REVIEWS

Transcriptional regulation of
photoreceptor development and
homeostasis in the mammalian retina
Anand Swaroop*, Douglas Kim*§ and Douglas Forrest‡
Abstract | In the developing vertebrate retina, diverse neuronal subtypes originate from
multipotent progenitors in a conserved order and are integrated into an intricate laminated
architecture. Recent progress in mammalian photoreceptor development has identified a
complex relationship between six key transcription-regulatory factors (RORβ, OTX2, NRL,
CRX, NR2E3 and TRβ2) that determine rod versus M cone or S cone cell fate. We propose a
step-wise ‘transcriptional dominance’ model of photoreceptor cell fate determination, with
the S cone representing the default state of a generic photoreceptor precursor. Elucidation
of gene-regulatory networks that dictate photoreceptor genesis and homeostasis will have
wider implications for understanding the development of nervous system function and for
the treatment of neurodegenerative diseases.

Leber’s congenital amaurosis

“Tamaso ma jyotir Gamaya ...” (lead us from darkness microcircuits for integration and processing of visual
A congenital form of early-onset to light) signals2–4 (FIG. 1a). The process of vision begins at the
blindness caused by mutations photoreceptors, which are unique sensory neurons that
at many genetic loci. In Sanskrit from the discussion of the life force ‘Prana’ are specialized to capture light quanta. The chemical
in Brihadaranyaka Upanishad (1.3.28) (800–2000 bc) output of photoreceptors is integrated and processed
by interneurons (bipolar, horizontal and amacrine
For thousands of years, humankind has sought light cells) and transmitted to visual centres in the brain
and vision to promote growth, survival and intellectual by ganglion cells. Abnormalities, dysfunction and/or
development. Darkness and the loss of sight are among death of retinal photoreceptors constitute the primary
*Neurobiology- our most basic fears. In humans, many higher-order cause of visual impairment or blindness in most reti-
Neurodegeneration and
neuronal functions including behaviour, learning, nal diseases, such as Leber’s congenital amaurosis (LCA),
Repair Laboratory (N-NRL),
Building 6/338, MSC 0610, memory and emotions are dictated, to a considerable retinitis pigmentosa and macular degeneration.
National Eye Institute, extent, by vision1. Importantly, almost 30% of the sen- In this Review, we synthesize current knowledge of
National Institutes of Health, sory input to the brain originates in the retina, which photoreceptor development and propose a framework
6 Center Drive, Bethesda, is referred to as the “window to the brain”1,2. Given the for the regulatory transcriptional networks that deter-
Maryland 20892, USA.
‡
Clinical Endocrinology
ease of diagnosis and quantification of visual defects, it mine photoreceptor cell fate in the mammalian retina.
Branch, National Institute of is not surprising that as many as 30% of approximately We briefly describe genetic defects that result in syndro-
Diabetes, Digestive and 20,000 entries in the Online Mendelian Inheritance mic or non-syndromic photoreceptor degeneration due
Kidney Diseases, Bethesda, in Man (OMIM) database (see the online links box) to aberrant differentiation, function or homeostasis, and
20892, Maryland, USA.
reveal some association with the eye, vision and/or discuss potential treatment strategies for neurodegenera-
§
Present address: Howard
Hughes Medical Institute, visual dysfunction. tive blinding diseases based on the current understanding
Janelia Farm Research The retina is an excellent model for studying com- of transcriptional networks.
Campus, 19700 Helix Drive, plex features of the nervous system, such as cell lineage
Ashburn, 20147, Virginia, and connectivity in general and visual function in par- Photoreceptors: types and organization
USA
Correspondence to A.S.
ticular. The vertebrate retina consists of six major types Mammalian photoreceptors are highly compartmental-
e-mail: of neurons (rod and cone photoreceptors, and horizon- ized for specialized functions and have an intimate rela-
[email protected] tal, bipolar, amacrine and ganglion cells) that exhibit tionship with the retinal pigment epithelium (RPE), which
doi:10.1038/nrn2880 laminar organization and form numerous parallel supports photoreception (FIG 1a,b). Cone photoreceptors

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a b c
Human
Retinal pigment
epithelium
Photoreceptor layer
C R R RPE
Outer limiting
membrane Disc
shedding
Outer nuclear layer

Outer plexiform layer OS OS

biogenesis
H H
B B B Transport
B
Inner nuclear layer IS Mouse
M Metabolism
Superior
A
(dorsal)

Inner plexiform layer

CB
Gene
regulation
Ganglion cell layer G
G G
Optic nerve fiber layer Synaptic Syn Inferior
Inner limiting membrane transmission (ventral)

Figure 1 | Functional circuitry of the retina. a | Organization of retinal circuits. Rod (R) and cone (C) photoreceptors
have cell bodies (CBs) in the outer nuclear layer and extend inner segments (IS), which contain metabolic machinery, and
outer segments (OS), which associate with retinal pigment epithelial (RPE) cells. Photoreceptor axons terminate in the
outer plexiform layer and synapse with horizontal (H) and bipolar (B) cells in the inner nuclearNature Reviews
layer, which | Neuroscience
also contains
Müller glial (M) and amacrine (A) cells. Bipolar cells relay signals to amacrine and ganglion (G) cells through synapses in the
inner plexiform layer. Ganglion cell axons project towards the optic nerve head and carry signals to the brain. End-feet of
Müller glia form the outer and inner limiting membranes. A representative cone pathway is shown in blue and a
representative rod pathway is shown in orange. b | Illustrations of rod and cone morphologies that include subcellular
locations of functions. c | A surface-view representation of cone distribution across the mammalian retina. In humans,
cones express S opsins (peak sensitivity to blue light), L opsins (peak sensitivity to red light) or M opsins (peak sensitivity to
green light) in a mosaic-like pattern. In mice, cones express S opsins and M opsins in opposing distribution gradients along
the superior (M opsin-high) to inferior (S opsin-high) axis. Syn, synaptic terminus.

Retinitis pigmentosa
An inherited progressive respond to bright light, mediate colour vision and per- the mouse) have two types of cone opsins that confer
degeneration of mit high resolution of visual images. Rod photorecep- dichromatic colour vision: s opsin (also known as blue-
photoreceptors, generally tors function only under conditions of low light and can sensitive opsin), which has peak sensitivity in the short
beginning in the peripheral respond to single light quanta, being a hundred-fold wavelength (ultraviolet or blue) region of the spectrum;
retina with rod cell dysfunction.
more sensitive than cones5 (see the Webvision website, and M opsin (also known as green-sensitive opsin),
Macular degeneration listed in Further information) which has peak sensitivity in the medium–long wave-
A progressive dystrophy Across the retina, rods and various cone subtypes are length (green) region of the spectrum. Duplication of
initially affecting arranged in an elegant laminar and non-random pattern an ancestral M opsin-like gene on the X chromosome
photoreceptors in the 5–6 mm
(mosaic)6–8 (FIG. 1c) that must be sequentially generated has provided humans and diurnal primates with an
area around the fovea (the
macula), which contains a with the right number of components, correct spatial additional opsin, L opsin (also known as red-sensitive
higher ratio of cones to rods organization and appropriate wiring to interneurons opsin), which is sensitive to longer wavelength (red)
than the peripheral retina. for serial and parallel processing of visual information9. light and, together with s opsin and M opsin, confers
Juvenile forms exhibit a In mice and humans, photoreceptors constitute over trichromatic colour vision11. Each human cone expresses
Mendelian inheritance pattern,
whereas age-related macular
70% of retinal cells, but rods outnumber cones by 30:1 only one of the three opsins, giving a mosaic-like pattern
degeneration is a complex in mice and 18–20:1 in humans6,7. A major difference over the retina12. In mice and other rodents, M opsins
multifactorial disease. between humans (and diurnal primates) and mice (and and s opsins are expressed in opposing gradients across
most other mammals) is the presence in humans of a the retina13. Cones in mid-retinal regions express vary-
Syndromic
thin, pit-like, cone-only region in the centre of the retina, ing amounts of both M and s opsin14,15, leading to the
Related to a pathology or
disease involving multiple called the fovea, which is responsible for highest visual concept of s- or M-dominant cone subpopulations in
organs. acuity. In humans, the density of rods increases from the some species16.
fovea to the periphery of the retina, with the highest rod
Retinal pigment epithelium density in the parafoveal region10. Multipotent progenitors and neuronal diversity
A polarized sheet of epithelial
cells between the choroidal
The mammalian retina has only one type of rod The proportions and distribution of photoreceptor sub-
capillaries and the visual pigment (rhodopsin, which has a peak spectral types in the mature retina are optimized to the precise
photoreceptor cells. sensitivity at ∼500 nm). Most mammals (including needs of a given species for capturing visual information

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 REVIEWS

in diverse habitats. The identification of genes and sig- and Fwk 10, respectively37,38. s opsin mRnA is pro-
nals that direct a dividing retinal progenitor cell (RPC) duced as early as Fwk 12, followed by rhodopsin,
to become a photoreceptor or another cell type has M opsin and L opsin transcripts at Fwk 15 (ReF. 38)
been the subject of intense investigations17–19. Classical (FIG. 3). At birth, opsin expression patterns are reason-
birth-dating studies using 3H-thymidine labelling, line- ably well established, but photoreceptors are still imma-
age tracing experiments, or cell-type specific antibodies ture. In the mouse, photoreceptor development is less
demonstrated a fixed chronological and overlapping advanced than in humans at birth, and the eyes of the
order of birth for all retinal neurons from common newborn pup remain closed for almost 2 weeks. Cone
pools of progenitor cells in the vertebrate retina20–24. genesis is essentially complete at birth (having started
The generation of functionally mature neurons from at embryonic day (E) 11), and s opsin is expressed at
multipotent RPCs proceeds through a series of steps late embryonic stages. In mice, rods are born both pre-
that increasingly restrict lineage choices and commit and postnatally. The peak of rod genesis occurs in the
cells to a particular fate25–27 (FIG. 2). How is this process first few postnatal days, accompanied soon thereafter
stringently controlled to be appropriate to the temporal by the expression of rhodopsin20,21 (FIG. 3). Expression
and spatial context? of M opsin by cones begins around postnatal day 6 and
Initially, it was proposed that the developmental lags behind that of other opsins. As photoreceptors
potency of uncommitted progenitors changes with mature, the opsin levels increase substantially, outer
developmental age28. According to a more detailed segments elaborate towards the RPE and synapses form
model29 both intrinsic control mechanisms of the pro- with horizontal and bipolar neurons.
genitor cells and the environment change over time, and
changes in competence generate different neurons. The From progenitors to photoreceptor precursors
progressive-restriction and competence models for gen- The earliest steps in photoreceptor development involve
Retinal progenitor cell
A proliferating cell that can give
erating cellular diversity provided a valuable framework signalling through the notch receptor 39,40. High levels of
rise to mature retinal cells. for subsequent developmental and molecular genetic notch seem to maintain the cycling of progenitor cells,
studies. A complex interplay of transcription factors and inhibition of notch drives progenitors to commit
Lineage tracing (including homeodomain proteins such as paired box to the cone or rod photoreceptor fate, depending on
An experimental method to
protein PAX6, retinal homeobox protein RX1, sIX3, the developmental stage, over other cell fates. notably,
identify the origin (progenitor)
of a differentiated cell. sIX6, visual system homeobox 2 (vsX2) and LIM– signalling molecules such as sonic hedgehog can also
homeobox protein LHX2) is implicated in an evolution- influence progenitor commitment by converging on
Competence arily conserved programme that establishes the anterior downstream targets of notch41.
The ability of a retinal neuroectodermal region for eye formation and main- notch signals through the bHLH transcription fac-
progenitor or precursor cell to
produce specific cell types.
tains the retinogenic potential of progenitor cells30–34. tors HEs1 and HEs5, which maintain the progenitor
The transient and quantitatively precise expression of pool by downregulating the expression of proneural
bHLH transcription factors regulatory transcriptional proteins, which are often bHLH proteins in the developing mouse retina39,40,42.
A family of transcription factors bHLH transcription factors and homeodomain transcription The molecular mechanisms that generate photorecep-
that contain a characteristic
factors18,19,26, mediates the acquisition of specific cell tor precursors from RPCs, including downregulation of
basic region and a
helix–loop–helix domain. fates and provides a starting point for the elucidation notch signalling, remain uncharacterized. The proneural
of regulatory pathways underlying neurogenesis. It has bHLH transcription factors (such as nEUROD1, neuro-
Homeodomain transcription emerged that many transcription factors (including genin 2, nEUROD4 (also known as MATH3), protein
factors PAX6, vsX2 and neurogenic differentiation factor 1 atonal homologue 7 (ATOH7; also known as MATH5)
A family of transcription factors
that contain a characteristic
(nEUROD1)) participate at an early point in cell fate and achaete-scute homologue 1 (AsCL1; also known
DNA recognition domain, specification as well as later in functional maturation and as MAsH1)) have redundant and poorly defined roles
called the homeodomain. They maintenance (homeostasis). in photoreceptor development 42,43. Their promiscuous
are often involved in patterning roles in early neuronal development make it difficult to
spatial domains of developing
The timing of photoreceptor genesis determine specific instructive or participatory function
tissues.
In mammals, the genesis of photoreceptors occurs in progenitor competence, lineage restriction and/or
Specification over a long temporal window 20,21,35,36 (FIG. 3). After exit photoreceptor fate specification (FIG. 2). Each step in early
The developmental process from final mitosis , functional maturation of a com- differentiation is probably regulated by combinatorial
that biases an immature cell to mitted photoreceptor precursor (rod or cone) can take and transient actions of several transcription factors and
adopt a particular fate; the
specified cell is not yet
weeks to months depending on the species (FIG. 3) . signalling proteins.
committed to the fate and The process of photoreceptor development can be
retains developmental divided into five major steps (FIG. 2): first, prolifera- Transcription factors in differentiation
plasticity. tion of multipotent RPCs; second, restriction of the Although we have only a limited understanding of the
competence of RPCs; third, cell fate specification and signals that prompt a progenitor to become a photore-
Final mitosis
The last mitotic division of a commitment to photoreceptor precursors during or ceptor precursor, we have better insights into subse-
cell. after final mitosis; fourth, expression of photorecep- quent events that direct a photoreceptor precursor to
tor genes, such as those for phototransduction and differentiate into a rod or a cone. Here, we discuss six
Photoreceptor precursor morphogenesis; and fifth, axonal growth, synapse transcription factors that dictate key steps of photore-
A post-mitotic cell that is not
yet differentiated and does not
formation and outer segment biogenesis. In humans, ceptor differentiation in the developing mammalian
have a mature functional all photoreceptors are generated prenatally. The first retina — namely, homeobox protein OTX2, cone–rod
phenotype of a rod or a cone. cones and rods are born around foetal week (Fwk) 8 homeobox protein (CRX), neural retina leucine zipper

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Transcription Dividing Lineage-restricted Post-mitotic Immature Functional Transcription

factors and multipotent progenitors photoreceptor photoreceptors photoreceptors regulatory
signalling progenitors precursors proteins
proteins

Non-photoreceptor Non-photoreceptor M
cell types cell types
TRβ2, RXRγ,
RORβ, COUP-TF1,
Cones
NEUROD1,
CRX and OTX2

S
Cone
PAX6, RX1, precursor
SIX3, SIX6,
LHX2, VSX2,
HES1, Notch 1
and others

RORβ, NRL,
Rods NR2E3, OTX2,
• Competence • Cell cycle exit Rod CRX, ASCL1,
restriction • Fate specification precursor NEUROD1 and
PIAS3
Proliferation • Photoreceptor fate commitment • Opsin expression and patterning
• Outer-segment formation
Photoreceptor subtype • Onset of phototransduction
specification
Terminal differentiation

Figure 2 | stages of photoreceptor development. Early in retinogenesis, multipotent retinal progenitor cells (RPCs)
Nature
divide and produce additional multipotent progenitors (thick circular arrow) or progenitor cells thatReviews
become| Neuroscience
restricted in
their competence to generate various cell types (thin circular arrow). Some of these proliferating cells become restricted
to a lineage that will give rise to at least one photoreceptor cell and possibly to non-photoreceptor cells. After cell cycle
exit, postmitotic precursors can remain plastic. During cell type specification of photoreceptors, precursors are directed
to become cones or rods that eventually express photopigments (M opsin and S opsin in cones, and rhodopsin in rods),
and form outer segments and synapses. The bar on the far left lists key transcription factors and signalling proteins that
maintain RPC multipotency and proliferation: paired box protein PAX6, retinal homeobox protein RX1, SIX3, SIX6,
LIM–homeobox protein LHX2, visual system homeobox 2 (VSX2), HES1 and Notch 1. The bar on the far right lists key
transcription regulatory proteins that are involved in cone and rod differentiation and maintenance (for cones: thyroid
hormone receptor β2 (TRβ2), retinoid X receptor-γ (RXRγ), nuclear receptor RORβ, COUP transcription factor 1
(COUP-TF1; also known as NR2F1), neurogenic differentiation factor 1 (NEUROD1), cone–rod homeobox protein (CRX)
and homeobox protein OTX2; for rods: RORβ, neural retina leucine zipper protein (NRL), photoreceptor-specific nuclear
receptor (NR2E3), OTX2, CRX, achaete-scute homologue 1 (ASCL1; also known as MASH1), NEUROD1 and E3
SUMO-protein ligase PIAS3).

protein (nRL), photoreceptor-specific nuclear receptor CRX and terminal differentiation of photoreceptors. The
(nR2E3), nuclear receptor RORβ and thyroid hormone transcriptional regulator CRX was initially identified as a
receptor β2 (TRβ2). photoreceptor-specific retinopathy gene, but is now known
Paired-type homeodomain to also be expressed in other mature retinal neurons46–50.
transcription factor OTX2 and commitment to a photoreceptor fate. The In Crx-deficient mice, photoreceptors are generated but
A DNA-binding transcription- paired-type homeodomain transcription factor OTX2 is a fail to express many phototransduction genes, resulting in
regulating protein that contains
a homeodomain with the
key regulator of the photoreceptor lineage. It is expressed a lack of outer segments and eventually retinal degenera-
characteristic amino acid during final mitosis in retinal progenitors and in early tion51. CRX is expressed early in postmitotic photoreceptor
residues of the homeodomain precursors (T. Furukawa, personal communication) that precursors and acts downstream of OTX2 (ReFs 44,45). Its
of the Drosophila become committed to the photoreceptor cell fate44,45. A exact role at this stage remains undefined. CRX enhances
melanogaster Paired
conditional knockout of Otx2 in immature retinal pro- the expression of photoreceptor-specific genes52,53 and
transcription factor.
genitors leads to almost complete loss of rods and cones, is important for terminal differentiation of rods and
Basic motif–leucine zipper as well as a loss of bipolar and horizontal cells in mice44,45. cones; however, CRX alone does not determine specific
transcription factor Retrovirus-mediated expression of Otx2 in RPCs of neo- photoreceptor cell fate51.
A transcription factor that natal mice can drive cells towards the rod cell fate44. As
contains a characteristic basic
motif for DNA binding and a
OTX2 is also expressed in other cell types throughout NRL and the specification of rod photoreceptor fate. The
leucine zipper domain for the brain and retina, it provides a necessary, but not a decision to be or not to be a rod is largely determined
dimerization. sufficient, signal to induce the photoreceptor cell fate. by nRL, a basic motif–leucine zipper transcription factor of

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a Mouse b Human

Cone precursors Rod precursor

Rod precursors Cone precursor

Photoreceptor birth
RPE

Rearrangement,
Formation and packing and
maturation of OS maturation of OS

ONBL

10 15 Birth 5 10 20 8 16 24 32 Birth 2 3

Embryoninc day Postnatal day Foetal week Postnatal year

S opsin S opsin
M opsin L and M opsin
Rhodopsin Rhodopsin
Figure 3 | Photoreceptor genesis and maturation in mice and humans. The relative numbers of cone and rod precursor
cells that are born over time are shown for mouse and human retinogenesis. a | In mice, cones are generated prenatally as
early as embryonic day (E) 11. Rods, which vastly outnumber cones, are generated both before Nature Reviews
and after | Neuroscience
birth, from around
E12 to postnatal day (P) 10. Expression of S opsin begins at ~E18, that of M opsin at ~P6 and that of rhodopsin at ~P2.
Photopigment levels increase substantially until after weaning in mice. As photoreceptors mature, outer segments (OS) and
synapses form. The inset panel shows double fluorescent detection of cone (thyroid hormone receptor β2-positive cells (red
staining)) and rod precursors (neural retina leucine zipper protein (NRL) -positive cells (green staining)) in mouse retina at
E18, superimposed on a phase contrast picture that reveals the outer neuroblastic layer (ONBL) (D. Sharlin, Alok Swaroop
and D. Forrest, unpublished data). Newly generated photoreceptors tend to reside near the edge of the retina.
b | In humans, cones and rods are generated around foetal week (Fwk) 8 and Fwk 10, respectively. Generation of cones is
completed prenatally, whereas that of rods continues into the early postnatal period. Expression of S opsin is observed at
~Fwk 12, and that of L opsin, M opsin and rhodopsin at ~ Fwk 15. Functional maturation of photoreceptors continues
postnatally. RPE, retinal pigment epithelium. Data are taken from ReFs 14,20,21,38,80,151,152.

the Maf subfamily 54. In mammals, nRL interacts with excess cones are non-functional and more primitive than
CRX and many other transcription factors and induces those in Nrl–/– mice. Nrl is not expressed in Rorb–/– mice,
the expression of rod-specific genes52,55,56. Loss of nRL indicating that Rorb lies upstream of Nrl in the photore-
in mice leads to a retina with no rods and an excess of s ceptor differentiation pathway. RORβ, like CRX, is also
opsin-expressing cones (s cones) because the precursors necessary for the formation of photoreceptor outer seg-
that were committed to become rods are converted to ments and contributes to the development of the inner
cones57. Ectopic expression of nRL in immature pho- nuclear and synaptic layers of the retina71. RORβ acts
toreceptor precursors results in loss of cone gene expres- synergistically with CRX to initiate s opsin transcription
sion and likely transformation to rods58. These studies in cones72.
establish an essential and instructive role of nRL in
determining rod cell fate. TRβ2, a mediator of M cone differentiation. The nuclear
receptor TRβ2 is a ligand-regulated transcription factor73
NR2E3, a suppressor of cone genes. The orphan nuclear expressed in new cones in the mammalian foetal
receptor nR2E3 (ReF. 59) is a direct transcriptional target retina74,75. In TRβ2-deficient mice, cones lack M opsin
of nRL60 and is expressed in postmitotic photoreceptors61. and express only s opsin74, indicating a crucial role for
nR2E3 primarily suppresses the expression of cone thyroid hormone signalling in opsin patterning and
genes, thereby irreversibly committing nRL-expressing maturation of the colour visual system. TRβ2 coordi-
precursors to a rod cell fate60,62–64. Mice carrying an L1 nates cone opsin patterning in response to the increasing
retrotransposon insertion65 in the Nr2e3 gene66 (rd7 thyroid hormone levels that accompany development 76.
mice) have rod-like photoreceptors that express many The preceding discussion of six key transcription fac-
cone genes63,64,67. nR2E3, together with nRL and CRX, tors suggests an intricate regulatory network that guides
also serves as a co-activator of rod genes, but cannot the determination of photoreceptor cell fate.
Nuclear receptor
substitute for nRL in rod gene activation63,68.
A ligand-regulated
transcription factor that The ‘transcriptional dominance’ hypothesis
includes members with known RORβ, a dual regulator in both rod and cone differ- Although a unified pathway cannot be put forward
ligands such as thyroid entiation. RORβ is an orphan nuclear receptor that is for all vertebrates17, we have synthesized the evidence
hormone receptor and retinoid expressed in all cell layers of the neural retina and in the from genetic studies of key transcription factors as well
X receptor, and those lacking a
known physiological ligand
pineal gland69,70. The retina of Rorb–/– mice lacks rods as earlier proposals that cones may differentiate into
such as retinoid-related orphan and features an excess of s cone-like photoreceptors, a common default form before diversifying into dis-
receptor. therefore resembling that of Nrl–/– mice71. However, the tinct cone subtypes13,14. We propose a ‘transcriptional

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Retinal progenitor The model predicts that the transcriptional control

signals exert a profound influence at two key points
?
during fate specification: first, the decision to form
OTX2 either a rod or a cone; and second, the decision for a
cone to differentiate with an s cone or M cone identity
(FIG. 4). It is likely that the commitment to produce a
functional rod or cone is dictated by context- and time-
Committed photoreceptor dependent combinations of cell-specific and widely
precursor expressed transcription factors. Modest changes in
the expression or activity of one or more transcriptional
• Inducing factors
• Amount of factor RORβ
regulators in response to extrinsic or cell–cell com-
• Activity munication signals might alter the overall outcome of
• Interactions differentiation. The model also predicts that, once the
• Post-translational CRX
modification NRL requisite numbers of committed cones are produced,
• Small molecules by as yet unknown counting mechanisms, the rod-
• Ligands NR2E3 inducing signals will overwhelmingly direct the differ-
entiation of precursors away from the default s cone
Cone precursor Rod precursor
state to the rod state.
The coordinated and precisely balanced action of
T3 the six key transcription factors (discussed above)
seems to be crucial as RPCs commit to a rod or cone
RXRγ
COUP-TFs photoreceptor cell fate. We propose that several of these
TRβ2 factors compete for dominance in a newborn photore-
RORα RORα ceptor precursor. These sometimes conflicting regu-
M opsin S opsin Rhodopsin
lators would determine the particular differentiation
M cone S cone Rod outcome of a given precursor cell at a specific stage
of development. several (if not all) of the six factors
Figure 4 | Transcriptional dominance model of photoreceptor cell fate
Nature Reviews | Neuroscience (OTX2, CRX, RORβ, nRL, nR2E3 and TRβ2) may be
determination. A generic photoreceptor is formed under the control of homeobox
protein OTX2 and other undetermined signals. This precursor is programmed to possess expressed at varying levels in a precursor cell (FIG. 4)
a ‘default’ S cone state under the control of OTX2 (and/or cone–rod homeobox protein and engage in a transcriptional contest for dominance
(CRX)) and nuclear receptor RORβ unless diverted into a rod or M cone state by (‘tug of war’) that will ultimately determine the cell
additional signals. The ‘NRL control box’ determines whether a precursor becomes a rod fate. In the default scenario, OTX2 biases the progeni-
or a cone, and the ‘thyroid hormone receptor β2 (TRβ2) control box’ prompts a cone to tor to a photoreceptor precursor fate, and CRX and
acquire an M opsin or an S opsin identity. Each control box is subject to modifying RORβ initially induce the expression of s opsin. Other
regulators. Induction of neural retina leucine zipper protein (NRL) and its target, pho- transcription factors including PR domain zinc finger
toreceptor-specific nuclear receptor (NR2E3), induces a rod state and suppresses cone protein 1 (also known as BLIMP1) are likely to help
genes, which consolidates the rod fate. Other factors involved in rod development stabilize the commitment of OTX2-expressing precur-
include E3 SUMO-protein ligase PIAS3, neurogenic differentiation factor 1 (NEUROD1),
sors to a photoreceptor fate77,78. In the photoreceptor
achaete-scute homologue 1 (ASCL1; also known as MASH1), myocyte enhancer factor
2C and retinoblastoma-associated protein, probably at both early and later stages of precursor, RORβ, presumably in cooperation with
differentiation. If NRL and NR2E3 fail to act, photoreceptor precursors follow the other specific factors, also induces nRL expression.
‘default’ pathway to S cones unless TRβ2 and its ligand triiodothyronine (T3) induce M When a certain threshold of nRL expression and/or
opsin and suppress S opsin expression in spatially restricted patterns in cone activity is achieved, nRL may recruit CRX and acti-
subpopulations over the retina. Other factors involved in M opsin and S opsin patterning vate nR2E3 expression. An nRL–nR2E3 complex
include retinoid X receptor-γ (RXRγ), RORβ, RORα, COUP transcription factors then suppresses s opsin and other cone genes, while
(COUP-TFs) and CRX. NEUROD1 and bone morphogenetic protein receptor type 1A–1B nRL–CRX-containing complexes initiate the rod dif-
contribute to inducing the expression of TRβ2 and COUP-TF, respectively. ferentiation programme. Below a ‘decisive threshold’
of expression and/or activity of nRL–nR2E3, the
precursor differentiates by default as a cone. In these
dominance’ model of photoreceptor cell fate determina- non-rod precursors, TRβ2 is crucial for the induction
tion (FIG. 4) that includes three fundamental attributes: of M opsin in cones and differential patterning of both
first, that all terminally differentiated photoreceptors M opsins and s opsins.
originate from a common postmitotic photoreceptor several lines of evidence suggest the existence of a
precursor that has the potential to form rods or any of common photoreceptor precursor. First, in the develop-
the cone subtypes; second, that a photoreceptor precur- ing retina of humans and rodents, s opsin is expressed
sor cell follows a default pathway to differentiate into before other cone opsins (FIG. 3). such observations in
an s cone unless additional regulatory signals direct the rat led to the proposal of a default pathway in which
the precursor to acquire a rod or M cone identity; and cones initially express s opsin before a subpopulation
third, that the acquisition of a specific fate by a pho- subsequently switches to an M opsin identity 79. In the
toreceptor precursor is established by a particular set human foetus, s cones cover almost 90% of the retina
of transcription-regulatory factors gaining dominance at Fwk 19, and this number decreases dramatically later
at a given developmental stage. in development, potentially reflecting a switch in opsin

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expression from a default s cone state to an M or L cone TRβ2 is a major regulator of M opsin and s opsin
identity 8. second, in TRβ2-deficient mice, cones lack patterning in mice but is expressed in cones in all reti-
M opsin and instead express s opsin74, consistent with nal regions, suggesting that other factors contribute to
a key role for TRβ2 in M cone versus s cone patterning. the regional variations in M opsin and s opsin distri-
Third, Nrl–/– mice possess s cones instead of rods, which bution. One such factor is the nuclear receptor retin-
supports the concept of the default s cone state57,80. oid X receptor-γ (RXRγ), which can heterodimerize
Furthermore, nR2E3 and RORβ were found to act on with TRβ2 and suppress s opsin expression in regions
the same transcriptional pathway as nRL in directing of the superior retina in mice84. Other possible pattern-
precursors to a rod state60,63,71. Fourth, nRL-positive pre- ing factors are COUP transcription factor (COUP-TF)
cursors selected from the retina of wild-type mice at E16 orphan nuclear receptors85, which help to ensure a
express s opsin transcripts58,80, which is consistent with normal distribution of s cones over the retina, and
the existence of an early photoreceptor precursor that bone morphogenic protein receptor, which may con-
has a propensity to follow a default s cone pathway. For trol COUP-TF expression 85. Factors that augment
a precursor to enter into rod differentiation, the s cone M and s opsin expression include RORα 86 and,
state must be actively turned off by the co-expression indirectly, nEUROD1, which enhances the expression
of nRL and nR2E3. some of the precursors begin to of TRβ2 and M opsin in cones87,88.
express nRL together with s opsin because they are not The action of TRβ2 requires the thyroid hormone
yet fully committed to a rod fate. There is likely to be an triiodothyronine (T3) at appropriate developmen-
‘interim’ state as the cone versus rod fate becomes fixed. tal stages76,89,90. In mice, the concentration of thyroid
At postnatal stages when cone genesis is complete and hormone in the circulation rises progressively during
only rods are produced, the default s cone state of the postnatal development and provides a temporal signal
photoreceptor precursors is likely to be transient under that induces the expression of M opsin in the second
the influence of strong nRL activity. Lastly, the model postnatal week (FIG. 3). In mice with congenital hypothy-
of a default s cone state is consistent with studies on the roidism, induction of M opsin expression is impaired76,
ontogeny and evolution of photoreceptors, which sug- whereas treatment with excessive T3 suppresses the
gest that the s cone represents an evolutionarily older induction of s opsin expression at early stages 89. At
form of photoreceptor 14,17,81. inappropriate neonatal stages, excessive T3 promotes
The transient nature of plastic photoreceptor precur- cone-specific cell death.
sors at different developmental stages may make it diffi-
cult to demonstrate their existence in vivo. However, the Selective expression of L opsin versus M opsin genes
model contains a testable prediction in that some (or all) in humans. A specialized mechanism of opsin pattern-
early photoreceptor precursors co-express some or many ing underlies human trichromacy and involves selec-
of the key transcription factors before their final commit- tive expression of L opsin or M opsin genes (OPN1LW
ment to form a rod or a cone. Other bHLH transcription or OPN1MW, respectively), which reside in a tandem
factors, such as ATOH7, AsCL1 and nEUROD1, are at cluster on the X chromosome82. In a given cone, expres-
least transiently expressed in some photoreceptor pre- sion of an L opsin or M opsin gene is selected by an
cursors and may contribute to the counting mechanism upstream locus control region (LCR), thereby ensuring
or the balance between the cone- and the rod-inducing their mutually exclusive expression91,92. studies in mice
programmes. carrying a miniature human L opsin–M opsin gene
cluster indicate that the pairing of the LCR with either
Cone opsin distribution patterns the L opsin or M opsin gene promoter is determined by
For cone-committed precursors that do not receive sig- stochastic events and not by individual sets of opsin-
nals to form rods, the next decision concerns the expres- specific transcription factors 93. Transcription factor
sion of s opsin versus M opsin (in most mammals) or complexes involved in LCR-mediated activation of the
s opsin versus either M opsin or L opsin (in humans, L opsin or M opsin genes have not been elucidated. In
as discussed below)82,83. The key questions are: how is human retinoblastoma cell lines that have some cone
an opsin gene selected for expression, and how does properties, assembly of regulatory complexes at the LCR
this exclude the expression of other opsins in that cone determines L opsin versus M opsin expression during
photoreceptor? final mitosis94. LCR-mediated selection of opsin gene
expression has also been observed in zebrafish, which
Selective expression of S opsin versus M opsin. The s opsin differentially express a tandem array of four green-
(Opn1sw) and M-opsin (Opn1mw) genes are located on sensitive RH2 opsin genes95. TRβ2 and RXRγ in human
different chromosomes (chromosome 6 and the X chro- foetal cones96 might participate in the activation of L
mosome, respectively, in mice), and each gene is sub- opsin and M opsin genes, analogously to their role in
ject to independent transcriptional regulation in cone M opsin induction in mice.
subpopulations. The mechanism of mutual exclusion of Other processes contribute to opsin patterning in the
s opsin and M opsin expression has not been elucidated, context of the growing eye. The retinal surface varies not
but a combination of intrinsic transcriptional controls, only in terms of cone opsin patterning but also in the
Superior retina
The dorsal region of the
external signals including thyroid hormone and signals density and distribution of cone and rod cells10. Thus, the
light-sensing tissue at the back that relate to spatial location on the retinal surface seem differential rates of generation of rods and cones20 and
of the eye. to have key roles. their programmed cell death in development 97,98 may

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REVIEWS

also determine the pattern of each cell type across the with potential binding sites for bHLH proteins, nuclear
retina. In the more local environment, cell–cell signal- receptors and other factors, such as myocyte enhancer
ling may control spatial arrangements between groups factor 2C, that are expressed in the developing and
of cell types99,100. mature retina (M.-A. Kautzmann, M. Akimoto and
A. swaroop, unpublished observations). Additional
Nuclear receptors regulatory factors that could promote rod development
nuclear receptors are ligand-regulated transcrip- via the RORβ–nRL pathway include taurine, activin A,
tion factors. They include TRβ2 and RXRγ, which sonic hedgehog, fibroblast growth factor and retinoblas-
have known ligands (thyroid hormone and retinoids, toma protein104–110. Retinoic acid increases the number
respectively), and orphan receptors (such as nR2E3, of cells expressing rod marker genes111,112, potentially by
COUP-TF, RORα and RORβ), which lack known physi- elevating Nrl expression113. By contrast, the signal trans-
ological ligands. The potential of these transcription ducer and activation of transcription (sTAT) signalling
factors to respond to ligands allows an integration of pathway, which is downstream of receptors for ciliary
transcriptional control with extrinsic signals that may neurotrophic factor (CnTF) and leukaemia inhibitory
be crucial for rod versus cone differentiation, cone factor, regulates nRL and CRX expression and therefore
opsin patterning or other functions during photore- rod differentiation114,115. CnTF is a negative regulator
ceptor differentiation62–64,71,84,86 (FIGs 3,4). some orphan of rhodopsin expression, yet it increases photoreceptor
receptors, such as RORβ, have a large carboxy-ter- cell survival116,117. How the cytokine and janus kinase
minal pocket that may bind hydrophobic molecules (JAK)–sTAT signalling pathways are integrated in the
with lower specificity and affinity than conventional transcriptional hierarchy of nRL–CRX–nR2E3 requires
hormonal ligands101. Thus, apart from the endocrine further investigation.
signal of thyroid hormone, other small molecules may Three key regulators of rod development — nRL,
provide paracrine or autocrine signals that locally CRX and nR2E3 — continue to be expressed at high
modify nuclear receptor activity during development. levels in the mature retina, suggesting that they have
Although some nuclear receptors such as nuclear important roles in rod homeostasis. We hypothesize
receptor subfamily 2 group E member 1 (nR2E1; also that these proteins auto-regulate and cross-regulate
known as TLX) may act early in neurogenesis in reti- each other, thereby maintaining their expression levels
nal progenitor cells102,103, nuclear receptors have a role throughout the life of rods.
predominantly at later stages of photoreceptor dif-
ferentiation. This provides an interesting contrast to GRNs during development and homeostasis. GRns118,119
the roles of bHLH, homeodomain and other types of have three key components — transcription factors,
transcription factors42 during earlier stages in retinal cofactors and/or modulators, and transcriptional tar-
neurogenesis. gets. In such networks, signalling molecules modulate
the expression and/or activity of transcription fac-
Gene-regulatory networks (GRNs) tors that act as key nodes and control multiple cellu-
We primarily discuss GRns pertaining to rod differ- lar targets. One approach for elucidating GRns is to
entiation and homeostasis because most studies have examine global expression profiles after disrupting
focused on the regulation of rod gene expression. key regulatory nodes. Computer-based analysis of
Early induction of nRL expression in photoreceptor cis-acting elements and chromatin immunoprecipitation
precursors80 seems to be a decisive step in determining (ChIP) studies have identified shared as well as specific
the rod versus cone outcome. We predict that a sub- transcriptional targets of nRL, CRX, TRβ2 and nR2E3
stantial fraction of photoreceptor precursors express (ReFs 60,62,120,121) . To elucidate GRns that under-
nRL at a given time in development in response to pre- lie photoreceptor development and homeostasis, the
vailing signals. The long period from final mitosis to approaches must be multi-pronged and include global
the functional maturation of rods (2–4 weeks in mice expression profiling (or RnA sequencing) of purified
and 5–10 weeks in humans) can be attributed to step- photoreceptors from different genetic backgrounds
wise changes in gene expression patterns (influenced (such as Nrl–/–, Crx–/–, Rorb–/– and rd7 mice), global target
by extrinsic factors) that eventually lead to the expres- analyses using sequencing of ChIP DnAs (ChIP-seq)
sion of a specific set of phototransduction genes and a at different stages of development, in vitro and in vivo
Cis-acting elements
functional rod photoreceptor. strategies for determining direct biological responses
DNA sequences that affect the (for example, in vivo electroporation122), and network
transcription of a gene and are Regulation of NRL expression. A major regulator of and pathway analyses.
present nearby, on the same rod development upstream of nRL is RORβ, which is Another important question is, how are nRL, CRX
chromosome.
widely expressed in retinal progenitors and in imma- and nR2E3 involved in assembling distinct transcrip-
Chromatin ture photoreceptors. The role of RORβ in driving a pre- tional complexes to generate quantitatively distinct and
immunoprecipitation cursor into the first phase of the rod state is probably precise gene expression patterns? In addition to unique
Often abbreviated as ChIP, mediated via induction of nRL expression71 in coop- combinations of cis-acting regulatory sequences, the
this is an experimental eration with other transcription factors. An approxi- strength of protein interactions and combinatorial syn-
technique used to identify
DNA sequences that bind
mately 3-kb genomic region 80 that lies immediately ergy (or antagonism) can produce quantitative differ-
to a specific DNA-binding upstream of the transcription start site of the mouse ences in the expression of genes that are regulated by a
protein in vivo. Nrl gene includes many conserved sequence elements similar set of transcription factors. nRL and CRX can

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Enhanced S cone syndrome recruit TATA box-binding protein123, CREB-binding proteins with different roles in photoreceptor func-
An inherited autosomal protein and sTAGA53 to generate transcriptional com- tion have been associated with retinal degeneration128
recessive retinal disease plexes. The strength and stability of interaction among (see the Retnet website listed in Further information).
associated with greater regulatory factors can be modulated by post-transla- Interestingly, distinct mutations can lead to a similar
sensitivity to blue light, night
blindness and eventual
tional modifications that permit the assembly of dis- clinical phenotype, or disease caused by similar muta-
photoreceptor degeneration. tinct complexes, which bind to enhancer sequences. For tions may have variable severity. Unlike the retina of
example, the human nRL protein of 237 amino acids many fish and amphibians129,130, the adult mammalian
contains 18 serine and 9 threonine residues and is highly retina cannot generate new photoreceptors; hence,
phosphorylated124. sUMOylation of nRL is also crucial dysfunction or death of photoreceptors results in the
for its function during photoreceptor development 125. loss of vision.
The modulation of nRL activity by kinases and other Mutations in nRL, nR2E3 and CRX cause varying
signalling molecules might therefore be a key mechanism retinal disease phenotypes (TABLe 1). All NRL muta-
for controlling the timing and level of expression of rod tions reported so far are missense or frameshift, and
genes. notably, disease-causing mutations in nRL affect most seem to have a dominant impact on transcrip-
its phosphorylation state and interaction with CRX and tionregulating activity 126,131. NR2E3 mutations, by
other transcriptional regulators126. Another modulator of contrast, result in phenotypes such as enhanced s cone
photoreceptor development is E3 sUMO-protein ligase syndrome (EsCs)132–135, which can be explained by the
PIAs3, which directly interacts with CRX and nR2E3; primary role of nR2E3 as a repressor of cone genes in
sUMOylation of nR2E3 by PIAs3 enhances its activity photoreceptor precursors63,136. CRX mutations gener-
as a repressor of cone genes127. ate an array of clinical phenotypes, from more severe
Although still a work in progress, a simplified congenital blindness in LCA to late-onset retinitis
GRn for rod differentiation and homeostasis is pigmentosa50,137. The retinal disease genes associated
illustrated in FIG. 5. with photoreceptor dysfunction or death (based on the
Retnet database; see Further information) are listed in
Retinal and macular diseases supplementary information s1 (table). A substantial
Photoreceptor function and homeostasis are strin- fraction of these disease genes seem to be direct or indi-
gently controlled at many levels — regulation of gene rect targets of nRL, CRX or nR2E3, as revealed by gene
expression, transport of selected proteins to the base expression profiles of knockout mice and/or ChIP-seq
of cilia, outer segment biogenesis and signal transmis- analysis 60,63,64,138 (H. Hao, D. Kim and A. swaroop,
sion (FIG. 1b,c). Mutations in over 100 genes that encode unpublished observations).
THRB mutations are associated with the syndrome of
resistance to thyroid hormone (see the OMIM entry for
THRB), but photoreceptor dysfunction has been unex-
M1 M2 M3 plored in this disease. Further studies of this syndrome
and other congenital thyroid disorders are warranted to
ascertain the requirement of human photoreceptors for
thyroid hormone.
NRL CRX NR2E3 NR1D1
Designing knowledge-based treatments
Molecular details of transcription-regulatory networks
underlying photoreceptor development, combined with
high throughput genomic and genetic technologies,
have facilitated the design of potential new treatment
strategies for retinal degenerative diseases.

PDEβ RHO GNAT1 PDEα S opsin Gene discovery. Disease-causing mutations have so far
Rod Cone
been identified in only approximately 50% of patients
Figure 5 | A simplified representation of rod gene-regulatory networks. Neural with inherited retinal degenerative diseases, owing
Nature Reviews | Neuroscience
retina leucine zipper protein (NRL) acts synergistically with cone–rod homeobox protein to their extensive clinical and genetic heterogeneity.
(CRX) and/or photoreceptor-specific nuclear receptor (NR2E3) in many distinct Comprehensive transcriptome analyses of photore-
complexes that bind to enhancer sequences and dictate the expression of various rod ceptors and the retina have led to the identification of
genes. The rhodopsin (RHO) promoter is likely to be regulated by additional factors, such genetic defects in retinal diseases, particularly for cases
as nuclear receptor subfamily 1 group D member 1 (NR1D1)68, to maintain high but in which the chromosomal region of a disease locus
precisely controlled levels in rods. Other promoters (such as that for rod cGMP-specific
has been determined. Positional cloning strategies are
3′,5′-cyclic phosphodiesterase subunit-β (PDEβ)) may bind the widely expressed
transcription factors SP1 and SP4 (ReF. 153), together with NRL. NRL and NR2E3 repress
inadequate in families with a low number of members
the activity of cone genes (such as S opsin), whereas CRX activates both rod and cone affected by retinal degenerative disease. The knowledge
genes. M1, M2 and M3 indicate modulators (such as fibroblast growth factor 2, retinoic of expression during photoreceptor development is also
acid and post-translational modifiers) that control or fine-tune the activity of key valuable in interpreting data from exome (capturing
transcription factors. Thin lines denote experiment-based suggested links, whereas thick and sequencing all exons) and whole-genome sequenc-
lines denote confirmed regulatory connections. GNAT1, guanine nucleotide-binding ing methods, which are now being used for discovering
protein G(t) subunit α1. novel disease mutations139,140.

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Table 1 | Clinical phenotypes caused by mutations in key transcriptional regulators of photoreceptor development
Gene Transcription factor type Human diseases Mouse mutant phenotypes
CRX Paired-type homeodomain Autosomal dominant cone–rod dystrophy; autosomal Lack of outer segments in rods and cones;
dominant and recessive Leber’s congenital amaurosis; circadian entrainment defect; impaired bipolar
autosomal dominant retinitis pigmentosa cell differentiation and maintenance
NRL Maf family basic leucine Autosomal dominant and recessive retinitis Complete loss of rods; transformation of rods
zipper pigmentosa into S cones, with shorter outer segments
than normal; enhanced S cone function;
photoreceptor layer rosettes
NR2E3 Orphan nuclear receptor Enhanced S cone syndrome; Appearance of rod photoreceptors expressing
autosomal dominant retinitis pigmentosa cone genes; photoreceptor layer rosettes;
retinal degeneration
OTX2 Paired-type homeodomain Microphthalmia (syndromic) Loss of rods and cones and increase in
amacrine-like neurons; loss of pinealocytes;
impaired bipolar cell differentiation and
maintenance; patterning abnormalities in the
midbrain and hindbrain
RORB Orphan nuclear receptor None discovered Loss of rods and over-production of S
cones; lack of all outer segments; retinal
disorganization; retarded induction of S opsin
expression in cones; circadian behaviour
abnormalities; abnormal gait
THRB Thyroid hormone-activated Resistance to thyroid hormone, typically autosomal Loss of M opsin and extended distribution of S
nuclear receptor dominant; endocrine and neurological dysfunction; opsin in cones across the retina instead of normal
rare colour vision abnormalities bias in the ventral region; endocrine dysfunction;
deafness
CRX, cone–rod homeobox protein; NR2E3, photoreceptor-specific nuclear receptor; NRL, neural retina leucine zipper protein; THRB, thyroid hormone
receptor β. Data are from the RetNet database (http://www.sph.uth.tmc.edu/retnet/) , the OMIM database (www.ncbi.nlm.nih.gov/omim) and from references
in the main text.

Determining gene-regulatory networks. Temporal studies have attempted to design stem cell-based
ChIP-seq analyses may allow identification of direct therapies for producing new photoreceptors. The
targets (the ‘targetome’) of the six key transcription fac- feasibility of one such approach was demonstrated
tors and help establish regulatory networks that dictate by transplantation of nRL-expressing committed
photoreceptor morphology and function. Elucidation rod precursors tagged with green flourescent pro-
of such gene-regulatory networks should have a major tein into wild-type and degenerating mouse retina143.
impact on delineating the pathways of photoreceptor The photoreceptor precursors differentiated into
degeneration (BOX 1). Proteomics analysis of the devel- mature rods and formed synaptic connections with
oping retina141 and of purified photoreceptors may interneurons, resulting in light responsiveness. The
also yield novel insights into regulatory pathways and age of the donor cells was important, as only the post-
provide disease-specific biomarkers. mitotic committed precursors — and not the dividing
cells or the mature photoreceptors — survived and
Gene- and small-molecule-based interventions. were integrated effectively. Many groups have now
Modulation of transcription factor activity can be used demonstrated successful differentiation of photore-
to control the expression of downstream targets and ceptors from human embryonic stem cells or from
consequently cellular phenotypes. For example, a short induced pluripotent stem cells and their transplanta-
37-amino-acid residue region of nRL that is sufficient tion into the mouse retina144–146. However, appropriate
for transactivation123 can be used in combination with a cell surface markers will be needed for sorting and
specific DnA-binding domain to modulate the expres- purification of photoreceptor precursors that can be
sion of downstream genes. Agonists of nR2E3 may be used for transplantation research in humans. CD73,
useful in retinal diseases including an enhanced s cone a recently identified marker of photoreceptor pre-
phenotype 142. Additionally, global patterns of gene cursors147 and other cell surface markers may assist
expression in photoreceptors at distinct stages of differ- in making the cell-based therapy feasible for retinal
entiation and ageing can be used as a reference point to neurodegeneration.
test the effects of candidate compounds for the treatment
of specific degenerative diseases. Conclusions
We have summarized the current status of research
Photoreceptor precursors and cell-based therapies. on the genesis of photoreceptors, with a focus on
Cell-based strategies to replace defective or dying cells transcription-regulatory factors that control the dif-
are among the promising approaches for the treatment ferentiation of rods and cones. We note that addi-
of diverse retinal degenerative diseases. numerous tional mechanisms (such as cellular interactions,

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Box 1 | Pathways of photoreceptor degeneration and targets for drug discovery

Leber’s congenital amaurosis,
retinitis pigmentosa and Many disease Convergent apoptotic Common apoptotic
genes triggers pathways
other Mendelian retinal
diseases constitute a
genetically heterogeneous
Rho
spectrum of clinical
phenotypes. Given that
mutations in over 100 genes Pre-apoptotic
can lead to photoreceptor Pde6b
signal 1
cell death and most are
limited to few patients and/
or families, interventions that Prph2 Pre-apoptotic Apoptotic
target these genes and signal 2 pathway 1
mutations are less likely to be Photoreceptor
developed than those that death
Cep290
target more common Pre-apoptotic Apoptotic
diseases because of cost and signal 3 pathway 2
efficiency considerations. Rpgr
Alternative approaches are
Pre-apoptotic
therefore needed to develop
signal 4
therapies for orphan
diseases. We postulate a gene 6

convergence of the impact of

••
mutations on a select group •
of ‘pre-apoptotic signals’ that
induce apoptotic pathways genen
(see the figure). Mouse
mutants with retinal
degeneration are excellent Time
models of human retinal
degenerative diseases
including those caused by mutations in rhodopsin (Rho), rod cGMP-specific 3′,5′-cyclic phosphodiesterase
Nature Reviewssubunit-β
| Neuroscience
(Pde6b), peripherin 2 (Prph2; also known as Rds), centrosomal protein of 290 kDa (Cep290) and X-linked retinitis
pigmentosa GTPase regulator (Rpgr) (left column). We suggest that temporal profiling of changes in gene expression in
mutant photoreceptors will allow the identification of a limited number of pre-apoptotic signalling molecules (middle
column). Targeting terminal effectors of apoptosis (right column) for therapy may be ineffective and lead to undesirable
side effects. As the photoreceptor degeneration process is associated with the remodelling of the inner retina, global
gene profiling of purified rods and cones80 should reduce the background noise149. Gene profiling or RNA sequencing,
followed by network or pathway analysis, should lead to identification of common pre-apoptotic signals. These can then
be targeted to screen small molecules to intervene in retinal degenerative diseases.

microenvironment and epigenetic modifications) designing treatment strategies for retinal degenerative
also influence the final pattern of the photoreceptor diseases, including the development of new drugs and
mosaic27,99,100, although we are only beginning to under- photoreceptor replacement therapy.
stand how such mechanisms guide the development of studies of RORβ, nRL and nR2E3 in mammals have
the mammalian retina. Despite considerable advances provided particularly strong support for the common
during the past decade, several important questions photoreceptor precursor hypothesis that is based on an
remain. What controls the timing, number and ratios s cone default state. However, it is worth mentioning that
of different photoreceptor cell types? What deter- a mutation of the zebrafish Tbx2b transcription factor
mines the expression of only one opsin in a human gene has been associated with excess rods and a loss of
cone photoreceptor? What sets the morphological cones that are sensitive to ultraviolet light 148. Although
and functional polarity of a photoreceptor? How is the fish have a more complex set of cones than mammals, it
spatial and functional relationship of rods and cones is possible that additional developmental plasticity from
with the RPE and interneurons organized in different rods to cones may be independent of the cone to rod
regions of the retina? How does a cone or a rod pho- plasticity directed by RORβ, nRL and nR2E3 discussed
toreceptor form synaptic connections with its cognate in this article.
bipolar or horizontal cell? How is the photoreceptor numerous challenges exist, yet we are confident
mosaic arranged? How is cone survival linked to rod that technological advances in genetics and high-
death? The answers to at least some of these questions throughput genomics will further our understand-
may lie in delineating GRns that determine the tem- ing of neurodevelopment and neurodegeneration of
poral sequence of events of photoreceptor develop- retinal cells and intensify the development of novel
ment and homeostasis. such insights will also help in treatment paradigms.

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134. Jacobson, S. G. et al. Nuclear receptor NR2E3 gene This report used NRL-positive rod photoreceptor transcription of rod-specific cGMP-phosphodiesterase
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enhanced S-cone syndrome, Goldmann-Favre cells. Proc. Natl Acad. Sci. USA 106, 16698–16703 standing scientist and a generous mentor and colleague. We
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retinal degeneration. Arch. Ophthalmol. 121, 145. Lamba, D. A., Gust, J. & Reh, T. A. Transplantation of Cogliati for productive discussions, and L. Ng, D. Sharlin, Alok
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