Biomedical subjects

Frank Schaeffel

Publications and source records attributed to Frank Schaeffel.

At least 19 recordsLinked to original sources

PubMed2005

Pupillographic evaluation of the time course of atropine effects in the mouse eye.

PURPOSE: The nonselective muscarinic antagonist atropine is currently the most potent drug against myopia development in both humans and animal models. However, the mechanism by which myopia is suppressed is still unknown, and the time course of its action is not well documented. Therefore, we have studied the duration of mydriasis in the mouse, a new model of myopia, after topical application of a single eye drop with different doses of atropine. METHODS: The light-induced pupil response of the C57BL/6 (B6) wildtype strain was studied in alert mice that were restrained by grasping their necks. A video image-processing program detected the pupil and measured its diameter at 25 Hz sampling rate. To stimulate, an arrangement of green LEDs, which was attached to the recording video camera, could be flashed for 40 ms by pressing a key on the keyboard. A single drop of atropine solution (1, 0.5, or 0.1%) was instilled in one eye and the recovery of the pupil responses was followed for at least 150 h. Both eyes were measured. RESULTS: 1) Under the defined stimulation conditions, untreated wildtype mice displayed a pupil constriction of 23.7 +/- 2.4%. 2) All doses of atropine caused complete suppression of the pupil responses in the treated eyes within 1 min. 3) The pupil responses of the fellow eyes remained unaffected and were not different from those in untreated animals. 4) The recovery from mydriasis was very slow and did not show clear differences with dose. The extrapolated duration of complete recovery was about 10 d (0.1%: 217 h; 0.5%: 230 h; 1%: 294 h). CONCLUSIONS: Atropine caused a longlasting suppression of the pupil responses in the mouse eye. That the duration of recovery was not obviously dose-dependent suggests that all doses used in this study were saturating the receptors in the iris musculature.

Animals↗

PubMed2005

Grating acuity at different luminances in wild-type mice and in mice lacking rod or cone function.

PURPOSE: The mouse eye has become an important model in vision research. However, it is not known how visual acuity changes with luminance. Therefore, grating acuity of mice was measured at different luminances in an automated optomotor paradigm. Furthermore, mutant mice lacking either rods (RHO-/- and CNGB1-/-) or cones (CNGA3-/-), or both, were studied to determine the rod and cone contribution to visual acuity. METHODS: Freely ranging individual mice were automatically tracked at a 25-Hz sampling rate with a self-programmed video system in a large rotating optomotor drum. The drum had a square-wave grating inside with adjustable spatial frequency. The angular speed of the mice with respect to the center of the drum and the angular orientation of the snout-tail body axis were analyzed. In addition, the motor activity of the wild-type mice was recorded at different luminances. RESULTS: The optomotor drum provided reliable data on visual input to the mouse's behavior and was convenient to use, since the experimenter's had only to place the mice individually in a Perspex cylinder. Optomotor grating acuity of the wild-type mice was limited to 0.3 to 0.4 cyc/deg. Maximum optomotor responses were obtained at 0.1 to 0.2 cyc/deg. The importance of visual input declined monotonically with decreasing luminance (30 cd/m2, 100%; 0.1 cd/m2, 76.4%; 0.005 cd/m2, 45.9%; and darkness, -9%). Mice lacking functional rods were able to resolve gratings up to 0.1 cyc/deg at 30 cd/m2. Surprisingly, mice lacking functional cones had an optomotor acuity that was similar to the wild-type. Double-knockout mice without rods and cones had no detectable grating acuity. CONCLUSIONS: Because the visual system of the mouse is more responsive at bright luminances, experiments in which visual input is important should be performed in photopic conditions (30 cd/m2 or even more). Apparently, spatial vision is governed by the rod system, which is not saturated in the mesopic or low photopic range. Mice lacking both rods and cones have no detectable grating acuity, indicating that the retinal melanopsin system does not contribute to spatial vision.

Animals↗

PubMed2004

Temporal changes of novel transcripts in the chicken retina following imposed defocus.

PURPOSE: Changes in retinal gene expression are one of the first steps in the signaling pathway underlying the visual control of eye growth. We tried to identify novel, yet unknown, genes, that alter their expression pattern following imposed defocus, wearing of diffusers, or during recovery from myopia. Sequences found earlier by differential display studies were applied to 5'-RACE and identified as 15 kDa selenoprotein P and prolidase. Moreover, we obtained more sequence information for a yet unidentified gene. We have studied the time course of expressions of these genes following lens or diffuser treatment. METHODS: Ten to 14 day old white leghorn chickens (4-7) were treated with a monocular +7 D or -7 D lenses for 2, 4, 6, or 24 h, or treated with monocular or binocular diffusers for 2, 4, or 6 h. Chickens of another group were allowed to recover from 4 days of diffuser wear for 4 h. Untreated chicks served as a control for contralateral eye effects. Following the extraction of retinal RNA, the relative expression of the three genes was determined by semi-quantitative real time PCR. RESULTS: We found a significant up regulation of selenoprotein P expression after 24 h of treatment with positive (+380%) or negative lenses (+387%) which was even more prominent in the contralateral untreated eyes (positive: +542%; negative: +786%). A rapid change in selenoprotein mRNA levels was induced by binocular diffuser wear for 2 h (+425%), whereas defocus blur in one eye led to an increase only after 6 h (+261%). There was a significant upregulation of prolidase mRNA after 24 h of treatment with positive (+75%) but not with negative lenses. Moreover, blur induced by diffusers resulted in a highly significant rise of prolidase mRNA levels after 4 h, both with monocular (142%) and binocular (106%) treatment. This is similar to what was found in the previous differential display (DD) screening of monocularly treated eyes. In contrast to the findings of the DD screening, the mRNA expression of the unknown gene remained unchanged both after hyperopic and myopic defocus. Again, blur induced by diffusers evoked the most prominent change after 6 h of binocular treatment. There were no significant alterations in the mRNA levels of the three investigated genes after 4 h of recovery from myopia that was induced by a 4 day period of diffuser treatment. CONCLUSIONS: The mRNA expression of selenoprotein P, prolidase, and of the not yet identified gene (sequence 3) is clearly altered by retinal image degradation imposed by diffuser wearing and, in part, by defocus imposed by spectacle lenses. However, none of the candidates are regulated by the sign of imposed defocus, suggesting a role in retinal contrast processing.

Animals↗

PubMed2004

Early transcriptional changes of retinal and choroidal TGFbeta-2, RALDH-2, and ZENK following imposed positive and negative defocus in chickens.

PURPOSE: Imposing defocus to the retina results in compensatory changes of axial eye growth. It is not clear which factors initially contribute to this process and whether they act on the post-translational, translational, or transcriptional level. We have measured early changes in mRNA levels, in response to imposed negative and positive defocus, of the transcription factor ZENK, the retinoic acid synthesis enzyme RALDH-2, and the growth factor TGFbeta-2. METHODS: Chickens 11 days of age were unilaterally treated with positive or negative spectacle lenses of 7 D power. After 0, 15, 30, and 120 min, mRNA was extracted from retina and choroid, and the concentration of the mRNAs of the three candidates was measured by quantitative real time PCR in both eyes. RESULTS: ZENK in the retina and RALDH-2 in the choroid displayed parallel signs of defocus dependent changes in mRNA levels after 15 or 30 min, respectively. ZENK mRNA levels were reduced in the retina after 15 min with both types of lenses but were then up regulated at 30 min with positive lenses and down regulated with negative lenses, similar to the previously observed changes in ZENK protein levels. Changes of RALDH-2 and TGFbeta-2 mRNA levels were confined to the choroid. Treatment with negative lenses resulted in a rapid (15 min) and persistent decrease in TGFbeta-2 mRNA concentration in the choroid. Negative lenses provoked parallel but less pronounced alterations in the open fellow eyes. CONCLUSIONS: Imposed defocus triggers extensive transcriptional changes of ZENK in the retina, and of TGFbeta-2 and RALHD-2 in the choroid. Changes in retina and choroid are rapid, show no phase delay with respect to each other, and can be considered, in the case of RALDH-2 and ZENK, as specific for the sign of imposed defocus. They occur prior to any morphological changes. This is consistent with a role in causing or controlling later changes in eye growth.

Aldehyde Oxidoreductases↗

PubMed2004

Localization and regulation of glucagon receptors in the chick eye and preproglucagon and glucagon receptor expression in the mouse eye.

Myopia is a condition in which the eye is too long for the focal length of cornea and lens. Analysis of the messengers that are released by the retina to control axial eye growth in the animal model of the chicken revealed that glucagon-immunoreactive amacrine cells are involved in the retinal image processing that controls the growth of the sclera. It was found that the amount of retinal glucagon mRNA increased during treatment with positive lenses and pharmacological studies supported the idea that glucagon may act as a stop signal for eye growth. Glucagon exerts its regulatory effects by binding to a single type of glucagon receptor. In this study, we have sequenced the chicken glucagon receptor and compared its DNA and amino acid sequence with the human and mouse homologues. After sequencing about 80% of the receptor, we found a homology between 79.4 and 75.6% on cDNA level. At the protein level, about 73% of the amino acids were identical. Moreover, the cellular localization and regulation of the glucagon receptor in the chick retina was studied. In situ hybridization studies showed that many cells in the ganglion cell layer and inner nuclear layer, and some cells in the outer nuclear layer, express the receptor mRNA. Injection of the glucagon agonist Lys17,18,Glu21-glucagon induced a down-regulation of glucagon receptor mRNA content. Since the mouse would be an attractive mammalian model to study the biochemical and genetic basis of myopia, and because recent studies have demonstrated that form deprivation myopia can be induced, the expression of preproglucagon and glucagon receptor genes were also studied in the mouse retina and were found to be expressed.

Amacrine Cells↗

PubMed2004

A paraxial schematic eye model for the growing C57BL/6 mouse.

PURPOSE: The mouse eye has potential to become an important model for studies on the genetic control of eye growth and myopia. However, no data are published on the development of its optical properties. We developed a paraxial schematic model of the growing eye for the most common laboratory mouse strain, the C57BL/6 mouse, for the age range between 22 and 100 days. METHODS: Refractive development was followed with eccentric infrared photorefraction and corneal curvature with infrared photokeratometry. To measure ocular dimensions, freshly excised eyes were immediately frozen after enucleation to minimize distortions. Eyes were cut with a cryostat down to the bisecting horizontal plane, until the optic nerve head became visible. The standard deviations were +/-10 microm for repeated measurements in highly magnified videographs, taken in several section planes close to the equator in the same eyes. To evaluate inter-eye and inter-individual variability, a total of 20 mice (34 eyes) were studied, with 3-4 eyes for each of the 9 sampling ages. Schematic eye models were developed using paraxial ray tracing software (OSLO, LT Lambda Research Corporation, and a self-written program). RESULTS: The measured refractive errors were initially +4.0+/-0.6 D at approximately 30 days, and levelled off with +7.0+/-2.5 D at about 70 days. Corneal radius of curvature did not change with age (1.414+/-0.019 mm). Both axial lens diameter and axial eye length grew linearly (regression equations: lens, 1619 microm +5.5 microm/day, R=0.916; axial length, 2899 microm +4.4 microm/day, R=0.936). The lens grew so fast that vitreous chamber depth declined with age (regression equation: 896 microm -3.2 microm/day, R=0.685). The radii of curvature of the anterior lens surface increased during development (from 0.982 mm at day 22 to 1.208 mm at day 100), whereas the radii of the posterior lens surface remained constant (-1.081+/-0.054 mm). The calculated homogeneous lens index increased linearly with age (from 1.568 to 1.605). The small eye artifact, calculated from the dioptric difference of the positions of the vitreo-retinal interface and the photoreceptor plane, increased from +35.2 to +39.1 D, which was much higher than the hyperopia measured with photorefraction. Retinal image magnification increased from 31 to 34 microm/deg, and the f/number remained < or =1 at all ages, suggesting a bright retinal image. A calculated axial eye elongation of 5.4-6.5 microm was sufficient to make the schematic eye 1 D more myopic. CONCLUSIONS: The most striking features of the mouse eye were that linear growth was slow but extended far beyond sexual maturity, that the corneal curvature did not increase, and that the prominent lens growth caused a developmental decline of the vitreous chamber depth.

Animals↗

PubMed2004

In vivo biometry in the mouse eye with low coherence interferometry.

PURPOSE: A major drawback of the mouse model of myopia is that the ocular dimensions cannot be measured in vivo, and that histological techniques post-mortem suffer from limited resolution. We have tested the potential of a newly developed technique, optical low coherence interferometry (OLCI), adapted for short measurement distances by Meditec, Carl Zeiss, Jena, Germany (the "ACMaster"). Using this technique, ocular biometry was performed in mice with normal vision and after deprivation of form vision. METHODS: Axial eye length, corneal thickness and anterior chamber depth were measured in 23 mice, aged 25-53 days, and standard deviations from repeated measurements in the same eyes, as well as intra-individual and inter-individual variability were determined in different age groups. The data were compared to those from a preceding study in which biometrical data were obtained from frozen sections [Vision Res. 44 (2004) 1857]. Refractions were measured by automated infrared photorefraction. Mice had either normal visual exposure or were monocularly deprived of form vision for 14 days. RESULTS: Using OLCI, axial length could be determined with an average standard deviation of 8.0 +/- 2.9 microm, corneal thickness with 3.5 +/- 2.1 microm, and anterior chamber depth with 10.6 +/- 12.3 microm. Neither axial length, nor corneal thickness, nor anterior chamber depth were significantly different in left and right eyes of individual mice that had normal visual experience (mean absolute difference between axial lengths: 17 +/- 18 microm, between corneal thickness 5.1 +/- 4.8 microm, and between anterior chamber depths 16.7 +/- 14.8 microm). Compared to the variability that was previously found in frozen sections, the variability of axial length measurements with OLCI was 2.7 times less. After two weeks of form deprivation, OLCI revealed a significant axial elongation in the occluded eyes, compared to the contralateral fellow eyes (+38 +/- 36 microm or 1.16%, p = 0.045, n = 7, paired t-test). In this sample, no accompanying myopic shift was observed in the occluded eyes but this observation is not unexpected given the inherently variable responses of mouse eye growth to visual deprivation. CONCLUSION: OLCI had sufficient resolution in living mice to detect axial length changes in vivo that were equivalent to a dioptric change of 2 D. Using this technique, it was confirmed that mouse eyes respond to form deprivation by axial elongation, similar to the eyes of other animal models. The lack of a myopic shift in this sample, despite the axial elongation, demonstrates that biometric data are particularly important when the mouse eye is used as a model to study myopia.

Animals↗

PubMed2004

Measurement of refractive state and deprivation myopia in two strains of mice.

PURPOSE: The mouse eye has a bright retinal image (f/number <1) but low optical quality (visual acuity about 0.5 cpd) that may render emmetropization unnecessary. However, this species is potentially a powerful model to study eye growth and myopia because its genome can be readily manipulated and has been completely sequenced. We have investigated how precisely eyes of mice can be refracted and tested whether deprivation myopia can be induced by frosted diffusers. METHODS: An automated eccentric infrared photorefractor was adapted to refract eyes of two mouse strains--C57BL/6 (B6) and DBA/2 (D2)--during Tropicamide cycloplegia without anesthesia. Axial lengths were measured in highly magnified video images of freshly excised eyes. Plastic hemispherical diffusers were applied between postnatal days and 29 and left attached for 7 or 14 days. RESULTS: (1) Trial lenses ranging from +10 to -10 D produced high correlations between the brightness slope in the pupil and applied lens power (r = 0.81 and r = 0.87), demonstrating reliable refraction. Five repeated measures in 12 eyes showed an average standard deviation of 3.0 D, equivalent to an axial length change <10 microm (derived from schematic eye modeling). (2) Deprivation produced a significant shift toward myopia, relative to untreated eyes, but only after 14 days and only in B6 mice (p = 0.02 with or p = 0.00038 without one outlier; N = 9). In contrast, DBA/2J were unaffected by occlusion, perhaps due to mutations that target eye, lens, or anterior segment. (3) Both eyes of untreated animals often had axial lengths that differed markedly. Surprisingly, we detected no significant correlation between refractive error and axial length after treatment. CONCLUSIONS: The infrared refraction technique is sufficiently sensitive to resolve equivalent changes in axial length of only +/- 10 microm in alert mice. Prolonged occlusion produces a significant myopic shift in B6 mice, but not in D2 mice. Even among isogenic B6 mice, the response is variable for reasons that presumably trace back to subtle developmental, environmental, and technical factors.

Animals↗

PubMed2004

Effects of quisqualic acid on retinal ZENK expression induced by imposed defocus in the chick eye.

PURPOSE: Expression of the transcription factor ZENK in glucagon amacrine cells of the chicken retina is enhanced after treatment with positive spectacle lenses and reduced after treatment with negative lenses. ZENK may, therefore, have an important role in emmetropization. To learn more about its regulation, we have studied its expression after retinal intoxication with quisqualic acid (QA, a glutamatergic excitotoxin). METHODS: Lenses of either +7 or -7 D power were placed in front of the eyes of young chickens 6 days after intravitreal QA injections. By this time, QA had caused severe damage to the retina. After 2 hours of lens wearing, changes in ZENK immunoreactivity were measured by means of double staining. In another experiment, lenses were worn for 4 days to study the residual function of emmetropization. RESULTS: QA injections caused a massive loss of cells in the inner nuclear layer and the ganglion cell layer but left the numbers of glucagon cells unchanged. Four of six QA-injected eyes became more myopic in response to wearing positive lenses, and all eyes with negative lenses also became myopic. QA caused a general reduction in ZENK expression, and there was no clear evidence that ZENK expression was still controlled by the sign of imposed defocus. CONCLUSIONS: After severe destruction of the inner retina by QA, retinal image processing appeared to be reduced to blur detection with no sign, causing myopia with both types of lenses. QA must remove synaptic input to the glucagon cells, which is necessary to transmit the information on the sign of imposed defocus.

Amacrine Cells↗

PubMed2004

Posttranscriptional regulation of the immediate-early gene EGR1 by light in the mouse retina.

Synaptic plasticity is modulated by differential regulation of transcription factors such as EGR1 which binds to DNA via a zinc finger binding domain. Inactivation of EGR1 has implicated this gene as a key regulator of memory formation and learning. However, it remains puzzling how synaptic input can lead to an up-regulation of the EGR-1 protein within only a few minutes. Here, we show by immunohistochemical staining that the EGR-1 protein is localized in synapses throughout the mouse retina. We demonstrate for the first time that two variants of Egr-1 mRNA are produced in the retina by alternative polyadenylation, with the longer version having an additional 293 base pairs at the end of the 3'UTR. Remarkably, the use of the alternative polyadenylation site is controlled by light. The additional 3'UTR sequence of the longer variant displays an even higher level of phylogenetic conservation than the coding region of this highly conserved gene. Additionally, it harbours a cytoplasmic polyadenylation element which is known to respond to NMDA receptor activation. The longer version of the Egr-1 mRNA could therefore rapidly respond to excitatory stimuli such as light or glutamate release whereas the short variant, which is predominantly expressed and contains the full coding sequence, lacks the regulatory elements for cytoplasmic polyadenylation in its 3'UTR.

Animals↗

PubMed2004

Stimulating human accommodation without changes in focus.

PURPOSE: Inspired by the finding in chickens that preferential stimulation of the ON retinal system suppresses myopia induced by negative spectacle lens wear and that stimulation of the OFF system suppresses the hyperopia induced by positive lens wear, we sought to determine whether stimulation of the ON-OFF retinal systems could drive directional accommodation responses in humans. If emmetropisation and accommodation use similar image processing algorithms, more accommodation would be expected with OFF stimulation. METHODS: Accommodation responses were measured while viewing a computer-generated pattern designed to stimulate the ON-OFF systems. The stimulus comprised a rectangular field (12 x 9.5 cm) on a black background filled with 196 discs (diameters: 0.4-1.0 cm). These were presented on an LCD monitor in a dark room at a viewing distance of 55 cm (1.8 D). Thirteen subjects aged 21-37 years took part. The individual discs had saw-tooth shaped temporal luminance profiles with the same time period but with random phases with respect to each other, so that the mean brightness of the stimulus was constant. To eliminate accommodation responses based on other cues (i.e. proximity) a 0.5 mm artificial pupil was used to open the accommodation loop. Refraction in the vertical pupil meridian was continuously recorded with an infrared photorefractor (the PowerRefractor). To verify that computer-based stimuli presented within our experimental design were effective in driving accommodation, previously studied stimuli were also tested: changes in size (looming) and incremental low pass filtering. RESULTS: Preferential stimulation of the ON or OFF subsystems produced a convincing depth illusion in all subjects (which was psychophysically confirmed in four subjects). Although the stimulus appeared to move in depth it did not produce accommodation responses that were consistent with that, i.e. the accommodation system did not appear to fluctuate in rhythm with the temporal oscillations of the stimulus. As the target appeared to loom it induced a greater accommodation response then when it appeared to recede. The looming target produced changes in the accommodation response in nine of 13 subjects that were consistent with its perceived change in proximity (although the target did not actually move in depth). Incremental low pass filtering produced non-directional drifts of accommodation in all subjects. Combinations of the stimuli (i.e. looming and low pass filtering, ON/OFF and looming) were not more effective stimuli to accommodation. After removal of the artificial pupil (closed loop conditions), accommodation was no longer induced with any of these stimuli. CONCLUSIONS: Although the preferential ON or OFF stimulation produced a pronounced illusion of motion in depth despite constant average brightness, proximal accommodation was induced in only one subject. Therefore, the ON/OFF stimulation appeared to have only minor input into proximal accommodation. Potential inputs into reflex accommodation need to be defined in further studies.

Accommodation, Ocular↗

PubMed2004

Effects of positive and negative lens treatment on retinal and choroidal glucagon and glucagon receptor mRNA levels in the chicken.

PURPOSE: It has been found in the chicken that the amount of retinal glucagon mRNA increases during treatment with positive lenses. Pharmacological studies support the idea that glucagon may act as a stop signal for visually induced eye growth. To gain more insight into the functional role of glucagon, the changes of glucagon and glucagon receptor mRNA concentrations in retina and choroid over time were studied. Furthermore, the abundance of glucagon and the glucagon receptor was studied in different fundal layers (retina, retinal pigment epithelium[RPE], choroid) and the blood. METHODS: Semiquantitative real-time RT-PCR was used to measure glucagon and glucagon receptor mRNA levels in retina and choroid after positive and negative lens treatment for 2, 6, or 24 hours, by unilateral -7- or +7-D lenses. Contralateral eyes served as the control, and completely untreated animals provided further reference data. Intravitreal colchicine injections (which are known to reduce the number of glucagon cells sharply) were used to verify that the related decline in glucagon mRNA could be measured by real-time RT-PCR. RESULTS: In the retina, treatment with -7-D lenses induced an initial upregulation of glucagon mRNA in both eyes, followed by a significant downregulation. The treatment with +7-D lenses showed a significant but transient downregulation in the control eye superimposed on a trend toward upregulation in the treated eye. However, the changes in glucagon mRNA expression were not confined to the lens-treated eyes but were also found, although sometimes to a lesser extent, in the non-lens-covered fellow eyes. There was evidence of a transient increase in glucagon receptor mRNA levels in lens-treated eyes after either -7- or +7-D lens wear. In the choroid, no effect of imposed defocus was detected. The injection of colchicine led to the destruction of approximately 75% of the glucagon amacrine cells but the mRNA level of retinal glucagon decreased by only approximately 50%. Glucagon receptor expression was found to be higher in the RPE than the retina and choroid whereas, in the blood, glucagon and glucagon receptor mRNA expression was below detection level. CONCLUSIONS: The observed bidirectional regulation of glucagon mRNA in correlation with the sign of imposed defocus supports the idea that glucagon may act as a stop-and-go signal for eye growth. This is in line with a previous proposal based on studies of changes of the glucagon peptide content.

Animals↗

PubMed2003

An evaluation of the lag of accommodation using photorefraction.

OBJECTIVE: The lag of accommodation which occurs in most human subjects during reading has been proposed to explain the association between reading and myopia. However, the measured lags are variable among different published studies and current knowledge on its magnitude rests largely on measurements with the Canon R-1 autorefractor. Therefore, we have measured it with another technique, eccentric infrared photorefraction (the PowerRefractor), and studied how it can be modified. METHODS: Particular care was taken to ensure correct calibration of the instrument. Ten young adult subjects were refracted both in the fixation axis of the right eye and from the midline between both eyes, while they read text both monocularly and binocularly at 1.5, 2, 3, 4 and 5 D distance ("group 1"). A second group of 10 subjects ("group 2"), measured from the midline between both eyes, was studied to analyze the effects of binocular vs monocular vision, addition of +1 or +2 D lenses, and of letter size. Spherical equivalents (SE) were analyzed in all cases. RESULTS: The lag of accommodation was variable among subjects (standard deviations among groups and viewing distances ranging from 0.18 to 1.07 D) but was significant when the measurements were done in the fixation axis (0.35 D at 3 D target distance to 0.60 D at 5 D with binocular vision; p<0.01 or better all cases). Refracting from the midline between both eyes tended to underestimate the lag of accommodation although this was significant only at 5 D (ANOVA: p<0.0001, post hoc t-test: p<0.05). There was a small improvement in accommodation precision with binocular compared to monocular viewing but significance was reached only for the 5 D reading target (group 1--lags for a 3/4/5 D target: 0.35 vs 0.41 D/0.48 vs 0.47 D/0.60 vs 0.66 D, ANOVA: p<0.0001, post hoc t-test: p<0.05; group 2--0.29 vs 0.12 D, 0.33 vs 0.16 D, 0.23 vs -0.31 D, ANOVA: p<0.0001, post hoc t-test: p<0.05). Adjusting the letter height for constant angular subtense (0.2 deg) induced scarcely more accommodation than keeping letter size constantly at 3.5 mm (ANOVA: p<0.0001, post hoc t-test: n.s.). Positive trial lenses reduced the lag of accommodation under monocular viewing conditions and even reversed it with binocular vision. CONCLUSIONS: After consideration of possible sources of measurement error, the lag of accommodation measured with photorefraction at 3 D (0.41 D SE monocular and 0.35 D SE binocular) was in the range of published values from the Canon R-1 autorefractor. With the measured lag, simulations of the retinal images for a diffraction limited eye suggest surprisingly poor letter contrast on the retina.

Accommodation, Ocular↗

PubMed2003

Molecular biology of myopia.

Experiments in animal models of myopia have emphasised the importance of visual input in emmetropisation but it is also evident that the development of human myopia is influenced to some degree by genetic factors. Molecular genetic approaches can help to identify both the genes involved in the control of ocular development and the potential targets for pharmacological intervention. This review covers a variety of techniques that are being used to study the molecular biology of myopia. In the first part, we describe techniques used to analyse visually induced changes in gene expression: Northern Blot, polymerase chain reaction (PCR) and real-time PCR to obtain semi-quantitative and quantitative measures of changes in transcription level of a known gene, differential display reverse transcription PCR (DD-RT-PCR) to search for new genes that are controlled by visual input, rapid amplification of 5' cDNA (5'-RACE) to extend the 5' end of sequences that are regulated by visual input, in situ hybridisation to localise the expression of a given gene in a tissue and oligonucleotide microarray assays to simultaneously test visually induced changes in thousands of transcripts in single experiments. In the second part, we describe techniques that are used to localise regions in the genome that contain genes that are involved in the control of eye growth and refractive errors in mice and humans. These include quantitative trait loci (QTL) mapping, exploiting experimental test crosses of mice and transmission disequilibrium tests (TDT) in humans to find chromosomal intervals that harbour genes involved in myopia development. We review several successful applications of this battery of techniques in myopia research.

Animals↗

PubMed2003

Interactions of genes and environment in myopia.

Myopia is a condition in which the eye is too long for the focal length of cornea and lens, and the plane of sharp focus ends up in front of the retina. Given that the growth of the length of the eye is normally controlled with extreme precision by an image-processing feedback mechanism in the retina, myopia can either be the result of inappropriate visual stimulation, genetically determined changes in the gain or offset of the feedback loops or of inappropriate responses of the target tissues. There is no doubt that an environmental component is involved and extended near work appears to be the major risk factor. However, there is also no doubt that myopia is inherited since myopic parents are much more likely to have myopic children, and myopia is far more frequent in Asian populations than in the USA or Europe, even if groups are compared that have performed similar amounts of near work. A number of systemic or ophthalmic diseases are associated with myopia, indicating that metabolic conditions may interfere either with the gains of the feedback loops or the responses of the target tissue, the sclera. Since there is still no therapy against myopia development, research is directed toward the identification of genes that control the axial elongation of the eye.

Animals↗

PubMed2002

Effects of longitudinal chromatic aberration on accommodation and emmetropization.

PURPOSE: Less accommodation was found when human subjects read in blue (peak at about 440 nm) than when they read in red light (above 600 nm; [Kroger & Binder, British Journal of Ophthalmology 84 (2000) 890]). On the other hand, emmetropization in chickens did not appear to compensate for the chromatic defocus (385 nm versus 665 nm; [Rohrer, Schaeffel & Zrenner, Journal of Physiology 449 (1992) 363]). The apparently contradictory result was studied in more detail in humans and chickens. METHODS: Accommodation was measured with an eccentric infrared photorefractor, the PowerRefractor, in human subjects reading under quasi-monochromatic illumination conditions. Chickens were refracted in quasi-monochromatic ambient illumination but with no particular fixation target. In a second experiment, they were also raised in monochromatic light for two days and subsequently refracted both in complete darkness, in monochromatic light, and in white light, both without and with cycloplegia. RESULTS: Consistent with the initial report by Kroger and Binder [British Journal of Ophthalmology 84 (2000) 890], accommodation in human subjects was found to shift in accordance with the chromatic aberration function. An immediate shift in accommodation tonus was also found in the chickens when they were refracted under red and in blue ambient illumination (average difference between refractions in both conditions: 1.26+/-0.54 D, p<0.001 paired t-test). This value is close to the chromatic focus difference between the two wavelengths (1.5 D [Mandelman & Sivak, Vision Research 23 (1983) 1555]). When chickens were raised in blue or red light for two days, and their refractions were subsequently measured in complete darkness, they showed also a difference in refractions (1.41+/-1.00 D; ANOVA: p<0.0012, post hoc t-test: at least p<0.05 among different groups). This difference was no longer significant when they were refracted in white light but became again significant when they were cyclopleged (0.57+/-0.58 D, p=0.039, unpaired t-test). The latter observation makes it unlikely that the difference resulted just from a shift in the resting tonus of accommodation. CONCLUSIONS: (1) Imposed chromatic defocus produces a shift in accommodation tonus in both humans and chickens which is, in the case of the chicken, followed by a shift in cycloplegic refractive state into the same direction, (2) the difference to the previous study by Rohrer, Schaeffel and Zrenner [Journal of Physiology 449 (1992) 363] can be explained from the fact that shorter wavelengths were used than in the present study, at which emmetropization was no longer functional and, (3) the small amplitude and the variability of the shifts in refraction do not allow clear statements about the role of the "lag of accommodation" in refractive development but they show that several cone types contribute to emmetropization.

Accommodation, Ocular↗

PubMed2002

Kappa and Hirschberg ratio measured with an automated video gaze tracker.

PURPOSE: To develop a fast automated procedure to measure kappa and the Hirschberg ratio for immediate use in a video gaze tracker. METHODS: Using the hardware platform of the PowerRefractor and a 200 mm lens, the pupil was imaged with a resolution of 57 pixels/mm, at a camera distance of 90 cm. Both the positions of the first Purkinje image and the edges of the pupil were located at 25 Hz sampling rate with subpixel resolution using video image processing software developed under Borland C++. Subjects fixated on a red spot on the left side of the monitor. If their fixation was stable (standard deviation <0.2 degrees in 25 subsequent measurements evaluated in 1 s), the fixation spot appeared automatically on the right side, and the procedure was repeated. Data on the angular position of the optical axis for both targets were stored and provided kappa and the Hirschberg ratio with a standard deviation of about 0.2 degrees or better. This enabled the system to track fixation with a resolution of about 0.2 degrees. RESULTS: (1) Kappa was +3.91+/-2.73 degrees (right eyes), -3.93+/-2.68 degrees (left eyes, mean +/- SD from 24 young adults). Kappa was highly correlated in both eyes (r = 0.8996), but there were significant asymmetries between both eyes in three subjects (delta up to 3 degrees). (2) The Hirschberg ratios were 12.93+/-1.23 degrees/mm = 22.56 delta/mm (right eyes) and 12.82+/-1.49 degrees/mm = 22.38 delta/mm (left eyes). They were also highly correlated in both eyes (r = 0.931). (3) Neither kappa nor the Hirschberg ratios were correlated to refractive errors (range +0.50 to -7.75 D, mean -1.73+/-2.29 D [spherical equivalents]). CONCLUSIONS: (1) The video gaze tracker measured fixation with an angular resolution high enough to display the eye position during reading of individual words on the computer screen. (2) The applicable Hirschberg ratio changed with the power of the spectacles of the subjects by about 3% per diopter. (3) In some subjects, there were significant differences in the geometry of both eyeballs, which may be important in the analysis of strabismus. (4) Consistent with the assumption of retinal stretching, the linear distance of the fovea from the optical axis appeared to increase in myopia.

Adult↗

PubMed2002

Peripheral refractive errors in myopic, emmetropic, and hyperopic young subjects.

To gain more insight into the relationship between foveal and peripheral refractive errors in humans, spheres, cylinders, and their axes were binocularly measured across the visual field in myopic, emmetropic, and hyperopic groups of young subjects. Both automated infrared photorefraction (the "PowerRefractor"; www. plusoptix.de) and a double-pass technique were used because the PowerRefractor provided extensive data from the central 44 deg of the visual field in a very convenient and fast way. Two-dimensional maps for the average cross cylinders and spherical equivalents, as well as for the axes of the power meridians of the cylinders, were created. A small amount of lower-field myopia was detected with a significant vertical gradient in spherical equivalents. In the central visual field there was little difference among the three refractive groups. The established double-pass technique provided complementary data also from the far periphery. At 45 deg eccentricity the double-pass technique revealed relatively more hyperopic spherical equivalents in myopic subjects than in emmetropic subjects [+/-2.73 +/- 2.85 D relative to the fovea, p < 0.01 (+/- standard deviation)] and more myopic spherical equivalents in hyperopic subjects (-3.84 +/- 2.86 D relative to the fovea, p < 0.01). Owing to the pronounced peripheral astigmatism, spherical equivalents (refractions with respect to the plane of the circle of least confusion) became myopic relative to the fovea in all three groups. The finding of general peripheral myopia was unexpected. Its possible roles in foveal refractive development are discussed.

Adult↗