salinetreated chicks (0.778 ± 0.197 mm, p = 0.007, t-test). The change in
refractive errors in chicks treated with nicotinic acid (mean ± SEM;
-8.35 ± 1.11D) were significantly less than that of the saline-treated
chicks (-10.81± 0.75D, p = 2.20E-06, t-test). There was no significant
difference in VCD between the groups wearing plano lenses (group
B: nicotinic acid vs. group D: saline; 0.398 ± 0.202mm, vs. 0.514±
0.151mm, p = 0.125, t-test). But there was significant difference in
refractive errors between the groups wearing plano lenses (group B:
nicotinic acid vs. group D: saline; -3.25 ± 0.30D, vs. -4.54 ± 0.35D, p
= 2.20E-09, t-test).
Conclusions: The nicotinic acid intake could retard the elongation of
VCD in lens-induced myopic chicks. Its effect on the normal ocular
growth is however not apparent.
Commercial Relationships: Hu XIAO, None; Panfeng Wang,
None; King Kit Li, None; Rachel Ka-man Chun, None; Thomas C
Lam, None; Chi Ho To, None
Support: PolyU research grants: GYK89, GU986; RGC GRF:
BQ29N
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Program Number: 2158 Poster Board Number: B0011
Presentation Time: 3:45 PM–5:30 PM
Form-deprived highly myopic chick eyes have lower than normal
corneal stiffness than emmetropic eyes
Byung Soo Kang1, Li Ke Wang2, Yong-Ping Zheng2, Chea-su Kee1.
1
School of Optometry, The Hong Kong Polytechnic University,
Hong Kong, Hong Kong; 2Interdisciplinary Division of Biomedical
Engineering, The Hong Kong Polytechnic University, Hong Kong,
Hong Kong.
Purpose: This study aimed to determine whether corneal stiffness
differed between normal and highly myopic eyes in the chick model
of myopia
Methods: Starting from day 5 post-hatching, the right eyes of 21
chicks (Gallus Gallus Domesticus) were covered with a translucent
occluder for 7 days to induce form-deprived myopia. At the end
of the treatment period, spherical equivalent (SE) refractive error
was measured under anesthesia (1.5% Isoflurane) by Hartinger
coincidence refractometry, and in-situ corneal stiffness (CS) was
measured using a custom-made air-jet optical coherent tomography
(OCT) system. Specifically, triplicate CS measurements were
obtained, with corneal deformation (in mm) assessed in response
to 5 cycles of increasing/decreasing air pressure (in N). A custom
algorithm calculated the slope of CS load-deformation curve (N/mm)
and the correlation between these parameters
Results: Compared to fellow untreated eyes, form-deprived eyes
developed significant myopia (mean±SEM: SE= -20.65 ± 1.41D
vs. -1.22 ± 0.26D; paired t-test, p<0.001) and exhibited reduced
corneal stiffness (mean±SEM: CS= 0.0203 ± 0.0008mm vs. 0.0239
± 0.0009mm; paired t-test, p<0.05). When data from both eyes were
used for correlation analyses, both SE (r=+0.462, p<0.002) and J45
astigmatic component (r=+0.351, p<0.05) were moderately correlated
with corneal stiffness. Expressing the corneal stiffness as percentage
of interocular difference in CS [100 %*( treated eye – fellow eye)
/ fellow eye], we found that in the 17 birds that showed a reduction
in corneal stiffness in the treated eyes (range= -0.35% ~ -46.65%),
eleven (64.7 %) of them had at least 15% reduction in corneal
stiffness (mean= -24.46%, one-sample t-test, T= -4.57, p<0.01)
Conclusions: Form deprivation induced high myopia and reduced
corneal stiffness. The correlation between SE and CS indicates that
the changes occurring in the biomechanical properties of the cornea
may be quantitatively related to those occurring in the sclera
Commercial Relationships: Byung Soo Kang, None; Li Ke Wang,
None; Yong-Ping Zheng, None; Chea-su Kee, None
Program Number: 2159 Poster Board Number: B0012
Presentation Time: 3:45 PM–5:30 PM
Regional Differences in Gene Expression with Imposed Defocus
in Chick RPE
Yan Zhang, Emily Eng, Christine F. Wildsoet. School of Optometry,
Univ of California, Berkeley, Berkeley, CA.
Purpose: We previously reported bidirectional gene regulation of
members of the Bone Morphogenetic Protein (BMP) family (2, 4, &
7) in chick retinal pigment epithelium (RPE) in response to as little as
2 h of imposed optical defocus. This study investigated whether there
were also regional differences in the effects of imposed defocus on
the expression of these genes in chick RPE.
Methods: 19-day old White-Leghorn chicks wore monocular +10
or -10 D lenses for 2 h. At the end of the treatment period, RPE
was isolated and divided into 3 circular zones using punches of 3
and 6 mm radius: central 3 mm zone (T3 or F3; T: treated eyes, F:
fellow controls), middle 3-6 mm zone (T6 or F6), and peripheral 6-9
mm circular zone (T9 or F9). RPE RNA was purified and reverse
transcribed to cDNA. qPCR was performed to examine the gene
expression of BMP2, 4, and 7. Expression levels were compared
between lens treated and fellow control eyes. Paired Student’s t test
was used for statistical analysis.
Results: The +10 D lens treatment induced up-regulation of both
BMP2 and BMP4 gene expression in the central and middle zones
(T3 & T6 regions compared to F3 & F6). For BMP2, gene expression
was highly up-regulated, by 129- and 28-fold (T3/F3, n = 3, p =
0.09; T6/F6, n = 4, p = 0.05). The equivalent values for BMP4 gene
expression were 13- and 10-fold (T3/F3, n = 3, p < 0.05; T6/F6, n =
4, p < 0.05). BMP7 expression was 3-fold up-regulated for the T3/
F3 comparison only. The most peripheral region (T9/F9) did not
show differential gene expression between treated and fellows for
any of the genes. As expected, the -10 D lens induced down- instead
of up-regulation; significant changes were recorded for both BMP2
and BMP4, for both T3/F3 and T6/F6 comparisons. BMP2 was
down-regulated by 32- and 12-fold for T3/F3 and T6/F6 comparison,
respectively (n = 4), but not for the T9/F9 comparison. Likewise,
BMP4 gene expression was down-regulated 12- and 3.6-fold in
central (T3/F3) and middle (T6/F6) regions respectively. BMP7 gene
expression did not change for any of these three regions.
Conclusions: This study provides evidence of regional variations in
the response of chick RPE to the same defocus, i.e. as imposed by
single vision lenses, generally decreasing with increasing eccentricity.
The significant up- or down-regulation of BMP gene expression in
central RPE points to a critical role of the central retina/RPE in early
stage of eye growth regulation.
Commercial Relationships: Yan Zhang, None; Emily Eng, None;
Christine F. Wildsoet, None
Support: NIH grants R01EY012392 (CFW), K08EY023609 (YZ),
K12EY017269 (YZ), T35EY007139 (EE)
Program Number: 2160 Poster Board Number: B0013
Presentation Time: 3:45 PM–5:30 PM
Retinal profile asymmetries in myopes and emmetropes
Christopher A. Clark, Ann E. Elsner, Bryan Haggerty, Joel A. Papay.
School of Optometry, University of Indiana, Bloomington, IN.
Purpose: Previous work has shown that peripheral refractive error
asymmetries exist between different locations in the retina and those
asymmetries are greater in myopes than emmetropes. This could be
due to a number of factors including physical restriction from the
optic nerve and differences in optical alignment between the optical/
visual axis. The purpose of this study is to investigate the source of
asymmetry.
Methods: Fifty-six subjects (refractive error +1.50 to -11.15)
had a battery of tests performed including axial length, corneal
topography, anterior chamber depth, peripheral refraction, peripheral
partial coherence interferometry, and SD OCT for retinal thickness.
Turning point location (TPL) was classified as the retinal location in
degrees where the retinal profile was at a minimum. Angle alpha was
measured as the distance from the apex of the corneal topography to
the visual axis. Statistics including repeated measures ANOVA were
performed with SPSS (IBM, Endicott, NY.)
Results: The TPL was greatest in high myopes with an average
displacement of three degrees temporal from the fovea compared
to approximately zero degrees for emmetropes (P = 0.045.) No
correlation was found in this study between angle alpha and either
the TPL or retinal profile asymmetry along the horizontal axis.
Asymmetry did increase with refractive error (P = 0.008.) Visual
inspection of the SD OCT images showed greater optic nerve head
tilt in subjects with higher asymmetries regardless of refractive error.
Conclusions: Retinal profile asymmetries increase with refractive
error which is consistent with previously reported data. These
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
asymmetries appear to be largely due to physical constriction by the
optic nerve rather than to do optical effects such as angle alpha.
Commercial Relationships: Christopher A. Clark, None; Ann E.
Elsner, None; Bryan Haggerty, None; Joel A. Papay, None
Support: NIH Grant K23EY022064
Program Number: 2161 Poster Board Number: B0014
Presentation Time: 3:45 PM–5:30 PM
Effects of 6-hydroxydopamine on refractive development and
form-deprivation myopia in C57BL/6 mice
Shi-Jun Weng, Xiao-Hua Wu, Yun-Yun LI, Kang-Wei Qian, Xiong-Li
Yang, Yong-Mei Zhong. Institute of Neurobiology, Fudan University,
Shanghai, China.
Purpose: In various species, reduced retinal dopamine (DA) levels
are thought to mediate the development of myopia. However, recent
evidence shows that in the C57BL/6 mouse, retinal DA levels are
unaltered when form-deprivation myopia is developed. Here, to
further explore the role of retinal DA in mouse eye growth, we
examined whether refractive development could be disturbed by
destroying retinal DA pathway in this mouse strain.
Methods: 6-hydroxydopamine (6-OHDA, 50 μg) was intravitreally
applied to the right eye using a micro-injector, and the left eye serves
as the control. Refractive errors were measured using an automated
eccentric infrared photorefractor, in animals raised in normal visual
environment, or in those with the injected eye wearing an occluder
for 4 weeks to induce form-deprivation myopia. The levels of retinal
DA and its primary metabolite 3,4-dihydroxyphenylacetic acid
(DOPAC) were assessed by HPLC analysis.
Results: Administration of 6-OHDA significantly reduced retinal
DA levels by 40-80%, and the effect lasted for at least 31 days.
With normal visual experience, the 6-OHDA-injected eyes became
markedly myopic relative to their fellow eyes (~6D of interocular
difference). Furthermore, in injected eyes, form-deprivation did not
induce further myopic shifts, nor did it cause further reduction in
retinal DA and DOPAC levels.
Conclusions: An intact retinal dopaminergic system, including
healthy dopaminergic amacrine cells and complete retinal DA
stores, is essential for both normal refractive development and the
generation of form-deprivation myopia in the C57BL/6 mouse. In this
mouse strain, refractive development could be interfered by reducing
retinal DA levels dramatically, even though the generation of formdeprivation myopia is not associated with retinal DA levels.
Commercial Relationships: Shi-Jun Weng, None; Xiao-Hua Wu,
None; Yun-Yun LI, None; Kang-Wei Qian, None; Xiong-Li Yang,
None; Yong-Mei Zhong, None
Support: Ministry of Science and Technology of China
(2011CB504602); the National Natural Science Foundation of China
(31171055, 31121061, 31100796); ARVO/Pfizer Collaborative
Research Fellowship to Shi-Jun Weng
Program Number: 2162 Poster Board Number: B0015
Presentation Time: 3:45 PM–5:30 PM
Identification of apolipoprotein A-I as a novel retinoic acid
binding protein
Jody A. Summers1, Angelica Harper1, Hanke Van-Der-Wel2,
Marcela Hermann3, Christopher M. West2. 1Cell Biology, University
of Oklahoma Health Science Center, Oklahoma City, OK;
2
Biochemistry, University of Oklahoma Health Science Center,
Oklahoma City, OK; 3Medical Biochemistry, Medical University of
Vienna, Vienna, Austria.
Purpose: All-trans-retinoic acid (atRA) may be an important
molecular signal in the postnatal control of eye size. Retinoids are
closely associated with atRA binding proteins, which are important
in regulating their transport, metabolism and biological activity. We
and others (Mertz and Wallman, Exp. Eye Res. 2000) have previously
identified a protein with an apparent Mr of 27 kD (p27) that
represents the major secreted atRA binding protein in chick choroids.
The goal of the current study was to identify p27 as we hypothesize
that p27 may play a role in the regulation of atRA activity during
visually guided ocular growth.
Methods: atRA binding proteins were initially identified from
conditioned medium of chick ocular tissues by photoaffinity 3H-atRA
labeling, SDS-PAGE and autoradiography. 3H-atRA binding proteins
were purified using a combination of anion exchange (Q-sepharose),
gel filtration (Superdex 200) columns and SDS-PAGE. Unlabeled
samples were processed in parallel and peak fractions of unlabeled
protein, corresponding to the elution position and mass of 3H-atRAlabelled protein were identified by mass spectrometry. The identify of
p27 was confirmed using immunoprecipitation of 3H-atRA-labelled
p27 from conditioned medium using anti-chick apolipoprotein A-I
antibodies.
Results: Following photoaffinity labeling of choroid and sclera
conditioned medium, radiolabeled proteins migrating at 60 kD
and 27 kD were detected by autoradiography. These proteins coeluted from Q-sepharose and Superdex 200. Mass spectrometry
analyses identified the 60 kD protein as serum albumin and the 27
kD protein as apolipoprotein A-I. Following immunoprecipitation
of 3H-atRA labeled proteins from sclera conditioned medium with
anti-chick apolipoprotein A-I, a single 27 kD band was detected on
autoradiograms.
Conclusions: Apolipoprotein A-I is the 27 kD 3H-atRA-binding
protein present in chick choroid and sclera conditioned medium. The
expression of this protein may play a role in the regulation of atRA
signaling in the choroid and sclera in postnatal ocular growth.
Commercial Relationships: Jody A. Summers, None; Angelica
Harper, None; Hanke Van-Der-Wel, None; Marcela Hermann,
None; Christopher M. West, None
Support: NIH Grant EY09391
Program Number: 2163 Poster Board Number: B0016
Presentation Time: 3:45 PM–5:30 PM
Transcriptome Analysis Indicates Adaptive Responses to
Physiological Stress in Recovery from FDM
Loretta Giummarra1, Nina Riddell1, Nathan Hall2, 3, Melanie
Murphy1, Sheila G. Crewther1. 1Psychological Science, La Trobe
University, Melbourne, VIC, Australia; 2Life Sciences Computation
Centre (LSCC), Victorial Life Sciences Computation Centre
(VLSCI), Melbourne, VIC, Australia; 3La Trobe University,
Melbourne, VIC, Australia.
Purpose: Form deprivation myopia (FDM) is associated with
dramatic increases in ocular volume, axial length, thinning of the
retina and choroid and hyperosmotic stress. Thus this study aimed to
assess the associated gene pathway changes using high throughput
RNA-sequencing and comprehensive bioinformatic analysis. Given
our previous ultrastructural and elemental microanalyses it was
hypothesized that profile changes would involve energy metabolism,
ionic solute changes and evidence of oxidative stress
Methods: Twelve male hatchling chicks were monocularly occluded
from days 4-11 after which chicks were given T=0hr, T=6hr or
T=24hr of normal vision. Four chicks were used as aged-matched
unoccluded controls. Biometrics were measured prior to tissue
collection. RNA was isolated from choroid/retina/RPE tissue and
prepared for sequencing on the Illumina HiSeq™ 1500. Raw reads
were mapped onto the chicken genome and counts determined for
each gene. Differential expression analysis was undertaken with
voom/EdgeR with an FDR of 0.05. Gene Set Enrichment Analysis
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
(GSEA) software was used to determine whether a priori defined
set of genes were significantly altered (FDR<0.25) during the
induction and recovery of FDM. Curated gene sets were obtained
from BioCarta, KEGG, and the Pathway Interaction Database and the
Reactome database.
Results: FD Chicks were ~20D myopic. Refractive normalization
began with removal of occlusion. GSEA analysis revealed an overall
suppression in genes associated with metabolism and ion homeostasis
at T=0hr. GSEA during the recovery period revealed an increase in
expression of genes associated with glucose metabolism, potassium
transport and hypoxia. These changes were positively correlated with
reduction in refraction.
Conclusions: Increased axial growth during FD is accompanied by
suppression of gene pathways associated with retinal metabolism
and refractive normalization. Removal of FD and reintroduction of
the normal visual environment with constantly changing luminance
levels requires upregulation of metabolic pathways and normalization
of ion distribution profiles across the eye. These results confirm our
previous work and build our understanding of the importance of
osmoadaptive pathways that use energy metabolism, ion transport, to
reduce hypoxia and restore osmotic homeostasis.
Commercial Relationships: Loretta Giummarra, None; Nina
Riddell, None; Nathan Hall, None; Melanie Murphy, None; Sheila
G. Crewther, None
Program Number: 2164 Poster Board Number: B0017
Presentation Time: 3:45 PM–5:30 PM
RNAseq gene expression analysis highlights the correlation
of changes in metabolic and structural pathways with axial
elongation during refractive compensation.
Nina Riddell1, Loretta Giummarra1, Nathan Hall4, 2, Melanie
Murphy1, David P. Crewther3, Sheila G. Crewther1. 1Psychological
Science, La Trobe University, Bundoora, VIC, Australia; 2La Trobe
University, Melbourne, VIC, Australia; 3Swinburne University,
Melbourne, VIC, Australia; 4Life Sciences Computation Centre
(LSCC), VLSCI, Melbourne, VIC, Australia.
Purpose: High throughput transcriptome studies in animal models
of refractive error have primarily analysed data at the single-gene
level. Results from these studies are disparate and a comprehensive
framework for understanding the biological cascades underlying
ocular growth regulation remains elusive. Thus, this study aimed to
identify characteristic biological features of refractive compensation
to myopic and hyperopic defocus in chick by correlating axial length
across lens-groups during defocus induction with expression of genes
in Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways.
Methods: Chicks were raised with ±10D lenses, or no lens.
Following biometric measurements at 1, 2, and 3 days, 3-4 chicks per
lens-group were euthanized and RNA extracted from the retina/RPE/
choroid. Libraries were sequenced on the Illumina HiSeq1500, raw
reads mapped to the chick genome, and counts determined for each
gene. Counts/million were imported into GSEA and expression of
KEGG pathways correlated with axial length phenotype across lensgroups at each time-point (FDR cut-off <.25).
Results: Refractive and axial length change was rapid during the
first day of defocus for both lens-types and slower over subsequent
days (particularly for plus lenses). Consistent with the initially rapid
change in ocular morphology, expression of structural pathways
including focal adhesion, tight junction, and vascular smooth muscle
contraction was positively correlated with axial length at 1 day.
Fatty acid metabolic and signalling pathways were also correlated
with axial length at this time. Although no structural pathways were
identified following 2 and 3 days of lens-wear when morphological
changes had slowed, metabolic pathways (such as oxidative
phosphorylation) were implicated at both time-points.
Conclusions: This study is the first to correlate ocular axial length
changes with pathway enrichment across a period of refractive
compensation to lenses. Results suggest that changes in structural
pathway expression are linked to periods of rapid axial growth
change. Perturbed metabolism was characteristic of all stages of
compensation, with implication of oxidative phosphorylation and
related pathways suggesting that growth changes elicit a shift in
energy homeostasis that may alter redox state and vulnerability to
later development of ocular pathologies.
Commercial Relationships: Nina Riddell, None; Loretta
Giummarra, None; Nathan Hall, None; Melanie Murphy, None;
David P. Crewther, None; Sheila G. Crewther, None
Program Number: 2165 Poster Board Number: B0018
Presentation Time: 3:45 PM–5:30 PM
Anti-Diuretic Hormone in the Regulation of Ocular Volume in
Compensation to Defocus
Melanie Murphy1, Loretta Giummarra1, Nina Riddell1, David P.
Crewther2, Vinh Nguyen1, Sheila G. Crewther1. 1Psychological
Science, La Trobe University, Melbourne, VIC, Australia; 2Centre for
Human Psychopharmacology, Swinburne University of Technology,
Melbourne, VIC, Australia.
Purpose: The hormone Arginine Vasopressin (AVP) is a
vasoconstrictor and anti-diuretic that is commonly associated with
stress. Our previous results show that AVP causes a myopic shift
in refractive compensation (RC) to +10D defocus (ARVO, 2013).
Further, environmental stress in the form of asymmetric flicker
impacts ocular growth (Crewther et al, 2006). Thus the current
experiment aimed to investigate whether AVP plays a role in RC to
defocus, and whether flicker affects this process.
Methods: Experiment 1: RNA was extracted from the retina/RPE/
choroid of chicks with + or -10D, or no defocus on days 5-7 posthatching (n = 3 per lens group, per day) and prepared for sequencing
on the Illumina HiSeq1500. Raw reads were mapped onto the chick
genome and counts determined for each gene. Counts per million
were imported into Pathway studio and GSEA conducted using the
Mann-Whitney U-test algorithm (p<.05).
Experiment 2: Chicks (n=360) were raised from day 5-9, with or
without asymmetric flicker in the 12 hr day cycle, with + or -10D
defocus (or non lens), following intravitreal injection of 5ml of either
PBS, AVP or the AVP receptor antagonist ([des-Gly9-β-Mercapto-β,
β cyclopentamethylenepropionyl1, O-Et-Tyr2,Val4,Arg8]Vasopressin) (in PBS) into the experimental eye. Fellow eyes
were injected with PBS. Retinoscopy and A-scan ultrasonography
was performed on day 9. Tissue was collected and prepared for
immunohistochemistry to examine AQP-4 and Kir4.1 expression.
Results: Experiment 1: RNAseq revealed sign-dependent changes in
AVP-related pathways over 3 days of rearing with defocus.
Experiment 2: Flicker alone induced a myopic shift in both lens
conditions. Flicker+AVP reduced hyperopia, axial elongation
and anterior chamber depth in +10D lenses. Flicker+AVP
antagonist reduced RC and ocular growth to -10D lenses.
Immunohistochemistry showed altered AQP-4 and Kir4.1 staining
across flicker conditions.
Conclusions: Results indicate that changes in AVP-related gene
expression occur concomitantly with changes in ocular volume
during the induction of RC. Further, AVP and its antagonist also
differentially interfered with the typical pattern of compensation
to lenswear. Physiological stress induced by flickering light further
influenced this. These results implicate stress-induced changes in the
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
rate of transretinal fluid movement in the development of refractive
error.
Commercial Relationships: Melanie Murphy, None; Loretta
Giummarra, None; Nina Riddell, None; David P. Crewther, None;
Vinh Nguyen, None; Sheila G. Crewther, None
Program Number: 2166 Poster Board Number: B0019
Presentation Time: 3:45 PM–5:30 PM
Hyperosmotic Stress and Osmo-Gene Adaptation During Early
Induction of Refractive Errors.
Sheila G. Crewther1, Nina Riddell1, Alan Marshall1, Loretta
Giummarra1, Melanie Murphy1, Melinda J. Goodyear1, David P.
Crewther2. 1Psychological Science, La Trobe University, Melbourne,
VIC, Australia; 2CHP, Swinburne University of Technology,
Melbourne, VIC, Australia.
Purpose: Why is myopia a common risk factor for most sight
threatening disorders? Our earlier biometric, ultrastructural
and elemental analyses of the chick form deprivation model
have provided evidence of severe physiological, oxidative and
hyperosmotic stress. More recently prolonged hyperosmotic stress
has been shown to lead to chronic inflammation in a number of
diseases (Brocker etal 2013). We hypothesized that perturbation
of axial growth during induction of refractive errors would also be
accompanied by hyperosmosis and osmoadaptative gene changes,
that should be demonstratable with elemental microanalysis (EDX)
and RNA seq respectively.
Methods: Chicks were raised with ±10D lenses, or no lens.
Following biometric measurements at 1, 2, and 3 days, 8 chicks per
lens group were euthanized. RNA was extracted from the retina/RPE/
choroid of 4. Four were used for scanning electron-microscopy and
EDX. Libraries were sequenced on the Illumina HiSeq1500. Counts
per million were imported into GSEA and expression of KEGG and
Reactome pathways during myopia/hyperopia induction compared to
age-matched no lens chicks (FDR cut-off <.25).
Results: Refractive compensation (RC) to -10D defocus continued
for 72hrs whereas RC to +10D was in near completion after 24hours.
EDX shows sodium and chloride ion distributions were greatly
upregulated in outer retina by -10D over the 72hrs but only at the
retino-vitreal border in +10D at 72hrs. Potassium profiles in RC
to +10D remained upregulated across the retina for 72 hrs with
concurrent up-regulation of reactome potassium channel pathways at
72hrs in RNAseq data. Consistent with altered osmotic and oxidative
stress, implicated pathways during refractive compensation included
those related to synthesis of small molecule osmolytes, structural
remodelling, inflammation, and metabolism.
Conclusions: The EDX results demonstrate that RC to optical
defocus is accompanied by hyperosmotic shifts in ion distribution
profiles across the entire posterior eye, while concurrent changes
in gene expression profiles were seen in metabolic and ion solute
processes. These pathways have previously been associated with
osmoadaptation and more severe disease states such as ARM and
diabetes. The findings suggest the need for further experimental
considerations of hyperosmotic changes as risk factors for severe
visual impairments and for development of therapeutics.
Commercial Relationships: Sheila G. Crewther, None; Nina
Riddell, None; Alan Marshall, None; Loretta Giummarra, None;
Melanie Murphy, None; Melinda J. Goodyear, None; David P.
Crewther, None
Program Number: 2167 Poster Board Number: B0020
Presentation Time: 3:45 PM–5:30 PM
Differences in the sensitivity to myopia-inducing stimuli of young
guinea pigs sourced from different colonies
Mariana Garcia2, David Hammond1, Christine F. Wildsoet2. 1Deakin
University, Geelong, VIC, Australia; 2Vision Science, University of
California, Berkeley, Berkeley, CA.
Purpose: To characterize the responses of guinea pigs sourced from
different breeding colonies to myopia-inducing stimuli using negative
lens and form deprivation paradigms.
Methods: English Short Hair guinea pig breeders were obtained from
a commercial vendor (Elm Hill Labs, Chelmsford, MA – designated
“Elm Hill” guinea pigs) and from a University-based breeding colony
(University of Auckland, NZ – designated “NZ” guinea pigs). Elm
Hill guinea pig pups were fitted with either negative lenses (-10, -5,
or 0 D) or diffusers at 10 days of age. NZ guinea pigs were fitted
with diffusers at 7 days of age. Both sets of animals were treated for
4 weeks. Ocular axial lengths were measured twice a week using
high frequency A-scan ultrasonography, cycloplegic refractions were
measured on treatment days 0, 14, and 28, and behavioral visual
acuity measured on treatment day 28.
Results: Elm Hill guinea pigs fitted with lenses exhibited minimal
interocular differences in axial length and refractive error after
28 days of treatment; likewise, form deprivation (FD) failed to
significantly affect the rate of ocular elongation or to induce a myopic
shift in refractive error. Overall changes in interocular difference
in axial length (treated minus control) were 0.03±0.1 mm for -10 D
lenses, -0.20±0.43 mm for -5D lenses, -0.01±0.21 mm for 0 D lenses,
and 0.07±0.16 mm for FD. Conversely, the NZ guinea pigs exhibited
a 0.17±0.12 mm increase in interocular differences in axial length
after 28 days of FD treatment.
Conclusions: A systematic study of the ocular growth responses of
young guinea pigs to myopia-inducing stimuli revealed significant
strain-related differences. These results point to genetically
determined differences in the sensitivity of emmetropization
mechanisms to visual manipulation, even within the same breed.
Finally, this works suggests that research groups wishing to work
with a guinea pig myopia model should carefully consider the source
of their animals.
Commercial Relationships: Mariana Garcia, None; David
Hammond, None; Christine F. Wildsoet, None
Support: EY012392
Program Number: 2168 Poster Board Number: B0021
Presentation Time: 3:45 PM–5:30 PM
Compensatory eye growth responded to the imposed defocus is
influenced by spatial content in chick
Man Pan Chin, Zhe Chuang Li, Allen Ming Yan Cheong, Ho Lung
Henry Chan. School of Optometry, The Hong Kong Polytechnic
University, Hong Kong, Hong Kong.
Purpose: Emmetopization is a visually guided eye growth, and the
compensatory eye growth is depending on the imposed defocus.
This study hypothesized if different spatial contents can affect the
compensatory eye growth responded to imposed defocus.
Methods: The right eyes of White Leghorn chicks from 10 to 12 days
old were glued with a cone-shaped lens system using Velco. Animals
were divided into six groups (n=8 to 12) for various levels of defocus
magnitude, including plano, -15D and -25D (lenses were attached
at the proximal end of the cone), with two different spatial stimulus
patterns at the other end of the cone: 1) high spatial frequency: 0.88
cycle/deg (0.4mm white/black checkers) with 100% of contrast, and
2) a lower spatial frequency: 0.28 cycle/deg (1.25mm white/black
checkers) with 100% of contrast. Axial ocular dimensions, including
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
anterior chamber depth, lens thickness, vitreous chamber depth
(VCD) and axial length, were obtained using A-scan ultrasound.
Measurement was carried out prior to fitting the lens system and on
the fourth day after treatment. Analysis of variance (ANOVA) was
used for statistical analysis.
Results: After 4 days of wearing the cones, the VCD of the right eye
increased with the imposed defocuses. Under the same magnitude
of defocus, chick eyes with low spatial stimulus had consistently
longer VCD than those with high spatial stimulus (p<0.05). Both
defocus and spatial stimulus showed significant main effect on VCD
percentage change (ANOVA, p<0.005). Significant group differences
of VCD percentage change were observed in Group -15D (low
vs high, 10.4±2.2 vs 7.25±3.35, p<0.05) and -25D (9.53±3.82 vs
4.93±4.79, p<0.05), but not in plano group. Similar trend was also
observed in axial length, but neither in anterior chamber depth nor
lens thickness.
Conclusions: Our results confirm our hypothesis that spatial
content in emmetropization can affect the compensatory eye growth
responded to same magnitude of imposed defocus. Effects of
different optical defocus on compensatory eye growth significantly
interacted with the spatial frequency of the visual stimulus. Further
studies are important to understand the mechanism of defocus and
spatial interaction in emmetropization.
Commercial Relationships: Man Pan Chin, None; Zhe Chuang
Li, None; Allen Ming Yan Cheong, None; Ho Lung Henry Chan,
None
Support: General Research Funds (PolyU5605/13M), Health and
Medical Research Fund (01121876) and PolyU Internal Grants
(G-YBBS, G-UA2E, Z-0GF, G-YM70)
Program Number: 2169 Poster Board Number: B0022
Presentation Time: 3:45 PM–5:30 PM
Effects of the relative strength of the more positive-powered
component in dual focus lenses on emmetropization in macaques
Baskar Arumugam1, 2, Li-Fang Hung1, 2, Chi-ho To3, Brien A. Holden2,
Earl L. Smith1, 2. 1College of Optometry, University of Houston,
Houston, TX; 2Vision CRC, Sydney, NSW, Australia; 3Hong Kong
Polytechnic University, Kowloon, Hong Kong.
Purpose: Dual focus lenses that impose relative myopic defocus
over a large part of the visual field can slow myopia progression in
children. Our aim was to determine how the relative surface area
devoted to the more positive-powered lens component influenced the
ability of dual focus lenses to alter refractive development.
Methods: Beginning at 3 weeks of age, infant rhesus monkeys
were reared with Fresnel lenses that had central 2mm zones of zero
power and concentric annular zones that had alternating powers of
+3.0D or 0D. The relative spatial widths of the annular zones were
varied from 1:1 (i.e., equal widths) to 1:4.5 (+3D:0D) between
treatment groups (n≥6 per group). The monkeys wore the treatment
lenses over both eyes continuously until 151±4.2 days. Comparison
data were obtained from monkeys reared with full field +3D lenses
over both eyes (FF+3D, n=6) and from 34 control monkeys reared
with unrestricted vision. Refractive status, corneal power and axial
dimensions were assessed every 2 weeks throughout the lens rearing
period.
Results: All of the dual focus lens designs produced relative
hyperopia. At the end of the treatment period, the median refractive
errors for the monkeys reared with dual focus lenses that had width
ratios of 1:1, 1:2, 1:3 and 1:4.5 were +5.25D, +5.19D, +4.31D, and
+4.28D, respectively, which were similar to the refractive errors
exhibited by animals reared with FF+3D lenses (+4.63D; p=0.22 to
0.94), but significantly more hyperopic than those found in agematched control monkeys (+2.50 D; p=0.0002 to 0.004). The average
vitreous chamber depths for the dual-lens-reared animals were also
not significantly different from those found in FF +3D lens-reared
monkeys (OD:+3D/pl 1:1; 9.31±0.34mm, 1:2; 9.44±0.60mm, 1:3;
9.74±0.38mm, 1:4.5; 9.55±0.25mm vs 9.58±0.32mm, respectively,
p=0.18 to 0.87). In addition, there were no significant differences
in either the median refractive errors (p=0.08 to 1.0) or the average
vitreous chamber depths (p=0.06 to 0.71) between the dual focus lens
groups.
Conclusions: The results demonstrate that even when the more
positive-powered zones make up only about 1/5th of a dual-focus
lens’ surface area, refractive development is dominated by relative
myopic defocus. Overall, the results emphasize that myopic defocus
distributed across the visual field evokes strong signals that can slow
eye growth in primates.
Commercial Relationships: Baskar Arumugam, None; Li-Fang
Hung, None; Chi-ho To, Inventor (P); Brien A. Holden, Zeiss (P);
Earl L. Smith, Zeiss (P)
Support: National Eye Institutes EY03611 and EY07551 and funds
from Vision CRC, Sydney, Australia
Program Number: 2170 Poster Board Number: B0023
Presentation Time: 3:45 PM–5:30 PM
Two-zone bifocal lenses with peripheral negative additions
control lens-induced hyperopia in young chicks
Huamao Miao1, Christine F. Wildsoet2. 1Department of
Ophthalmology, Eye and ENT Hospital of Fudan University,
Shanghai, China; 2Center for Eye Disease & Development, School of
Optometry, University of California Berkeley, Berkeley, CA.
Purpose: It has been proven that bifocal lenses designed with relative
positive additions can slow ocular elongation and thus myopia
progression. This study addressed a related question of whether
similar designed lenses with relative negative additions can control
hyperopia using young chicks as an animal model.
Methods: To induce hyperopia, chickens wore monocular +5
diopter (D) single vision lenses (SVLs) from 7 days of age for 3
days; the lenses were then switched for either +10 D SVLs or 2-zone
concentric bifocal lenses (BFLs), which were worn for 5 days. BFLs
had a central zone power of +10 D and one of 3 peripheral zone
powers (plano, +5 or +8 D, corresponding to additions of -10, -5 or
-2 D, respectively). For all BFL designs, both 2.5 and 4.5 mm central
zone diameters (CZDs) were tested. Central refractive errors and
ocular axial parameters were measured using static retinoscopy and
high frequency A-scan ultrasonography.
Results: At the last time point, the control group (i.e., wearing +10
D SVLs) was most hyperopic (+9.37 D), with the group wearing
2.5 mm CZD BFLs with the highest (-10 D) addition being least
hyperopic (+4.28 D). For both the 2.5 and 4.5 mm CZDs, there
were trends towards decreasing induced hyperopia with increasing
negative add power, with this dose effect being significant for 2.5
mm CZD lenses. Induced changes in vitreous chamber depth and
optical axial length (relative shortening) as well as choroid thickening
followed trends consistent with induced refractive errors.
Conclusions: Our study explored the possible application of BFLs
as a treatment to control hyperopia. Our data provide evidence that
2-zone concentric BFLs incorporating peripheral negative additions
could restrain lens-induced hyperopia progression in young chicks,
and treatment effects increasing with both add power and larger
peripheral zones. Further studies are warranted to examine whether
mammals and primates show similar beneficial effects.
Commercial Relationships: Huamao Miao, None; Christine F.
Wildsoet, None
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Support: NIH/NEI Grants R01 EY012392 (CFW) & exchange
program fund for doctoral student, Graduate Medical Education,
Fudan University (HMM)
Program Number: 2171 Poster Board Number: B0024
Presentation Time: 3:45 PM–5:30 PM
Evaluation on the changes of 90 degree visual field defects
accompanied peripheral retinal degenerative lesions in high
myopic eye——the roles in following-up and prophylaxis in
retinal detachment
Yining Shi1, Yanming Chen2, 3, Ji Liu3. 1Department of Ophthalmology,
Shaanxi Provincial People’s Hospital, Xian, China; 2Department
of Ophthalmology, China Medical University, Shenyang, China;
3
Department of Ophthalmology and Visual Science, Yale University
School of Medicine, New Haven, CT.
Purpose: To observe the relationship between peripheral retinal
degenerative changes in high myopic eyes and 90 degree visual fields
defects, and the influence with the refractive and aging factors. We
performed a retrospective, observational clinical study to provide a
safer prophylactic treatment of retinal detachment (RD).
Methods: 360 cases of high myopia were tested by LVP, OCTOPUS
101 perimeter. and compared with the 161 fellow eyes of high
myopia with RD, 118 eyes of high myopia with RD, 41 fellow eyes
of low myopia with RD, 54 eyes of low myopia with RD, and 108
normal eyes.High risk RD eyes were treated by defined equatorial
pan-retinal photocoagulation, at meantime detected the visual field
changes and posterior vitreous detachment(PVD) developing, posttreated complications.
Results: The 90 degree mean light sensitivity (MS) of high myopia
eyes was (21.34±5.40)dB (Fig. 1). The MS in high myopia was
declined with aging and severity of myopia, the linear regression
formula was R=32.981-0.161ages+0.468 diopters.There was definite
co-relationship with 2 kinds of visual defect and the degenerations:
1. The peripheral absolute defects, related with lattice degeneration;
2. The peripheral MS declining, with white-without-pressure.
Following 32 to 74 months after the defined equatorial pan-retinal
photocoagulation,there were complete PVD formation observed
under split lamp with 90°Volk pre-set lens (Fig. 2), and there were no
significant changes between the MSs pre- and post treatment.
Conclusions: There are linear relations with MS and aging, myopic
degree. The MS may indicate the peripheral degenerative lesions and
its degrees, played a roles in following-up and prophylaxis in retinal
detachment.The defined equatorial pan-retinal photocoagulation is a
safer, effective intervene for prophylaxis of retinal detachment and
forming iatrogenous PVD.
Commercial Relationships: Yining Shi, None; Yanming Chen,
None; Ji Liu, None
Program Number: 2172 Poster Board Number: B0025
Presentation Time: 3:45 PM–5:30 PM
Identification of Elements of the BMP2 Signaling Pathway in
Cultured Chick Scleral Fibroblast
Emilia A. Zin, Yan Zhang, Christine F. Wildsoet. University of
California, Berkeley, Berkeley, CA.
Purpose: Previous studies have indicated the role of Bone
Morphogenetic Protein 2 (BMP2) in eye growth regulation. This
study investigated the role of BMP2 in scleral remodeling using a
chick scleral fibroblast culture model to look for evidence of BMP2
signaling.
Methods: Primary chick scleral fibroblast (CSF) were cultured
on 24-well plates or chamber slides in DMEM/F-12 medium with
10% FBS and 1% penicillin-streptomycin at 37 °C in a 5% CO2
incubator. CSF total RNA was collected and purified using RNeasy
Mini Kit and then subjected to cDNA synthesis and real-time PCR
semi-quantification. Gene expression was examined for BMP2,
BMP receptors (BMPR1A, 1B, -2), SMAD1, -5, and -9, and the
localization of relevant proteins, BMP2, BMPR1A, BMPR2,
SMAD1, -4, and -5, phosphorylated SMAD1 (p-SMAD1), and
p-SMAD1/5 in CSF and 293T cell lines was investigated using
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
immunocytochemistry. Protein expression was also validated with
Simon automated western blot.
Results: Cultured CSF showed detectible expression of BMP2,
BMP receptors, and SMAD 1, 5, 9 genes and immunohistochemistry
confirmed the expression of BMPR1A, BMPR2, SMAD1, -4, and
-5 proteins in CSF as well as 293T cells. Western blot analyses
confirmed expression of the p-SMAD1/5 protein in both CSF and
293T cells, while SMAD1 and p-SMAD1 proteins were only detected
in 293T cells.
Conclusions: That cultured chick scleral fibroblasts express many
of the components of the BMP2 signaling pathway, at both gene and
protein levels, points to their likely involvement in scleral remodeling
and ocular growth. These results provide a foundation for future
in vivo studies into the role of BMP2 in ocular (scleral) growth
regulation.
Commercial Relationships: Emilia A. Zin, None; Yan Zhang,
None; Christine F. Wildsoet, None
Support: NIH grant R01EY012392, NIH grant K08EY023609, NIH
grant K12EY017269
Program Number: 2173 Poster Board Number: B0026
Presentation Time: 3:45 PM–5:30 PM
Growth and completion of emmetropization in the normally
developing chick eye
Zheng Shao1, 2, Marsha Kisilak1, Elizabeth L. Irving2, Melanie C.
Campbell1, 2. 1Physics and Astronomy, University of Waterloo,
Waterloo, ON, Canada; 2School of Optometry and Vision Science,
University of Waterloo, Waterloo, ON, Canada.
Purpose: Normal emmetropization has long been assumed
to result in zero refractive error, but recently this has been
questioned. In normally growing chick eyes, we are interested in
objectively determining when emmetropization is complete. We
are also interested in whether growth during and following normal
emmetropization differs.
Methods: From literature values of chick eye parameters, functions
were fit to MOR (mean ocular refraction or spherical equivalent) and
optical axial length (OAL; anterior cornea to anterior retina) vs. age.
Dioptric length (K’) and eye power (F) were derived up to day 75
using our previously reported method to calculate eye power. Pupil
size data were also used to calculate the angular and linear retinal
blurs (EB and LRB) due to defocus.
Results: Eye power and K’ decrease exponentially with age at
slightly different rates until power and K’ reach almost equal values
about day 35. Subsequently, power and K’ decrease almost identically
with age. This gives an initial rapid exponential decrease in MOR,
which reaches a relatively stable value of 1.0 D of hyperopia beyond
day 35. The completion of emmetropization is defined as the first
time point beyond which MOR remains relatively stable, estimated as
between 30 and 35 days. EB and LRB decrease almost exponentially
until day 35. After emmetropization is complete, MOR changes little,
EB remains almost constant while LRB increases slowly from about
day 45 to the end of available measurements on day 75, in agreement
with predictions of an almost uniformly expanding eye model. The
radius of the blur on the retina is larger than cone spacing prior to
completion of emmetropization, and approaches cone spacing as
emmetropization is completed.
Conclusions: Concurrent variations in eye power and length
combine to produce the smaller, more rapid changes in MOR during
emmetropization. The time point at which emmetropization is
complete can be defined as the first time point after which MOR and
angular retinal blur are stable. Emmetropization appears to be driven
by an active reduction of EB to a value close to cone resolution.
After emmetropization is complete, the subsequent change in retinal
blur is consistent with a slow, almost uniform ocular expansion.
However, after the age when normal emmetropization is complete,
an emmetropization response to additional imposed defocus blur has
been observed.
Commercial Relationships: Zheng Shao, None; Marsha Kisilak,
None; Elizabeth L. Irving, None; Melanie C. Campbell, None
Support: NSERC Canada 35321
Program Number: 2174 Poster Board Number: B0027
Presentation Time: 3:45 PM–5:30 PM
Adaptive optics measurements of cone density in chick eyes
during lens-induced myopia
Marsha Kisilak, Laura Emptage, Ian Andrews, Melanie C. Campbell.
University of Waterloo, Waterloo, ON, Canada.
Purpose: In vivo measurements of cones in the chick eye, an animal
model of myopia, are desirable as a marker of retinal changes
during axial length increases. In vivo images allow longitudinal
measurements of the angular cone spacing in the growing chick eye
and during lens-induced myopia. We can then compare measured
densities with models of retinal changes during eye growth and
myopia development.
Methods: Four Ross Ross chicks were acquired on the day of
hatching. Axial length was measured using A scan ultrasound and
aberrations and defocus were measured in a custom built HartmannShack aberrometer. Eyes were imaged in an adaptive optics corrected
scanning laser ophthalmoscope modified for small animal use (2.5
mm diameter pupil). After this, the right eyes were goggled with
-15D lenses. Measurements were repeated on days 7 and 14. All
measurements were taken close to the optical axis and the anatomical
position of the area centralis. Angular cone densities were measured
directly. Linear cone spacings on the retina were calculated from
published schematic eye models modified for measured eye lengths
and cone packing properties were assessed. Paired t-tests were
performed to compare between days and between treated and control
eyes.
Results: By day 14 goggled eyes were on average 15D myopic.
Cones were successfully imaged on all days. The angular density of
cones was not significantly different between control and goggled
eyes (p > 0.2) on any day. As seen in previous control birds, angular
density was not significantly different between days 0 and 7 (p =
0.1) in control eyes, after which it significantly increased (p < 0.02).
Goggled eyes showed no significant change in angular density with
growth. The calculated linear distance between cones increased
significantly from 6.4 microns on day 0 to 7.0 microns on day 14 in
control eyes and did not differ from goggled values of 6.2 microns
on day 0 (before goggling) and 8.3 microns on day 14. On average,
cones were 38% hexagonally packed across all days and for both
control and treated eyes.
Conclusions: Average cone spacings in control eyes on day 14
were within 10% of some literature values. Results for control eyes,
showing initial uniform expansion followed by either cone migration
or optic pole elongation are consistent with our previous data. Eyes
with lens-induced myopia expand uniformly relative to control eyes.
Commercial Relationships: Marsha Kisilak, None; Laura
Emptage, None; Ian Andrews, None; Melanie C. Campbell, None
Support: NSERC Canada 35321
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Program Number: 2175 Poster Board Number: B0028
Presentation Time: 3:45 PM–5:30 PM
Peripheral Wavefront Aberration in Myopia with and without
Orthokeratology Lenses
Young Sik Yoo1, Kyung-Sun Na1, Choun-Ki Joo1, Geunyoung Yoon2.
1
The Catholic University of Korea, Seoul, Korea (the Republic of);
2
University of Rochester, Rochester, NY.
Purpose: Peripheral refractive error degrades the quality of retinal
images and has been hypothesized as a potential factor to stimulate
the development of refractive error. Various contact lens designs
based on the hypothesis have shown the efficacy of controlling
myopia progression. The aim of the study was to evaluate the impact
of orthokeratology lens (OK lens) on the peripheral wavefront
aberration in myopic eyes.
Methods: We conducted a cross-sectional study to evaluate the
effect of OK lens on the peripheral aberration profile of myopic
subjects in adolescents. Study subjects were divided into two groups;
one was OK lens group and the other was myopic patients who
did not experience the OK lens correction. A custom-developed
Shack-Hartmann aberrometer was used to measure ocular wavefront
aberrations along different horizontal retinal eccentricities with ten
degree step across the central 30 degrees of visual field. The study
subjects maintain their natural foveal fixation while the aberrometer
is rotated around the eye for the off-axis measurements. Wavefront
refraction for each retinal eccentricity was quantified over 4 and 6
mm pupils.
Results: The mean refractive error was -1.92D ± 0.83 in the OK
lens group and -5.84D ± 2.47 in the naked eye group, respectively.
Hyperopic defocus at the peripheral visual field in the myopic eyes
increases with increasing amounts of forveal refractive error. These
effects varied with the degree of the corrected power after a treatment
of OK lens. Significant difference (p < 0.05) in the value of defocus
was found at the peripheral retinal eccentricity in the OK lens group
compared to the naked eye group. In the analysis of high order
aberration, values of on-axis and off-axis showed a different tendency
along with the symmetry of each aberration.
Conclusions: OK lens treatment is found to be effective in reducing
the degree of peripheral hyperopic defocus in myopic eyes.
Commercial Relationships: Young Sik Yoo, None; Kyung-Sun Na,
None; Choun-Ki Joo, None; Geunyoung Yoon, None
Program Number: 2176 Poster Board Number: B0029
Presentation Time: 3:45 PM–5:30 PM
Hyperoxic Myopia: A Prospective Study of Twelve Divers with
Six Hours of Exposure to 1.35 ATM PO2 for Five Consecutive
Days
Jonathan W. Brugger1, 2, Anita Gupta1, Barbara Shykoff2, John
Florian2. 1Ophthalmology, New York Eye & Ear, New York, NY;
2
Navy Experimental Diving Unit, Panama City Beach, FL.
Purpose: Hyperoxic myopia is a phenomenon associated with
prolonged exposure to an increased partial pressure of oxygen (PO2)
resulting in a myopic shift of refractive error. This has been described
in patients undergoing hyperbaric oxygen therapy and in divers
exposed to high PO2. The mechanism of action for hyperoxic myopia
is not understood. This prospective study collected ocular data in
healthy divers exposed to 1.35 ATM PO2 at the Navy Experimental
Diving Unit to better characterize hyperoxic myopia PO2 thresholds
and the mechanism of action.
Methods: Twelve healthy healthy U.S. Navy Divers participated in
five consecutive days of exposure to 100% Oxygen via surfacedsupplied, open-circuit MK20 breathing apparatuses at the bottom of
a 15-foot pool (PO2 of 1.35 ATM) for 6 hours. Prior to diving, and
three days after the last dive, subjects had an ocular examination
consisting of visual acuity (VA), autorefraction, intraocular pressure
(IOP), biometry, and corneal topography. Before and after every dive,
subjects had VA, and autorefraction. IOP was measured on the first,
third, and fifth day.
Results: Two of the twelve divers had subjective symptoms of blurry
vision 2-3 days after the last dive. The first diver had a myopic shift
of -0.50 diopters OS via autorefraction and VA change from 20/16 to
20/20-2. The other diver had a myopic shift of approximately -0.25
diopters OU via autorefraction with a VA shift from 20/30-1 to 20/100
OD and 20/20-1 to 20/40 OS. Both subjects had no significant changes
in IOP, topography, and biometry measurements and both had
spontaneous resolution of their myopia over two to three weeks with
no residual symptoms.
Conclusions: Two healthy divers exposed to an increased PO2
(1.35ATM for 30 hours in 5 days) developed symptomatic myopia
with no changes in corneal topography and biometry (axial length,
lens thickness, aqueous depth). With no appreciable changes in eye
structure, a change in refractive index of the lenticular crystalline lens
is likely responsible for the myopic shift. Hyperoxic myopia is a risk
for those conducting intense diving with a PO2 between 1.3-1.6 ATM
and warrants additional studies to better define risk factors, recovery
time, mechanism of action, and PO2 thresholds.
Commercial Relationships: Jonathan W. Brugger, None; Anita
Gupta, None; Barbara Shykoff, None; John Florian, None
Program Number: 2177 Poster Board Number: B0030
Presentation Time: 3:45 PM–5:30 PM
Malondialdehyde in high myopia.
Amparo Navea1, Francisco Bosch-Morell2, 1, Salvador Mérida
Donoso2. 1Oftalmología Médica, FISABIO, Valencia, Spain; 2Instituto
de Ciencias Biomédicas, Universidad CEU Cardenal Herrera,
Valencia, Spain.
Purpose: Malonyldialdehyde (MDA), a secondary product of lipid
peroxidation is widely used as an indicator of lipid peroxidation.
Lipids and lipid-soluble compounds are essential constituents of the
cells and tissues that comprise the eye. Simultaneously, lipids are
also crucial targets of the attack by reactive oxygen species such as
oxygen free radicals. The role of lipid peroxidation, a process under
which oxidants such as free radicals attack lipids containing carboncarbon double bond(s), especially polyunsaturated fatty acids, has
been described in several ocular pathologies in the past decades.
The aim of this work is to establish, if any, the relationship between
myopia and oxidative damage in subretinal fluid of myopic patients
with retinal detachment.
Methods: Protein content and MDA was evaluated in subretinal
fluid of 71 myopic and no myopic patients with retinal detachment.
Samples were collected in three different groups attending to myopia
degree of the subjects: group 1, non-myopic, group 2, low myopia
(patients with less than 6 dioptries) and group 3, high myopia
(patients with more than 6 dioptries).
Results: Similar data were obtained for groups 1 and 2 (group 1: 0,20
± 0,09 mM MDA and 9,24 ± 4,54 mg protein/ml; group 2: 0,22 ± 0,06
mM MDA and 9,26 ± 4,29 mg protein/ml). However high myopia
patients displayed statistically significant higher values (p<0,001)
of both components: MDA (0,39 ± 0,10 mM) and proteins (17,47 ±
4,55 mg/ml). One of the most remarkable result was the high positive
correlation obtained (r=0,87) when representing individual data pairs
of MDA and myopia degree of myopic patients.
Conclusions: These results ratify the direct contribution of oxidative
stress in retinal detachment. They also suggest that myopia may play
a role (qualitative and quantitative), that deteriorate the natural course
of ocular diseases that involve oxidative stress.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Commercial Relationships: Amparo Navea, None; Francisco
Bosch-Morell, None; Salvador Mérida Donoso, None
Support: CEU-SANTANDER PRCEU-UCH 13/17
Program Number: 2178 Poster Board Number: B0031
Presentation Time: 3:45 PM–5:30 PM
Collagen crosslinking using genipin diminishes cyclic softening of
tree shrew sclera during lens-induced myopia development
Alexander Levy2, Sarah M. Baldivia2, Rafael Grytz1. 1Ophthalmology,
University of Alabama at Birmingham, Birmingham, AL;
2
Biomedical Engineering, University of Alabama at Birmingham,
Birmingham, AL.
Purpose: To assess the effect of exogenous crosslinking using
genipin on the cyclic softening response of the remodeling tree shrew
sclera during monocular -5 diopter (D) lens wear.
Methods: Cyclic tensile tests were performed on 2-mm wide scleral
strips, first at physiological loads (50 cycles, 0-3.3 g, 30 sec/cycle)
and subsequently after 10 minutes rest at supra-physiological loads
(50 cycles, 0-33.3 g, 60 sec/cycle) conditions. Two scleral strips
were obtained from each eye of two juvenile tree shrews exposed to
4 days of monocular -5 D lens wear to induce axial elongation and
myopia. The scleral strips of the control eye were mechanically tested
immediately after enucleation or after 24 hours incubation at 37°C
in PBS. The scleral strips of the lens treated eye were tested after 24
hours incubation in PBS or PBS supplemented with genipin at a low
cytotoxicity concentration (1mM). Cyclic softening was defined as
the incremental strain increase from one to the next cycle. This value
was averaged over both animals and cycles 5 to 50. Cycles that led to
tissue failure were excluded.
Results: At both loading conditions (physiological / supraphysiological loads), the average incremental strain increase (%
per cycle) was nearly identical in the fresh (0.03 /0.2) and PBS
incubated tissue of the control eye (0.03/0.26). The cyclic softening
was approximately four times higher in the sclera of the myopic
eye (0.14/0.81) and two orders of magnitude lower after genipin
crosslinking (0.001/0.004).
Conclusions: Results indicate that cyclic tensile loading leads to
continued softening of the juvenile tree shrew sclera. The softening
rate increases during lens-induced myopia and is diminished after
genipin crosslinking. This finding suggests that axial elongation in
myopia may be due to a remodeling mechanism that increases the
cyclic softening response of the sclera, which can be inhibited by
scleral crosslinking using genipin.
Cyclic tensile tests at supra-physiological loads of three scleral strips
of one animal showing the increased cyclic softening leading to tissue
failure in the treated sclera versus the control and the diminished
cyclic softening after genipin crosslinking.
Commercial Relationships: Alexander Levy, None; Sarah M.
Baldivia, None; Rafael Grytz, None
Support: NIH grant EY003909 (P30); EyeSight Foundation of
Alabama; Research to Prevent Blindness
Program Number: 2179 Poster Board Number: B0032
Presentation Time: 3:45 PM–5:30 PM
Immunolesioning of glucagonergic amacrine cells in the chick
retina
Diane Nava1, 2, Tatiana Lupashina2, Bhavna Antony1, Li Zhang3,
Michael D. Abramoff3, Christine F. Wildsoet1, 2. 1Vision Science
Graduate Group, UC Berkeley, Berkeley, CA; 2Center for Eye
Disease and Development, Berkeley, CA; 3Department of Electrical
and Computer Engineering, University of Iowa, Iowa City, IA.
Purpose: Neurotoxins have been used in myopia research to ablate
inner retinal cells in order to study their contributions to eye growth
and refractive error regulation, although those used in past studies
have been relatively unselective. The purpose of this study is to
investigate immunolesioning as a potential tool to selectively ablate
glucagon amacrine cells (GACs) in the chick and to compare its
selectivity to previous methods.
Methods: A Saporin immunotoxin conjugated to a primary antiglucagon antibody was injected intravitreally in the left eyes of 7-day
old chicks as a 10uL solution in one of 5 concentrations (0.125, 0.25,
0.5, 0.75 or 1 uM).
TUNEL staining (Roche) was used to determine the distribution of
apoptotic cells and immunohistochemistry on vertical sections used
to assess changes in the glucagonergic cell population.
Optical coherence tomography imaging was used to investigate
changes in the retina and choroid, and flash electroretinograms
(ERGs) were recorded to assess changes in retinal function 4, 6 and 9
days after injection.
Results: The maximum loss of GACs was seen with the 1 uM
concentration and for concentrations lower than 1 uM, the central
retina seems to be more affected than peripheral retina, where GAC
immunoreactivity in the IPL/INL boundary was more apparent than
in the center.
The 1uM concentration significantly attenuated the photopic
negative response of the flash ERG at both 4 and 7 days (p=0.0236
& p=0.0393), with no significant effect on b-wave and a-wave
amplitudes. The peak of the flash ERG at approximately 200 ms was
also significantly attenuated at 4 days after injection (p=0.03), but not
at later time points.
With the 1 uM concentration, total retinal thickness was not
significantly reduced in injected eyes at any time point, while
choroidal thickness underlying the area centralis was significantly
increased compared to values for the contralateral eyes at both 6 and
9 days post injection (p=0.0475 & p=0.04097 respectively).
Conclusions: The above immunotoxin conjugate, injected
intravitreally, appears to more selectively lesion GACs in the
chick retina than previously tested neurotoxins, as evidenced by
histological as well as retinal structural and functional data, and thus
represents a suitable tool for investigating the role of GACs in eye
growth regulation. The finding of choroidal thickening in lesioned
eyes is a novel finding that warrants further investigation.
Commercial Relationships: Diane Nava, None; Tatiana
Lupashina, None; Bhavna Antony, None; Li Zhang, None;
Michael D. Abramoff, University of Iowa (P); Christine F.
Wildsoet, None
Support: NIGMS 3R25GM090110-04S1
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Program Number: 2180 Poster Board Number: B0033
Presentation Time: 3:45 PM–5:30 PM
Ciliary Muscle Cell Changes During Guinea Pig
Emmetropization
Andrew D. Pucker1, Ashely R. Carpenter2, Hugh J. Morris1, Andrew
J. Fischer3, Kirk M. McHugh2, Donald O. Mutti1. 1Optometry,
Ohio State University, Columbus, OH; 2Center for Molecular and
Human Genetics, Nationwide Children’s Hospital, Columbus, OH;
3
Department of Neuroscience, Ohio State University, Columbus, OH.
Purpose: To establish normal morphological parameters and to
characterize ciliary muscle (CM) cell changes with age during guinea
pig emmetropization.
Methods: Three pigmented guinea pig eyes were collected at three
different ages (n = 9 eyes). Mean refractive error was determined
with retinoscopy by two trained examiners. Eyes were then
enucleated, hemisected, and fixed with paraformaldehyde. Temporal
eye segments were then embedded in OCT compound and 30 mm
serial sections were collected; the two most temporal slides of each
eye were then labeled with anti-α-smooth muscle actin antibodies
(smooth muscle) and Draq5 nuclear stain. Sections were then
visualized with a fluorescent microscope (Leica Microsystems) and
analyzed with Stereo Investigator (MBF Bioscience) to determine
the mean CM cross-sectional area, nuclei number, and cell crosssectional area.
Results: Guinea pigs displayed emmetropization as refractive error
decreased from +9.08 ± 2.75 D at 1-day-old to +2.91 ± 0.88 D at
90-days-old. The mean CM length and CM cross-sectional area both
significantly increased with age, from 0.42 ± 0.035 mm to 0.90 ±
0.095 mm (p = 0.003) and from 0.037 ± 0.005 mm2 to 0.057 ± 0.015
mm2 (p = 0.011), respectively. The mean cross-sectional area covered
by each CM cell did not change (p = 0.82), which is consistent with
the marginal increase in the mean CM cell number from 72.0 ± 7.4
cells to 105.5 ± 19.2 cells per section (p = 0.059; all by JonckheereTerpstra test).
Conclusions: Guinea pig CM undergoes morphologic changes during
development in the first 90 days of life characterized by significant
increases in cross-sectional area and length while the mean area
occupied by each cell does not significantly change. These data
suggest that the CM grows via cell proliferation rather than through
hypertrophy. These normative data will be useful when contrasted
with potential CM changes during myopia induction experiments.
Commercial Relationships: Andrew D. Pucker, None; Ashely
R. Carpenter, None; Hugh J. Morris, None; Andrew J. Fischer,
None; Kirk M. McHugh, None; Donald O. Mutti, None
Support: NIH/NEI: K08EY023264
measure the axial length (AL), height, width, volume and surface
area of the internal eye at birth. Eye shape was assessed by an index
of oblateness, calculated as 1–(AL/width) or 1–(AL/height). Oblate
eyes had oblateness >+0.01, spherical eyes had oblateness between
+0.01 and -0.01, and prolate eyes had oblateness <-0.01. Cycloplegic
autorefraction and optical biometry (IOLMaster) were performed 3
years later.
Results: In total, 346 eyes of 173 children were analysed. The
majority were male (94 children, 54%) and of Malay (43%) or
Chinese (43%) origin. Most eyes were prolate at birth. At three years,
the mean AL was 21.74±0.68mm (range 19.77-23.84), representing
a mean increase from birth of 4.47±0.94mm (1.71–7.20). The mean
spherical equivalent refraction(SER) was 0.91±0.80D (-2.40 to
+3.47) and only a small proportion of eyes was myopic (8 eyes,
3.6%). After multivariate adjustment, eyes with longer AL at birth
had smaller increases in AL at 3 years (p<0.001). Eyes with larger
baseline volumes and surface areas had smaller increases in AL at
3 years (p<0.001 for both). Eyes which were more oblate at birth
had greater increases in AL at 3 years (p<0.001). Using width to
calculate oblateness, prolate eyes had smaller increases in AL at 3
years compared to oblate eyes (p<0.001), and, using height, prolate
and spherical eyes had smaller increases in AL at 3 years compared to
oblate eyes (p<0.001 for both). There were no significant associations
between eye size and shape at birth and SER, corneal curvature or
myopia at 3 years.
Conclusions: Eyes that are longer, larger and have prolate or
spherical shapes at birth exhibit smaller increases in AL over the
first 3 years of life. Eye size and shape at birth influence subsequent
eye growth but not the development of refractive error, suggesting
adequate compensatory mechanisms to maintain emmetropia for at
least the first 3 years of life.
Commercial Relationships: Laurence S. Lim, None; Sharon
Chua, None; Pei Ting T. [email protected], None; Shirong Cai,
None; Yap Seng Chong, None; Kenneth Kwek, None; Marielle
Fortier, None; Cheryl Ngo, None; Anqi Qiu, None; Seang-Mei
Saw, None
Support: This work is supported by the Translational Clinical
Research (TCR) Flagship Program on Developmental Pathways
to Metabolic Disease funded by the National Research Foundation
(NRF) and administered by the National Medical Research
Council (NMRC), Singapore- NMRC/TCR/004-NUS/2008.
Additional funding is provided by the Young Investigator Award
at the National University of Singapore (NUSYIA FY10 P07) and
Singapore Ministry of Education Academic Research Fund Tier 2
(MOE2012-T2-2-130).
Program Number: 2181 Poster Board Number: B0034
Presentation Time: 3:45 PM–5:30 PM
Eye size and shape in newborn children and its relation to axial
length and refraction at three years
Laurence S. Lim1, 2, Sharon Chua2, Pei Ting T. [email protected],
Shirong Cai2, Yap Seng Chong2, Kenneth Kwek3, Marielle Fortier3,
Cheryl Ngo2, Anqi Qiu2, Seang-Mei Saw2. 1Ophthalmology, Singapore
National Eye Center, Singapore, Singapore; 2National University
of Singapore, Singapore, Singapore; 3KK Women’s and Children’s
Hospital, Singapore, Singapore.
Purpose: Eye shape has been postulated to be a risk factor for
refractive error. The purpose of this study is to determine if eye shape
and size at birth are associated with refractive error and eye size 3
years later.
Methods: A subset of 173 full-term newborn infants from the
Growing Up in Singapore Towards healthy Outcomes (GUSTO)
birth cohort underwent magnetic resonance imaging (MRI) to
Program Number: 2182 Poster Board Number: B0035
Presentation Time: 3:45 PM–5:30 PM
Normative ocular biometric values for the adult mouse, rat,
rabbit, dog, pig, non-human primate, and human
Joshua S. Eaton1, Andrea D. Rodrigues2, Craig B. Struble3, Sara M.
Thomasy4, Christopher J. Murphy4, 1. 1Ocular Services on Demand,
Madison, WI; 2Non-Clinical Development - Toxicology, Allergan,
Irvine, CA; 3Covance Laboratories, Madison, WI; 4Department of
Surgical and Radiological Sciences, University of California - Davis,
Davis, CA.
Purpose: Confidence in knowledge of biometric dimensions of
the ocular chambers and structures is essential in pharmacokinetic/
dynamic modeling and preclinical development of therapeutic
compounds and devices. Furthermore, knowledge of comparative
biometric values between animal models and humans can impact
dosing methods, concentration delivered, and techniques employed
in nonclinical studies. Consideration of comparative differences also
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
improves accuracy in prediction of human safety risk. Our objective
was to summarize available normative data for the human and six
common animal models - mouse, rat, rabbit, dog, pig, and non-human
primate (NHP).
Methods: References citing normative ocular biometric values in
all seven species were collected by searching online publication
databases, textbooks, and journals available to the authors. Biometric
parameters researched included: axial globe length (AGL), anterior
chamber depth (ACD), axial lens diameter (ALD), vitreous chamber
depth (VCD), and vitreous chamber volume (VCV). Available data
were assembled and evaluated. For scarce or unavailable values,
additional data was generated to establish normative values. All data
are presented as mean ± SD.
Results: Reported values were obtained using both in vivo and ex
vivo techniques as well as calculation and/or estimation. Parameters
with values of higher intraspecies variability (>25% coefficient of
variation) included: ACD in the mouse (0.50 ± 0.17); ALD in the rat
and rabbit (3.55 ± 1.08 and 6.18 ± 1.88 mm, respectively); VCD in
the mouse and NHP (0.61 ± 0.16 and 8.72 ± 3.58 mm, respectively);
and VCV in the mouse, rat, dog, and NHP (0.01 ± 0.01, 0.03 ± 0.02,
2.63 ± 0.81, and 2.20 ± 0.89 ml, respectively). Relative to other
laboratory species, fewer values were reported for AGL in the rat;
ACD in the rat, rabbit, pig, and NHP; VCD in the rabbit, pig, and
NHP; and VCV in the rat, dog, and NHP.
Conclusions: Reliable normative ocular biometric values are critical
for use of animal models in eye research and preclinical ocular
drug and device development. This review summarizes published
data, highlighting parameters of greater variability and identifying
data gaps for species commonly used in the development of new
therapeutics.
Commercial Relationships: Joshua S. Eaton, None; Andrea D.
Rodrigues, None; Craig B. Struble, None; Sara M. Thomasy,
None; Christopher J. Murphy, None
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].