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Regulation of Terminal
Differentiation Programs
in the Nervous System
Oliver Hobert
Howard Hughes Medical Institute and Department of Biochemistry and Molecular
Biophysics, Columbia University Medical Center, New York, NY 10032;
email: or38@columbia.edu

Annu. Rev. Cell Dev. Biol. 2011. 27:68196


The Annual Review of Cell and Developmental
Biology is online at cellbio.annualreviews.org
This articles doi:
10.1146/annurev-cellbio-092910-154226
c 2011 by Annual Reviews.
Copyright 
All rights reserved
1081-0706/11/1110-0681$20.00

Keywords
neuronal identity, transcriptional regulation, terminal selector,
regulon

Abstract
The generation of individual neuron types in the nervous system is a
multistep process whose endpoint is the expression of neuron type
specic batteries of terminal differentiation genes that determine the
functional properties of a neuron. This review focuses on the regulatory mechanisms that are involved in controlling the terminally differentiated state of a neuron. I review several case studies from invertebrate and vertebrate nervous systems that reveal that many terminal
differentiation features of a neuron are coregulated via terminal selector
transcription factors that initiate and maintain terminal differentiation
programs.

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Contents

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INTRODUCTION . . . . . . . . . . . . . . . . . .
COREGULATION OF TERMINAL
DIFFERENTIATION GENES
THROUGH TERMINAL
SELECTORS . . . . . . . . . . . . . . . . . . . . .
Coregulation of Neuronal Terminal
Differentiation Batteries in
Caenorhabditis elegans . . . . . . . . . . . .
Terminal Selector Transcription
Factors Coregulate Terminal
Differentiation Batteries . . . . . . . . .
Coregulation and Terminal
Selectors in Vertebrates . . . . . . . . .
MAINTENANCE OF THE
DIFFERENTIATED STATE . . . . .
PARALLEL REGULATORY
ROUTINES . . . . . . . . . . . . . . . . . . . . . .
COMBINATORIAL CODES OF
TERMINAL SELECTOR
FUNCTION . . . . . . . . . . . . . . . . . . . . . .
DIVERSIFICATION
DOWNSTREAM OF
TERMINAL SELECTORS . . . . . . .
COREGULATION OUTSIDE
THE NERVOUS SYSTEM . . . . . . .
THE HOURGLASS TOPOLOGY
OF REGULATORY
PROGRAMS . . . . . . . . . . . . . . . . . . . . . .
FUTURE CHALLENGES . . . . . . . . . . .

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INTRODUCTION

Terminal
differentiation genes:
effector genes that
dene the functional
properties of a mature,
nondividing neuron
throughout its life

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The enormous diversity of cell types in the


nervous system was rst appreciated through
studies conducted by the founder of modernday neuroscience, Santiago Ramon
y Cajal
(1911). Since the days of Ramon
y Cajal, understanding how this diversity is programmed
into a developing organism has been one of
the most fascinating goals of biology. Early
studies, such as those by Hans Spemann, employed the physical manipulation of embryonic
structures to obtain valuable insights into how
neural structures initially are induced on a
morphological level (Hamburger 1981). The
Hobert

availability of tools to visualize the vast diversity


of cell types in the nervous system (e.g., molecular markers and neuron typespecic staining
procedures) has, over the past few decades,
focused developmental neurobiology on efforts
to decipher the molecular mechanisms that
lead to the generation of distinct neuronal
cell types at later stages of development. In
vertebrate systems, these attempts have largely
concentrated on identifying and manipulating
the activity (i.e., removing or ectopically
expressing) of regulatory factors such as transcription factors or signaling proteins. These
studies produced a wealth of information about
the molecular cascades and networks that act
during development to commit neurons to
specic fates (Guillemot 2007, Jessell 2000).
However, much remains to be learned about
the precise nature of the regulatory programs
that control the terminal step of neuronal differentiation and that ensure the maintenance
of the differentiated phenotype.
In the nal step of neuronal differentiation,
individual neuron types are instructed to
express neuron typespecic gene batteries,
which are composed of terminal differentiation
genes (also sometimes called effector genes or
structural genes) that code for proteins that
dene the differentiated properties of a neuron
throughout its lifetime. These proteins include
enzymes that synthesize neurotransmitters,
transporters that package neurotransmitters,
synaptically localized neurotransmitter receptors, ion channels that determine the resting
potential of a neuron, molecules that dene
structural features such as dendritic arborization patterns, and synaptic adhesion molecules.
These proteins form a molecular signature that
uniquely denes the core identity of a neuron.
Even though components of the expression
programs of adult neurons are exible and
responsive to specic external conditions, the
core identity features of a neuron show little
variation during its postembryonic life. None
of these core identity features, i.e., the products
of terminal differentiation genes, are necessarily exclusive to a specic neuron type; rather,
a unique combination of genes that themselves

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may be more broadly expressed denes a


specic neuron type (Hobert et al. 2010).
With this argument in mind, the challenge
of understanding neuronal cell type specication and neuronal differentiation lies in
understanding how the expression of these
terminal differentiation genes is regulated.
In other words, what are the cis-regulatory
elements and trans-acting factors that control
the expression of such terminal differentiation
genes? Rather than comprehensively reviewing
all studies dealing with the regulation of terminal differentiation genes, I highlight in this
review several relatively recent, exemplary studies, conducted in invertebrates and vertebrates,
that have begun to reveal what are perhaps
phylogenetically conserved, general features
and principles of the regulatory mechanisms
that drive terminal neuron differentiation.

COREGULATION OF TERMINAL
DIFFERENTIATION GENES
THROUGH TERMINAL
SELECTORS
How to study the regulation of terminal differentiation genes? Early studies conducted in
the 1980s and 1990s attempted to dissect the
regulatory control regions of terminal differentiation genes with reporter genes expressed in
cell lines or primary cell cultures. Such studies
were hard to interpret because the complement
of regulatory factors often differs between a heterologous ex vivo context and a cell undergoing
differentiation in its normal organismal context.
The most incisive way to conduct studies on
the regulatory architecture of gene expression
programs is to carry out studies in vivo, ideally
within an animal, using transgenic approaches.

Coregulation of Neuronal Terminal


Differentiation Batteries in
Caenorhabditis elegans
The experimental amenability of the nematode
Caenorhabditis elegans, as well as the precisely
known cellular composition of its nervous
system, which contains only 302 neurons, has
made this system an excellent choice for prob-

ing the cis-regulatory mechanisms that control


the expression of terminal differentiation
genes in the nervous system at single-neuron
resolution (Hobert 2010). Such studies have
been enabled by green uorescent protein ( gfp)
reporter gene technology developed initially in
C. elegans (Chale et al. 1994). In this approach
terminal differentiation genes are fused to
gfp, and regulatory elements necessary and
sufcient to drive expression in specic neuron
types are identied through standard mutational analysis (e.g., Wenick & Hobert 2004).
This type of cis-regulatory analysis has revealed
what appears to be a core principle of terminal
neuron differentiation, that of coregulation.
Take, for example, the battery of genes
that denes the neurotransmitter phenotype
of individual neuron types. In GABAergic
motor neurons, the expression of genes
coding for the GABA-synthesizing enzyme
unc-25/GAD (glutamic acid decarboxylase) and
the vesicular GABA transporter (VGAT)/unc47 is controlled by a similar cis-regulatory
motif that consists of a K50 homeodomain
binding site (Eastman et al. 1999). The
C. elegans genes encoding enzymes involved
in acetylcholine biosynthesis, transport, and
degradation are coexpressed in cholinergic
motor neurons, and all contain one or more
copies of another, dened cis-regulatory motif
(P. Kratsios and O. Hobert, submitted). The
genes encoding the dopamine-synthesizing enzymes cat-2/TH (tyrosine hydroxylase), bas-1,
and bas-4, as well as the dopamine transporters
cat-1/Vmat (vesicular monoamine transporter)
and dat-1/DAT (dopamine transporter), also
all share a common cis-regulatory signature
composed of ETS and Hox binding sites, which
are required to drive expression in all C. elegans
dopaminergic neurons (Flames & Hobert
2009; O. Hobert et al., unpublished data).
These cis-regulatory motifs may occur in
multiple copies and are often, but not always,
in close proximity (<1 kb) to the genes they
regulate. In some cases they have also been
found many kilobases upstream of a gene that
they regulate or within introns. Regardless, the
existence of shared cis-regulatory motifs or
www.annualreviews.org Terminal Neuron Differentiation

Coregulation:
regulation of a set of
distinct genes (a gene
battery) through a
similar cis-regulatory
signature

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Autoregulatory
module

Terminal
selector(s)

Single input
module (SIM)

Others

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Signaling
proteins

Ion channels

Neurotransmitter
pathway

Autoreceptors

Neurotransmitter
receptors
Neurotransmitters

Figure 1
Terminal selector regulons. A terminal selector directly controls the expression of terminal differentiation genes, representative
examples of which are depicted schematically here. Direct regulation is mediated via cis-regulatory motifs (black boxes) located in
proximity to members of the terminal gene batteries. The terminal selector can be a single protein, a heterodimer of two proteins that
cooperatively bind a specic motif, or a more complex assembly of separate trans-acting factors that bind to distinct cis-regulatory
elements that together form a specic cis-regulatory signature. An example of the last is an assembly of ETS and two distinct
homeodomain-binding sites that controls expression of a terminal gene battery in Caenorhabditis elegans dopaminergic neurons
(O. Hobert et al., unpublished data).

signatures (i.e., specic combinations of distinct cis-regulatory motifs) suggests that within
a specic neuron type, each gene battery is
controlled by a common trans-acting factor or
a combination thereof (Figure 1).
Selector: any
regulatory factor that
controls the identity of
a developing region or
organ at any stage of
development
Terminal selector: a
transcription factor (or
a combination of
several) that directly
initiates and maintains
expression of terminal
differentiation genes
Regulon: a set of
coregulated genes and
the transcription
factor(s) that directly
controls these genes

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Terminal Selector Transcription


Factors Coregulate Terminal
Differentiation Batteries
Classic forward genetic screens have identied
trans-acting factors that operate through
the cis-regulatory motifs shared by terminal
differentiation genes. For example, the Pitx
homeobox gene unc-30 coregulates the GABA
synthesis/transport pathway via the K50
homeodomain site (Eastman et al. 1999, Jin
et al. 1994). The ETS factor ast-1 and the
homeobox gene ceh-43 coregulate dopamine
synthesis/transport (Flames & Hobert 2009;
O. Hobert et al., unpublished data). The
EBF-type transcription factor unc-3 coreguHobert

lates the cholinergic gene battery in ventral


nerve cord motor neurons (P. Kratsios & O.
Hobert, submitted). A heterodimer of two
homeodomain transcription factors, ttx-3 (ortholog of mouse Lhx2/9) and ceh-10 (ortholog
of mouse Chx10), regulates the cholinergic
gene battery in a central interneuron (Wenick
& Hobert 2004). Because of their direct
regulation of terminal identity features of a
neuron, these transcription factors have been
termed terminal selectors (Hobert 2008).
Together with its downstream target genes,
a terminal selector constitutes a regulon
(Figure 1). In systems biology terminology,
terminal selectors are wired in what is called
a single input module (SIM) (Alon 2007;
Figure 1).
The coregulatory principle extends beyond the neurotransmitter phenotype. Terminal selectors also coordinately regulate
other identity features of neurons, including
neurotransmitter receptors, sensory receptors,

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Table 1 Examples of invertebrate and vertebrate terminal selectors and their targets. See text for references

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Terminal
selector

Type of transcription
factor

Terminal neuron
identity controlled by
terminal selector

Selected examples of direct target genes

CHE-1

C2H2 zinc nger

C. elegans gustatory
neuron class ASE

Putative chemoreceptors of the GCY family,


neuropeptides of the FLP family,
cyclic-nucleotide gated ion channels, itself
(autoregulation), many others

TTX-3/CEH-10

LIM/Prd homeodomains

C. elegans cholinergic
interneuron class AIY

Choline acetyltransferase, choline transporter,


neuropeptide receptors, neuropeptides, Ig
domain proteins, ion channels, itself
(autoregulation), many others

UNC-86/MEC-3

POU/LIM
homeodomains

C. elegans glutamatergic
touch sensory neurons

Touch receptor channel subunits, specialized


tubulin mec-7, extracellular matrix proteins,
vesicular glutamate transporter, itself
(autoregulation), many others

UNC-30

Pitx-type homeodomain

C. elegans GABAergic
motor neurons

Glutamic acid decarboxylase, vesicular GABA


transporter, GABA reuptake transporter,
acetylcholine receptor, Ig domain protein,
potassium channel, neuropeptides of the FLP
family, itself (autoregulation), many others

AST-1/CEH-43

ETS/homeodomain

C. elegans dopaminergic
neuron

Tyrosine hydroxylase, aromatic amino acid


decarboxylase, vesicular dopamine transporter,
dopamine reuptake transporter, dopamine
autoreceptor, ion channels

UNC-3

EBF-type

C. elegans A/B-type
cholinergic motor
neurons

Choline acetyltransferase, choline transporter,


neuropeptide receptors, neuropeptides, Ig
domain proteins, signaling proteins, many others

Pet-1

ETS domain

Mouse serotonergic
neurons

Tryptophan hydroxylase, 5-HT1a receptor,


aromatic amino acid decarboxylase, serotonin
transporter, itself (autoregulation), many others

Crx

Homeodomain

Mouse retinal
photoreceptors

Retinol-binding protein, rhodopsin, G protein


Gnat1, phosphodiesterase, vesicular glutamate
transporter, kinesin, itself (autoregulation), many
others

Nurr1/Pitx-3

C4 zinc
nger/homeodomain

Mouse midbrain
dopaminergic neurons

Tyrosine hydroxylase, aromatic amino acid


decarboxylase, vesicular dopamine transporter,
dopamine reuptake transporter, dopamine
autoreceptor D2R, receptor tyrosine kinase Ret,
many others

ion channels, cytoskeletal proteins, and cell


surface adhesion molecules (Figure 1; see
Table 1 for examples). That is, the expression of these genes is lost upon knockout of
the terminal selector, and these genes contain
binding sites for the terminal selector. Take,
for example, the MEC-3 and UNC-86 terminal
selectors, two homeodomain proteins that are

coexpressed in touch sensory neurons, where


they heterodimerize to cooperatively activate
downstream target genes (Xue et al. 1993) including mechanosensory receptors, associated
subunits, cytoskeletal proteins, and extracellular matrix components (Duggan et al. 1998).
Transcriptome proling of touch receptor neurons has revealed many more genes expressed in
www.annualreviews.org Terminal Neuron Differentiation

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touch sensory neurons that contain functional


binding sites for MEC-3/UNC-86 (Zhang
et al. 2002). Similarly, the transcriptome of the
gustatory neuron ASE contains hundreds of
terminal differentiation genes with functional
binding sites for the CHE-1 zinc-nger transcription factor (Etchberger et al. 2007).
As mentioned above, distinct neuron types
share specic terminal differentiation genes.
Accordingly, the cis-regulatory regions of
terminal differentiation genes expressed in
distinct neuron types typically contain an
assembly of multiple binding sites for distinct
terminal selectors. For example, the EBF-type
factor unc-3 and the ttx-3/ceh-10 homeodomain
heterodimer are terminal selectors for two
distinct types of cholinergic neurons, A/Band AS-type ventral cord motor neurons
(unc-3) and the AIY interneurons (ttx-3/ceh10), respectively (Wenick & Hobert 2004;
P. Kratsios & O. Hobert, submitted). Each
transcription factor regulates expression
of the cholinergic gene batteryi.e., the
enzymes and transporter for acetylcholine
independently through distinct cis-regulatory
elements located in the regulatory control regions of cholinergic pathway genes
(Figure 2). In addition, unc-3 and ttx-3/ceh-10
regulate the expression of terminal differentiation genes that distinguish A/B-type motor
neurons and AIY interneurons from one
another (Wenick & Hobert 2004; P. Kratsios
& O. Hobert, submitted).
Taken together, many terminal identity
features of individual neuron types in C. elegans
are controlled via a coregulatory strategy that
relies on shared cis-regulatory motifs and a
small number of cognate terminal selector
transcription factors. Importantly, the observation of coregulation is not a mere matter of
course. Transcriptome proling of individual
neurons has revealed that mature neuron types
coexpress a multitude of transcription factors.
For example, the C. elegans gustatory neuron
ASE differentially expresses 68 transcription
factors when compared with another sensory
neuron class (Etchberger et al. 2007). Similarly, other motor neuron and sensory neuron

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Hobert

types also differentially express many dozen


transcription factors (Fox et al. 2005, Smith
et al. 2010). The abundance of transcription
factors therefore provides, in theory, ample opportunity for independent, piecemeal
control of the many terminal differentiation
genes that a neuron may express. However, as
genetic analysis of the individual transcription
factors expressed in a given neuron type has
revealed (Table 1), control of terminal differentiation genes is heavily biased toward a small
number of transcription factors, the terminal
selectors.

Coregulation and Terminal Selectors


in Vertebrates
Does terminal differentiation in the vertebrate
nervous system follow a similar regulatory logic
in that many terminal differentiation genes
are coregulated by common cis-regulatory
motifs and trans-acting factors? Regulatory
control regions are more difcult to study in
vertebrates because cis-regulatory elements
may be distantly located and spaced over larger
intervals. In addition, time and cost represent a
signicant barrier to high-throughput analysis
of regulatory elements in transgenic animals.
Nevertheless, over the past few years, several
studies have provided evidence that the terminal differentiation of vertebrate neurons is
mediated via the coordinate regulation of gene
batteries by a small number of transcription
factors, thereby supporting the terminal
selector concept. The most notable examples
are the homeodomain transcription factor Crx,
which controls rod and cone photoreceptor
differentiation (Blackshaw et al. 2001, Corbo
et al. 2010, Hsiau et al. 2007, Livesey &
Cepko 2001, Swaroop et al. 2010); the Pet-1
ETS domain transcription factor, which
drives serotonergic neuron differentiation
(Hendricks et al. 1999, 2003; Liu et al. 2010);
and the Nurr1 and Pitx3 transcription factors,
which control midbrain dopaminergic neuron
differentiation (Smidt & Burbach 2009).
Crx. A combination of transcriptome analysis
in wild-type and mutant cells, bioinformatic

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Cholinergic A/B-type
motor neurons

Cholinergic AIY
interneuron

Cho

Cho

ChAT

ChAT
AcCh

CEH10

AcCh

VAChT

TTX-3

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ChT

VAChT

UNC-3

ChT

A/B-type
motorneurons

UNC-3

AIY
interneurons

Other cholinergic
neurons

CEH- TTX-3
10
?

Cholinergic gene battery

Figure 2
Terminal differentiation genes shared by distinct neuron types are controlled by distinct terminal selectors.
(a) Genes coding for proteins involved in acetylcholine (AcCh) synthesis, transport, and reuptake
(cholinergic pathway genes) are expressed in distinct neuronal cell types (example shown here: Caenorhabditis
elegans AIY interneurons and A/B-type ventral cord motor neurons) in which they are under control of
distinct terminal selectors, namely the TTX-3/CEH-10 homeodomain heterodimer in AIY (Wenick &
Hobert 2004) and the EBF-type transcription factor UNC-3 in ventral cord motor neurons (P. Kratsios &
O. Hobert, submitted). ChAT, choline acetyltransferase; Cho, choline; ChT, choline transporter; VAChT,
vesicular acetylcholine transporter. (b) The cis-regulatory regions of cholinergic pathway genes, which are
expressed in several distinct neuron types, contain a collection of distinct terminal selector binding sites.
Terminal selector motifs may occur in multiple copies, and they also may be interspersed (not shown), which
may complicate the isolation of discrete and separable elements for expression of terminal differentiation
genes in individual neuron types.

analysis, and chromatin immunoprecipitation


(ChIP) studies has revealed over the past few
years a picture of Crx gene function that is
remarkably similar to the C. elegans terminal
selectormediated,
coregulatory
strategy
discussed above: Crx protein binds directly to
the cis-regulatory control regions of hundreds
of terminal differentiation genes, including,
for example, genes coding for intracellular
signaling proteins involved in phototransduction as well as genes coding for proteins
that determine specic structural aspects of
photoreceptors (Corbo et al. 2010, Hsiau et al.

2007). Loss of Crx results in a failure of these


genes to be expressed. Intriguingly, a C. elegans
ortholog of Crx, the ceh-36 gene, also acts as
a terminal selector for a sensory neuron (Kim
et al. 2010, Lanjuin & Sengupta 2004) and
appears to regulate a similar set of sensory
transduction genes, which suggests deep
conservation of these regulons (E. Serrano &
O. Hobert, unpublished data).
One intriguing observation made when
examining terminal differentiation of retinal
photoreceptors has been that target genes are
not always turned on synchronously but may be
www.annualreviews.org Terminal Neuron Differentiation

Chromatin
immunoprecipitation
(ChIP): a technique
that assesses binding of
a transcription factor
to cis-regulatory
control regions in
genomic DNA in vivo

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deployed at different stages during the differentiation process (Blackshaw et al. 2004). Both
early and late photoreceptor genes can, nevertheless, be direct targets of Crx (Corbo et al.
2010). The onset of expression of early versus
late genes might be determined by the specic
architectural features of the cis-regulatory
elements that control their expression, such as
the number and afnity of Crx sites and their
spacing and orientation relative to other adjacently bound transcription factors. Although
the rules of such cis-regulatory grammar are
not known in detail for any system, the organ
selector gene, pha-4, in C. elegans offers an
informative example. During foregut development, the afnity of the binding sites for pha-4
determines whether target genes are expressed
early or late (Gaudet & Mango 2002).

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CB27CH27-Hobert

Pet-1. Mouse Pet-1, an ETS domain factor,


is another example of a vertebrate terminal
selector (Hendricks et al. 1999, 2003; Liu et al.
2010). Pet-1 expression uniquely denes most if
not all serotonergic neurons in the central nervous system. Either constitutive or temporally
controlled removal of Pet-1 results in severe
differentiation defects of serotonergic neurons;
they fail to express the synthesis and transport
machinery for serotonin, fail to grow axons to
their correct targets, and display defects in their
ring properties. As a consequence, Pet-1decient mice display several behavioral decits
(Liu et al. 2010). Pet-1 is a terminal selector
because it directly controls the expression of a
multitude of terminal features of serotonergic
neurons, as shown by ChIP conducted in vivo
and reporter gene assays conducted in vitro
(Hendricks et al. 1999, 2003; Liu et al. 2010;
E. Deneris, personal communication). Direct
targets include the rate-limiting enzyme of
serotonin synthesis, tryptophan hydroxylase,
and serotonin transporter. As is the case for
Crx, Pet-1 controls the expression of genes
expressed at different postmitotic stages of
serotonergic neuron terminal differentiation.
Nurr1 and Pitx3. The orphan nuclear receptor Nurr1 and the homeobox factor Pitx3
688

Hobert

are terminal selectors for midbrain dopaminergic neurons (Smidt & Burbach 2009). Both
proteins are required for the expression of
many terminal differentiation genes, they are
sufcient to induce dopaminergic fate in an in
vitro differentiation model, and recent ChIP
analysis demonstrates that they bind directly to
cis-regulatory control regions of terminal differentiation targets ( Jacobs et al. 2007, 2009a,b;
Martinat et al. 2006). On the molecular level,
the mechanistic basis of the cooperation
between Nurr1 and Pitx3 lies in Nurr1 being
a transcriptional repressor through association with a corepressor protein. In midbrain
dopaminergic neurons, the only site of Pitx3 expression in the central nervous system, Pitx3 is
capable of displacing this corepressor, thereby
allowing Pitx3 and Nurr1 to jointly activate
target gene expression ( Jacobs et al. 2009b).
Etv1. The role of the C. elegans ETS domain
transcription factor ast-1 as a terminal selector of dopaminergic neurons appears to be conserved in vertebrates, as the knockout of Etv1, a
homolog of ast-1, results in the failure of olfactory bulb dopaminergic neurons to terminally
differentiate (Flames & Hobert 2009). As in
C. elegans, mouse Etv1 directly controls a key
terminal differentiation marker of dopaminergic neurons, tyrosine hydroxylase (Cave et al.
2010, Flames & Hobert 2009). However, the
differentiation defect observed in Etv1 mutant
mice does not appear to extend equally to all terminal features of olfactory bulb dopaminergic
neurons (Cave et al. 2010). This may be because
ast-1, and likely Etv1 as well, requires a specic
set of cofactors that can partially compensate for
loss of ast-1 (O. Hobert et al., unpublished data).
Other examples. Several other postmitotically expressed vertebrate transcription factors
are required for the expression of terminal features of a neuron and, consequently, have been
referred to as selector genes in the literature.
However, in all of these cases it remains to
be shown whether these transcription factors
directly control terminal differentiation genes
or whether they instead act further upstream in

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a regulatory hierarchy. These factors include


the Tlx1 and Tlx3 homeobox genes that drive
glutamatergic fate in the embryonic dorsal
spinal cord (Cheng et al. 2004) and Gata2, a
driver of midbrain GABAergic neurons (Kala
et al. 2009). Mouse Runx1 and Brn3A are
also reasonable candidates to directly control
terminal features of somatic sensory neuron
subtypes (Liu & Ma 2011).

MAINTENANCE OF THE
DIFFERENTIATED STATE
One critical, key feature of invertebrate and vertebrate terminal selectors is their sustained expression throughout the life of a neuron. This
observation suggests that terminal selectors
may not only initiate the expression of terminal differentiation genes but also maintain their
expression. Conrming the sustained function
of terminal selectors, genetic approaches in
C. elegans have revealed that postembryonic
removal of a terminal selector results in the
loss of the differentiated properties of a neuron
(Etchberger et al. 2009, Flames & Hobert
2009). Similarly, removal of mouse Pet-1
in the adult nervous system through a Cre
recombinasemediated strategy results in a progressive loss of serotonergic neuron function
and loss of expression of terminal differentiation genes such as tryptophan hydroxylase (Liu
et al. 2010). A loss of dopaminergic terminal
fate markers is also observed upon removal of
Nurr1 in adult mice (Kadkhodaei et al. 2009),
further corroborating the importance of sustained terminal selector expression.
Direct transcriptional autoregulation ensures the sustained expression of terminal
selectors (Figure 1). That is, both C. elegans
terminal selectors and mouse terminal selectors
such as Pet-1 or Crx contain binding sites
for themselves in their own cis-regulatory
control regions. Mutating these binding sites
in the context of transcriptional reporters that
monitor terminal selector expression does not
affect initiation of their expression but does
affect maintenance of expression (Bertrand &
Hobert 2009). Moreover, genetic removal of

terminal selectors affects the maintenance of


their own expression. This has been observed
repeatedly in C. elegans (Etchberger et al. 2007,
Way & Chale 1989, Wenick & Hobert 2004)
and was also demonstrated recently in the
mouse through removal of Pet-1 in the adult
brain (Liu et al. 2010).

PARALLEL REGULATORY
ROUTINES
In C. elegans, the knockout of a given terminal
selector results in loss of the expression of many
terminal identity components of a neuron, but
several molecular identiers remain unaffected.
For example, sensory neurons continue to express panneuronal and pansensory features
upon removal of the che-1 gene, a terminal
selector for gustatory neuron specication
(Uchida et al. 2003), or upon removal of ast-1,
a terminal selector for dopaminergic neurons
(Flames & Hobert 2009). Similarly, loss of the
mouse terminal selector Pet-1 does not affect
the overall neuronal identity of serotonergic
neurons; rather, it affects their neuron type
specic properties (Hendricks et al. 2003).
Components of terminal neuron identity that
are not affected upon loss of these terminal
selectors may be regulated by another terminal
selector(s) acting in parallel. For example, in
C. elegans, and possibly other organisms as
well, pansensory neuron features, i.e., many
or most components of all ciliated apparatus
of all sensory neurons, are coregulated via a
common cis-regulatory motif (the X box) and
its cognate transcription factor DAF-19, a
phylogenetically conserved RFX-type factor
(Swoboda et al. 2000; Figure 3). Similarly, on
the basis of the bioinformatically determined
presence of shared cis-regulatory motifs in
many panneuronally expressed genes in invertebrates and vertebrates, several researchers
have speculated (though not experimentally
conrmed) that a coregulatory strategy may
also control panneuronal features (Hadley
et al. 2006, Liu et al. 2009, Ruvinsky et al.
2007). Taken together, several parallel-acting
regulons appear to control gene expression
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ASE sensory neuron

CHE-1

X1

X2

X3

X4

DAF-19

X5

X6

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Neuron-type specific
identity (terminal selector)

X1

X2

X3

X4

X5

X6

Pan-sensory identity
(terminal selector)

X1

X2

X3

X4

X5

X6

Pan-neuronal identity
(?)

ASE motif
X box

N1 box

Figure 3
Multiple terminal selector regulons dene the terminal molecular signatures of a neuron. As exemplied
with the C. elegans ciliated gustatory neuron class ASE, the molecular signature of a terminally differentiated
neuron can be broken down into several distinct regulons, each controlled via a distinct, cognate terminal
selector. One is controlled by the terminal selector CHE-1, which controls ASE-specic identity features
(see Table 1 for examples) (Etchberger et al. 2007). The second regulon is controlled by the RFX-type
transcription factor DAF-19, which controls expression of the core components of the ciliated structures of
the ASE (as well as all other sensory neurons) via an X box (Swoboda et al. 2000). The third regulon is as yet
hypothetical and may be composed of a battery of coregulated panneuronal features that are unaffected by
che-1 and daf-19. Coregulation of panneuronal features is presumed to exist on the basis of bioinformatic
identication of a shared motif, the N1 box, in panneuronally expressed genes (Ruvinsky et al. 2007).

programs in terminally differentiated neurons


(Figure 3). However, regulation of these
parallel routines may not be strictly separated;
points of intersection may exist. For example,
the C. elegans MEC-3/UNC-86 terminal
selectors also regulate the expression of the
panneuronal SNAP-25/ric-4 gene (Hwang &
Lee 2003). Furthermore, mouse Crx regulates
the expression of ciliary genes (Corbo et al.
2010, Hsiau et al. 2007), perhaps in conjunction
with an RFX factor (Piasecki et al. 2010).

COMBINATORIAL CODES
OF TERMINAL SELECTOR
FUNCTION
Most known terminal selectors are expressed
in several distinct neuron types. In analogy to
transcription factors that act earlier in neuronal
development ( Jessell 2000), the specicity of
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their activity as terminal selectors in individual neuron types is determined by combinatorial codes of terminal selectors. For example,
the ttx-3 LIM homeobox gene is a terminal
selector for a cholinergic interneuron class in
C. elegans called AIY, but is also expressed in
several sensory neurons, where it has no apparent terminal selector function (Altun-Gultekin
et al. 2001). However, AIY neurons express another gene, the ceh-10 Prd homeobox gene,
which is itself expressed in several distinct neuron types (Altun-Gultekin et al. 2001). But
the expression of ttx-3 and ceh-10 overlaps exclusively in AIY, where the two proteins heterodimerize to cooperatively activate downstream terminal differentiation genes (Wenick
& Hobert 2004).
In vertebrates, the specicity of the more
broadly expressed terminal selector for midbrain dopaminergic neurons, Nurr1, is ensured

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HLH-2
HLH-3
REF-2 HLH-16

REF-2
bHLHs
TTX-3

TTX-3
CEH-10

Wnt
POP-1

TTX-3
ceh-10

TTX-3

AIY
interneuron

Time

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Multiple, transient
regulatory inputs

AIY
motif

TTX-3
CEH-10
Battery

SMDD motor
neuron

b
ttx-3

Wnt
TTX-3

7:57

Terminal
selector(s)

TF

CEH-10
Terminal diff.
battery
ttx-3
ceh-10

X1

X2

X3

X4

X5

TF
X6

Terminal gene battery

Autoregulation
(lock in)

Figure 4
The hourglass topology of terminal selector gene function. (a) An example from C. elegans that illustrates the complex nature of the
regulatory strategy that turns on the two terminal selectors ttx-3 and ceh-10 in the AIY interneuron class (Bertrand & Hobert 2009;
V. Bertrand & O. Hobert, unpublished data). Once turned on via a multitude of transient regulatory inputs [i.e., the REF-2 and basic
helix-loop-helix (bHLH) factors are only transiently expressed; the Wnt signal is also transient], ttx-3 and ceh-10 initiate and maintain
the expression of terminal differentiation genes and ensure their own maintenance through autoregulation. All of this is achieved by a
cis-regulatory element called the AIY motif (red box) that is bound cooperatively by TTX-3 and CEH-10. Because of the lack of the
Wnt signal, ceh-10 is not turned on in the sister of the AIY interneuron, the SMDD motor neuron; thus, ttx-3 cannot maintain its own
expression, and the AIY gene battery is not expressed in SMDD. (b) Terminal selectors may be bottlenecks in an hourglass that link
complex upstream regulatory events to the coordinated expression of terminal gene batteries. In other developmental contexts, terminal
selectors have also been termed input/output genes (Stern & Orgogozo 2008). Terminal selectors are not regulatory endpoints;
however, they may regulate the expression of downstream regulatory factors to control the expression of specic subroutines that could,
e.g., diversify gene expression proles within a specic neuron class. Terminal selectors may cooperate with downstream regulators to
turn on terminal genes, thereby constituting a feed-forward motif (Alon 2007).

by the midbrain-specic Pitx3 transcription


factor ( Jacobs et al. 2009b). As mentioned
above, Pitx3 serves to remove a corepressor
from Nurr1 and subsequently binds with
Nurr1 to cis-regulatory control regions of
terminal differentiation genes.

DIVERSIFICATION
DOWNSTREAM OF
TERMINAL SELECTORS
Even though terminal selectors directly regulate many terminal features of a neuron, they
are not regulatory endpoints (Figure 4). Some
targets of terminal selectors are transcription
factors themselves, and these may further diversify individual neuron classes. Two examples,
one in invertebrates and one in vertebrates,

illustrate this point. First, the C. elegans ASE


gustatory neuron class is composed of two
neurons, the left ASE neuron (ASEL) and the
right ASE neuron (ASER). Many of this classs
terminal features, such as neurotransmitter
phenotype and chemoreceptor expression, are
controlled directly by the terminal selector
che-1 (Etchberger et al. 2007). But the two
ASE neurons subtly differ in their ability to
sense and respond to chemical cues (Ortiz et al.
2009, Suzuki et al. 2008). A complex regulatory
network that contains the cog-1 homeobox gene
as its key component controls these differences.
The cog-1 homeobox gene promotes ASER
neuron fate and is expressed only in ASER
(Chang et al. 2003). Its expression is induced directly via che-1 (OMeara et al. 2009), but cog-1
expression subsequently is switched off in ASEL
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via the miRNA lsy-6 (which is also a target of


che-1) ( Johnston & Hobert 2003, Sarin et al.
2007). Even though it is not yet understood
how lsy-6 expression is restricted to ASEL, the
key points are that the regulatory factors that
distinguish ASEL and ASER are under control
of a terminal selector and that they act to diversify what would be an otherwise bilaterally
symmetric terminal differentiation program.
Second, a conceptually similar scenario is
observed in the vertebrate retina. As mentioned
above, the Crx homeobox gene is a terminal
selector of rod and cone photoreceptors, as it
initiates and maintains the expression of hundreds of terminal differentiation genes. In rod,
but not in cone, photoreceptors, Crx directly
turns on expression of the Nrl leucine zipper
transcription factor, which is essential for determining rod differentiation (Corbo et al. 2010,
Swaroop et al. 2010). Crx and Nrl then cooperate to control the expression of terminal features of rod photoreceptors (Corbo et al. 2010).
This is a particularly nice example of a terminal selector that is wired into feed-forward
loops (Alon 2007) in which it activates a downstream regulator and then cooperates with this
regulator to control target genes. Similar feedforward loops are also apparent in the ASE
gustatory example mentioned above (Hobert
2006).

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CB27CH27-Hobert

COREGULATION OUTSIDE
THE NERVOUS SYSTEM
Terminal selectors are not restricted to the
nervous system. The coregulation of terminal differentiation genes by a small number
of transcriptional drivers has many precedents
(Davidson 2006). For example, the mouse Pax5
homeobox gene is a terminal selector for a
specic B cell type in the immune system
(Cobaleda et al. 2007), and shavenbaby is a
terminal selector for a specic type of epidermal cells in Drosophila (Chanut-Delalande
et al. 2006). In fact, grouping functionally
related genes into regulons is an ancient
and broadly applied gene regulatory strategy. Bacteria, for example, respond to specic
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environmental conditions by activating batteries of coregulated genes via so-called global


regulators, the bacterial equivalent of terminal selectors (e.g., Yoshida et al. 2003). In
metazoans, functionally closely related genes
are also often coregulated. For example, genes
that code for proteins involved in lysosomal biogenesis in vertebrates are coregulated
by the transcription factor TFEB (Sardiello
et al. 2009); the y transcription factor DIMM
coregulates genes involved in the neuropeptidergic secretory pathway (Hamanaka et al.
2010); and the C. elegans homeobox gene ceh6 coregulates genes involved in controlling
excretory cell function via a shared cisregulatory motif, the octamer (Armstrong &
Chamberlin 2010, Mah et al. 2010). What is
conceptually novel about the coregulatory principle of several of the neuronal terminal selectors described above is the apparent breadth
of the spectrum of target genes. Rather than
falling into dened, functionally tightly linked
categories (such as neurotransmitter pathway
genes), target genes of at least some terminal selectors rather broadly dene multiple aspects of
a neurons identity, as schematized in Figure 1.

THE HOURGLASS TOPOLOGY


OF REGULATORY PROGRAMS
How is terminal selector expression initiated?
Bertrand & Hobert (2009) describe the molecular mechanism that turns on the ttx-3/ceh10 terminal selector complex that drives AIY
interneuron specication, which reveals three
possibly generalizable concepts (Figure 4).
First, initiation of terminal selector gene expression is separable from maintenance of
expression. That is, a terminal selector locus contains cis-regulatory initiator elements
that are separable from the maintenance elements, which are controlled via autoregulation.
Second, regulatory factors that initiate terminal selector gene expression may often act
only transiently. Autoregulation of terminal
selectors therefore serves as a mechanism to
lock in an initial, transient regulatory state. In
other words, cis-regulatory elements of terminal

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selectors read out transient regulatory states.


Third, transient initiation inputs are complex,
as they integrate a complex series of several regulatory events including extrinsic signals (Bertrand & Hobert 2009). Other C. elegans terminal selectors also appear to require
complex regulatory inputs for their initiation
( J. Etchberger & O. Hobert, unpublished data).
Taken together, in the two cases examined in
detail here, the multilayered regulatory hierarchy that controls terminal differentiation gene
expression can be considered to have an hourglass topology in which several distinct inputs
are integrated at the level of terminal selector
expression, the node of the hourglass. Downstream of the terminal selector, the network
again fans out to turn on many terminal differentiation genes (Figure 4).
Shavenbaby, a terminal selector for a specic
type of Drosophila skin cells called trichomes,
also follows the hourglass/bow-tie logic. A
combination of transient signaling inputs controls the expression of shavenbaby; once turned
on, it directly controls the expression of several terminal differentiation genes in trichomes
(Chanut-Delalande et al. 2006). The case of
shavenbaby is particularly illuminating because
it has revealed how such an hourglass topology
can serve as a substrate for evolutionary change.
Alterations in the cis-regulatory architecture
of the shavenbaby locus, that is, alterations in
the response of shavenbaby to signaling pathways, have resulted in the co-option of the
shavenbaby-controlled regulon in distinct types
of epidermal cells in distinct Drosophila species,
thereby explaining the evolutionary changes in
the patterns of specialized hair cells in different Drosophila species (McGregor et al. 2007,
Sucena et al. 2003). The evolution of distinct
cell types in the nervous system similarly may be
driven by the recruitment of neuronal terminal
selectors into novel cellular contexts through

modulation of their cis-regulatory inputs. Another driver for evolutionary change is mutations in the cis-regulatory regions of terminal
differentiation genes themselves, which lead to
the loss of or recruitment of a terminal differentiation gene into a terminal selector regulon
(Erwin & Davidson 2009).

FUTURE CHALLENGES
In spite of promising leads, a strong need
remains to further map regulons within individual neuron types in simple and accessible
model systems such as C. elegans. Such mapping
should entail not only genes expressed in specic neuron types but also broadly expressed
neuronal genes. In vertebrates, the identication of terminal selector regulons will remain
more challenging. The greater genomic complexity of vertebrates complicates the search
for discrete cis-regulatory elements through
transgenic reporter gene approaches because a
wider distribution of multiple terminal selector
binding sites over larger genomic intervals
may render the isolation of discrete functional
units difcult. A recent cis-regulatory analysis
of gene expression in dopaminergic neurons in
zebrash is consistent with such a complication
(Fujimoto et al. 2011). In such cases it may be
more useful to identify regulons not from the
cis-regulatory angle but from the trans-acting
angle through ChIP of transcription factors
whose maintained expression and genetic
requirement for terminal differentiation make
them good terminal selector candidates (such
as the Gata2, Tlx, or Runx genes mentioned
above). The in-depth analysis of the abovementioned vertebrate terminal selectors Crx,
Pet-1, Nurr1, and Pitx-3, which has involved
loss-of-function analysis, transcriptome analysis, bioinformatics, and chromatin immunoprecipitation, sets the stage for such future analysis.

DISCLOSURE STATEMENT
The author is not aware of any afliations, memberships, funding, or nancial holdings that might
be perceived as affecting the objectivity of this review.
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ACKNOWLEDGMENTS
Work from my laboratory cited in this review is funded by the NIH (R01NS039996-05;
R01NS050266-03) and the Howard Hughes Medical Institute. I thank members of the Hobert
lab, Evan Deneris, and Joseph Corbo for comment on this manuscript.
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Contents

Volume 27, 2011

Annu. Rev. Cell Dev. Biol. 2011.27:681-696. Downloaded from www.annualreviews.org


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Looking Back
Martin Raff p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 1
Membrane Protein Insertion at the Endoplasmic Reticulum
Sichen Shao and Ramanujan S. Hegde p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p25
Control of Organelle Size: The Golgi Complex
Debrup Sengupta and Adam D. Linstedt p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p57
Dynamin: Functional Design of a Membrane Fission Catalyst
Sandra L. Schmid and Vadim A. Frolov p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p79
The Role of Atg Proteins in Autophagosome Formation
Noboru Mizushima, Tamotsu Yoshimori, and Yoshinori Ohsumi p p p p p p p p p p p p p p p p p p p p p p p 107
Principles of Unconventional Myosin Function and Targeting
M. Amanda Hartman, Dina Finan, Sivaraj Sivaramakrishnan,
and James A. Spudich p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 133
Force Generation, Transmission, and Integration during Cell
and Tissue Morphogenesis
Thomas Lecuit, Pierre-Francois Lenne, and Edwin Munro p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 157
Degrading Devices: Invadosomes in Proteolytic Cell Invasion
Stefan Linder, Christiane Wiesner, and Mirko Himmel p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 185
Membrane-Anchored Serine Proteases in Vertebrate Cell
and Developmental Biology
Roman Szabo and Thomas H. Bugge p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 213
Wound Repair: Toward Understanding and Integration of Single-Cell
and Multicellular Wound Responses
Kevin J. Sonnemann and William M. Bement p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 237
Transmembrane Collagen Receptors
Birgit Leitinger p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 265
Cooperation Between Integrins and Growth Factor Receptors
in Signaling and Endocytosis
Johanna Ivaska and Jyrki Heino p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 291
viii

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Regulation of Integrin Activation


Chungho Kim, Feng Ye, and Mark H. Ginsberg p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 321
The Ins and Outs of the Epithelial to Mesenchymal Transition
in Health and Disease
M. Angela Nieto p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 347
Morphogen Gradients: From Generation to Interpretation
Katherine W. Rogers and Alexander F. Schier p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 377

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Limb Regeneration: A New Development?


Eugen Nacu and Elly M. Tanaka p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 409
Aerobic Glycolysis: Meeting the Metabolic Requirements
of Cell Proliferation
Sophia Y. Lunt and Matthew G. Vander Heiden p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 441
Cyclin-Dependent Kinases in Brain Development and Disease
Susan C. Su and Li-Huei Tsai p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 465
Epithelial Progenitor Cells in Lung Development, Maintenance,
Repair, and Disease
Jason R. Rock and Brigid L.M. Hogan p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 493
Gli Proteins in Development and Disease
Chi-chung Hui and Stephane Angers p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 513
Mechanisms of T Cell Development and Transformation
Ute Koch and Freddy Radtke p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 539
Developmental and Pathological Angiogenesis
Alicia S. Chung and Napoleone Ferrara p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 563
The Causes and Consequences of Polyploidy in Normal Development
and Cancer
Teresa Davoli and Titia de Lange p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 585
The Coupling of X-Chromosome Inactivation to Pluripotency
Jane Lynda Deuve and Philip Avner p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 611
The Role of MeCP2 in the Brain
Jacky Guy, Hel`ene Cheval, Jim Selfridge, and Adrian Bird p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 631
Neurogenesis at the BrainCerebrospinal Fluid Interface
Maria K. Lehtinen and Christopher A. Walsh p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 653
Regulation of Terminal Differentiation Programs
in the Nervous System
Oliver Hobert p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 681

Contents

ix

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Role of Leucine-Rich Repeat Proteins in the Development


and Function of Neural Circuits
Joris de Wit, Weizhe Hong, Liqun Luo, and Anirvan Ghosh p p p p p p p p p p p p p p p p p p p p p p p p p p p 697
Optogenetic Control of Cells and Circuits
Gero Miesenbock p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 731
Sensory Perception and Aging in Model Systems: From the Outside In
Nancy J. Linford, Tsung-Han Kuo, Tammy P. Chan, and Scott D. Pletcher p p p p p p p p p p 759
Indexes
Annu. Rev. Cell Dev. Biol. 2011.27:681-696. Downloaded from www.annualreviews.org
by Monash University on 09/02/13. For personal use only.

Cumulative Index of Contributing Authors, Volumes 2327 p p p p p p p p p p p p p p p p p p p p p p p p p p p 787


Cumulative Index of Chapter Titles, Volumes 2327 p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 790
Errata
An online log of corrections to Annual Review of Cell and Developmental Biology articles
may be found at http://cellbio.annualreviews.org/errata.shtml

Contents

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