Vision Res., Vol. 37, No. 19, pp.

2661-2673, 1997
© 1997 Elsevier Science Ltd. All rights reserved
Pergamon Printed in Great Britain
PII: S0042-6989(97)00114-4
0042-6989/97 $17.00 + 0.00

Flicker Parameters are Different for Suppression

of Myopia and Hyperopia
HARTMUT N. SCHWAHN,*t FRANK SCHAEFFEL*
Received 22 August 1996; in revised form 29 January 1997

Axial eye growth rates in the chicken are controlled by local retinal image-processing circuits.
These circuits quantify the loss of contrast for different spatial frequencies and promote axial
eye growth rates in correlation with the amount of retinal image degradation ("deprivation
myopia"). They also distinguish whether the plane of focus lies in front of or behind the retina. How
the sign of defocus is detected still remains unclear. Cues from chromatic aberration are not
important.
In an attempt to isolate retinal circuits controlling the development of myopia or hyperopia,
young chickens were raised in flickering fight of different frequencies (12 and 6 Hz) and duty cycles
(4-75 %) produced by rotating chopper disks. The effects of flickering light on refractive errors and
change in axial growth rates induced by translucent occluders or defocnsing lenses were measured
by infrared retinoscopy and A-scan ultrasound, respectively. Retinal electrical activity was
evaluated by flicker ERG after matching flicker parameters and stimulation brightness at retinal
surface. Changes in retinal and vitreal dopamine content caused by flicker in occluded and normal
eyes were determined by HPLC-ECD.
Strikingly, suppression of myopia occurred for similar flicker parameters, whether induced by
translucent occluders ("deprivation") or negative lenses ("defocns"). The degree to which myopia
was suppressed was correlated with the duration of flicker dark phase and with the ERG amplitude.
In contrast, suppression of hyperopia did not correlate with these parameters. We conclude that
two different retinal circuits with different temporal characteristics are involved in the processing
of hyperopic defocus/deprivation and of myopic defocus, the first one dependent on flicker ERG
amplitude. However, we did not find any correlation between the rate of dopamine release and the
degree of inhibition of deprivation myopia in flickering light. © 1997 Elsevier Science Ltd

Flicker light Myopia Hyperopia Electroretinogram Chicken

INTRODUCTION minimum. Apparently, however, the proposed image

Axial eye growth and refractive development in higher
vertebrates are controlled by signals derived from retinal
image processing; the role of the brain is unclear and
possibly marginal (Diether & Schaeffel, 1997; Wildsoet
& Wallman, 1995). There is experimental evidence that
the retina can compile information on the average image
quality (i.e. the high spatial frequency content) because
translucent eye occluders produce myopia ("deprivation
myopia"; Wallman et al., 1978) in correlation with the
loss of image contrast (Bartmann & Schaeffel, 1994). It
is tempting to speculate that a role of deprivation
myopia is to reduce hyperopia in neonates and to tune
refractive state so that the average "deprivation" is at a
*Department of Experimental Ophthalmology, University Eye Hospi-
tal, Rrntgenweg 11, D-72076 Ttibingen, Germany.
JTo whom all correspondence should be addressed [Tel (+49 7071) 29
85926; Fax (+49 7071) 29 5777; Email schwahn@uni-tuebin-
gen.de].
2661
processor in the retina not only quantifies the amount of
deprivation, but also determines the relative position of
the plane of focus, including the sign of defocus imposed
by spectacle lenses (Schaeffel et al., 1988; Irving et al.,
1992). Input from the accommodation feedback loop is
not necessary (Schaeffel et al., 1990; Troilo & Wallman,
1991; Schwahn & Schaeffel, 1994; Diether & Schaeffel,
1997; Schmid & Wildsoet, 1996a). There is even
evidence that the sign of defocus is detected locally
within the eye. This is suggested by experiments showing
a local compensation for local refractive errors imposed
by lens segments that defocus only parts of the visual
field (Diether & Schaeffel, 1997). How can the sign of
defocus be determined locally within the eye without
input from the brain? This is still a puzzling enigma.
Although it would have been a plausible explanation,
chromatic aberration occurring in the eye's optical
system does not provide cues for the sign of defocus
(Schaeffel & Howland, 1991; Rohrer et al., 1992;
Wildsoet et al., 1993).
2662 H.N. SCHWAHN and F. SCHAEFFEL
13ms
4%
!10 ms
12%
~1 ms
12 HZ 25 %
~42 ms
5oo m
•3ms
75%
~--
~42 ms
6 Hz f 25%: 813 ms
50 %
FIGURE I. Angular extent of the black and transparent sections of the rotating chopper disc for the generation of flickering
light. Duty cycles reflect the ratio of light to dark periods and are given in percent. Durations are given in msec and are valid for
the disk's speed of 6 rotations per second.
A possible approach to learn more about the retinal
image-processing involved is to study the effects of
flickering light stimulation on both deprivation myopia
and lens-induced refractive errors. There is experimental
evidence (Schaeffel et al., 1995) that the underlying
image processing occurs, at least in part, by pharmaco-
logically different mechanisms; they may also have
different sensitivity to spatial and temporal stimulation.
Both, defocus-induced and deprivation-induced myopia
share some common features; they can both be
suppressed by stroboscopic light (Gottlieb et al., 1987;
Rohrer et al., 1995; Vingrys et al., 1991) or by reserpine
(Schaeffel et al., 1995), a neurotoxin that depletes retinal
dopamine stores. Intermittent periods of clear vision
reduce both occluder- and negative lens-induced myopia.
"Clear vision periods" of about the same duration are
necessary for a 95% suppression of myopia: 130 min for
deprivation myopia (Napper et al., 1995) and less than
180 min for lens-induced myopia (Schmid & Wildsoet,
1996a). On the other hand, both types of induced myopia
have been claimed to show different frequency tuning for
suppression by stroboscopic light (Schmid & Wildsoet,
1996b). Also, deprivation myopia develops normally
after optic nerve section (Troilo et al., 1987) whereas
lens-induced myopia is reduced by this intervention
(Wildsoet & Wallman, 1995). Hyperopia induced by
positive lenses, on the other hand, does not require an
intact optic nerve (Wildsoet & Wallman, 1995) and is not
suppressed by reserpine (Schaeffel et al., 1995).
i
:
1 rotation(1/6 sec)
i
Rather than just varying the flicker frequency as in
previous studies with xenon-lamp stroboscopes (extre-
mely short light pulses), we have applied flickering light
of defined brightness, frequency and duty cycle to
chickens treated with image-degrading occluders or
defocusing lenses. To characterize retinal responses to
flicker, electroretinograms (ERGs) were recorded at
comparable duty cycles and frequencies with the retinal
brightness carefully matched to those in the lens- and
occluder experiments.
Dopamine released from retinal amacrine cells has
been proposed to be a messenger linking retinal image
processing and eye growth, since its release and retinal
content drop during lens-induced defocus and deprivation
(Stone et al., ! 989; Rohrer et al., 1993; Ohngemach et al.,
1997). Dopamine release, however, can also be triggered
in some species by stimulation with flickering light
(Dong & McReynolds, 1992; Kirsch & Wagner, 1989;
Boelen et al., 1996) and, in a frequency-dependent
fashion, by electrical stimulation (Dubocovich & Hens-
let, 1986). Possibly the loss of high spatial contrast
during lens and occluder treatment might cause a
decrease in the activity of retinal cells leading to the
reported drop in the rate of dopamine release. However,
in eyes deprived from clear vision, amacrine cells might
be stimulated by flickering light to release dopamine,
which in turn could compensate the drop in dopamine
levels, mimicking a period of clear vision. We have,
therefore, measured retinal dopamine content and
 MYOPIA AND HYPEROPIA SUPPRESSEDBY DIFFERENTFLICKER LIGHT 2663

TABLE 1. Groups of chickens, treatment and number

Rearing condition
Treatment
Flicker Flicker Diurnal
Group frequency duty cycle fight cycle Treatment Age Number
1 12 Hz 4% 12F-12D occl. day 9--day 16 6
2 12 Hz 12% 12F-12D eccl. day 9--day 16 7
3 12 Hz 50% 12F-12D eccl. day 9--day 16 9
4 12 Hz 75% 12F-12D eccl. day 9--day 16 6
5 6 Hz 25% 12F-12D eccl. day 9--day 16 5
6 6 Hz 50% 12F-12D eccl. day 9-day 16 5
7 no flicker 12N-12D eccl. day 9---day16 9
8 12 Hz 12% 12F-12D +8 D day lO--day 15 6
9 12 Hz 12% 12F-12D -8 D day 10--day 15 6
10 12 Hz 50% 12F-12D +8 D day 10--day 15 8
11 12 Hz 50% 12F-12D -8 D day 10.-day 15 10
12 6 Hz 25% 12F-12D +8 D day 10--day 15 5
13 6 Hz 25% 12F-12D -8 D day 10-day 15 5
14 6 Hz 50% 12F-12D +8 D day 10--day 15 5
15 6 Hz 50% 12F-12D -8 D day 10--day 15 5
16 no flicker 12N-12D +8 D day 10-day 15 5
17 no flicker 12N-12D -8 D day 10-day 15 5
19 12 Hz 4% 9N-3F-12D eccl. day 10--day 17 7
20 12 Hz 50% 12F-12D eccl. day 10--day 17 6
21 12 Hz 50% 9N-3F-12D eccl. day 10--day 17 6
F, hours of flicker illumination; N, hours of normal incandescentlight; D, hours of darkness.

dopamine release in flickering light of different duration
(12 and 3 hr) and its effect on deprivation myopia.
MATERIAL AND METHODS
Animals
Male white leghorn chickens were obtained from a
local chicken farm (Suppingen, Germany) at day 1 post-
hatching. All chicks were kept under a 12/12 hr light/dark
cycle at 28°C for the first 10 days and at 22°C thereafter.
They had access to food and water ad libitum. Chickens
were raised and treated according with the ARVO
resolution for the use of laboratory animals and treatment
was approved by the University commission for animal
welfare (AK 2/91).
Housing and light stimulation
On day 6, groups of 6 to 14 chickens were transferred
from standard cages illuminated by 60 Watt incandescent
light bulbs to a hemispherical plastic dome (diameter
1 m). The dome was covered inside with highly reflective
white paint and was used as an integrating sphere to
provide uniform illumination. A 1 5 0 W xenon-lamp,
mounted on an optical bench, produced a light beam that
was focused on a rotating chopper disc (diameter 15 cm)
and was then projected via a convex lens and a mirror
onto a diffusing plate (5 × 5 cm) that covered a hole cut in
the top of the dome. The dome was set up in a darkroom
and shielded from the lamp's stray light so that, except
for the flickering light, no light from external sources
could enter. A small electrical fan equipped with a light
trap was installed at the dome to ensure sufficient
ventilation.
To obtain the different flicker parameters, the dark and
light sectors of the chopper disc were varied in angular
extent, resulting in different duty cycles. Doubling the
number of light and dark sectors on the chopper disc
provided twice the flicker frequency (from 6 to 12 Hz)
but left the light--dark transition times (6 msec) un-
changed. Only for the 4% duty cycle at 12 Hz, did the
rotation speed have to be doubled, resulting in only a
3 msec, rather than a 6 msec transition time. The timing
of the different flicker stimuli used in this study is
schematically illustrated in Fig. 1.
To compensate for the decrease in the total amount of
light with decreasing duty cycles (e.g. at 4% duty cycle a
one light flash would require 12.5-times as much light
energy than a light flash at 50% duty cycle in order to
deliver the same energy per cycle), we placed an
adjustable aperture on the optical bench to control the
intensity of the beam. To provide a similar average
illuminance in the dome, light was measured by a slowly
integrating photometer ( t = 2 . 5 s e c ) and the lamp's
aperture was adjusted until the photometer showed
similar readings for all flicker protocols and for constant
light for all flicker protocols. As a result, the shortest light
pulses (4% duty cycle at 12 Hz) with a peak illuminance
of 1500 lux and about 150 lux for a 75% duty cycle at
12 Hz and 50% at 6 Hz produced an average illuminance
level (_+0.2 log units) equivalent to normal (constant)
illumination.
Treatment protocols
The treatment of the different experimental groups is
summarized in Table 1. To exclude effects of adaptation
to flicker during the treatment periods with occluders or
2664 H.N. SCHWAHNand F. SCHAEFFEL
lenses, all groups except 18 and 21 were exposed to
flicker light 1 day before the treatment began. Control
groups 7, 16 and 17 were kept under continuous
illumination during a 12 hr day (8 a.m. to 8 p.m.; below
referred to as "normal illumination" or "normal light").
Groups 1-6, 8-15 and 18 and 20 received flickering light
in a diurnal cycle of 12 hr light/dark. Groups 19 and 21
were kept under continuous illumination during the day
for 9 hr and were then exposed to flicker light for the
remaining 3 hr only (5 p.m. to 8 p.m.) before the light
was turned off. Occluders and lenses ( ___8 D power) were
used as previously described (Schaeffel et al., 1994a).
Only one eye was treated, whereas the contralateral eye
served as individual control. During the day, occluders
and lenses were checked hourly for proper attachment
and cleanness. If occluders or lenses were lost more than
once, the animals were excluded from the study. Previous
studies in our laboratory showed that a single loss of a
lens or a occluder of less than 1 hr during the whole
experiment (5-7 days) did not significantly reduce the
effect on refractive development. However, we consid-
ered a repeated loss as a condition of intermittent lens-/
occluder-wear and expected a reduction in deprivation
myopia (Napper et al., 1995) or lens-induced myopia
(Schmid & Wildsoet, 1996a).
Electroretinograms
The ERG recording device consisted of an optical
bench and a computer system which controlled stimula-
tion and data sampling. Contact lens electrodes (Henke's
type) specially designed for chickens were used (Medical
Workshop, Gronningen, Holland). The recording device
and procedures are described in detail in a previous paper
(Schaeffel et al., 1991). Brightness was controlled by
neutral density wedges actuated by stepping motors.
White Ganzfeld stimuli (about 120 deg angular extent in
the central visual field of the chicken; Schaeffel et al.,
1991) were projected onto the eye of the anesthetized
animal in Maxwellian view. Flickering light of different
duty cycles and frequency was produced by an electronic
shutter which was controlled by the computer. ERG-
responses were low-pass filtered (cut-off at 400 Hz,
Bessel function, 10th order) and digitized to 12-bit
precision. After flicker stimulus onset, recording was
delayed for 10 sec until a stable response amplitude to the
flicker stimulation was attained. Twenty sweeps were
averaged and the absolute maximal voltage difference
between peaks and troughs was determined and plotted as
"ERG-modulation" in Fig. 6. Flicker electroretinograms
[Fig. 6(A)] were recorded from two chickens. Since the
recordings were very consistent and reproducible
(Schaeffel et al., 1991), data from only one animal are
shown.
Matching stimulus brightness for the ERG to the
brightness in the dome
To compare the results of the flicker ERGs to results
obtained in the flicker-rearing experiments, it may be
very critical that flicker stimulation of the retina occurred
at the same brightness in both the ERG recording sessions
and in flicker-rearing. Although 150 W xenon-lamps with
the same spectral composition were used in both
experiments, it was difficult to compare the intensity of
a light beam projected onto the eye's cornea in
Maxwellian view to the illuminance levels measured in
the dome (since different physical units apply). To bypass
this problem, we employed the following procedure: a
photometric photocell (United Detectors Technology w/
AP-10) acted as an "artificial retina" in an artificial eye of
the chicken adjusted to emmetropia (focal length 15 ram,
pupil diameter 6 mm, f/# 2.5; lateral stray light in the
artificial eye was excluded by making its walls from
black cardboard). (1) If the "artificial chicken eye" was
moved through the dome (average illuminance level
about 100 lux, see above), the readings of the photocell
ranged from 9.0 mV (pointed to ground, from 8 cm
above) to 19.0 mV (pointed at illuminated diffuser at the
ceiling of the dome). We considered this reading as
equivalent to the amount of light that fell on the chickens'
retinas when they were foraging in the dome. (2) If the
same artificial eye was placed in the stimulation beam of
the ERG recording device and was illuminated in
Maxwellian view, as in the respective ERG experiment,
the photocell gave a reading of 8.9 mV. Therefore, it
seems unlikely that the difference in retinal image
brightness in the ERG-measurements and the experi-
ments in the dome would exceed a factor of 2
(0.3 log units).
Measurement of refractive state and ocular dimensions
At the end of the occluder treatment (after 7 days) or
lens treatment (after 5 days), refractive state and ocular
dimensions were determined by automated infrared
photoretinoscopy (Schaeffel et al., 1994b) and A-scan
ultrasound (Schaeffel & Howland, 1991), respectively. In
some experimental groups (2, 3, 7), corneal radius of
curvature was determined, in addition, by infrared
photokeratometry as previously described (Schaeffel &
Howland, 1987).
Preparation of tissue samples and measurement of
catecholamines
Retinal and vitreal catecholamine levels were deter-
mined in groups 18-21 (Table 1) by high pressure liquid
chromatography with electrochemical detection (HPLC-
ECD), as described earlier (Bartmann et al., 1994). On
the last day of the experiments, after the measurements of
refractions and ocular dimensions, chicks were exposed
to 3-3.5 hr of flickering light. They were then removed
from the dome one by one, killed by decapitation and
their eyes instantly excised. The eyeballs were cut into
halves in the equatorial plane. The gelatinous vitreous
could be readily removed from the posterior segment by a
pair of tweezers. Subsequently, the retina was carefully
scraped from the posterior eye cup with a blunt hook,
leaving the retinal pigment epithelium in place. Samples
of vitreous and retina were frozen in liquid air and stored
at 80°C. Catecholamine content was referenced to wet
 MYOPIA AND HYPEROPIA SUPPRESSED BY DIPFERENT FLICKER LIGHT 2665

10 retinal catecholamine levels fluctuate over the year,

experiments with groups 18-21 were done within
1 month. Data in the text are mean values +__1 standard
8 ...............................................* ~ .................................................. deviation (SD). Flicker parameters are expressed as
"frequency/duty cycle", i.e. "12/4" for 12 Hz at a duty
7
cycle of 4%. In the figures, data are given as mean
value___standard error of the mean (SEM), except Fig.
2(A) and Fig. 2(B), where standard deviations are plotted.

p RESULTS
3
Effects of flickering light on the development of the
2 anterior segment of the eye
1 Previous work (Gottlieb & Wallman, 1987; Wildsoet
& Pettigrew, 1988; Vingrys et al., 1991; Squires et al.,
0 1992) has shown that the growth of anterior segment of
4% 12% 50% 75% 25% 50% no the eye is independently controlled from that of the
duty cycle flicker posterior segment. Therefore, we have compared the
FIGURE 2. Refractions of non-occluded fellow eyes of chickens kept corneal radius of curvature in groups 2 and 3, which were
in flickering light for 12 hr a day for 6-8 days. Significance levels refer kept under permanent flicker illumination during the day,
to comparisons with chicks that were kept in normal light during the and corneal curvature in the animals kept in normal light
day (black bar on the right). ( * * P < 0 . 0 1 ; * P < 0 . 0 5 ; Dunnett's
(group 7). Corneas were significantly flatter in the chicks
ANOVA).
raised under flickering light (Table 2). In normal
illumination, occluded eyes had significantly steeper
weight rather than protein content since the vitreous corneas than the open fellow eyes. Steepening of corneas
contained virtually no protein (Ohngemach et al., 1997). in occluded eyes has been reported previously (Gottlieb
Conversion of the retinal content per wet weight to et al., 1987; Hayes et al., 1986). Strikingly, this
content per mg protein can be readily done because difference vanished in animals kept under flickering
retinas in young chicks contain 103 + 8 mg protein per light (Table 2).
1000 mg wet weight (n = 169 eyes, unpublished observa- In all flicker groups, except for group 1 (12/4; Table 3)
tion). flicker-rearing alone had an inhibitory effect on axial
elongation, even if the eyes were not covered by
occluders or lenses (Tables 3 and 4). Interestingly, there
Data analysis and statistics were no significant differences in vitreous chamber depth
To evaluate the effects of flickering light on refractive to the groups kept under normal light (data not shown).
development with occluders or lenses, we used one-way Thus, the differences in axial eye length did not reflect
analysis of variance (ANOVA) after Dunnett. Paired two- changes in vitreous chamber depth but are rather due to
tailed t-tests were employed to assess the significance of flattening of corneas accompanied by shallower anterior
interocular differences in individual chicks, both for chambers. Indeed, except for group 4 (12/75; Table 3), all
ocular dimensions and measurements of biogenic amines. chickens raised in flickering light were more hyperopic in
Unpaired two-tailed t-tests were applied to compare axial their uncovered control eyes than the ones raised in
length and refraction data from different groups. Since normal light (Fig. 2; Tables 3 and 4).

TABLE 2. Mean values + SD of corneal curvature in flicker and non-flicker groups

Corneal radius of curvature (mm) Significance*

Rearing
condition Occluded (O) Control (C) O vs C F/O vs N/O F/C vs N/C F/M vs N/M

12 Hz/12% (F) 3.38 +__0.09 (7) 3.36 + 0.07 (7) n.s. P < 0.001 P < 0.01
12 Hz/50% (F) 3.32 -t-_0.03 (9) 3.31 ± 0.02 (9) n.s. P < 0.001 P < 0.01
mean (M)
3.34 +_ 0.08 (32) P < 0.01

no flicker (N) 3.09 -t- 0.03 (9) 3.18 ± 0.04 (9) P < 0.01
mean (M)
3.13 _ 0.06 (18)
See also Fig. 2.
*Significance levels for two-tailed t-tests. O vs C: occluded eyes vs uncovered fellow eyes, paired t-test. Unpaired t-tests in the following: F/O vs
N/O: occluded eyes, flicker group vs occluded eyes, non-flicker group; F/C vs N/C: uncovered eyes, flicker groups vs uncovered eyes, non-
flicker group; F/M vs N/M: all eyes, flicker group vs all eyes non-flicker group.)
2666 H . N . SCHWAHN and F. SCHAEFFEL

TABLE 3. Mean values + SD of axial length and refractive error of occluded and uncovered eyes of monocularly occluded chicks raised under
flicker of different frequency and duty cycles

Treatment Axial length (ram) Refractive error (D)

Occluder Occluded Control Difference Significance* Occluded Control Difference Significance*

12 Hz/4% 9.60 __+0.55 9.46 _ 0.38 0.15 _+ 0.08 (6) P < 0.01 1,5 _+ 4.9 5.3 _+ 1.9 -3.8 ___ 2.4 (6) P < 0.01
12 Hz/12% 9.01 _+ 0.26 8.87 _ 0,14 0.16 ___ 0.11 (7) P < 0.01 2.2 __+ 5.6 5.5 ___ 2.1 -3.2 ___ 1.4 (7) P < 0.01
12 Hz/50% 9.44 __+ 0.44 9.00 ___ 0.20 0.43 _+ 0.09 (9) P < 0.01 -3.1 _+ 5.4 7.8 _ 3.0 -10.9 ___ 2.1 (9) P < 0.05
12 Hz/75% 9.93 _+ 0.21 9.09 + 0.11 0.84 + 0.08 (6) n.s, -12.3 -t- 2.1 2.9 _ 1.1 -15.2 _+ 1.2 (6) n.s.
6 Hz/25% 9.38 -t- 0.18 9.26 -t- 0.23 0.12 ___ 0.10 (5) P < 0.01 6.0 _ 0.9 6.7 _ 0.8 -0.7 _-4-0.5 (5) P < 0.01
6 Hz/50% 9.53 _+ 0.19 9.29 __. 0.25 0.24 + 0.03 (5) P < 0.01 0.8 + 0.9 4.8 __+0.8 -4.0 _ 0.3 (5) P < 0.01
No flicker 10.06 + 0.28 9.14 -t- 0.50 0.91 + 0,10 (9) -13.4 _ 2.5 3.1 + 1.0 -16.5 -t- 1.1 (9)
Interocular differences are given as mean values _+SEM.
*Significance levels of Dunnett's ANOVA on interocular differences: effect o f occluder deprivation on flicker groups vs occluder effect in the
non-flicker group.

TABLE 4. Mean values _+SD of axial length and refractive error of experimental eyes covered by negative ( - 8 D) or positive (+8 D) lenses and
the uncovered fellow eyes of chicks raised under flicker of different frequency and duty cycles

Treatment Axial length (mm) Refractive error (D)

Negative lenses Experimental Control Difference Significance* Experimental Control Difference Significance*

12 Hz/12% 8.91 _+ 0.07 8.85 _+ 0.10 0.06 _ 0.03 (5) P < 0.01 4.3 _+ 2.1 6.1 _ 1.0 -1.8 _ 0.7 (5) P < 0.05
12 Hz/50% 9.05 + 0.23 8.84 ___ 0.17 0.21 __+0.04 (10) P < 0.01 -0.2 + 3.3 6.4 -t- 1.8 -6.6 ___ 1.1 (10) n.s.
6 Hz/25% 9.02 _+ 0.16 9.04 _+ 0.11 -0.02 + 0.07 (5) P < 0.01 2.7 _+ 1.4 5.2 + 1.4 -2.5 ___ 0.9 (5) P < 0.05
6 Hz/50% 9.06 + 0.07 9,01 + 0.05 0.05 _+ 0.05 (5) P < 0.01 4.6 _+ 5.4 6.6 ___ 2.8 -1.9 + 1.7 (5) P < 0.05
No flicker 9,83 _+ 0.17 9.47 _+ 0.14 0.36 _ 0.02 (5) -4.8 _+ 3.0 3.3 _ 2.4 -8.1 _+ 1.5 (5)

Positive lenses

12Hz/12% 9.00+0.14 8.98__+0,16 0.02+0.07(6) P<0.01 2.9+2.9 6.1 + 1.4 -3.2 _+ 1.2(6) P<0.01
12 Hz/50% 8.70 _+ 0.20 8,82 _ 0.19 -0.11 + 0.04 (8) n.s. 8.5 ± 2.4 6,4 ___ 1.1 2.1 ___ 1.0 (8) P < 0.05
6 Hz/25% 8.77 + 0.13 8.89 _ 0.09 -0.12 + 0.04 (5) n.s. 9.0 + 2.4 6.0 + 2.9 3.1 + 0.8 (5) n.s.
6 Hz/50% 8.87 + 0.13 8.93 _+ 0.08 -0.06 + 0.03 (5) P < 0.05 12.2 + 0.8 8.7 + 2.1 3.5 ___ 1.0 (5) n.s.
No flicker 9.07 _+ 0.09 9.36 _ 0.07 -0.29 + 0.02 (5) 9.4 _ 1.7 3,0 _+ 1.4 6.4 + 0.9 (5)
Interocular differences are given as mean values _+SEM.
*Significance levels for Dunnett's ANOVA on interocular differences: effect of lens-imposed defocus on flicker groups vs lens effect in the non-
flicker group.

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E ~-12
t~ 0.2
-16

0.0 -18
4% 12% 50% 75% 25% 50% no 4% 12% 50% 75% 25% 50% no
duty cycle flicker duty cycle flicker
FIGURE 3. (A) Interocular differences in axial length of monocularly occluded chicks that were raised under flicker during the
day, with different parameters (gray bars; frequency/duty cycle: 12/4, 12/12, 12/50, 12/75, 6/25, 6/50) and under normal light
during the day (black bar). Significance levels refer to the group raised in normal light (**P < 0.01; Dunnett's ANOVA). (B)
Interocular differences in the refractions of the same animals in (A) (**P < 0.01; *P < 0.05; Dunnett's ANOVA).
 MYOPIA AND HYPEROPIA SUPPRESSED BY DIFFERENT FLICKER LIGHT
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12% 50% 25% 50%
duty cycle
FIGURE4. (A) Interoculardifferencesin axial lengthin chicksraisedwithnegativelenses (-8 D) on one eye.Plot as in Fig. 3
(**P < 0.01; Dunnett's ANOVA). (B) Interocular differencesin refractions in the same animals as in (A) (**P < 0.01;
*P < 0.05; Dunnett'sANOVA).
To facilitate the separation of the effects of flicker-
raising on deprivation myopia development or lens-
induced refraction changes (i.e., the increase/inhibition of
vitreous chamber growth) from the general hyperopic
shift in flickering light due to flatter corneas, interocular
differences between experimental and uncovered eyes
rather than absolute axial lengths and refractions are
plotted in the subsequent figures. Absolute refractions
and axial lengths are given in Tables 2, 3 and 4.
Effects of flicker light on the development of deprivation
myopia
There was a significant suppressive effect of flickering
light on deprivation myopia. The occluded eyes of groups
raised in flickering light showed less myopic refractions
than the ones raised in normal light (Table 3). This
suppressive effect on the development of deprivation
myopia, however, was clearly dependent on the flicker
parameters [Fig. 3(A) and Fig. 3(B)]. Note that at both
6/25 and 6/50, which are equivalent to light pulse
durations of 42 and 83 msec, the suppression of
deprivation myopia was significantly more effective than
in 12/50 flicker, which has the same light pulse duration
as 6/25, and that the same duty cycle as 6/50.
Effects offlickering light on the compensation of imposed
defocus by spectacle lenses
Negative lenses. Under flickering light, negative lenses
were less effective in producing axial myopia than in
normal light [compare gray columns in Fig. 4(A) and Fig.
4(B) with black columns). The reduction of the lens-
effect was significant in all cases, except 12/50, for both
axial lengths and refractions (Table 4). Superimposed on
the suppressive effect of flickering light, there was a
general shift towards more hyperopic refractions in both
2667
B
E
-10
-12
no 12% 50% 25% 50% no
flicker duty cycle flicker
eyes (Dunnett, P < 0.01; except 12/50). Axial lengths
were consistently shorter in the flickering light than in
normal light (Dunnett, P < 0.01).
Positive lenses. Positive lens-treated eyes of flicker-
reared chicks had absolute refractions that were not
different from lens-treated eyes of chick raised normal
light (except 12/12; Table 4). Considering a general
hyperopic shift that was also found in the untreated
fellow eyes, flicker indeed reduced the effect of positive
lenses on refractive development [Fig. 5(A, B)]. How-
ever, in contrast to the negative lens experiments, the
reduction was significant only in the 12 Hz flicker groups.
In 12/12 flickering light, there was even a tendency to
reverse the sign of the change in refraction (interocular
difference: 12/12: -3.2 +_ 1.2D vs normal light:
+6.4 _+ 0.9 D; Table 4). As in the negative lens-treated
groups, both positive lens-treated and open eyes had
significantly shorter axial lengths than the eyes of chicks
raised in normal light (Dunnett, P < 0.01).
In summary, flickering light suppressed lens-induced
as well as deprivation-induced refractive errors. How-
ever, there may be a tendency that 6 Hz flicker was more
effective on negative lens-induced myopia (Fig. 4),
whereas 12Hz flicker seemed more competent in
suppressing the effect of positive lenses (Fig. 5). Note,
that flicker of 12/12 was most effective in suppressing the
effects on refractive development of both positive and
negative lenses, as well as occluders.
Global retinal activity under various duty cycles
Electroretinograms were recorded in response flicker-
ing light of different duty cycles and frequencies, with the
stimulation intensities matched to retinal image bright-
ness. For technical reasons, stimulation was not possible
with exactly the same duty cycles and frequencies as in
 2668 H• N. SCHWAHN and F. SCHAEFFEL

0.05
A B
E o.oo
tS -o.os
~D
I=
!l -o.lo
~ -o.15
w
c
--
3
.0.20
:!i!iii!!iiiiiiiiiiiiiiiiiiil;i"
:
= -0.25 iiiiiiiiiiiiii!iiiiiiiiiiiiiiiiiiii

~
' u -0.30

-0.35

-0.40
12% 50% 25% 50% no 12% 50% 25% 50% no
duty cycle flicker duty cycle flicker
FIGURE 5. (A) Interocular differences in axial length in chicks raised with positive lenses (+8 D) in one eye. Plot as in Fig. 3
(**P < 0.01; *P < 0.05; Dunnett's ANOVA). (B) Interocular differences in refractions in the same animals as in (A)
(**P < 0.01; *P < 0.05: Dunnett's ANOVA).

150
.~ ...... ~6t25
A. 140
B ,~ a duration of light stimulus
• duration of dark phase 90
"'l. ,
125 x ~x x flicker duty cycle
• "e" - ~ 1 6 / 5 0 [] __ Ka= 0.8~3 80
f- ,\ ,•
> 120 X X
• i 0
i" \ 70
•=1~.100 • e t~Jt.2 \
i:: r '~100
0
om E 6o ~"
0s
75 ~ 8o
"O
[] o x
0
E ~ 60 40 ~
t~ 50
rr 30
40 =o • •o x o
W
20
25
i l / I ~ X 0
20 x [] o
o° o 10
interpolated
I I i P 0 0
20 40 60 80 100 0 50 100 150

duty cycle [%1 E R G - r n o d u l a t i o n [t~V]

FIGURE 6. (A) Amplitudes in the flicker electroretinogram (ERG) during white Ganzfeld stimulation for different stimulus
frequencies and duty cycles (filled symbols). Since for technical reasons exactly the same stimulus parameters as in the previous
experiments could not be delivered by the ERG recording apparatus, the respective flicker ERG amplitudes were obtained from
the amplitude traces by linear interpolation (open squares). In the boxed sample traces of original ERG responses to flicker
stimulation of 6.5, l0 or 15 Hz are shown for different duty cycles, with the shortest light pulse duration (smallest duty cycle)
the uppermost traces. (B) IfERG data from different flicker frequencies were pooled, the inter-stimulus intervals (filled squares)
remained correlated to the flicker ERG amplitudes but not duty cycles (crosses) or light pulse durations (open squares)•

the occluder and lens experiments. However, the required Comparable ERG amplitudes were generated for a duty
flicker ERG amplitude data could be determined by linear cycle of 25% at 15 Hz and a duty cycle of 65% at 6.5 Hz.
interpolation of the actually measured ERG data [Fig. Which flicker stimulus parameter was the most
6(A)]. Flicker modulation of ERG amplitude increased important factor to determine the amplitude in the
with decreasing duty cycle (shorter light pulses) for all ERG? For that, we plotted duty cycle, light pulse
flicker frequencies tested [Fig. 6(A), see insets]. Given duration and its complement, the duration of the dark
the same duty cycle, ERG modulation was about twice as phase between two light pulses, further denominated as
high at 6.5 Hz stimulation than at 15 Hz stimulation• dark phase (DP), against the ERG amplitude [Fig. 6(B)].
 MYOPIA AND HYPEROPIA SUPPRESSED BY DIFFERENT FLICKER LIGHT 2669

TABLE 5. Mean values _ SD of axial length and refractive error of occluded and uncovered fellow eyes of monocularly occluded chicks raised
under flicker of different parameters and different duration (12 and 3 hr)
Treatment Axial length (mrn) Refractive error (D)
Occluder Occluded Control Difference Significance* Occluded Control Difference Significance*

2 Hz/4% 12 hr 9.65 ___0.45 9.50 -t- 0.12 0.15 +__0.07 (7) P < 0.001 -1.9 _+ 3.6 2.6 __ 1.5 -4.5 _ 1.6 (6) P < 0.01
12 Hz/50% 12 hr 9.83 ___0.27 9.43 + 0.18 0.40 _+ 0.06 (6) P < 0.01 -5.2 ___6.9 4.6 ___2.1 -9.8 _+ 2.1 (6) P < 0.05
12 Hz/4% 3 hr 10.36 ___0.42 9.61 + 0.14 0.75 _ 0.15 (7) n.s, -10.3 ___3.0 3.2 +__1.7 -13.5 ___ 1.3 (7) n.s.
12 Hz/50% 3 hr 10.05 ___0.37 9.42 ___0.14 0.63 + 0.08 (6) P < 0.05 -9.4 _ 2.9 2.8 + 1.6 -12.2 ___ 1.2 (7) n.s.
no flicker 10.06 _ 0.28 9.14 __ 0.50 0.91 + 0.10 (9) -13.4 ___2.5 3.1 + 1.0 -16.5 __+0.9 (9)
Interocular differences are given as mean values _ SEM.
*Significance levels of Dunnett's ANOVA on interocular differences: effect of occluder deprivation on flicker groups vs occluder effect in the
non-flicker group.

1.0

0.9

E 0.8 ,a" , - 2
£ ¢0
•~ 0.7 =..4
¢11
_e 0.6 -6

N 0.5
tl .E
t=
-o- 0.4 o-10
u
o g[
¢=
0.3
~ -12

~0.2 "1:;-14

0.1 -16

0.0 -18
4% 50% 4% 50% no 4% 50% 4% 50% no
d u t y cycle flicker d u t y cycle flicker
FIGURE 7. (A) lnterocular differences in axial eye length of monocularly deprived chicks raised under different flicker
treatment: 12/4 and 12/50 flicker, either 12 hr a day (light bars) or for only 3 hr with normal illumination for the remaining 9 hr
(dark bars). Significance levels refer to comparison of the flicker groups with the group raised in normal light during the day
(**P < 0.01; *P < 0.05; Durmett's ANOVA). (B) Interocular differences in refractions in the same animals as in (A). Only
12 hr of flicker could inhibit the effect of occluders on refractive development (compared with the respective differences found
in normal illumination (black bar) **P< 0.01; *P < 0.05; Dunnett's ANOVA).

F o r p o o l e d E R G d a t a f r o m all flicker frequencies, there d e v e l o p e d , to a slightly lesser extent at 12/50 (signifi-

was a high correlation o f the E R G a m p l i t u d e to the D P
(R = 0 . 9 4 8 , n = 2 5 , P < 0 . 0 0 1 ) but not to the other
parameters.
Dopamine release during flicker stimulation
T o study the r e l a t i o n s h i p b e t w e e n flicker, retinal
d o p a m i n e release, and rescue f r o m refractive errors, w e
used t w o p a r a d i g m s : flicker for 12 hr p e r d a y and for only
3 hr a d a y (see M e t h o d s ) . W e r e p e a t e d e x p e r i m e n t s with
t w o flicker parameters, 12/50 and 12/4, that s u p p r e s s e d
d e p r i v a t i o n m y o p i a to different degrees. A s f o u n d in the
p r e v i o u s e x p e r i m e n t s (groups 1, 3; Fig. 3), d e p r i v a t i o n
m y o p i a was s u p p r e s s e d b y 12 hr flicker p e r d a y with 12/4
and 12/50 [Table 5; Fig. 7 ( A ) and (B)]. H o w e v e r , in
groups 19 and 21, w h i c h w e r e treated with flickering light
for o n l y 3 hr at the e n d o f the day, d e p r i v a t i o n m y o p i a
cance level: P < 0.05).
Retinal d o p a m i n e content was d e c r e a s e d in the
o c c l u d e d e y e s o f the groups raised in n o r m a l i l l u m i n a -
tion, as well as in the o c c l u d e d e y e s o f the groups that
e x p e r i e n c e d flicker light o f different duration (12 hr/3 hr)
[Fig. 8(B)]. B e c a u s e the d e v e l o p m e n t o f d e p r i v a t i o n
m y o p i a m a y be influenced b y the release o f d o p a m i n e
into the interstitial space (as m e a s u r e d in the vitreous)
rather than the overall retinal d o p a m i n e content (Ohnge-
m a c h et al., 1997), w e d e t e r m i n e d also the vitreal
d o p a m i n e levels. E x c e p t for 12/4 in the 3 hr flicker
r e g i m e n there was a d e c r e a s e in d o p a m i n e release [Fig.
8(A)]. A p p a r e n t l y , d e p r i v a t i o n m y o p i a c o u l d b e sup-
p r e s s e d b y 12/4 flickering light (12 hr a d a y ) without
p r e v e n t i n g the d e c l i n e in retinal d o p a m i n e content and
d o p a m i n e release, M o r e o v e r , in the groups treated with
3 hr o f 12/4 flicker a d a y there was no significant d r o p in
2670 H.N. SCHWAHN and F. SCHAEFFEL

100%

80%
A .
• 300h

¢11 60% | non-occluded fellow eye i

5.5 o~ 200A
40% • 3.5 - .¢_
I
10%
20%
i 12hours | " l ~
0% 0% .............. , , ! i i
70 . "s6 • II 90 i
-20% 7O
-10%

-40% a

0
~ -20%
-60% " "1~ ,#i;'~!Fi'0~i
-30%
-80%
vitreous retina
- 100% - - -40%
4% 50% 4% 50% no 4% 50% 4% 50% no
duty cycle flicker duty cycle flicker

FIGURE 8. (A) Vitreal dopamine content in monocularly deprived chicks raised under different flickering lights (12/4 or 12/50
flicker, applied either for 12 or 3 hr a day). Changes in vitreal dopamine content are expressed as percentages of the vitreal
content in the non-occluded fellow eyes. Boxed values represent the absolute content of dopamine [ng/1 g wet weight]: upper
value: occluded eye; lower value: non-occluded fellow eye. Significance levels refer to the effect of occluder deprivation on the
experimental eyes vs contralateral control eyes (*P < 0.05, paired t-test). (B) Retinal dopamine contents. Plots as in (A)
(*P < 0.05, paired t-test).

retinal dopamine content [Fig. 8(B)], despite the fact that
deprivation myopia developed (Fig. 7). In fact, there was
a considerable increase in vitreal dopamine content in the
occluded eye in the 12/4 group (P < 0.01), whereas in the
12/50 (3 hr) flicker group, as in all other groups, vitreal
dopamine was decreased in the occluded eye [Fig. 8(A)].
DISCUSSION
We have found that the development of both depriva-
tion-induced and defocus-induced refractive errors can be
suppressed by flickering light. Flicker-rearing also
caused a hyperopic shift, which was mainly due to
corneal flattening, even in uncovered eyes. However, this
effect did not account for the suppression of deprivation
myopia and lens-induced myopia but was rather super-
imposed upon all refractive changes observed in this
study. In order to compare effects of flicker on the
outcome of deprivation myopia and lens-induced myopia
or hyperopia, we calculated a measure of "suppression of
refractive errors" which was given as the ratio of
experimentally induced refractive errors (interocular
differences) in normal light (N) to those in flicker (F):
suppression = (1 - F ) . 100%. (Negative percentages re-
fer to an enhancement of refractive errors and suppres-
sion by more than 100% represents a reversal of the sign
of the induced refractive error.)
What parameters of flicker light might account for the
suppression of refractive errors?
In a normal visual environment (with a wide variety of
viewing distances) hyperopic eyes without accommoda-
tion experience a blurred image all the time, whereas in
myopic eyes the retinal image is defocused only for
vision at a distance. If the image is defocused or blurred
over a certain coherent time period, a "blur detector"
within the retina might trigger increased growth in the
sclera in case of occluder deprivation, as well as under
negative lens treatment, as suggested by Diether &
Schaeffel (1997). A quite trivial explanation could then
account for the suppression of experimental refractive
errors in flicker: with short light pulses there might be
less integration time for a presumed retinal image
processor ("blur detector") to detect changes in image
quality and shifts in the focal plane. Since flicker of 6/25
has the same exposure time per light pulse as 12/50 thus,
one could expect similar suppression of refractive errors
under both flicker conditions. However, in contrast to
lens-induced hyperopia, where 6/25 and 12/50 flicker
indeed lead to a similar reduction, 6/25 flicker was
significantly more competent in reducing both lens-
induced and deprivation myopia treatment than 12/50
(both P < 0.01, t-test). Hence, light pulse duration does
not seem to be the major critical parameter for the
suppression of myopia.
What else might be the nature of this blur detector?
Since most retinal ganglion cells show transient
responses to light stimuli, the retina is considered as a
sensory organ that is best adapted to detect changes in the
visual environment, Thus, no neural activity will be
generated if the retinal image does not change, which is
the case with a constantly blurred image caused by
negative lenses or occluders. With clear vision, retinal
activity returns to normal levels and inhibits exaggerated
scleral growth (Gottlieb & Wallman, 1987; Wallman,
1990). In other words, the "signal" within the retina that
finally leads to myopia might just be reduced retinal
 MYOPIA AND HYPEROPIA SUPPRESSED BY DIFFERENT FLICKER LIGHT 2671

200%

A Ill : ,i

,o0% ................................. .............................. ,00%

J
¢ o i
0% e, • • , • = 0% -- i & "1 J ~ o, ,
A &
A O A o
0 0

-50% -50%
0 25 50 75 100 125 150 0 20 40 60 80 100 120 140 160
dark phase duration [m~c] ERG-Modulation [pV]
FIGURE 9. Suppression of both deprivation- and lens-induced myopia correlates with the flicker ERG amplitudes. In contrast,
suppression of lens-induced hyperopia is not correlated with the ERG, suggesting different retinal processing for myopia and
hyperopia. Suppression of refractive errors due to flicker: (1 - ~) • 100% (F refractive errors (interocular differences) in flicker,
N under normal light). Significance levels of correlation for deprivation myopia (n = 38): r = 0.709, P < 0.001; for lens-induced
myopia (n = 25): r = 0.512, P < 0.01; for lens-induced hyperopia (n -- 24): r = 0.162, n.s.)

activity. Considering the result shown in Fig. 6(B), where focal plane, whereas with negative lenses scleral growth
the amplitude of flicker ERG modulation, that serves as a is stimulated. Our data suggest that the separation of both
crude measure of overall retinal activity, correlates with processes occurs at a very early stage, i.e. at different
the dark phase duration of flickering light but not with neuronal circuits in the retina, and that both circuits may
light pulse duration, we found it appropriate to look for release different factors that control choroid or sclera,
correlation of this parameter to the flicker-effects. respectively. The experiments with reserpine and apo-
Indeed, we found correlation of the suppression of both morphine suggest that only the mechanism that controls
deprivation myopia and negative lens-induced myopia scleral growth in deprivation and lens-induced myopia
with dark phase duration and therefore with the involves dopamine (Schaeffel et al., 1995; Schmid et al.,
interpolated flicker ERG amplitudes of the corresponding 1997).
flicker parameters (Fig. 9A). In contrast, the suppression Unfortunately, we can only speculate about the cellular
of positive lens-induced hyperopia does not correlate origin of the flicker ERG responses. In the single-flash
with the dark phase duration and flicker ERG amplitude. ERG the amplitude of the positive going b-wave of the
This suggests that "deprivation" and "hyperopic defocus" light ON-response is believed to reflect retinal activity
might trigger similar retinal processes, supporting the postsynaptic to the photoreceptors: a change in the
"retinal activity model" of myopia. membrane potential of the MUller cells, as a consequence
of depolarizing bipolar cells which extrude potassium
Possible retinal mechanisms for deprivation myopia and into the extracellular space (Dick & Miller, 1978). In the
lens-induced myopia or hyperopia single-flash ERG, the amplitude of the b-wave decreased
while the amplitude of the d-wave, which is the light-
Although we still do not know how the sign of defocus
OFF response, increased with increasing flash duration
is detected, we do know that there are probably two
(Sieving, 1993). In recent models (Bush & Sieving, 1996)
different retinal circuits. The data agree with earlier
the superimposition of depolarizing and hyperpolarizing
findings that reserpine, a dopamine uptake blocker,
activity (resulting in b-wave and parts of the d-wave) also
suppressed negative lens-induced and deprivation myopia
resembles the amplitude of the periodic flicker ERG. We
but not positive lens-induced hyperopia (Schaeffel et al.,
must consider that this sum electrical activity, as
1995). The two different circuits may, therefore, have a
measured on the cornea, is determined mostly by retinal
different accessibility to reserpine treatment, but may also
cells reaching perpendicular to the retinal surface (i.e.
have a different pharmacology. Apomorphine, a non-
bipolar cells, photoreceptors and Mtlller cells); cells that
selective dopamine agonist, which suppressed deprivation
extend horizontally within the retinal plane, like the
myopia (Rohrer et al., 1993) also inhibited lens-induced
amacrine cells, do not contribute to the ERG.
myopia, but not lens-induced hyperopia (Schmid et al.,
1997). Wallman et al. (1995) have shown that the target
tissues for compensation of positive and negative lenses Frequency tuning of the suppression of deprivation
are also different: with positive lenses, the choroid myopia and lens-induced myopia
increases in thickness to move the retina towards the We did not find significant differences in the suppres-
2672 H.N. SCHWAHN and F. SCHAEFFEL
sion of deprivation myopia and negative lens-induced
myopia with the flicker frequencies tested (6 and 12 Hz).
Schmid & Wildsoet (1996b) reported slightly different
suppression functions using stroboscopic flicker at 10, 15
and 20 Hz. In their experiments, deprivation myopia
declined linearly with increasing frequency but lens-
induced myopia was significantly suppressed only at
20 Hz. The latter result is different from ours, since we
obtained suppression already at 6 Hz. It is possible that
our experimental conditions are not comparable, since
Schmid & Wildsoet (1996b) added the flickering light to
an ambient background illumination of 105 lux. ERG
recordings under these conditions are not available. They
would have been most useful to determine whether there
was a similar correlation of "retinal activity" to
suppression of myopia, as in our study.
Role o f dopamine
Flickering light releases dopamine from retinal sources.
This was measured in various species (Kramer, 1971;
Kirsch & Wagner, 1989), but this has not been
demonstrated previously in chick. Rohrer et al. (1995)
hypothesized that deprivation myopia might be sup-
pressed by flicker, because intermittent illumination at
certain frequencies may counteract the drop in dopamine
release which occurs normally during deprivation and
lens treatment (Ohngemach et al., 1997). However, our
measurements show that retinal dopamine content
(intracellular DA stores) does not necessarily correlate
with the amount of deprivation myopia [Fig. 7, Fig. 8(B)].
The occluded eyes of the group treated with only 3 hr of
flicker per day showed no decrease in retinal dopamine
but still developed deprivation myopia. Even with the
most severe decrease of retinal dopamine from occlusion,
no myopia developed in the 12/4 12 hr flicker group. If the
release of dopamine from the retina, rather than retinal
content were important for eye growth control (Ohnge-
mach et al., 1997), we would expect that vitreal content is
increased under flickering light in the occluded eyes when
deprivation myopia is suppressed. An increase in
dopamine release was only found in the 12/4 3 hr flicker
group [Fig. 8(A)]. In this group, however, deprivation
myopia developed. Taken together, the available data still
reveal no clear picture of how dopamine is involved in
experimentally induced myopia. Probably signals carried
by other retinal neurotransmitters are also important (e.g.
Seltner & Stell, 1995; Seko et al., 1995).
Rohrer et al. (1993) found that intravitreal injection of
apomorphine, a non-selective dopamine agonist, was
competent to suppress deprivation myopia in chickens.
The effect of apomorphine was blocked by spiperone, an
antagonist of D2-receptors. If the "dopamine release"-
hypothesis of flicker was correct, our observations
suggest further experiments with flicker together with
simultaneous administration of a D2-antagonist, whereby
the rescuing effect of flickering light on myopia should be
reduced or blocked.
CONCLUSIONS
How the sign of imposed defocus is detected by local
retinal circuits is still not clear. Because the refractive
errors induced by positive and negative lenses are
suppressed most effectively by different flicker para-
meters, we propose that two different neural circuits are
involved at the retinal level. Deprivation myopia and
negative lens-induced myopia both are suppressed by
flicker with a similar dark phase duration (DP). The
suppression of myopia correlates with the DP and flicker
ERG amplitude. Suppression of hyperopia does not
correlate with DP or flicker ERG amplitude. Taken
together, the finding that reserpine did not prevent
compensation of positive lenses and choroidal thicken-
ing, and our observations on flicker-reared chicks we
suggest that the development of myopia and hyperopia
probably utilizes different retinal circuits with different
pharmacology.
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Acknowledgements~This study was supported by the German
Research Council (SFB 307, TP A7). We thank Dr. Norbert Benda
from the Institute of Medical Biometry, Tiibingen, for advice on the
statistics and Prof. Dr. E. Zrenner for general support.