STEM CELLS AND DEVELOPMENT

Volume 18, Number 1, 2009

CONCISE REVIEW
Mary Ann Liebert, Inc.
DOI: 10.1089/scd.2008.0169

Wnt-Signaling in Retinal Development and Disease

Eleonora M. Lad,1 Samuel H. Cheshier,2 and M. Yashar S. Kalani2

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The Wnt-signaling pathway is a known regulator of stem cell maintenance, cellular proliferation and differ-
entiation, and cancer development in various tissues. Wnt proteins play a central role during various stages of
retinal development; retinal field establishment, retinal and hyaloid vasculogeneis, cornea and lens develop-
ment, eye field formation, and maintenance of retinal stem cell and neuronal specification in many species are
Wnt-regulated processes. Uncontrolled Wnt signaling may cause retinal diseases such as familial exudative
vitroretinopathy, retinitis pigmentosa, and Norries disease, further underscoring the importance of the Wnt-
signaling pathway in the retina. This review summarizes major developments and discoveries regarding the
role of the Wnt-signaling pathway as it pertains to retinal development and disease.

Introduction
T he Wnt-signaling pathway is a key regulator of em-
bryogenesis, tissue patterning and regeneration, and
cancer development [1]. Wnt signals have been shown to be
key regulators of various stages of retinal development, in-
cluding both not limited to retinal field establishment, main-
tenance of retinal stem cells, vasculogeneis in the retina and
formation of the ciliary body. Given the central role of the
pathway in stem cell regulation and cell-fate decisions, it
is not surprising that unchecked activation of the pathway
results in the development of retinal diseases such as fa-
milial exudative vitroretinopathy, Norries disease, and
retinitis pigmentosa. The identification of the role of Wnt
signaling in the development program and pathologies of
the retina has made it an attractive target for pharmaceutical
and cell-based therapies of the retina.
The Wnt-signaling pathway was discovered by Nusse
and Varmus in the early 1980s [1,2]. Wnts are a large fam-
ily of lipid-modified secreted glycoproteins that signal by
binding to various membrane proteins resulting in a change
in the expression of genes important for cell-to-cell interac-
tion, cell cycle control, proliferation, and self-renewal [1,3].
A search of the human genome has identified 19 Wnt genes
to date [4].
Nearly 30 years of biochemical and cell biological inves-
tigations have identified several key players for the propa-
gation of the Wnt signal from the membrane to the nucleus.
Mechanistic studies have classified the Wnt signal mediated
via -catenin as a canonical signal, while other mechanisms
not involving -catenin have been termed noncanonical-
signaling pathways. Several excellent reviews have
addressed the biochemical studies that have resulted in our
extensive yet preliminary understanding of these pathways
[1,5]. Wnt proteins may signal via either or both the canon-
ical or noncanonical pathways. It is generally believed that
the Wnt1 class of proteins (Wnt1, 3, 3a, and 8) predominantly
signal through the canonical-signaling pathway, whereas
the Wnt5a class (Wnt4, 5a, and 11) mainly activate various
noncanonical pathways [5]; although more recent evidence
suggests that this is a gross oversimplification of this differ-
ential signaling concept [5,6].
Canonical Wnt Pathway
The canonical pathway, the signaling pathway proceeding
via -catenin, is the best understood of the Wnt pathways.
In the absence of Wnt proteins (off state), a cytoplasmic
destruction complex composed of adenomatous polyposis
coli (APC) and AXIN1 causes casein kinase 1 and glycogen
synthase kinase 3 to phosphorylate -catenin, leading to the
ubiquitination and proteasomal degradation of -catenin [7]
(Fig. 1A).
When Wnt proteins bind to one of several transmembrane
frizzled (Fz) receptors and form a ternary complex with low-
density lipoprotein (LDL)-receptor-related proteins 5 and 6
(LRP5 and LRP6) the signal is propagated from the cell sur-
face to the cytoplasm where the phosphoprotein Disheveled
is activated (Fig. 1B). Recruitment of Disheveled leads to

1
Department of Ophthalmology and 2Department of Developmental Biology, Stanford University School of Medicine, Stanford,
California.

7
 8 LAD, CHESHIER, AND KALANI

DKK LRP6
Frizzled
P
b-cat
U Proteosome
P
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b-cat

b-cat
P
APC
GSK3b
Axin

No Transcription

FIG. 1. Canonical Wnt pathway. (A) In

the off state (absence of Wnt), a cyto-
plasmic destruction complex composed
of APC and AXIN1 results in phosphory-
B lation of -catenin by casein kinase 1 and
Wnt GSK-3. This leads to ubiquitination and
proteasomal degradation of -catenin,
LRP6

Frizzled resulting in lack of transcription of Wnt

target genes. (B) During activation of
Wnt pathway, Wnt binds extracellular Fz
receptors and LRP5 and LRP6 leading to
activation of phosphoprotein Disheveled,
GSK-3- inhibition and binding of AXIN1
b-cat

b-cat

Dishevelled
to LRP5/6. This results in degradation
of AXIN, breakdown of the destruction
complex and stabilization of -catenin.
APC -catenin is then able to translocate to
the nucleus, where it interacts with TCF/
GSK3b
LEF family members to induce gene
Axin transcription. The equilibrium arrows
illustrate that -catenin exists in several
states, many of which are in equilibrium
b-cat TCF Transcription within the cell. Various signals, including
the Wnt signal, can alter the equilibrium
within the cell resulting in the recruit-
ment of -catenin to one of its many roles
within the cell.

translocation of AXIN1 and the destruction complex to the
membrane, which is followed by binding of AXIN1 to LRP5/6.
As a result, AXIN1 is degraded, the destruction complex is
disassembled and -catenin phosphorylation and breakdown
ceases [8]. Activated Disheveled also results in inhibition of
GSK3. With cytoplasmic accumulation, -catenin is now
free to take part in various cellular processes including trans-
location to the nucleus, where it interacts with the TCF/LEF
family of transcription factors and results in transcription of
a genetic network important for regulation of the cell cycle
and cell-fate decisions [7] (Fig. 1B) -catenin can also partake
in various cell adhesion complexes important for cell-cell con-
tact and communication (for review please see ref. 3).
Given the importance of the Wnt-signaling pathway, it is
not surprising that evolution has provided several key steps
for the regulation of the Wnt signal. Indeed the signal can
be interrupted at the cell surface via activation of alternate
pathways, interruption of the ternary complex, or binding of
ligands to the Wnt protein by various factors such as sfrps
[911], WIF-1 [12], and the DKK family members [13,14] or
modulated at the cytoplasmic and nuclear level by factors
such as the AXIN family of genes [15], p53 [16], and other sig-
naling pathways such as members of the TGF- [17,18] inter-
acting at the level of -catenin or LEF/TCF [19]. For further
details about the canonical pathway, please refer to excel-
lent reviews by Nusse et al. [1,2] (or visit the Wnt Homepage
 Wnt-SIGNALING IN RETINAL DEVELOPMENT AND DISEASE
(http://www.stanford.edu/~rnusse/wntwindow.html) for
up-to-date information about major discoveries involving
the Wnt pathway.
Noncanonical Wnt Pathway
The noncanonical Wnt pathway is independent of
-catenin signaling. The best understood noncanonical Wnt
pathway is the planar polarity signaling in Drosophila and in
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epithelial tissue polarity in vertebrates [20]. There is ample
evidence that the noncanonical Wnt pathway regulates gas-
trulation in Drosophila [21] and a variety of biological pro-
cesses in vertebrates such as establishment of cochlear hair
morphology [22], heart formation [23], dorsoventral pattern-
ing [24], neuronal migration, and cancer [19]. Noncanonical
Wnt signaling may involve many different players in-
cluding calcium, JNK, small and heterotrimeric G proteins,
Fz, Disheveled, RhoA, receptor tyrosine kinases and others.
Several excellent reviews address the complexity of the non-
canonical Wnt-signaling pathways identified thus far [5]. A
great simplification of the noncanonical model of Wnt sig-
naling suggests that binding of the secreted Wnt protein
to various membrane proteins at the cell surface results in
change in the genetic program of the cell through activa-
tion of various cytoplasmic and nuclear pathways but not
through -catenin (Fig. 2).
Wnt Signaling in Retinal Development
The Wnt/-catenin pathway plays a central role during
various stages of retinal development: retinal field estab-
lishment, maintenance of retinal stem cell progenitors and
neuronal specification, retinal vasculogenesis and the devel-
opment of the cornea, and the lens (See Table 1 for important
regulators in various anatomical locales) [1,25]. A number of
Wnt
LRP6
Ryk Ror2 FRL1 Frizzled
? FIG. 2. Noncanonical Wnt pathway.
Transcription
9
studies have analyzed the expression of Wnt and Fz recep-
tors during various stages of retinal development in the
chick, mouse and Xenopus [2629]. Despite many proposed
roles for Wnt proteins and members of the Wnt pathway in
retinal development, there are still controversies as to the
exact mechanistic contribution of the pathway [1]. While
several groups have identified Wnts as important regula-
tors of self-renewal and stem cell maintenance for retinal
progenitors, others have implicated Wnts and members of
the signaling pathway as important regulators of differen-
tiation. It is very likely that different members of the Wnt
family may regulate both self-renewal and differentiation.
An excellent avenue for the study of these Wnt-mediated
processes is now made possible with the purification of the
protein by Nusse et al.
Retinal fi eld establishment
In the development of the eye, the groups of cells that
will give rise to this organ are initially part of a bilateral do-
main termed eye field that is localized within the anterior
neural plate. The eye field can be distinguished by the ex-
pression of eye specification network of genes ET, Rx1, Pax6,
Six3, Lhx2, tll, and Optx2 (also known as Six6) [3033]. Using
cocktails containing Otx2, Zuber et al. induced ectopic eye
formation outside the nervous system. Their data suggested
that Optx2 and tll first prime the anterior neural plate for
future eye formation, then ET, Rx1, Pax6, Six3, and Lhx2
form a network of transcription factors that specify the fate
of the tissue that will give rise to the eye field during eye
development [30].
Key experiments by Rasmussen et al. first highlighted
the essential role of the Wnt pathway of the establishment
of the retinal field. Rasmussen et al. have shown in Xenopus
that Fz3 regulates eye development by affecting expression
-catenin independent pathways are acti-
vated by binding of Wnts to membrane
proteins [Fz, receptor tyrosine kinases
(RTKs), etc.], which results in various intra-
cellular changes, not directly mediated
through -catenin. Examples of intracellu-
lar-signaling partners (marked with a ?
mark because many of these players have
yet to be identified) include calcium, small
GTPases of the Rho family, the JNK cas-
cade, and many others. These events lead
to alterations in cell polarity, cytoskeletal
structure, and cell behavior.
 10
Table 1. Members of the Wnt Family and Other
Components of the Signaling Pathway Have Been
Shown to Be Critical for the Development of
Various Components of the Visual System
in Several Species [26,28,34,4142,44]
Location Signaling component
GCL MWnt-2b
CFz-1, 2, 7, 8, 9
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MFz-3, 4, 6, 7
MSfrp-1
INL MWnt-2b, 5a, 5b
CFz-1, 2, 3, 8, 9
MFz-3, 4, 6, 7
MSfrp-1, 2, 3
CMZ XFz-5
PRL CFz-2
MFz-4
MSfrp-1
RPE CFz-2
Lens MWnt-5a, 5b, 7a, 7b, 8a, 8b
MFz-1, 3, 4, 6
CWnt-2b
Important and well-accepted members of the signaling
pathway are represented here along with the anatomical
locale with which they are associated. GCL = ganglion
cell layer, INL = inner nuclear layer, PRL = photoreceptor
layer, CMZ = ciliary margin zone, RPE = retinal pigment
epithelium. C = Caenorhabditis elegans, and M = mouse.
of Pax6 [34]. During development, Fz3 expression is at first
ubiquitous but later becomes restricted to the developing
nervous system, mainly in an area of the anterior neural
plate that will become the early eye field [35]. Further, Fz3
continues to be expressed in the developing optic vesicle
[35]. Overexpression of Fz3 during Xenopus development
promoted formation of complete ectopic eyes and ectopic
expression of Pax6, Rx, and Otx2 [34]. The ectopic eyes had
a laminar structure similar to that of endogenous eyes and
contained differentiated retinal cells. Fz3 overexpression was
present near the midline close to the midbrain-hindbrain
junction and in the roof of the fourth ventricle, where there
is a high level of expression of Wnt-1, Wnt-3a, and Wnt-10
during normal Xenopus development [36,37]. Targeted over-
expression of a soluble inhibitory, dominant-negative form of
Fz3 was found to block normal eye development and endog-
enous expression of the same genes; this effect was rescued
by coexpression of Fz3 again suggesting that the presence of
Fz3 is necessary and sufficient for eye formation. Blockade
of Fz3 signaling by overexpression of Kermit, a protein that
binds to the C-terminal intracellular region of Fz3, also pre-
vented normal eye development. Taken together, the above
data suggest that various Wnt proteins may be activating the
Fz3 receptor at these sites resulting in eye specification and
promotion of eye development. The Wnt-Fz signal through
Fz3 receptor is both necessary and sufficient to regulate nor-
mal development of the eye in Xenopus [34]. It is important
to note that to date there is very little understanding of the
specificity of various Wnt proteins for the receptors that they
signal through and it is possible for one Wnt to activate sig-
naling through several receptors.
LAD, CHESHIER, AND KALANI
Extending the role of the Wnt signaling in the develop-
ment of the eye field in other species Cavodeassi et al. investi-
gated the differential and exclusionary role of two opposing
Wnt signals in the development of the eye field of zebrafish.
Early during the development of the eye field in zebrafish,
activation of the canonical Wnt pathway through interaction
of Wnt8b with its receptor Fz8a results in specification of
the caudal diencephalic fate and inhibition of eye develop-
ment whereas the activation of a noncanonical pathway via
interaction of Wnt11 and Fz5 promotes eye development [38].
Activation of the noncanonical Wnt pathway via binding of
Wnt11 to Fz leads to release of calcium in the cell, activa-
tion of small guanosine triphosphatases of the Rho fam-
ily, and activation of the JNK cascade. These events result
in changes in the cytoskeletal structure and in cell polarity
and cell behavior [38]. This study highlights the elegant dif-
ferential role of Wnt proteins as activators and repressors of
the same developmental program by differential interaction
with receptors at the cell surface.
Upon establishment of the eye field, the optic vesicle,
originating from the ventral diencephalon and invaginating
within the ectoderm forming a bilayered optic cup, leads
to the development of the early structures of the eye. The
outer layer of the optic cup is destined to form the retinal
pigmented epithelial (RPE) layer and the inner layer will
give rise to the retina [28]. In the early optic vesicle (stage
10 in the chick), several of the Fz family receptors (Fz 4 and
5) are expressed in the distal optic vesicle that differentiates
into the neural retina [26] again highlighting the fact that
the Wnt pathway is an important regulator of not only the
specification of the eye field, but also the later steps in the
commitment and differentiation of various appendages of
the eye.
Further evidence in support of the notion that members
of the Wnt family are important for the process of commit-
ment is the presence of various agonists and antagonists of
the Wnt pathway during various steps of the development of
the nervous system. Several Sfrp molecules (Sfrp-1, -2, and
-4) are present throughout the neural ectoderm from stage
4 in the chick [11] and from day E9.5 in the mouse [39]. At a
later time when the neuroblast layer of the neural retina is
formed (from gastrula stage in Xenopus, from stage 10 up
to stage 32 in the chick, and from day E18.5 in the mouse),
several Wnt molecules (Wnt-3, -5a, -5b, and -7b), Fzs (Fz-2
through -7) and Sfrp molecules (Sfrp-1 and 2) are expressed
in the outer neuroblast layer [26,28,3941]. Wnt-2b and -13 are
present in the ciliary margin zone (CMZ), which will give
rise to the iris and ciliary body [28], and in the RPE. In these
regions, Wnt-2b is expressed in a peripheral-to-central gradi-
ent, in conjunction with a high level of activity of the Wnt/-
catenin promoter starting at stage 16 in the chick [28,42,43].
The specific localization of Wnt-2b to the CMZ may be due
to the modulatory, inhibitory effect of Sfrp-2 on Wnt expres-
sion. Starting on day E12.5 in the mouse, Fz-3 through -7 and
Sfrp-1 and -2 are also expressed in the CMZ, and Fz-4 was
also found in the RPE [28]. Sfrp-1 and -2 were found in high
level in the astrocyte precursor cells in the optic disc area at
E14.5 [28]. Analysis of expression of Wnt, Fz, and Sfrp expres-
sion in the neural retina in various animal models suggest
that there is significant overlap between these molecules.
Thus, it is likely that Wnts bind the Fzs and Sfrps with differ-
ent affinities within the neural retina or that the activation of
Fz receptors results in different downstream effects [28].
 Wnt-SIGNALING IN RETINAL DEVELOPMENT AND DISEASE
Again taken together the above observations suggest
that different Wnts are involved in the processes of commit-
ment and specification of the various cell types of the eye
and the nervous system. Differential signaling, whether it be
through activation of the canonical or noncanonical pathway
or through antagonism of the pathway by other signaling
pathways or peptide antagonists of the Wnt pathway results
in the formation of gradients necessary for the specification
of the lineage of the various layers and cell types present in
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the eye.
Retinal progenitor cell maintenance
Gradients are important for various developmental pro-
cesses such as dorsoventral patterning and formation of the
neural axis. During development many signaling molecules
produce gradients and it is the production of the various gra-
dients that produces the diversity of the cell types present in
an organism. In the eye, during the stage of retinoblast com-
mitment within the central retina, Wnt, Fz, and Sfrp expres-
sion changes to a peripheral-to-central gradient that reflects
retinal differentiation patterns present in the adult eye [28].
Retinal histogenesis occurs from the center to the periph-
ery, such that peripheral retina contains the least-committed
retinoblasts, while the central retina contains the mature
retinoblasts, the postmitotic, differentiated retinal precur-
sors and the earliest differentiated retinal neurons [28]. It
is thought that the differential expression of Wnt molecules
in the peripheral retina and central retina correlates with
a potential role for these molecules in retinoblast differen-
tiation. The presence of Wnt-5a and Fz-4 in the peripheral
retina suggests a role for these molecules in maintenance
of multipotent progenitors and not in specifying cell func-
tion, a process that takes place in central retina [28]. Wnt/-
catenin reporter is also expressed in a peripheral-to-central
gradient, being present at robust levels in the retinoblasts in
peripheral margin and RPE, but not in the central retina and
central RPE. Conversely, Wnt-11, a marker of the noncanoni-
cal Wnt pathway is expressed in the central retina [44,45].
This supports a role for the noncanonical Wnt pathway,
which does not involve Wnt/-catenin reporter activation, in
specification and differentiation of the neural retina. Again
these observations suggest that differential activation of sig-
naling pathways and developmental programs by members
of the Wnt family are important for tissue patterning and
development in the eye.
Tissue remodeling is an important mechanism for the
repair and maintenance of all tissues. The various compo-
nents of the eye are continuously remodeled. The process of
retinoblast maintenance and remodeling are Wnt dependent
processes. At the stage of retinoblast maintenance, Wnt-3 and
-5a are expressed in the RPE and Wnt-2b and -5a are present
in the peripheral CMZ [27,28,42]. Wnt-2b has been shown to be
important in maintenance and proliferation of neuroblasts.
Overexpression of Wnt-2b into the developing optic vesicle
of chicks induced markers of the CMZ and inhibited matura-
tion of the neural retina [43]. It was found that, in vitro, Wnt-
2b-stabilized cytoplasmic -catenin (meaning prevented its
degradation) [43], induced neuroepithelial phenotype and
inhibited differentiation [46]. In contrast, the signaling cas-
cade mediated by the Delta ligand and Notch receptor is an
important factor that maintains the undifferentiated state of
retinal progenitor cells [4749]. Wnt2b (formerly known as
11
Wnt13) was shown to inhibit neuronal differentiation even
when the Delta/Notch-signaling pathway was blocked and
to downregulate messenger RNA expression of Notch1 [50].
Thus, Wnt2b maintains progenitor cells in the undifferen-
tiated state by preventing their differentiation mediated
through Notch [50]. These data implicated Wnt-2b and the
canonical Wnt/-catenin pathway in the proliferation and
multipotency of the chick embryonic CMZ.
Inoue et al. investigated whether Wnt signaling could
induce the proliferation of adult retinal stem cells from
the mammalian CMZ [51]. Indeed, they demonstrated that
Wnt3a increased the size of the spheres derived from murine
CMZ and increased the number of cells expressing Ki-67,
a marker of cell division, and bromodeoxyuridine (BrdU).
Wnt3a resulted in maintenance of the multipotent state of
the CMZ cells, which expressed several retinal markers
under differentiating culture conditions [51]. Wnt3a also
enhanced formation of secondary spheres which accumu-
lated -catenin in the nucleus [51]. Stimulation of the Wnt
pathway through inhibition of GSK-3, a key enzyme in the
canonical Wnt-signaling cascade that destabilizes -catenin
[52,53] increased expression of Ki-67 in the sphere-derived
cells. These experiments implicated a role of canonical Wnt
pathway in self-renewal of retinal stem cells. These effects
of Wnt signaling were accelerated by exogenous fibroblast
growth factor (FGF)2 and decreased by a FGF receptor inhib-
itor, suggesting that FGF2 and Wnt3a had a strong additive
effect on proliferation of retinal stem cells [51].
Wnt3a also promoted proliferation of Mller gliaderived
retinal progenitors in the photoreceptor-damaged retina [54].
Retinal injury resulted in nuclear accumulation of -catenin,
cyclin D1 upregulation, and Wnt/-catenin reporter activity.
Activation of Wnt cascades by GSK-3 inhibitors promoted
retinal regeneration, while inhibition of the Wnt signaling
with Dkk-1 attenuated retinal regeneration [54]. The Wnt3a-
mediated regeneration of retinal cells was also present in rd
mice, a model of retinal degeneration [54]. These data sug-
gested that Wnt/ catenin pathways mediate a process of
retinal repair and that application of Wnt or GSK-3 inhibi-
tors may promote retinal neuron regeneration.
Cell fate specifi cation
All seven cell types of the vertebrate retina (retinal gan-
glion cells, amacrine cells, bipolar cells, horizontal cells, rod,
and cone photoreceptors and Mller glial cells) originate
from a single population of naive retinal progenitor cells fol-
lowing a distinct temporal birth order [55]. Specification and
differentiation of the individual cell types require specific
regulatory factors that act to determine cell fate [56,57]. Wnt
proteins and members of the pathway have been implicated
in various steps of retinal development. A recent review by
Van Raay and Vetter nicely outline the expression patterns
of various Wnts and signaling components in the develop-
ing retina [29].
Studies of Van Raay et al. performed in Xenopus showed
that Fz5 receptor acts through the canonical -catenin path-
way, and not through the noncanonical Wnt pathway, to
regulate retinoblast proliferation and neural differentiation
[58]. Fz5 exerts its effects by regulating the expression of the
neural competence factor Sox2. Inhibition of either Fz5 or
canonical Wnt-signaling pathway did not impact the level
of retinal progenitor markers but inhibited Sox2 expression,
 12
which led to reduced cellular proliferation and failure of acti-
vation of proneural genes needed for retinal neurogenesis.
Blockage of Sox2 had a similar effect, suggesting that Sox2
expression is necessary for differentiation of retinoblast into
neural progenitors. Inhibition of either Fz5 or Sox2 expres-
sion caused the retinoblasts to become Mller glial cells [58].
The study of Van Raay et al. showed that activation of Fz5 via
the Wnt canonical pathway is needed for differentiation of
retinal progenitors along a neural fate.
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In addition to adopting a specific cell fate, differentiating
retinal cells must also establish spatial identity by migrating
to the correct position or layer within the retina [59,60].
Cellular adhesion plays a key role in sorting different cell
types into a specific cellular organization during develop-
ment [6163]. Recent studies have shown that mutations in
proteins involved in cell adhesion result in retinal lamina-
tion defects [6469]. -catenin therefore plays a central role
in retinal lamination by bridging adherens junctions to the
cytoskeletal network and participating in the canonical Wnt-
signaling pathway. Fu et al. have investigated the function
of -catenin in mouse retinal neurogenesis [70]. Deletion
of the -catenin gene in the developing retina resulted in a
disorganized retina but did not affect the formation of any
of the retinal cell types. Activation of the nuclear function
of -catenin in the retina did not lead to retinal progenitor
cell overproliferation in this model. These results indicated
that -catenin is a central player in retinal development by
affecting retinal lamination rather than cell proliferation or
differentiation, thus supporting the view that these are ge-
netically distinct processes [70].
Using gene transfer, Cho et al. showed that activation of
the canonical Wnt pathway in the developing chick eye can
result in differentiation of retinal progenitor cells into cells
of ciliary body and iris [71]. In contrast to the studies of Kubo
et al. who found that expression of Wnt2b led to the prolifer-
ation of retinal cells in vitro [43,50], expression of Wnt2b or
of a constitutively active form of -catenin in vivo inhibited
expression of retinal progenitor gene and differentiation
of retinal neurons. Instead, Wnt2b or constitutively active
-catenin expression led to the conversion of retinal cells into
cells expressing markers of the ciliary body and iris, tissues
derived from the optic cup. Loss-of-function of Wnt2b using
expression of DN-Lef1 or a fusion of Lef1 with the engrailed
repressor resulted in inhibition of peripheral marker expres-
sion and iris hypoplasia without affecting retinal tissues.
These data suggested that canonical Wnt pathway is neces-
sary for peripheral eye development and for specifying the
fate of the ciliary body and iris [71]. In agreement with the
study of Cho et al., Liu et al. have recently shown that phar-
macological activation of canonical Wnt/-catenin pathway
was sufficient and necessary for inducing development of
the peripheral retina, the ciliary margin, which will give rise
to the ciliary body and iris [72]. Activation of this pathway
using lithium (Li+) induced expression of markers of the
ciliary margin Otx1 and Msx1 in retinal explants in vitro.
Stabilization of -catenin expression in the peripheral retina
led to upregulation of these markers, downregulated expres-
sion of markers of neural retina and inhibited neurogenesis.
Conversely, inactivation of -catenin in vivo resulted in
decreased ciliary margin gene expression and reduced size
of the ciliary margin, ciliary body, and iris. These data sug-
gested that activation of canonical Wnt pathway promoted
LAD, CHESHIER, AND KALANI
development of peripheral eyecup fates at the expense of the
development of neural retina [72].
The generation of retinal vasculature
The nervous system and the system of vessels carrying
blood throughout an organism share much in their develop-
ment, patterning, and sprouting. Given the frequent associa-
tion of nerves and blood vessels, it is not surprising to note
that similar developmental queues are responsible for forma-
tion of both organs. (An excellent review by Tessier-Lavigne
highlights important similarities between these develop-
mental processes and the reader is encouraged to refer to
this review for further detail [73].) In search of a molecular
cause for familial exudative vitreoretinopathy, Samuels et al.
[74] identified mutant Fz4 receptor as the culprit behind the
disrupted retinal angiogenesis. Further work by Nathans
et al. [75] has showed that Norrin and Fz4 control the for-
mation of retinal blood vessels and that this ligand-receptor
combination are necessary for the proper vasculogenesis
in the retina. They have shown that unregulated signaling
promotes an uncontrolled program of vasculogenesis and
cell death resulting in the formation of familial exudative
vitreoretinopathy.
Development of the lens and cornea
Many early studies suggest that members of Wnt and
TGF- family are important in the regulation and develop-
ment of the lens and cornea. Lyu and Joo showed that Wnt
signaling enhances FGF2-triggered lens fiber cell differenti-
ation [76]. Lang et al. have shown that -catenin is essential
for lens morphogenesis and signaling suppression of lens
fate in the periocular ectoderm [77]. From a pathological
standpoint Sundin et al. have shown that extreme hyper-
opia, a defect in the lens is the result of null mutations in
MFRP a gene coding for a member of Fz-related proteins
[78]. Recent studies by Mukhopadhyay et al. and Gage et al.
have shown that Dkk2 plays an essential role in the corneal
fate of the ocular surface epithelium further highlighting
the importance of the Wnt pathway to the development of
the organs of the eye [79,87]. More recent work by Cain et al.
implicates differential requirement for -catenin in epithe-
lial and fiber cells during lens development [86]. The authors
observed that -catenin was required in lens epithelium
and during early fiber differentiation but appeared to be
redundant in differentiated fiber cells. Using a CRE-LOX
model they observed that complete loss of -catenin results
in abnormal formation of the epithelial layer with loss of
E-cadherin and Pax6 expression. Fiber cell differentiation is
also disrupted as characterized by poor cell elongation, de-
creased -crystallin expression, epithelial cell cycle arrest at
G(1)-S transition and premature cell cycle exit.
Wnt Signaling in Retinal Disease
Based on the results of numerous recent studies [2628,58],
it is evident that Wnt-signaling pathways play a central role
in retinal field establishment, maintenance of retinal stem
cell progenitors, and retinal specification in the developing
retina and in homeostasis in the adult retina. Given the cen-
tral role of the pathway on the development, maintenance
 Wnt-SIGNALING IN RETINAL DEVELOPMENT AND DISEASE
and regeneration of the retina, it is not surprising that dereg-
ulation of many components of the Wnt-signaling pathway
may result in retinal disease. For example, a loss-of-function
mutation in Fz-4 and the coreceptor LRP5 have been linked
to familial exudative vitroretinopathy, characterized by
microophthalmia, failure of retinal angiogenesis, and com-
pensatory retinal neovascularization, development of leaks,
bleeding, and scarring. As a result, patients develop retinal
detachment and blindness [74]. Sfrp-1, -2, -3, and -5 were
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shown to be upregulated in retinitis pigmentosa patients,
in which photoreceptors degenerate by apoptosis [80,81].
Mfrp, a gene encoding a Fz-related protein, is mutated in the
mouse retinal degeneration 6 [89].
Patients with mutations in the APC tumor suppressor
gene develop congenital hypertrophy or hyperplasia of the
retinal pigmented epithelium and even failure of ventral ret-
inal development known as retinal coloboma [82]. Mice with
targeted APC gene disruption were characterized by the
same ocular abnormalities [82,83]. Nadauld et al. have re-
cently shown that APC-deficient zebrafish harbored develop-
mental abnormalities of the retina similar to those present in
mice and humans with APC mutation. Their data supported
a dual role for APC in regulation of canonical Wnt/-catenin
signaling and retinoic acid signaling within the eye. APC
regulation of canonical Wnt/-catenin signaling appeared
to have a central role in lens development. APC control of
retinoic acid production via retinol dehydrogenase 5 was
specific to the retina and was required for retinal differen-
tiation, suggesting that that retinoic acid deficiency may be
responsible for congenital hypertrophy or hyperplasia of the
retinal pigmented epithelium [84].
The Danish ophthalmologist Norrie described the first case
of a rare congenital blindness syndrome in 1927 that bares his
name today [85]. The mutation mapping to Norrin, a secreted
protein, was shown by Nathans et al. to act as a Wnt ligand
and through interaction with the Fz4 receptor to activate the
Wnt-signaling pathway [75]. The persistence of fetal vascula-
ture as well as degenerative and proliferative changes in the
neuroretina and vitreous result in progressive atrophy of the
eyes, clearly demonstrating another incidence where aberra-
tions in the Wnt pathway result in retinal pathology.
In the setting of injury, Wnts may serve a protective role.
Yi et al. have recently shown that Wnt3a protects photorecep-
tors [88]. The results of this study may well be interpreted as
an upregulation of self-renewal of stem cells in the setting of
injury and require further investigation.
Conclusion
Since their discovery in the early 1980s, Wnt proteins and
their associated signaling pathways have generated a great
deal of interest from scientists and physicians. Notably, Wnt
molecules have an important role in promoting self-renewal
of stem cells, in maintaining their stemness, and in their
expansion and proliferation. Due to these properties, Wnt
molecules have become attractive targets for potential ther-
apeutic options and for development of stem cellderived
regeneration strategies.
The expression patterns of numerous Wnts, Fz, and Sfrp
molecules, as well as the activity of the Wnt pathway strongly
suggest that Wnt signaling plays an important role during
retinal development. Future studies will certainly add to this
13
wealth of information and provide valuable insights into the
roles that Wnt-signaling pathways have during retinogen-
esis, as well as in potential future cell-based therapies for
retinal regeneration and disease.
Acknowledgments
M.Y.S.K. is a fellow of the Howard Hughes Medical
Institute and the P&D Soros Society and a John and Shirley
Hanbery scholar.
References
1. Logan CY and R Nusse. (2004). The Wnt signaling pathway in
development and disease. Annu Rev Cell Dev Biol 20:781810.
2. Nusse R and HE Varmus. (1982). Many tumors induced by the
mouse mammary tumor virus contain a provirus integrated in
the same region of the host genome. Cell 31:99109.
3. Nelson WJ and R Nusse. (2004). Convergence of Wnt, beta-
catenin, and cadherin pathways. Science 303:14831487.
4. de Iongh RU, HE Abud and GR Hime. (2006). WNT/Frizzled
signaling in eye development and disease. Front Biosci
11:24422464.
5. Gordon MD and R Nusse. (2006). Wnt signaling: multiple path-
ways, multiple receptors, and multiple transcription factors.
J Biol Chem 281:2242922433.
6. Mikels AJ and R Nusse. (2006). Purified Wnt5a protein activates
or inhibits beta-catenin-TCF signaling depending on receptor
context. PLoS Biol 4:e115.
7. Moon RT, B Bowerman, M Boutros and N Perrimon. (2002).
The promise and perils of Wnt signaling through beta-catenin.
Science 296:16441646.
8. He X, M Semenov, K Tamai and Zeng X. (2004). LDL receptor-
related proteins 5 and 6 in Wnt/beta-catenin signaling: arrows
point the way. Development 131:16631677.
9. Dennis S, M Aikawa, W Szeto, PA dAmore and J Papkoff. (1999).
A secreted frizzled related protein, FrzA, selectively associates
with Wnt-1 protein and regulates wnt-1 signaling. J Cell Sci
112:38153820.
10. Uren A, F Reichsman, V Anest, WG Taylor, K Muraiso, DP Bottaro,
S Cumberledge and JS Rubin. (2000). Secreted frizzled-related
protein-1 binds directly to Wingless and is a biphasic modulator
of Wnt signaling. J Biol Chem 275:43744382.
11. Galli LM, T Barnes, T Cheng, L Acosta, A Anglade, K Willert,
R Nusse and LW Burrus. (2006).: Differential inhibition of
Wnt-3a by Sfrp-1, Sfrp-2, and Sfrp-3. Dev Dyn 235:spc1.
12. Hsieh JC, L Kodjabachian, ML Rebbert, A Rattner,
PM Smallwood, CH Samos, R Nusse, IB Dawid and J Nathans.
(1999). A new secreted protein that binds to Wnt proteins and
inhibits their activities. Nature 1999;398:431436.
13. Bafico A, G Liu, A Yaniv, A Gazit and SA Aaronson. (2001). Novel
mechanism of Wnt signalling inhibition mediated by Dickkopf-1
interaction with LRP6/Arrow. Nat Cell Biol 3:683686.
14. Li L, J Mao, L Sun, W Liu and D Wu. (2002). Second cysteine-
rich domain of Dickkopf-2 activates canonical Wnt signaling
pathway via LRP-6 independently of dishevelled. J Biol Chem
277:59775981.
15. Tan C, P Costello, J Sanghera, D Dominguez, J Baulida, AG de
Herreros and S Dedhar. (2001;). Inhibition of integrin linked
kinase (ILK) suppresses beta-catenin-Lef/Tcf-dependent tran-
scription and expression of the E-cadherin repressor, snail, in
APC-/- human colon carcinoma cells. Oncogene 20:133140.
16. Sadot E, B Geiger, M Oren and A Ben-Zeev (2001). Down-regulation
of beta-catenin by activated p53. Mol Cell Biol 21:67686781.
17. Yu X, S Hoppler, S Eresh and M Bienz. (1996). Decapentaplegic, a
target gene of the wingless signalling pathway in the Drosophila
midgut. Development 122:849858.
 14
18. Kim JS, H Crooks, T Dracheva, TG Nishanian, B Singh, J Jen and
T Waldman. (2002). Oncogenic beta-catenin is required for bone
morphogenetic protein 4 expression in human cancer cells.
Cancer Res 62:27442748.
19. Moon RT, AD Kohn, GV De Ferrari and A Kaykas. (2004). WNT
and beta-catenin signalling: diseases and therapies. Nat Rev
Genet 5:691701.
20. Zallen JA. (2007). Planar polarity and tissue morphogenesis.
Cell 129:10511063.
21. Jenny A and M Mlodzik. (2006). Planar cell polarity signaling:
Downloaded by COLUMBIA UNIVERSITY LIBRARY from online.liebertpub.com at 08/24/17. For personal use only.
a common mechanism for cellular polarization. Mt Sinai J Med
73:738750.
22. Kelly M and P Chen. (2007). Shaping the mammalian auditory
sensory organ by the planar cell polarity pathway. Int J Dev Biol
51:535547.
23. Tzahor E. (2007). Wnt/beta-catenin signaling and cardiogen-
esis: timing does matter. Dev Cell 13:1013.
24. Meinhardt H. (2006). Primary body axes of vertebrates: gen-
eration of a near-Cartesian coordinate system and the role of
Spemann-type organizer. Dev Dyn 235:29072919.
25. Wilson SW and C Houart. (2004). Early steps in the develop-
ment of the forebrain. Dev Cell 6:167181.
26. Fuhrmann S, MR Stark and S Heller. (2003). Expression of
Frizzled genes in the developing chick eye. Gene Expr Patterns
3:659662.
27. Jin EJ, LW Burrus and CA Erickson. (2002). The expression pat-
terns of Wnts and their antagonists during avian eye develop-
ment. Mech Dev 116:173176.
28. Liu H, O Mohamed, D Dufort and VA Wallace. (2003).
Characterization of Wnt signaling components and activation
of the Wnt canonical pathway in the murine retina. Dev Dyn
227:323334.
29. Van Raay TJ and ML Vetter. (2004). Wnt/frizzled signaling dur-
ing vertebrate retinal development. Dev Neurosci 26:352358.
30. Zuber ME, G Gestri, AS Viczian, G Barsacchi and WA Harris.
(2003). Specification of the vertebrate eye by a network of eye
field transcription factors. Development 130:51555167.
31. Hill RE, J Favor, BL Hogan, CC Ton, GF Saunders, IM Hanson,
J Prosser, T Jordan, ND Hastie and V van Heyningen. (1991).
Mouse small eye results from mutations in a paired-like homeo-
box-containing gene. Nature 354:522525.
32. Mathers PH, A Grinberg, KA Mahon and M Jamrich. (1997). The
Rx homeobox gene is essential for vertebrate eye development.
Nature 387:603607.
33. Oliver G, R Wehr, NA Jenkins, NG Copeland, BN Cheyette,
V Hartenstein, SL Zipursky and P Gruss. (1995). Homeobox genes
and connective tissue patterning. Development 121:693705.
34. Rasmussen JT, MA Deardorff, C Tan, MS Rao, PS Klein and ML
Vetter. (2001). Regulation of eye development by frizzled signal-
ing in Xenopus. Proc Natl Acad Sci USA 98:38613866.
35. Shi DL, C Goisset and JC Boucaut. (1998). Expression of Xfz3, a
Xenopus frizzled family member, is restricted to the early ner-
vous system. Mech Dev 70:3547.
36. Wolda SL and RT Moon. (1992). Cloning and developmental
expression in Xenopus laevis of seven additional members of
the Wnt family. Oncogene 7:19411947.
37. Wolda SL, CJ Moody and RT Moon. (1993). Overlapping expres-
sion of Xwnt-3A and Xwnt-1 in neural tissue of Xenopus laevis
embryos. Dev Biol 155:4657.
38. Cavodeassi F, F Carreira-Barbosa, RM Young, ML Concha,
ML Allende, C Houart, M Tada and SW Wilson. (2005). Early
stages of zebrafish eye formation require the coordinated activ-
ity of Wnt11, Fz5, and the Wnt/beta-catenin pathway. Neuron
47:4356.
39. Leimeister C, A Bach and M Gessler. (1998). Developmental
expression patterns of mouse sFRP genes encoding mem-
bers of the secreted frizzled related protein family. Mech Dev
75:2942.
LAD, CHESHIER, AND KALANI
40. Deardorff MA and PS Klein. (1999). Xenopus frizzled-2 is
expressed highly in the developing eye, otic vesicle and somites.
Mech Dev 87:229233.
41. Borello U, V Buffa, C Sonnino, R Melchionna, E Vivarelli and
G Cossu. (1999). Differential expression of the Wnt putative
receptors Frizzled during mouse somitogenesis. Mech Dev
89:173177.
42. Jasoni C, A Hendrickson, H Roelink. (1999). Analysis of chicken
Wnt-13 expression demonstrates coincidence with cell division
in the developing eye and is consistent with a role in induction.
Dev Dyn 215:215224.
43. Kubo F, M Takeichi and S Nakagawa. (2003). Wnt2b con-
trols retinal cell differentiation at the ciliary marginal zone.
Development 130:587598.
44. Heisenberg CP, M Tada, GJ Rauch, L Saude, ML Concha,
R Geisler, DL Stemple, JC Smith, SW Wilson. (2000). Silberblick/
Wnt11 mediates convergent extension movements during
zebrafish gastrulation. Nature 405:7681.
45. Pandur P, M Lasche, LM Eisenberg and M Kuhl. (2002). Wnt-11
activation of a non-canonical Wnt signalling pathway is
required for cardiogenesis. Nature 418:636641.
46. Nakagawa S, S Takada, R Takada and M Takeichi. (2003).
Identification of the laminar-inducing factor: Wnt-signal from
the anterior rim induces correct laminar formation of the neural
retina in vitro. Dev Biol 260:414425.
47. Dorsky RI, DH Rapaport and WA Harris. (1995). Xotch inhibits
cell differentiation in the Xenopus retina. Neuron 14:487496.
48. Bao ZZ and CL Cepko. (1997). The expression and function
of Notch pathway genes in the developing rat eye. J Neurosci
17:14251434.
49. Austin CP, DE Feldman, JA Ida Jr. and CL Cepko. (1995). Vertebrate
retinal ganglion cells are selected from competent progenitors
by the action of Notch. Development 121:36373650.
50. Kubo F, M Takeichi and S Nakagawa. (2005). Wnt2b inhibits dif-
ferentiation of retinal progenitor cells in the absence of Notch
activity by downregulating the expression of proneural genes.
Development 132:27592770.
51. Inoue T, T Kagawa, M Fukushima, T Shimizu, Y Yoshinaga,
S Takada, H Tanihara and T Taga. (2006). Activation of canon-
ical Wnt pathway promotes proliferation of retinal stem
cells derived from adult mouse ciliary margin. Stem Cells
24:95104.
52. Sato N, L Meijer, L Skaltsounis and P Greengard, AH Brivanlou.
(2004). Maintenance of pluripotency in human and mouse
embryonic stem cells through activation of Wnt signaling by a
pharmacological GSK-3-specific inhibitor. Nat Med 10:5563.
53. Dann CE, JC Hsieh, A Rattner, D Sharma, J Nathans and DJ
Leahy. (2001). Insights into Wnt binding and signalling from
the structures of two Frizzled cysteine-rich domains. Nature
412:8690.
54. Osakada F, S Ooto, T Akagi, M Mandai, A Akaike and M
Takahashi. (2007). Wnt signaling promotes regeneration in the
retina of adult mammals. J Neurosci 27:42104219.
55. Livesey FJ and CL Cepko. (2001). Vertebrate neural cell-fate
determination: lessons from the retina. Nat Rev Neurosci
2:109118.
56. Hatakeyama J and R Kageyama (2004). Retinal cell fate determi-
nation and bHLH factors. Semin Cell Dev Biol 15:8389.
57. Mu X and WH Klein. (2004). A gene regulatory hierarchy for ret-
inal ganglion cell specification and differentiation. Semin Cell
Dev Biol 15:115123.
58. Van Raay TJ, KB Moore, I Iordanova, M Steele, M Jamrich, WA
Harris and ML Vetter. (2005). Frizzled 5 signaling governs the
neural potential of progenitors in the developing Xenopus ret-
ina. Neuron 46:2336.
59. Hinds JW and PL Hinds. (1974). Early ganglion cell differentia-
tion in the mouse retina: an electron microscopic analysis uti-
lizing serial sections. Dev Biol 37:381416.
 Wnt-SIGNALING IN RETINAL DEVELOPMENT AND DISEASE
60. Hinds JW and PL Hinds. (1979). Differentiation of photorecep-
tors and horizontal cells in the embryonic mouse retina: an
electron microscopic, serial section analysis. J Comp Neurol
187:495511.
61. Steinberg MS and PM McNutt. (1999). Cadherins and their con-
nections: adhesion junctions have broader functions. Curr Opin
Cell Biol 11:554560.
62. Steinberg MS. (1970). Does differential adhesion govern self-
assembly processes in histogenesis? Equilibrium configura-
tions and the emergence of a hierarchy among populations of
Downloaded by COLUMBIA UNIVERSITY LIBRARY from online.liebertpub.com at 08/24/17. For personal use only.
embryonic cells. J Exp Zool 173:395433.
63. Steinberg MS. (1978). Cell-cell recognition in multicellular
assembly: levels of specificity. Symp Soc Exp Biol 32:2549.
64. Pujic Z and J Malicki. (2004). Retinal pattern and the genetic basis
of its formation in zebrafish. Semin Cell Dev Biol 15:105114.
65. Erdmann B, FP Kirsch, FG Rathjen and MI More (2003).
N-cadherin is essential for retinal lamination in the zebrafish.
Dev Dyn 226:570577.
66. Horne-Badovinac S, D Lin, S Waldron, M Schwarz, G Mbamalu,
T Pawson, Y Jan, DY Stainier and S Abdelilah-Seyfried (2001).
Positional cloning of heart and soul reveals multiple roles for
PKC lambda in zebrafish organogenesis. Curr Biol 11:14921502.
67. Jensen AM and M Westerfield. (2004). Zebrafish mosaic eyes is
a novel FERM protein required for retinal lamination and ret-
inal pigmented epithelial tight junction formation. Curr Biol
14:711717.
68. Malicki J, H Jo and Z Pujic. (2003). Zebrafish N-cadherin,
encoded by the glass onion locus, plays an essential role in ret-
inal patterning. Dev Biol 259:95108.
69. Wei X and J Malicki. (2002). nagie oko, encoding a MAGUK-
family protein, is essential for cellular patterning of the retina.
Nat Genet 31:150157.
70. Fu X, H Sun, WH Klein and X Mu. (2006). Beta-catenin is essen-
tial for lamination but not neurogenesis in mouse retinal devel-
opment. Dev Biol 299:424437.
71. Cho SH and CL Cepko. (2006). Wnt2b/beta-catenin-mediated
canonical Wnt signaling determines the peripheral fates of the
chick eye. Development 133:31673177.
72. Liu H, S Xu, Y Wang, C Mazerolle, S Thurig, BL Coles, JC Ren,
MM Taketo, D van der Kooy, VA Wallace. (2007). Ciliary margin
transdifferentiation from neural retina is controlled by canoni-
cal Wnt signaling. Dev Biol 308:5467.
73. Carmeliet P and M Tessier-Lavigne. (2005). Common mecha-
nisms of nerve and blood vessel wiring. Nature 436:193200.
74. Robitaille J, ML MacDonald, A Kaykas, LC Sheldahl, J Zeisler,
MP Dube, LH Zhang, RR Singaraja, DL Guernsey, B Zheng, LF
Siebert, A Hoskin-Mott, MT Trese, SN Pimstone, BS Shastry,
RT Moon, MR Hayden, YP Goldberg and ME Samuels. (2002).
Mutant frizzled-4 disrupts retinal angiogenesis in familial exu-
dative vitreoretinopathy. Nat Genet 32:326330.
75. Xu Q, Y Wang, A Dabdoub, PM Smallwood, J Williams, C
Woods, MW Kelley, L Jiang, W Tasman, K Zhang and J Nathans.
(2004). Vascular development in the retina and inner ear: con-
trol by Norrin and Frizzled-4, a high-affinity ligand-receptor
pair. Cell 116:883895.
76. Lyu J and CK Joo. (2004). Wnt signaling enhances FGF2-triggered
lens fiber cell differentiation. Development 131:18131824.
77. Smith AN, LA Miller, N Song, MM Taketo and RA Lang. (2005).
The duality of beta-catenin function: a requirement in lens mor-
phogenesis and signaling suppression of lens fate in periocular
ectoderm. Dev Biol 285:477489.
78. Sundin OH, GS Leppert, ED Silva, JM Yang, S Dharmaraj, IH
Maumenee, LC Santos, CF Parsa, EI Traboulsi, KW Broman,
15
C Dibernardo, JS Sunness, J Toy and EM Weinberg. (2005).
Extreme hyperopia is the result of null mutations in MFRP,
which encodes a Frizzled-related protein. Proc Natl Acad Sci
USA 102:95539558.
79. Mukhopadhyay M, M Gorivodsky, S Shtrom, A Grinberg, C
Niehrs, MI Morasso and H Westphal. (2006). Dkk2 plays an
essential role in the corneal fate of the ocular surface epithe-
lium. Development 133:21492154.
80. Jones SE, C Jomary, J Grist, HJ Stewart, MJ Neal. (2000).
Modulated expression of secreted frizzled-related proteins in
human retinal degeneration. Neuroreport 11:39633967.
81. Jones SE, C Jomary, J Grist, HJ Stewart and MJ Neal. (2000).
Identification by array screening of altered nm23-M2/PuF
mRNA expression in mouse retinal degeneration. Mol Cell Biol
Res Commun 4:2025.
82. Kermane A, S Tachfouti, H El Moussaif, Z Mohcine. (2004).
[Association of choroidal coloboma, congenital hypertrophy of
retinal pigmented epithelium and familial adenomatous poly-
posis: case report]. Bull Soc Belge Ophtalmol 5964.
83. Marcus DM, AK Rustgi, D Defoe, SE Brooks, RS McCormick, TP
Thompson, W Edelmann, R Kucherlapati and S Smith. (1997).
Retinal pigment epithelium abnormalities in mice with ade-
nomatous polyposis coli gene disruption. Arch Ophthalmol
115:645650.
84. Nadauld LD, S Chidester, DN Shelton, K Rai, T Broadbent, IT
Sandoval, PW Peterson, EJ Manos, CM Ireland, HJ Yost and DA
Jones. (2006). Dual roles for adenomatous polyposis coli in reg-
ulating retinoic acid biosynthesis and Wnt during ocular devel-
opment. Proc Natl Acad Sci USA 103:1340913414.
85. Berger W, A Meindl, TJ van de Pol, FP Cremers, HH Ropers,
C Doerner, A Monaco, AA Bergen, R Lebo, M Warburg,
L Zergollern, B Lorenz, A Gal, EM Bleeker-Wagemakers and
T Meitinger. (1992). Isolation of a candidate gene for Norrie dis-
ease by positional cloning. Nat Genet 1:199203.
86. Cain S, G Martinez, K Turner, RJ Richardson, MI Kokkinos,
HE Abud, J Huelsken, ML Robinson and RU de Iongh.
(2008). Differential requirement for beta-catenin in epithelial
and fiber cells during lens development. Dev Biol. ahead of
print.
87. Gage PJ, M Qian, D Wu and KI Rosenberg. (2008). The canon-
ical Wnt signaling antagonist DKK2 is an essential effector
of PITX2 function during normal eye development. Dev Biol
317:310324.
88. Yi H, RE Nakamura, O Mohamed, D Dufort and AS Hackam.
(2007). Characterization of Wnt signaling during photoreceptor
degeneration. Invest Ophthalmol Vis Sci 48:57335741.
89. Kameya S, NL Hawes, B Chang, JR Heckenlively, JK Naggert
and PM Nishina. (2002). Mfrp, a gene encoding a frizzled
related protein, is mutated in the mouse retinal degeneration 6.
Hum Mol Genet 11:18791886.
Address reprint requests to:
Dr. M. Yashar S. Kalani
Stanford University School of Medicine
Department of Developmental Biology
279 Campus Drive, Room B273
Stanford, CA 94305
Email: kalani@stanford.edu
Received for publication June 22, 2008; accepted after
revision August 08, 2008.
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