The refractive development of the eye of the American kestrel (Falco sparverius): a new avian model

Summary Most measures of avian visual performance are carried out on commonly available domestic species such as the chicken, and most of the data on avian induced refractive error deals with chickens. Raptors are predatory birds in which good visual resolving ability is particularly important. Behavioral studies indicate that the eyes of raptors have two to three times the resolving ability of the human eye. The domestic chicken is precocial at hatching whereas most raptors are semi-altricial. This study was an effort to determine if the effect of early visual deprivation on the refractive development of the chicken eye can be reproduced in the American kestrel, a species which is not domesticated and in which the need for acute vision is particularly important.Visual deprivation was achieved by unilaterally applying translucent plastic goggles over the eyes of kestrels two days after hatching. Refractive error was measured using a retinoscope and trial lenses. Ocular growth was monitored by A-scan ultrasonography, and frozen ocular sections of sacrificed birds. The effect of the experimental manipulation on the contralateral control eye and body weight was evaluated each day over a 42-day period. The goggles did not significantly affect the normal changes in body weight or the normal pattern of ocular growth and refractive development in the untreated eyes. An analysis of the refractive state changes as a result of form deprivation was made each week for 6 weeks after hatching on both the treated and untreated eyes in a separate group of experimental birds. Visual form deprivation caused a significant myopic shift in refractive error and a significant increase in the vitreous chamber depth in the treated eyes at 3 and 6 weeks of age. However, the amount of myopia produced is much less than that induced in chicks, and in certain cases hyperopia is produced. The kestrels recover from myopia and hyperopia within 10 days of goggle removal, after 3 to 4 weeks of deprivation.This study is the first indication that chickens may not be a representative bird model for studying form deprivation myopia. First, myopia is not always produced in kestrels in response to form deprivation. Second, kestrels are severely myopic at hatching and therefore, the direction of emmetropization is opposite to that found in hatchling chicks.     

How can squint change the spacing of ocular dominance columns?

The pattern of ocular dominance columns in primary visual cortex of mammals such as cats and macaque monkeys arises during development by the activity-dependent refinement of thalamocortical connections. Manipulating visual experience in kittens by the induction of squint leads to the emergence of ocular dominance columns with a larger size and larger column-to-column spacing than in normally raised animals. The mechanism underlying this phenomenon is presently unknown. Theory suggests that experience cannot influence the spacing of columns if the development proceeds through purely Hebbian mechanisms. Here we study a developmental model in which Hebbian mechanisms are complemented by activity-dependent regulation of the total strength of afferent synapses converging onto a cortical neurone. We show that this model implies an influence of visual experience on the spacing of ocular dominance columns and provides a conceptually simple explanation for the emergence of larger sized columns in squinting animals. Assuming that during development cortical neurones become active in local groups, which we call co-activated cortical domains (CCDs), ocular dominance segregation is controlled by the size of these groups: (1) Size and spacing of ocular dominance columns are proportional to the size sigma of CCDs. (2) There is a critical size sigma* of CCDs such that ocular dominance columns form if sigmasigma*. This critical size of CCDs is determined by the correlation functions of activity patterns in the two eyes and specifies the influence of experience on ocular dominance segregation. We show that sigma* is larger with squint than with normal visual experience. Since experimental evidence indicates that the size of CCDs decreases during development, ocular dominance columns are predicted to form earlier and with a larger spacing in squinters compared to normal animals.     

On emmetropization

The growth of the ocular elements, such as axial length, corneal power, and lens power, is represented by a set of first order, linear, coupled differential equations. Since the rate of growth and the size of the ocular elements vary from one individual to the next, the parameters which arise in these differential equations are assumed to be Gaussian random variables. These assumptions provide a basis for understanding the controlled growth of the eye. They predict that the distribution of refractive error is normal at birth and leptokurtic in adulthood. They also predict that the variance of refractive error will decrease with age provided the second order moments of the random variables obey some rather general conditions.     

Genomewide scan in Ashkenazi Jewish families demonstrates evidence of linkage of ocular refraction to a QTL on chromosome 1p36

The development of refractive error is mediated by both environmental and genetic factors. We performed regression-based quantitative trait locus (QTL) linkage analysis on Ashkenazi Jewish families to identify regions in the genome responsible for ocular refraction. We measured refractive error on individuals in 49 multi-generational American families of Ashkenazi Jewish descent. The average family size was 11.1 individuals and was composed of 2.7 generations. Recruitment criteria specified that each family contain at least two myopic members. The mean spherical equivalent refractive error in the sample was −3.46D (SD=3.29) and 87% of individuals were myopic. Microsatellite genotyping with 387 markers was performed on 411 individuals. We performed multipoint regression-based linkage analysis for ocular refraction and a log transformation of the trait using the statistical package Merlin-Regress. Empirical genomewide significance levels were estimated through gene-dropping simulations by generating random genotypes at each of the 387 markers in 200 replicates of our pedigrees. Maximum LOD scores of 9.5 for ocular refraction and 8.7 for log-transformed refraction (LTR) were observed at 49.1 cM on chromosome 1p36 between markers D1S552 and D1S1622. The empirical genomewide significance levels were P=0.065 for ocular refraction and P<0.005 for LTR, providing strong evidence for linkage of refraction to this locus. The inter-marker region containing the peak spans 11 Mb and contains approximately 189 genes. Conclusion: We found genomewide significant evidence for linkage of refractive error to a novel QTL on chromosome 1p36 in an Ashkenazi Jewish population.     

A detailed paraxial schematic eye for the White Leghorn chick

We studied the normal ocular development of the chick (Gallus gallus domesticus, White Leghorn) up to 15 days of age using both longitudinal and cross-sectional methods. The change in refractive error, corneal curvature and axial ocular distances were used to construct schematic eyes. Equations are presented which allow prediction of refractive error changes associated with changes in vitreous chamber depth. The mean refractive error was +3.2 D at hatching, which reduced by 66% over the first 3 days and stabilized by 11 days of age. The lens thickened and the anterior chamber deepened from hatching, but vitreal elongation and corneal flattening were delayed until after the first 3 days, suggesting that normal eye growth may be initially inhibited or inactive during an initial emmetropization period, and subsequently activated in response to myopic defocus arising from the continually expanding lens. Finally, when compared with published data on other chick strains, we find differences in the degree of hyperopia at hatching due to differences in lens thickness. However, the rate of ocular and vitreal expansion and the developmental changes in corneal power are similar, making the schematic eyes presented here generally applicable to different strains of chickens.     

Identification of MFRP and the secreted serine proteases PRSS56 and ADAMTS19 as part of a molecular network involved in ocular growth regulation

Precise regulation of ocular size is a critical determinant of normal visual acuity. Although it is generally accepted that ocular growth relies on a cascade of signaling events transmitted from the retina to the sclera, the factors and mechanism(s) involved are poorly understood. Recent studies have highlighted the importance of the retinal secreted serine protease PRSS56 and transmembrane glycoprotein MFRP, a factor predominantly expressed in the retinal pigment epithelium (RPE), in ocular size determination. Mutations in PRSS56 and MFRP constitute a major cause of nanophthalmos, a condition characterized by severe reduction in ocular axial length/extreme hyperopia. Interestingly, common variants of these genes have been implicated in myopia, a condition associated with ocular elongation. Consistent with these findings, mice with loss of function mutation in PRSS56 or MFRP exhibit a reduction in ocular axial length. However, the molecular network and cellular processes involved in PRSS56- and MFRP-mediated ocular axial growth remain elusive. Here, we show that Adamts19 expression is significantly upregulated in the retina of mice lacking either Prss56 or Mfrp. Importantly, using genetic mouse models, we demonstrate that while ADAMTS19 is not required for ocular growth during normal development, its inactivation exacerbates ocular axial length reduction in Prss56 and Mfrp mutant mice. These results suggest that the upregulation of retinal Adamts19 is part of an adaptive molecular response to counteract impaired ocular growth. Using a complementary genetic approach, we show that loss of PRSS56 or MFRP function prevents excessive ocular axial growth in a mouse model of early-onset myopia caused by a null mutation in Irbp, thus, demonstrating that PRSS56 and MFRP are also required for pathological ocular elongation. Collectively, our findings provide new insights into the molecular network involved in ocular axial growth and support a role for molecular crosstalk between the retina and RPE involved in refractive development.     

Adenomatous Polyposis Coli Mutation Leads to Myopia Development in Mice

Myopia incidence in China is rapidly becoming a very serious sight compromising problem in a large segment of the general population. Therefore, delineating the underlying mechanisms leading to myopia will markedly lessen the likelihood of other sight compromising complications. In this regard, there is some evidence that patients afflicted with familial adenomatous polyposis (FAP), havean adenomatous polyposis coli (APC) mutation and a higher incidence of myopia. To clarify this possible association, we determined whether the changes in pertinent biometric and biochemical parameters underlying postnatal refractive error development in APC^Min mice are relevant for gaining insight into the pathogenesis of this disease in humans. The refraction and biometrics in APC^Min mice and age-matched wild-type (WT) littermates between postnatal days P28 and P84 were examined with eccentric infrared photorefraction (EIR) and customized optical coherence tomography (OCT). Compared with WT littermates, the APC^Min mutated mice developed myopia (average -4.64 D) on P84 which was associated with increased vitreous chamber depth (VCD). Furthermore, retinal and scleral changes appear in these mice along with: 1) axial length shortening; 2) increased retinal cell proliferation; 3) and decreased tyrosine hydroxylase (TH) expression, the rate-limiting enzyme of DA synthesis. Scleral collagen fibril diameters became heterogeneous and irregularly organized in the APC^Min mice. Western blot analysis showed that scleral alpha-1 type I collagen (col1α1) expression also decreased whereas MMP2 and MMP9 mRNA expression was invariant. These results indicate that defective APC gene function promotes refractive error development. By characterizing in APC^Min mice ocular developmental changes, this approach provides novel insight into underlying pathophysiological mechanisms contributing to human myopia development.     

Identification of apolipoprotein A-I as a "STOP" signal for myopia

Good visual acuity requires that the axial length of the ocular globe is matched to the refractive power of the cornea and lens to focus the images of distant objects onto the retina. During the growth of the juvenile eye, this is achieved through the emmetropization process that adjusts the ocular axial length to compensate for the refractive changes that occur in the anterior segment. A failure of the emmetropization process can result in either excessive or insufficient axial growth, leading to myopia or hyperopia, respectively. Emmetropization is mainly regulated by the retina, which generates two opposite signals: "GO/GROW" signals to increase axial growth and "STOP" signals to block it. The presence of GO/GROW and STOP signals was investigated by a proteomics analysis of the retinas from chicken with experimental myopia and hyperopia. Of 18 differentially expressed proteins that were identified, five displayed an expression profile corresponding to GO/GROW signals, and two corresponded to STOP signals. Western blotting confirmed that apolipoprotein A-I (apoA-I) has the characteristics of a STOP signal both in the retina as well as in the fibrous sclera. In accordance with this, intraocular application of the peroxisome proliferator-activated receptor alpha agonist GW7647 resulted in up-regulation of apoA-I levels and in a significant reduction of experimental myopia. In conclusion, using a comprehensive functional proteomics analysis of chicken ocular growth models we identified targets for ocular growth control. The correlation of elevated apoA-I levels with reduced ocular axial growth points toward a functional relationship with the observed morphological changes of the eye.     

Animal Models of Myopia: Learning How Vision Controls the Size of the Eye

As they grow up, approximately 25% of children in the United States become myopic (nearsighted). A much smaller fraction become significantly hyperopic (farsighted), while the majority develop little or no refractive error and are emmetropic. The causes of refractive error, especially myopia, have been the subject of debate for more than a century. Some have held that myopia is primarily an inherited disorder, and others, that myopia is caused by protracted near work and, especially, by accommodation during protracted near work. It has not been possible, based solely on clinical observations, to resolve the relative roles of heredity versus environment in the development of refractive error. In the mid-1970s, several animal models were developed to study the mechanisms underlying refractive error. Using animal models, it was found that the visual environment exerts a powerful influence on refractive state by controlling the axial length of the eye during the postnatal developmental period. Although several species have been examined, three have emerged as primary models and have played complementary roles: tree shrews (mammals closely related to primates), chicks, and monkeys. Each has advantages and disadvantages. Collectively, research on animal models has provided evidence on three issues, namely that (1) the visual environment can produce refractive error; (2) an emmetropization mechanism normally guides eyes to low refractive error; and (3) under-accommodation, rather than excessive accommodation, may cause myopia. Two decades of research on animal models have provided criteria that may be used to evaluate the usefulness of additional species as models of emmetropization.     

Studies of translocation catalysis

There is a symbiotic relationship between the evolution of fundamental theory and the winning of experimentally-based knowledge. The impact of the General Chemiosmotic Theory on our understanding of the nature of membrane transport processes is described and discussed. The history of experimental studies on transport catalysed by ionophore antibiotics and the membrane proteins of mitochondria and bacteria are used to illustrate the evolution of knowledge and theory. Recent experimental approaches to understanding the lactose-H⁺ symport protein ofEscherichia coli and other sugar porters are described to show that the lack of experimental knowledge of the three-dimensional structures of the proteins currently limits the development of theories about their molecular mechanism of translocation catalysis.     

Biochemical systems theory: operational differences among variant representations and their significance

An appropriate language or formalism for the analysis of complex biochemical systems has been sought for several decades. The necessity for such a formalism results from the large number of interacting components in biochemical systems and the complex non-linear character of these interactions. The Power-Law Formalism, an example of such a language, underlies several recent attempts to develop an understanding of integrated biochemical systems. It is the simplest representation of integrated biochemical systems that has been shown to be consistent with well-known growth laws and allometric relationships--the most regular, quantitative features that have been observed among the systemic variables of complex biochemical systems. The Power-Law Formalism provides the basis for Biochemical Systems Theory, which includes several different strategies of representation. Among these, the synergistic-system (S-system) representation is the most useful, as judged by a variety of objective criteria. This paper first describes the predominant features of the S-system representation. It then presents detailed comparisons between the S-system representation and other variants within Biochemical Systems Theory. These comparisons are made on the basis of objective criteria that characterize the efficiency, power, clarity and scope of each representation. Two of the variants within Biochemical Systems Theory are intimately related to other approaches for analyzing biochemical systems, namely Metabolic Control Theory and Flux-Oriented Theory. It is hoped that the comparisons presented here will result in a deeper understanding of the relationships among these variants. Finally, some recent developments are described that demonstrate the potential for further growth of Biochemical Systems Theory and the underlying Power-Law Formalism on which it is based.     

Incorporating experimental design and error into coalescent/mutation models of population history

Coalescent theory provides a powerful framework for estimating the evolutionary, demographic, and genetic parameters of a population from a small sample of individuals. Current coalescent models have largely focused on population genetic factors (e.g., mutation, population growth, and migration) rather than on the effects of experimental design and error. This study develops a new coalescent/mutation model that accounts for unobserved polymorphisms due to missing data, sequence errors, and multiple reads for diploid individuals. The importance of accommodating these effects of experimental design and error is illustrated with evolutionary simulations and a real data set from a population of the California sea hare. In particular, a failure to account for sequence errors can lead to overestimated mutation rates, inflated coalescent times, and inappropriate conclusions about the population. This current model can now serve as a starting point for the development of newer models with additional experimental and population genetic factors. It is currently implemented as a maximum-likelihood method, but this model may also serve as the basis for the development of Bayesian approaches that incorporate experimental design and error.     

A Probabilistic Palimpsest Model of Visual Short-term Memory

Working memory plays a key role in cognition, and yet its mechanisms remain much debated. Human performance on memory tasks is severely limited; however, the two major classes of theory explaining the limits leave open questions about key issues such as how multiple simultaneously-represented items can be distinguished. We propose a palimpsest model, with the occurrent activity of a single population of neurons coding for several multi-featured items. Using a probabilistic approach to storage and recall, we show how this model can account for many qualitative aspects of existing experimental data. In our account, the underlying nature of a memory item depends entirely on the characteristics of the population representation, and we provide analytical and numerical insights into critical issues such as multiplicity and binding. We consider representations in which information about individual feature values is partially separate from the information about binding that creates single items out of multiple features. An appropriate balance between these two types of information is required to capture fully the different types of error seen in human experimental data. Our model provides the first principled account of misbinding errors. We also suggest a specific set of stimuli designed to elucidate the representations that subjects actually employ.     

Adequate Examination of the Eye

A concise outline of the methodology of an adequate examination of the eye is presented so that the ocular findings may be used properly in establishing a diagnosis. An accurate ocular history is important. Visual acuity at a distance and at the reading point should be measured in such a way as to avoid the introduction of error. A pinhole disc can be used to differentiate a refractive error from organic pathology. Clinical methods of performing the visual field examination, estimating the lacrimal tear production, and determining intraocular pressure are described and illustrated.     

Refractive index measurement in viable cells using quantitative phase-amplitude microscopy and confocal microscopy.

BACKGROUND: The refractive index (RI) of cellular material provides fundamental biophysical information about the composition and organizational structure of cells. Efforts to describe the refractive properties of cells have been significantly impeded by the experimental difficulties encountered in measuring viable cell RI. In this report we describe a procedure for the application of quantitative phase microscopy in conjunction with confocal microscopy to measure the RI of a cultured muscle cell specimen. METHODS: The experimental strategy involved calculation of cell thickness by using confocal optical sectioning procedures, construction of a phase map of the same cell using quantitative phase microscopy, and selection of cellular regions of interest to solve for the cell RI. RESULTS: Mean cell thickness and phase values for six cell regions (five cytoplasmic and one nuclear) were determined. The average refractive index calculated for cytoplasmic and nuclear regions was 1.360 +/- 0.004. The uncertainty in the final RI value represents the technique measurement error. CONCLUSIONS: The methodology we describe for viable cell RI measurement with this prototype cell has broad generic application in the study of cell growth and functional responses. The RI value we report may be used in optical analyses of cultured cell structure and morphology.     

Density‐dependent offspring interactions do not explain macroevolutionary scaling of adult size and offspring size

Most life forms exhibit a correlated evolution of adult size (AS) and size at independence (SI), giving rise to AS–SI scaling relationships. Theory suggests that scaling arises because relatively large adults have relatively high reproductive output, resulting in strong density‐dependent competition in early life, where large size at independence provides a competitive advantage to juveniles. The primary goal of our study is to test this density hypothesis, using large datasets that span the vertebrate tree of life (fishes, amphibians, reptiles, birds, and mammals). Our secondary goal is to motivate new hypotheses for AS–SI scaling by exploring how subtle variation in life‐histories among closely related species is associated with variation in scaling. Our phylogenetically informed comparisons do not support the density hypothesis. Instead, exploration of AS–SI scaling among life‐history variants suggests that steeper AS–SI scaling slopes are associated with evolutionary increases in size at independence. We suggest that a positive association between size at independence and juvenile growth rate may represent an important mechanism underlying AS–SI scaling, a mechanism that has been underappreciated by theorists. If faster juvenile growth is a consequence of evolutionary increases in size at independence, this may help offset the cost of delayed maturation, leading to steeper AS–SI scaling slopes.     

The role of growth factors in the embryogenesis and differentiation of the eye.

The vertebrate eye is composed of a variety of tissues that, embryonically, have their derivation from surface ectoderm, neural ectoderm, neural crest, and mesodermal mesenchyme. During development, these different types of cells are subjected to complex processes of induction and suppressive interactions that bring about their final differentiation and arrangement in the fully formed eye. With the changing concept of ocular development, we present a new perspective on the control of morphogenesis at the cellular and molecular levels by growth factors that include fibroblast growth factors, epidermal growth factor, nerve growth factor, platelet-derived growth factor, transforming growth factors, mesodermal growth factors, transferrin, tumor necrosis factor, neuronotrophic factors, angiogenic factors, and antiangiogenic factors. Growth factors, especially transforming growth factor-beta, have a crucial role in directing the migration and developmental patterns of the cranial neural-crest cells that contribute extensively to the structures of the eye. Some growth factors also exert an effect on the developing ocular tissues by influencing the synthesis and degradation of the extracellular matrix. The mRNAs for the growth factors that are involved in the earliest aspects of the growth and differentiation of the fertilized egg are supplied from maternal sources until embryonic tissues are able to synthesize them. Subsequently, the developing eye tissues are exposed to both endogenous and exogenous growth factors that are derived from nonocular tissues as well as from embryonic fluids and the systemic circulation. The early interaction between the surface head ectoderm and the underlying chordamesoderm confers a lens-forming bias on the ectoderm; later, the optic vesicle elicits the final phase of determination and enhances differentiation by the lens. After the blood-ocular barrier is established, the internal milieu of the eye is controlled by the interactions among the intraocular tissues; only those growth factors that selectively cross the barrier or that are synthesized by the ocular tissues can influence further development and differentiation of the cells. An understanding of the tissue interactions that are regulated by growth factors could clarify the precise mechanism of normal and abnormal ocular development.     

Association between Childhood Strabismus and Refractive Error in Chinese Preschool Children

PurposeTo investigate the association between concomitant esotropia or concomitant exotropia and refractive error in preschool childrenMethodsA population-based sample of 5831 children aged 3 to 6 years was selected from all kindergartens in a representative county (Yuhuatai District, Nanjing, Jiangsu Province) of Nanjing, China. Clinical examinations including ocular alignment, ocular motility, visual acuity, optometry, stereopsis screening, slit lamp examination and fundus examination were performed by trained ophthalmologists and optometrists. Odd ratios (OR) and 95% confidence intervals (95% CI) were calculated to evaluate the association of refractive error with concomitant esotropia and concomitant exotropia.ResultsIn multivariate logistic regression analysis, concomitant esotropia was associated independently with spherical equivalent anisometropia (OR, 3.15 for 0.50 to <1.00 diopter (D) of anisometropia, and 7.41 for > = 1.00 D of anisometropia) and hyperopia. There was a severity-dependent association of hyperopia with the development of concomitant esotropia, with ORs increasing from 9.3 for 2.00 to <3.00 D of hyperopia, to 180.82 for > = 5.00 D of hyperopia. Concomitant exotropia was associated with astigmatism (OR, 3.56 for 0.50 to 1.00 D of astigmatism, and 1.9 for <0.00 D of astigmatism), myopia (OR, 40.54 for -1.00 to <0.00 D of myopia, and 18.93 for <-1.00 D of myopia), and hyperopia (OR, 67.78 for 1.00 to <2.00 D of hyperopia, 23.13 for 2.00 to <3.00 D of hyperopia, 25.57 for 3.00 to <4.00 D of hyperopia, and 8.36 for 4.00 to <5.00 D of hyperopia).ConclusionsThis study highlights the close associations between refractive error and the prevalence of concomitant esotropia and concomitant exotropia, which should be considered when managing childhood refractive error.