Developmental Biology: Cécile Milet, Anne H. Monsoro-Burq

Developmental Biology 366 (2012) 2233
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Developmental Biology
journal homepage: www.elsevier.com/developmentalbiology
Review
Neural crest induction at the neural plate border in vertebrates

Ccile Milet, Anne H. Monsoro-Burq
Institut Curie; INSERM U1021; CNRS, UMR 3347, F-91405 Orsay, France
Universit Paris Sud-11, F-91405 Orsay, France
a r t i c l e
i n f o
Article history:
Received for publication 13 January 2012
Accepted 13 January 2012
Available online 25 January 2012
Keywords:
Neural crest
Neural plate border
AP2a
Pax3
Zic1
Hairy2
Msx1
Xenopus embryo
Vertebrate gene regulatory network
a b s t r a c t
The neural crest is a transient and multipotent cell population arising at the edge of the neural plate in
vertebrates. Recent ndings highlight that neural crest patterning is initiated during gastrulation, i.e. earlier
than classically described, in a progenitor domain named the neural border. This chapter reviews the dynamic
and complex molecular interactions underlying neural border formation and neural crest emergence.
2012 Published by Elsevier Inc.
Introduction
The neural crest, a key feature of vertebrate embryos, is a population
of highly multipotent and migratory progenitors. Neural crest cells give
rise to the many types of neurons and glia of the enteric, sensory and autonomous peripheral nervous system. They also differentiate into a variety of pigment cells such as melanocytes or melanophores, iridophores
and xanthophores. Moreover, they form chromafn cells, participate in
heart outow tract formation in amniotes, and are major progenitors of
craniofacial mesenchyme and skeleton (Le Douarin and Kalcheim,
1999). Neurocristopathies are a large group of birth defects resulting
from alterations in the complex succession of events required for the
induction, proliferation, migration and nal differentiation of neural
crest derivatives.
The neural crest originates from the border between the non-neural
ectoderm and the neurectoderm
Neural crest (NC) cells have been dened morphologically, as they
emerge from the edge of the neural folds (His, 1868; Le Douarin and
Kalcheim, 1999). At trunk levels, neural crest cells exit from the closing
dorsal neural tube via a stereotypical epithelium-to-mesenchyme transition (see Theveneau and Mayor, 2012this issue) (Ahlstrom and Erickson,
2009; Cheung et al., 2005; Serbedzija et al., 1989). At head levels, the
Corresponding author at: Institut Curie, UMR 3347, Batiment 110, Centre Universitaire,
F-91405 Orsay Cedex, France.
E-mail address: anne-helene.monsoro-burq@curie.fr (A.H. Monsoro-Burq).
0012-1606/$ see front matter 2012 Published by Elsevier Inc.
doi:10.1016/j.ydbio.2012.01.013
cranial neural crest forms before neural folds fusion, resulting in two bilateral masses of premigratory neural crest cells lying on both sides of the
neural plate. Soon thereafter, cranial neural crest cells initiate migration
toward the craniofacial areas, either as individuals or via collective migration recently described by live imaging (Theveneau et al., 2007, 2010; for
further details on migration events, see Theveneau and Mayor, 2012this
issue).
At both head and trunk locations, premigratory neural crest progenitors can be dened prior to any morphological change, by their specic
expression of characteristic molecular markers that distinguish them
from the adjacent ectoderm and neurectoderm (also see Labonne and
Sauka-Spengler, this issue on neural crest specication). Many of
these molecules are involved in the cellular process underlying the
epithelial to mesenchymal transition. Among the earliest and most
specic neural crest markers, Snai1, Snai2 (previously called Snail
and Slug) and SoxE family members are common to all vertebrate species examined (see references in Table 1).
Using both morphological and molecular analyses to identify neural
crest derivatives, lineage tracing was performed at neural plate and
neural tube stages, to follow the progeny of single or small groups of
cells from neurulation until organogenesis. Most classical analyses
were performed in the closing trunk neural tube and demonstrate the
common lineage between dorsal neural tube and neural crest, in avians,
amphibians and mammals during neurulation (Ahlstrom and Erickson,
2009; Bronner-Fraser and Fraser, 1988; Collazo et al., 1993; Serbedzija
et al., 1994). However, when such labeling is performed on small groups
of cells at the end of neurulation in the trunk of avian embryos, i.e. after
neural tube closure and just prior to cell emigration, the neural crest
C. Milet, A.H. Monsoro-Burq / Developmental Biology 366 (2012) 2233
lineage seems segregated from the roof plate lineage (Krispin et al.,
2010). Earlier mapping around the neural plate during avian gastrulation provides evidence for an area posterior and lateral to the neural
23
plate where progenitor for epidermis, placodes, neural crest and dorsal
neural tube are still intermingled (Basch et al., 2006; Fernandez-Garre
et al., 2002; Streit, 2002). In the early frog gastrula, neural crest
Table 1
List of the main neural border and neural crest markers in various vertebrates.
Each ectoderm subdomain (i.e. the non-neural ectoderm, the placodes, the neural border and neural crest, the neural plate and neural tube) can be characterized by the co-expression of
a group of markers. Although these markers are mostly conserved in vertebrates, some species-specic changes have been recorded and indicate the need for a careful characterization of
each model organism. The white boxes indicate genes without a described homolog in a given species; or that have not been implicated in NC development in the given
species. Reference links: Aoki et al., 2003; Aruga et al., 2002; Avilion et al., 2003; Barrionuevo et al., 2008; Bell et al., 2000; Bellefroid et al., 1998; Bellmeyer et al., 2003; Bulfone et al., 1993;
Byrd and Meyers, 2005; Cheng et al., 2000; Cheung and Briscoe, 2003; Davidson and Hill, 1991; Dottori et al., 2001; Dy et al., 2008; Essex et al., 1993; Feledy et al., 1999; Hill et al., 1989;
Honore et al., 2003; Inoue et al., 2004; Jen et al., 1996; Jiang et al., 1998; Jostes et al., 1990; Kee and Bronner-Fraser, 2001a; Kee and Bronner-Fraser, 2001b; Kos et al., 2001; Kuhlbrodt et al.,
1998; Liu and Harland, 2003; Mansouri et al., 1996; Marin and Nieto, 2004; Martin and Harland, 2001; Martinsen and Bronner-Fraser, 1998; Matsunaga et al., 2001; Mayor et al., 1995;
Mitchell et al., 1991; Myat et al., 1996; Nagai et al., 1997; Nakata et al., 1997; Nakata et al., 1998; Nakata et al., 2000; Nieto et al., 1994; Niss and Leutz, 1998; O'Donnell et al., 2006; Pera and
Kessel, 1999; Perez-Alcala et al., 2004; Robinson and Mahon, 1994; Saint-Germain et al., 2004; Salzberg et al., 1999; Sanchez-Guardado et al., 2011; Sasai et al., 2001; Sauka-Spengler et al.,
2007; Sawai et al., 1990; Schepers et al., 2000; Shen et al., 1997 Smith et al., 1992; Su et al., 1991; Suzuki et al., 1991; Suzuki et al., 1997; Thomas et al., 2008; Tour et al., 2001;
Uwanogho et al., 1995; Warner et al., 2003; Williams et al., 1995; Yang et al., 1998. Abbreviations: NC: neural crest; pNC: premigratory neural crest; mNC: migrating neural crest;
NP: neural plate; NT: neural tube; NB: neural border; HH Hamburger and Hamilton stage. Xenopus stages refer to Nieuwkoop and Faber stage table.
A- NEURAL BORDER SPECIFIERS (AND SOME ADJACENT NEURAL AND ECTODERM MARKERS)
Xenopus
Chick
Mouse
Lamprey
Human
Ectoderm lateral and anterior

to the neural plate in the
subcephalic pocket at HH st. 8
Neural folds corresponding to

pNC location atE 8.5
Ectoderm of early gastrula

(E3), ectoderm and NP border
(E4)
mNC in rostral sections,

branchial arches and cranial
mesenchyme at CS12
Luo et al., 2003
Shen et al., 1997
Mitchell et al.,1991
Sauka-Spengler et al., 2007;

Nikitina et al., 2008
Thomas et al., 2008
Neural fold at st12.5
Hindbrain at HH st. 10
Neural plate/ectoderm
Delta A
Not described in human early

neural development
Glavic et al., 2004
Myat et al., 1996
Williams et al., 1995
Sauka-Spengler et al., 2007
pNC in early neurula
ap2
Delta/notch
dlx3
dlx5
Anterior region of the

prospective NP at st12
pNC at HH st. 10
Branchial arches
DlxA, B, and C: Ventro-lateral

ectoderm, excluded from NP
(E4)
Feledy et al., 1999
Pera and Kessel, 1999
Robinson and Mahon, 1994
Not described in chick early

neural development
Lateral edges of the neural

plate at E7,75
Central part of the epiblast,

excluding the primitive streak
and Hensen's node at HH st. 4
Gastrula (E6.5), neuroectoderm and underlying

mesoderm (E7.5-E8.5), mNC
(pharyngeal arches)
Niss and Leutz, 1998
Bulfone et al., 1993, Byrd and

Meyers, 2005
No described homologue in
chick
mouse
Not described in chick early

neural development
Ectoderm at the end of

gastrulation (E7.5)
No chick homologue
Ectoderm except in prospective

NP at mid to late gastrula
Luo et al., 2001
NB and lateral ectoderm at st 12
gbx2
Tour et al., 2001; Li et al. 2009
Hairy2
Neural fold at st12.5-13
iro3
meis3
Salzberg et al., 1999
msx1
msx2
pax7
serrate
sox2

neural development
Hairy1/2A and B

neural development
Not described in lamprey early

neural development

neural development
Not described in mouse early

neural development
MeisA and B

neural development
Bellefroid et al., 1998
Sanchez-Guardado et al., 2011
confined dorsally to the lateral

edge of the neural plate at
st14
NC
NC at E9.5
MsxA: Ectoderm of gastrula

(E3.5), ectoderm and NP
border (E4)
Suzuki et al., 1997
Suzuki et al., 1991
Hill et al., 1989
Sauka -Spengler et al., 2007;

dorsal mesoderm in gastrula
not described in chick early

neural development
tip of the neural folds at E8,

cephalic NC
Su et al., 1991
pax3
lamprey
Bellefroid et al., 1998

NP at early mid neurula

neural development
Yang et al., 1998
Glavic et al., 2004

Prospective NP in gastrula

neural development
NT emigrating cells in culture,

late C10 to late C12
Thomas et al., 2008
not described in human early
neural development
Davidson and Hill, 1991
Lateral domains of the

prospective NP at st11.5
NT at HH st. 10
Dorsal part of the

neuroepithelium at E8.5
Pax3/7: Ectoderm of early

gastrula (E3), NP border (E4)

late C10 to late C12, pNC of
dorsal NT at C12
Bang et al., 1999
Matsunaga et al., 2001
Goulding et al., 1991

Thomas et al., 2008, Betters

et al., 2010
Brain (st14), paraxial

mesoderm (st18)
Bilaterally symmetric oblique

bands lateral to Hensens node
in epiblast at HH st. 4+
Alar plate of the neural tube

(E8.5)
Maczkowiak et al., 2010
Basch et al., 2006
Neural fold at st12.5-13
Neural fold HH st. 10
Glavic et al., 2004
Myat et al., 1996
End of gastrulation :
prospective neurectoderm.
Neural epithelium at HH st. 10
Mizuseki et al., 1998
Uwanogho et al., 1995
Jostes et al., 1990, Mansouri

et al., 1996
not described in mouse early
neural development
Dorsal NT at CS12
Thomas et al., 2008
lamprey

neural development
Anterior neurectderm at E77.5, neural tube and branchial

arches at E9.5
SoxB1: NP border (E4)

Avilion et al., 2003
Thomas et al., 2008
24
Table 1 (continued)
B- EARLY NEURAL CREST MARKERS

Xenopus
zic1
zic2
zic3
id4
snail1
snail2
sox5
sox 8
sox 9
sox 10
zic 5
Lamprey
ZicA: Ectoderm of early
gastrula (E3), NP (E4)
Mizuseki et al., 1998, Nakata

et al., 1998
Aruga et al., 2002, Warner et

al., 2003
Nagai et al., 1997

Prospective NP in early
gastrula
NT HH st. 13
Embryonic mesoderm and

ectodermal cells in gastrula
(E7), neurectoderm (E7.25),
dorsal NT and dorsal
mesoderm derivatives (E9.5)
Nakata et al., 1998
Warner et al., 2003
Nagai et al., 1997
Prospective NP in gastrula
NT HH st. 13, dorsal

epithelium st23
Embryonic mesoderm and

ectodermal cells in gastrula
(E7), dorsal NT and dorsal
Nakata et al., 1997
Warner et al., 2003
Nagai et al., 1997
pNC at st14 (early neurula)
N-Myc: dorsal NT and NC

derivatives at E3.5
Not described in mouse NC
Bellmeyer et, al., 2003
Sawai et al., 1990
Presumptive NC region in late

gastrula
pNC and mNC at HH st. 6 to 10
Human
neural development

neural development

neural development
N-myc: Ectoderm of early

gastrula (E3), ectoderm
including the NP (E4)
Not described in human NC

pNC and mNC at E9.5-E10.5
FoxD-A: pNC, mNC at E6 (NT

formation)

Sasai et al., 2001
Kos et al., 2001
Dottori et al., 2001
Thomas et al., 2008
mNC
Cranial neural folds and mNC

in 5 somite stage embryo
mNC at E8.5
Id: Ectoderm of early gastrula

(E3), ectoderm and NP border
(E4)
Liu and Harland, 2003
Martinsen and Bronner-Fraser,

1998
Jen et al., 1996

Migrating NC
Border of neural plate at HH

st. 5-. Dorsal NT at HH st 8.
Olfactory, lens and otic
placodes at stage 12
mNC at E8.5
Liu and Harland, 2003
Kee et al., 2001b
Jen et al., 1996
Ectoderm at the gastrula

stages
Epiblast ectoderm at HH st. 5.Dorsal NT at HH st. 10. mNC

at HH st. 12
Not described in mouse NC
Liu and Harland,2003
Kee et al., 2001a
Gastrula st 11,5
mNC at HH st. 21
Cephalic NC at E9.5
SnailA: Open NP and border at

E4
Essex et al., 1993
Marin and Nieto, 2004
Smith et al., 1992
Gastrula st 12,5
pNC, mesoderm (5 somite

stage)
Not in pNC, experssed in mNC
lamprey

Not described in lamprey NC
id2
id3
Mouse
dorsal NT HH st. 10, NC HH

st. 23
c-myc
foxd3
Chick
Embryonic mesoderm in
gastrula (E7), neurectoderm
(E7.25), dorsal NT and dorsal
End of gastrulation :
prospective neurectoderm.
NP, NB
Mayor et al., 1995
Nieto et al., 1994
Jiang et al., 1998
migrating NC
Neural folds HH st. 7
NC
Martin and Harland, 2001
Perez-Alcala et al., 2004
Dy et al., 2008
Gastrula st 11,5
Dorsal NT at HH st. 11-13
mNC (branchial arches, dorsal

root ganglia) at E10
SoxE1, 2 and 3: pNC, mNC at

E6 (NT formation)
O'Donnell et al., 2006
Bell and Briscoe, 2000
Schepers et al., 2000
Gastrula st 12
Prospective NC at HH st. 10-12
Thomas et al., 2008
NT emigrating cells in culture

late C10 to late C12; pNC of
dorsal NT, branchial arches
and cranial mesenchyme,
mNC in rostral sections at CS12
NC at 5-6 somites stage
Saint-Germain et al., 2004
Cheung and Briscoe, 2003
Barrionuevo et al., 2008
Thomas et al., 2008, Betters

et al., 2010
Mid-neurula st 14
Prospective NC from HH st.7-8

to NC derivatives HH st. 26
pNC at E8.5
pNC of dorsal NT, mNC in

rostral sections at CS12
Aoki et al., 2003, Honore et

al., 2003
Cheng et al., 2000
Kuhlbrodt et al., 1998
pNC at late gastrula (stage

11.5-13)
chick
Nakata et al., 2000
Betters et al., 2010
NP at E8, dorsal NT
ZicA: Ectoderm of early

gastrula (E3), NP (E4)
Inoue et al., 2004

progenitors were mapped across an ectodermal area and the lateral part
of the neural plate, lying adjacent to the dorsallateral marginal zone
(Steventon et al., 2009). Similar fate maps were obtained for mouse
(Tam, 1989) and chick embryos (Ezin et al., 2009). This posterior and
lateral ectoderm area is the site of key patterning events that will dene
the neural crest lineage from these adjacent fates and constitutes the
domain named the neural border (or the neural plate border) (Fig. 1).
The neural border (NB) could thus be dened as the broad competence domain, established between neural and non-neural ectoderm,
within which subtle and sequential cell autonomous and non-cellautonomous molecular interactions will progressively dene the neural
crest fate from adjacent dorsal neural fates. This chapter will describe
the current state of knowledge regarding neural border patterning during the course of vertebrate gastrulation and neurulation, the process of
25
anterior
Ectoderm
high BMP
Preplacodal
field
low Wnt / low BMP
Otx2
Anterior NF
Neural border
Medial NP
Midline
Late gastrula
high Wnt / medium BMP

Gbx2
AP2 (border / ectoderm)
high Wnt / low BMP
Gbx, Cdx
posterior
anterior
Ectoderm
keratins
Preplacodal
field
eya1, six1
Anterior NF
anf1,
sox2
Neural border
pax3
Medial NP
pax3/snail2
Midline
Mid-neurula
posterior
Fig. 1. Schematic representation of the ectoderm at late gastrula and mid-neurula stages.The non-neural ectoderm (yellow) surrounds the neural border and the pre-placodal ectoderm (pink and green respectively), which themselves surround the neural plate (blue). At gastrulation stage, the main secreted inducers and key patterning transcription factors
expressed in these areas are indicated. At neurula stage, each domain can be dened by expression of a few characteristic markers (one is indicated, see text and Table 1 for additional information).
neural crest induction from neural border progenitors and link these
molecular events to the neural crest gene regulatory network.
A combination of secreted signals patterns the neural border
during gastrulation and neurulation
Molecular characterization of the neural border during gastrulation and
neurulation
The earliest detection of specic neural crest markers is reported
at mid-to-late gastrula stage in frog, and at 57 somites stage in
chick and mouse head neural folds stage (see references in Table 1).
This suggests that neural border patterning occurs prior to these
stages, namely during early- or mid-gastrulation. At similar stages,
neural patterning has also started (Puelles et al., 2005). In agreement
with its location at the prospective edge between neural and nonneural ectoderm, the neural border can be rst identied as the
overlap between two types of more broadly expressed markers:
genes also expressed in the lateral ectoderm (e.g. tfap2, hairy2,
msx1, msx2, dlx5) and genes also expressed in the more medial neural
plate (e.g. zic1, zic2, gbx2, meis3) (Fig. 1, see references in Table 1). More
specic neural border expression is described for pax3 and pax7 paralogs whose expression is initiated at early gastrula stage and can be
traced later to the dorsal neural fold and neural crest in both in frog
and chick embryos (Bang et al., 1999; Basch et al., 2006; de Croze
et al., 2011; Monsoro-Burq et al., 2005). Although mouse and human
pax3 and pax7 have been described in the dorsal neural tube, there is little information about their expression in the open neural plate (Betters
et al., 2010; Gerard et al., 1995; Goulding et al., 1991; Terzic and SaragaBabic, 1999). In summary, during gastrulation, the prospective neural
border encompasses a broad lateral domain (rather than a dened
line), comparable to the neural plate edge described at a similar stage
(Puelles et al., 2005; Rodriguez-Gallardo et al., 2005).
As the neural folds thicken and elevate, the neural border of vertebrate embryos is characterized by the enhanced co-expression of pax3/
7, msx1/2, zic1/2, tfap2a, iroquois1 (Xiro1) and hairy2, while it is outlined
by the expression of serrate and delta, as reported in chick, frog, zebrash,
lamprey, mouse and human (see references in Table 1). Simultaneously,
many of these NB genes are either down regulated or excluded from the
immediately adjacent ectoderm and the medial neural plate (e.g. msx1,
ap2a, hairy2, zic1, zic2). In contrast, the non-neural lateral ectoderm is
characterized by specic markers, such as dlx3 and ectodermal keratins
(Fuchs, 1995; Luo et al., 2001). Anteriorly, between the neural plate
(expressing pan-neural markers such as sox2 and sox3) (Iwafuchi-Doi
et al., 2011; Mizuseki et al., 1998; Rogers et al., 2008) and the nonneural ectoderm, the preplacodal markers (e.g. six1, eya1) mark the prospective placodal eld (Schlosser and Ahrens, 2004; Streit, 2002) (Fig. 1).
At mid-neurula stage, NC markers' expression is enhanced within the
neural border and overlaps with some NB markers such as pax3/7. In
contrast, other early NB markers are soon excluded from the prospective
neural crest domain (e.g. msx1) (Monsoro-Burq et al., 2005). In summary, the neural border can be dened molecularly as early as gastrulation,
26
by the overlap in expression of several neural border genes, a domain

within which early neural crest markers will appear.
Tissue interactions control early neural crest patterning
Explants recombination in vitro and grafting experiments in vivo have
explored how the neural crest is induced in the embryo. These
experiments have uncovered the critical role of ectodermneurectoderm
and mesodermectoderm interactions during neural crest formation.
Moreover, heterochronic recombination experiments have explored the
timing of neural crest patterning.
The ectoderm induces neural crest when apposed to a nonpatterned neural plate fragment (Liem et al., 1995). Similarly, an
early ventral neural plate implanted into lateral ectoderm in vivo,
leads to snai2 induction at the edge between the two tissues, with
both neural and ectoderm-derived cells expressing the NC markers
(Mancilla and Mayor, 1996; Selleck and Bronner-Fraser, 1995). The
effect of such experiments on neural border gene expression has
not been reported. Recombination of the non-neural ectoderm (or
early ectoderm taken from the blastocoele roof in amphibians, the
animal cap) and prospective paraxial/intermediate mesoderm (i.e.
the dorsal lateral marginal zone in amphibians) also lead to neural
crest induction followed by melanocyte differentiation in the ectoderm
(Bonstein et al., 1998; Marchant et al., 1998). Similar recombinations
have resulted in pax3 and msx1 induction using chick inducing tissues
and Xenopus responding ectoderm (Bang et al., 1999). In contrast, the
anterior neural fold or the notochord prevents either anterior or medial
snai2 expression in the neural plate (Carmona-Fontaine et al., 2007;
Rufns et al., 1998). However, notochord grafts performed after neural
tube closure do not prevent neural crest emigration (Artinger and
Bronner-Fraser, 1992), consistent with commitment to neural fold
fate after gastrulation.
In addition, explants cultured in vitro and heterochronic recombination experiments in vivo show that Xenopus neural border ectoderm is
committed to form snai2-positive cells by the end of gastrulation
(around Nieuwkoop and Faber stage 12) and that competence to respond to inducing cues is lost at the same stage (Mancilla and Mayor,
1996). This suggests that the neural border is induced and determined
prior to this stage. In chick embryos, explantation of the prospective
(pax3/7+) neural border ectoderm shows that these cells are also determined to form NC cells by gastrulation stage 4+ (Basch et al., 2006).
These experiments have opened the path for the identication of
the molecular signals responsible for neural border and neural crest
induction, as it has been assumed that similar secreted molecules
may act during both processes.
During gastrulation in mouse, Otx2 is required for expression of the

canonical Wnt antagonist dickkopf-1 (dkk1) in the anterior visceral
endoderm, the mouse head organizer (Zakin et al., 2000). Dkk1 is essential for head formation in vertebrates; it allows placode formation and
represses posterior (i.e. neural crest) fates at the anterior neural border
(Carmona-Fontaine et al., 2007; Fossat et al., 2011; Niehrs et al., 2001).
The negative feedback loop established between the anterior otx2 positive domain and the posterior gbx1/2-positive domain establishes a
sharp MHB as early as E7.5 in mouse embryos (Millet et al., 1999;
Sunmonu et al., 2011). In the posterior domain, Gbx2 represses isthmus
markers (engrailed2, wnt1, fgf8) and activates posterior neural platespecic genes, both in the medial (cad2) and lateral (pax3) domains
(Li et al., 2005, 2009; Tour et al., 2002) (Fig. 1).
Mediolateral patterning involves a combination of secreted signals at the
neural border
Within the posterior gbx2-positive domain, another complex
combination of secreted molecules establishes neural border patterning,
as gastrulation movements place the prospective paraxial/intermediate
mesoderm under the lateral neural plate (Schroeder, 1970; Steventon
et al., 2009). Neural induction, primed by FGF signals at the end of blastula stage, is controlled by simultaneous inhibition of Smad1 and Smad2
signaling (Chang and Harland, 2007; Delaune et al., 2005; Londin et al.,
2005; Wilson et al., 2000). A medial (low) to ventral (high) BMP signaling gradient, visualized by phospho-Smad1-5-8 staining, indicates the
establishment of a zone of intermediate BMP activity at the level of the
prospective neural border at the onset of gastrulation in chick, sh and
frog embryos in vivo (Faure et al., 2000, 2002; Nguyen et al., 1998).
Later, as the neural folds elevate, increased BMP4/7 mRNA expression and high BMP signaling mark the neural folds in chick, mouse,
frog and sh (Barth et al., 1999; Faure et al., 2002; Monsoro-Burq
and Le Douarin, 2001; Sakai et al., 2005; Wu et al., 2011). At
gastrula-stages, BMP4 appears to act as a morphogen in a graded
fashion by providing initial positional information in the eld of
the ectoderm, which denes main ectodermal fates: neural (low
activity); neural crest/placodes (intermediate activity); and ectoderm/cement gland (high activity).
However, the experimental modulation of BMP signaling in vivo
or in ectoderm isolated in vitro, is an inefcient neural crest inducer
Neural plate patterning cues position the neural crest-forming neural

border along anteriorposterior and mediallateral axes
Early anteriorposterior patterning denes the boundary between neural
crest-forming and neural crest-free neural border
Anterior-to-posterior neural patterning denes the extent of the
neural crest forming neural border along the body axis. Classical fatemapping experiments have shown that the neural crest originates
posterior to the mid-diencephalon (reviewed in Le Douarin, 2004).
During gastrulation, anterior and posterior brain domains are established by the mutual antagonism between otx2 and gbx1/2 transcription
factors, dening the prospective midbrainhindbrain boundary (MHB)
(Heimbucher et al., 2007; Martinez-Barbera et al., 2001; Tour et al.,
2002). This results from the combined action of Wnts, FGFs, and retinoic
acid, that impose posterior fates to the forming neural plate (Kudoh et
al., 2002; Ribes et al., 2009; Takemoto et al., 2006). Studies in zebrash,
mouse and frog embryos show that retinoic acid and FGF8 regulate the
early neural expression of gbx2; and that Wnt8 signals, diffusing from
the lateral mesendoderm, directly activate gbx1/2 (Li et al., 2009; Liu
et al., 1999; Rhinn et al., 2003, 2005, 2009; von Bubnoff et al., 1996).
Fig. 2. Neural border and neural crest patterning is a highly dynamic process which involves reiterated action of secreted signals. Several signaling pathways (BMP, WNT,
FGF, Notch) cooperate and dene the posterior neural border and the neural crest. Recently described novel modulators of these signals are activated during gastrulation or
neurulation. Collectively, their action is critical to achieve correct spatial and temporal
patterning.
(LaBonne and Bronner-Fraser, 1998). Wnts and FGFs potentiate NC

induction and its further development, both in vivo and in neuralized
ectoderm, leading to the two-signal model of neural crest induction
(Chang and Hemmati-Brivanlou, 1998; LaBonne and Bronner-Fraser,
1998; Saint-Jeannet et al., 1997; Villanueva et al., 2002). Similar
rules control neural border formation (de Croze et al., 2011; Hong
et al., 2008; Luo et al., 2003; Monsoro-Burq et al., 2005; Nichane et
al., 2008b; Sato et al., 2005; Tribulo et al., 2003). Multiple Wnts are
expressed in the ectoderm; some being restricted to the non-neural
ectoderm from where they diffuse towards the neural border (wnt6
in chick and wnt10a in Xenopus) (Garcia-Castro et al., 2002;
Garriock et al., 2007). Several FGFs are expressed in the paraxial/
intermediate mesoderm (Kimelman et al., 1992; Monsoro-Burq et
al., 2003; Moon, 1993) (Fig. 2). Thus far, the respective role of Wnts
and FGFs secreted from the mesoderm has been a matter of debate.
The requirement for Wnt signals at multiple steps of NB/NC
development makes it particularly difcult to evaluate the action of
other signals independently of Wnt activity (de Croze et al., 2011;
Lewis et al., 2004; Monsoro-Burq et al., 2005; Sato et al., 2005). In
addition, there are indications that wnt8 is activated by FGFs in the
mesoderm and reciprocally that Wnts activate fgf3/fgf8 in the
ectoderm (Hong et al., 2008; Zhao et al., 2008) (Fig. 2).
There are several lines of evidence suggesting that FGFs participate in NC induction. First, although FGFs seem to be weaker NC inducers than canonical Wnt signals, when applied after neural and
mesoderm induction, they display clear neural crest inducing activity
on isolated ectoderm. Basic FGF (bFGF/FGF2) also induces melanocytes in mid-gastrula ectoderm cells grown in vitro, while an earlier
treatment, at late blastula stage, activates neural fates (Kengaku and
Okamoto, 1993). Secondly, the FGF8 isoform-a (FGF8a), which does
not induce mesoderm but efciently promotes posterior neural
fates, is sufcient to induce some NC markers (foxd3, sox9 but not
snail2) in isolated blastula ectoderm (Fletcher et al., 2006; MonsoroBurq et al., 2003). The mechanism of neural crest induction by FGFs
seems to involve Wnt signals since Wnt8 is a target of FGF8 in the mesoderm, and FGFs induce neural crest in a Wnt-dependent manner in
neuralized ectoderm (Hong et al., 2008; LaBonne and Bronner-Fraser,
1998). These observations suggest that FGFs would activate Wnt signals within the mesoderm, and that in turn, Wnts would induce neural border and neural crest in the overlying ectoderm (Hong et al.,
2008).
There are, however, several strong indications of a parallel Wntindependent activation of neural border/neural crest by FGF signals.
First, in the explants recombination assay between dorsallateral
mesoderm and blastula ectoderm, blocking response to Wnts in the
ectoderm still allows efcient NC induction (Monsoro-Burq et al.,
2003). It is only when extremely high doses of Wnt inhibitors are
used that neural crest induction is lost, suggesting a non-specic
effect (Steventon et al., 2009). Secondly, FGF receptors are expressed
at the neural border and blocking FGFR signaling in the ectoderm
prevents NC induction in the mesoderm/ectoderm recombinant
assay. Furthermore, FGF-dependent Stat3 activity is essential in the
prospective NC cells (Hongo et al., 1999; Monsoro-Burq et al., 2003;
Nichane et al., 2010; Zhao et al., 2008). Finally, while Wnt and FGF
pathways may have common modulators such as the transmembrane
protein Lrig3, Wnt/Lrig3 and FGF/Lrig3 interactions display different
roles in neural crest induction (Zhao et al., 2008). Resolution of
these differences will likely occur within the next few years, thanks
to the availability of new tools and markers for analyzing discrete
steps of neural border and neural crest formation.
Fine-tuned modulation of the secreted signals allows further neural crest
induction
The precise timing of signal activity is a key parameter for neural
border and neural crest induction, highlighted by recent studies of
novel signaling modulators. In relationship to the two phases of
27
BMP signaling mentioned above, initial BMP inhibition is required

for the rst phase of NB/NC induction during gastrulation, while
further NC specication requires simultaneously increased BMP and
Wnts (Patthey et al., 2009; Steventon et al., 2009).
The timing of BMP activity also varies along the anteriorposterior
axis (Tucker et al., 2008). This dynamic activity is tightly regulated, in
part by newly identied molecules, which shape the initial BMP
gradient and rene neural border positioning (Fig. 2). For example,
a dynamic negative BMP regulation has been demonstrated for the
novel secreted antagonist Tsukushi (Ohta et al., 2004). At gastrula
stage, the BMP gradient is shaped by BMP antagonists such as Noggin,
Chordin, Follistatin, Dan, Cerberus and Tsukushi, secreted from the
node/organizer (Harland, 2000; Ohta et al., 2004). At early neurula
stage, tsukushi is secondarily activated by BMP signals around the
anterior neural border and participates in modulating BMP signaling,
by a complex, dose-dependent effect on Notch signaling, which in
turn, affects bmp4 expression at the neural border (Glavic et al.,
2004; Kuriyama et al., 2006). By this activity, Tsukushi synergizes
with Wnt signals and allows mid-neurula stage hairy2, msx1,
snail2 and sox9 expression at the neural border (Kuriyama et al.,
2006).
Simultaneously, positive BMP regulators are also needed for
neural border patterning. For example, Snw1 (also known as skiinteracting protein SKIP), which encodes a highly conserved putative
nuclear protein, was isolated in a large-scale functional screening
conducted to identify novel modulators of the neural crest fate in
Xenopus embryos (Wu et al., 2011). Snw1/SKIP is expressed dorsally
during gastrulation and enriched in the neural and neural crest tissue
at neurulation. The neural border does not form in Snw1/SKIP
morphants (lack of msx1 expression) while ectoderm and neural
plate markers are expressed in adjacent ectoderm territories. Snw1
morphants exhibit decreased phospho-Smad signals after (but not
during) gastrulation, indicating that Snw1/SKIP is essential to
promote BMP signaling and neural plate border sharpening during
neurulation (Wu et al., 2011). Another novel regulator of BMP signals, Tumor Necrosis Factor-Receptor associated Factor-4 (TRAF4),
isolated as a target of the ubiquitin ligase Smurf1, is strongly
expressed in mid-neurula stage premigratory neural crest, and potentiates both BMP and Nodal signals at the neural border (Kalkan
et al., 2009).
Similarly, several modulators of Wnt and FGF pathways have been
implicated in NC and NB formation. In addition to its action upstream
of BMP receptors, Snw1/SKIP is also an essential partner of the
canonical Wnt pathway by interacting with the transcription factor
LEF1 and repressing Wnt targets (Wang et al., 2010). In the anterior
neural plate, attenuation of Wnt signaling has also been reported for
the secreted Wnt antagonist Dkk1, and for Kremen when bound to
Dkk1 and the Wnt co-receptor LRP6 (Davidson et al., 2002; Fossat
et al., 2011; Glinka et al., 1998). Similarly, a tight control of Wnt
signaling around the neural plate or in trunk neural tube patterning,
involves various Wnt negative regulators, such as SFRPs, Kcdt15, or
ephrins (Dutta and Dawid, 2010; Misra and Matise, 2010). In contrast,
in the posterior neural border, activation of Wnt signaling is achieved
by Kremen, by Dkk1-independent LRP6 signaling activation (Hassler
et al., 2007). Similarly, the scaffolding transmembrane protein
TEM198 stimulates LRP6 signaling and is essential for Wnt signaling
in neural crest formation, while the metalloprotease ADAM13
stimulates Wnt signaling via cleavage of ephrinb2 (Liang et al.,
2011; Wei et al., 2010). Finally, the transmembrane protein Lrig3,
acts downstream of neural border formation and differentially
modulates Wnt and FGF/FGFR signaling in a context-dependent manner (Zhao et al., 2008).
These results highlight that ne-tuned signaling activities are
essential in controlling neural border positioning, and that these signals are highly dynamic both in space and time. These observations
underline the importance of a careful stage-specic analysis, especially
28
when these regulations are compared between different vertebrate

species.
Neural border speciers crosstalk to induce neural crest
In 2004, Meulemans and Bronner proposed a model primarily based
on expression patterns, grouping actors of neural crest development
according to their putative timing of activity at the neural border.
Hence, secreted signaling molecules act upstream of neural border
formation while stabilizing the neural border which expresses genes
that activate downstream NC specic genes. In turn, the latter genes
control EMT and migration (Meulemans and Bronner-Fraser, 2004).
This hierarchical view has since proven a very useful framework for
validation of the proposed interactions (Betancur et al., 2010). It has
dened the notion of neural border and neural crest speciers
compared to neural border/crest markers. While a marker (better a
combination of markers) is useful to identify a given tissue, a specier
has been shown to be expressed in this tissue, and to play an essential
function in its formation. Perturbed expression of a NB specier will
affect both neural border formation and downstream neural crest
development. Altered NC speciers expression does not affect neural
border formation or marker expression, but rather impacts the
premigratory neural crest and its later development.
Cross-regulation between the NB speciers has been explored
mainly in sh, lamprey and Xenopus embryos, as these vertebrate
models allow numerous types of experimental manipulations at
blastula and gastrula stages, using morpholino knock-down or
analysis of mutants. These studies have identied three sequential
steps upstream of neural crest formation: neural border induction
and maintenance, and NC speciers' induction.
Neural border speciers cooperate and stabilize the neural border
Neural border gene expression is initiated during early gastrulation,
in the posterior half of the developing neural plate (de Croze et al.,
2011; Nikitina et al., 2008). Morpholino-mediated knockdown
experiments have explored which factors initiate neural border
induction. When AP2 is depleted, neither pax3, hairy2 nor msx1
expression occurs at the edge of the neural plate of gastrula embryos,
while ap2 expression remains normal when all these other NB speciers
are depleted (de Croze et al., 2011). This indicates that AP2 is essential
in initiating the neural border. In addition, Wnt signals activate ap2,
pax3 and msx1 expression as immediate-early targets. However, this
induction depends on the presence of AP2, which directly binds to
pax3 regulatory elements (de Croze et al., 2011). A parallel observation
shows that Wnt signals directly activate gbx2; in turn, Gbx2 is essential
for early onset of the neural border by Wnts (Li et al., 2009). Additional
direct regulations involve Pou-domain factors Brn1/2 and Pbx/Hox
genes on pax3 promoter to dene neural expression and its most
anterior level of expression, respectively (Pruitt et al., 2004). Thus,
within the posterior neural domain, cooperation between Wnt signals,
Gbx2 and AP2 is responsible for neural border initiation; these factors
cooperate with additional transcription factors to directly control pax3
induction.
In a second step, as soon as neurulation starts, NB speciers
cooperate with each other, resulting in the maintenance of a proper
neural border development. Hence, AP2 depletion also prevents
later expression of the NB specier, indicating that neural border
development is not just delayed but blocked, in favor of neural formation (zic1 and sox2 are expanded) (de Croze et al., 2011; Li and
Cornell, 2007; Nikitina et al., 2008). In addition, in Xenopus embryos,
when either Msx1, Zic1, Pax3 or Hairy2 are depleted, hairy2, ap2, or
msx1 are lost, while pax3 and/or zic1 expression are increased (de
Croze et al., 2011; Monsoro-Burq et al., 2005; Nichane et al., 2008a;
Nikitina et al., 2008; Sato et al., 2005). Some species-specic
differences are observed in these regulatory relationships, since pax3
or zic1 can be either increased of diminished in lamprey embryos subjected to similar knockdown experiments (Nikitina et al., 2008).
Species-specic differences can also exist in the differential deployment
of paralog genes (pax3 or pax7) as well as in redundancy between duplicated genes (ap2a and ap2c in sh) (Basch et al., 2006; Li and Cornell,
2007; Maczkowiak et al., 2010).
As a whole, these results indicate that the neural border is not
patterned and maintained normally after perturbation of any of the
NB speciers. Consequently all these manipulations result in the
lack of neural crest induction.
Neural border speciers cooperate to induce premigratory neural crest
NB speciers' activity is tightly regulated, both positively and
negatively to activate NC speciers. For example, Nichane and
colleagues have provided a detailed view of the complex interactions
taking place between the NB speciers Hairy2, Id3 and Stat3 in neural
crest induction. By loss and gain of function experiments followed by
NC markers detection (snail2, foxd3, sox10), they showed that Id3 is
required for Hairy2 to expand the neural crest, and that Hairy2 and
Stat3 act synergistically in NC induction (Nichane et al., 2008a,
2010). In particular, Hairy2 plays a dual role in NC specication:
through cell-autonomous mechanisms involving Hairy2 DNAbinding activity, Hairy2 promotes undifferentiated NC progenitor
survival and maintenance, while a non-cell-autonomous mechanism
involves Hairy2 binding to FGFR4/Stat3 complex leading to id3
activation. In turn, Id3 promotes neural crest proliferation and
differentiation (Kee and Bronner-Fraser, 2005; Light et al., 2005).
Stat3 plays a central role in regulating this balance. At low levels,
Stat3 activates hairy2, which will prevent id3 activation, via bmp4
inhibition (Nichane et al., 2008a, 2010). Conversely, at high activity
levels, Stat3 induces id3, which in turn down regulates hairy2 and
Stat3 signaling by disrupting the Stat3-Hairy2-FGFR4 ternary
complex (Nichane et al., 2010). Thus, low Stat3 activity maintains
prospective NC cells undifferentiated; high Stat3 activity induces NC
specication; respectively, Hairy2 and Id3 function as positive and
negative feedback regulators of Stat3 activation, allowing ne-tuned
control of the system.
Other NB speciers have been shown to cooperate in NC
induction. In particular, Pax3 and Zic1 play a key role in premigratory
NC induction. Pax3 and Zic1 co-expression induces ectopic ventral
snai2 expression in whole embryos, as well as snai2 and foxd3 expression in animal caps (Monsoro-Burq et al., 2005; Sato et al., 2005).
Over expressing Zic1 (or Pax3) cannot compensate for Pax3 (or Zic1
respectively) depletion (Sato et al., 2005). Balanced Pax3 and Zic1
levels are critical for allowing neural crest fate choice, at the expense
of other ectoderm fates (placodes or hatching gland in Xenopus embryos) (Hong and Saint-Jeannet, 2007). In parallel, Msx1 activates
pax3 cell-autonomously, while Pax3 activity is necessary for Msx1
to induce snai2, and Pax3 gain-of-function compensates for Msx1
knockdown (Monsoro-Burq et al., 2005). Finally, downstream of
Pax3 and Zic1 cooperation, AP2 plays another role: it is both required
to allow NC marker induction by Pax3/Zic1 in explants, and sufcient
to compensate for Pax3 or Zic1 knockdown in snai2 and foxd3
induction in vivo (de Croze et al., 2011). Together, these results
indicate hierarchical (i.e. epistatic) relationships between the NB
speciers. Future work will further validate these as direct or indirect interactions (Betancur et al., 2010).
NB speciers are essential mediators of secreted factors in NC induction
By their cooperative response, the NB speciers integrate the
multiple developmental cues provided by the tissues surrounding
the neural plate. Hairy2 and Msx1 act downstream of FGF and BMP
signaling to induce NB (pax3) and NC markers' expression (snai2,
foxd3) while Wnt signaling induces NC in Hairy2 and Msx1 morphant
embryos (Nichane et al., 2008a) (Monsoro-Burq et al., 2005). In

parallel, AP2, Pax3 and Zic1 are essential for Wnt, FGF and BMP
signaling to induce NC (de Croze et al., 2011; Monsoro-Burq et al.,
2005; Sato et al., 2005). In addition, several combinations between a
given NB specier and a secreted signal have been validated for
neural border and neural crest induction in animal caps (e.g. Msx1
and Noggin, AP2 and Chordin/Wnt and AP2 and Noggin; Pax3 and
Wnt or BMP, Zic1 and Wnt or BMP etc.) (de Croze et al., 2011; Luo
et al., 2003; Monsoro-Burq et al., 2005; Sato et al., 2005). These
experiments indicate that, BMP levels must be tightly balanced, and
that such assays result into the activation of the whole complement
of NB speciers followed by neural crest induction.
29
affects the expression of the NC speciers sox9, foxd3 and snai2 but
not dorsal neural tube markers. Changes in H3K9me3, H3K36me3
and JMJD2A occupancy on NC specier genes (sox10, snail2) were
observed during chick development coincident with NC specication.
JmjD2A is responsible for derepressing sox10 by H3K9me3 demethylation (Strobl-Mazzulla et al., 2010).
Chromatin structure regulation is known to provide spatiotemporal control of stem cell maintenance, specication and differentiation. The discovery of a developmental role for epigenetic regulators in
controlling neural crest specication opens exciting avenues for novel
research.
Conserved evolutionary function of the neural border speciers
Downstream regulators of NC fate and secondary use of signaling

pathways
Downstream of Pax3 and Zic1, several novel effectors and a reiterated
activation of the Wnt, FGF, BMP and Notch pathway are necessary to
allow further NC development. The TALE homeodomain Meis proteins
are important candidates for coordinating hindbrain neural and neural
crest patterning. Meis3 is a co-factor of Hox proteins (Nakamura et al.,
1996) expressed in the neural plate and regulated by mesodermal
Wnt3a (Elkouby et al., 2010; Gutkovich et al., 2010). Loss of function
experiments have established that Meis3 acts downstream of Pax3/
Zic1 and is essential for the expression of a subset of NC genes and NC
formation (Gutkovich et al., 2010; Maeda et al., 2002). In addition,
Meis3 directly activates FGF3 and FGF8 and interacts with retinoic acid
signaling in the neurectoderm, promoting further secondary signaling
necessary for NC induction (Aamar and Frank, 2004; Dibner et al.,
2004; Elkouby et al., 2010). In parallel, Lrig3 also acts downstream of
Pax3 and Zic1 in neural crest specication (Zhao et al., 2008).
Furthermore, dissociated AC experiments showed that after initial
induction of Msx1 and Pax3 by Wnt, Pax3 and Zic1 cooperate with the
canonical Wnt pathway to induce snail2 and foxd3 expression in a
non-cell-autonomous manner (Monsoro-Burq et al., 2005; Sato
et al., 2005). Downstream of Hairy2/Stat3 interactions, high Stat3
signaling activates Delta/Notch signaling which results in bmp4/id3
expression modulation and NC specication (Nichane et al., 2008a,
2008c, 2010). As described previously, modulation of several other
signaling pathways is involved in NC formation at the neurula stage.
These include the modulation of BMP and Wnt pathways by Tsukushi,
requirement of Snw1/SKIP for BMP signaling, Nodal and BMP signals
potentiated by TRAF4, modulation of Wnt signaling by positive and
negative regulators. In conclusion, neural border patterning sets the
scene for NC specication, by the induction and maintenance of
expression of essential transcription factors and signaling molecules
that will, in turn, activate NC speciers' gene expression.
Epigenetic regulation: jumonji and CDH7 control timing and access to
neural crest speciers' promoters
Recently, several epigenetic regulators have been found to play a
crucial role in NC induction, in Xenopus, chick and human. CHD7, an
ATP-dependent chromatin remodeler, is essential for multipotent
migratory neural crest formation, both in human embryonic stem
cells (hESCs) in vitro and in Xenopus embryos. CHD7, together with
BRG1 (a SWI/SNF family member), occupies distal regulatory
elements controlling the expression of critical NC speciers (SOX9
and TWIST) in human cells. Furthermore, they act synergistically in
vivo in the Xenopus embryo to regulate NC gene expression and migration (but not neural border formation) (Bajpai et al., 2010).
Another chromatin regulator, JumonjiD2A, regulates NC formation
in chick embryos. JMJD2A, a Histone Lysine Demethylase, is a subunit
of the PRC2 complex. Fine-tuned, dynamic regulation of H3K27me3
by JMJD2A and PRC2 balances stem cell and differentiation status in
mouse ESCs (Shen et al., 2009). In chick, JmjD2A loss of function
Although the function of NB speciers on NC formation during

human gastrulation cannot be directly studied, some NB speciers
are found mutated in neurocristopathies. Interestingly, human Zic3
can promote NC formation in Xenopus embryos (Chin et al., 2007)
and one of the mutated human alleles tested in this study has lost
its NC-inducing properties when over expressed. Pax3 is mutated in
the Waardenburg syndrome (WS type I and III), is responsible for
the mouse Splotch phenotype (Epstein et al., 1991; Tassabehji et al.,
1992), and is expressed in human premigratory NC (Betters et al.,
2010). However, no functional analysis of Pax3 on NC formation in
humans has been performed thus far.
Thanks to the development of protocols allowing hESCs manipulation,
it is now possible to induce the in vitro differentiation of hESCs into NC
cells (for a review see Chimge and Bayarsaihan, 2010; and Milet and
Monsoro-Burq, 2012this issue). Several genes involved in early NC specication are expressed in neuralized hESCs and in the derived NC-like
cells: ap2, pax3 and msx1 (Pomp et al., 2005) (Curchoe et al., 2010). This
model is of great interest to test the role of known NB specier genes
on the onset of human NC specication, or the role of genes that are
found mutated in neurocristopathies. For example, CHD7, important for
sox9 and twist induction, is mutated in the CHARGE syndrome (Bajpai
et al., 2010).
Conclusion and perspectives
In the last few years, studies of the earliest steps of neural crest
formation have revealed the complexity of the signaling and
transcriptional events taking place at the edge of the neural plate, as
early as gastrulation starts. This region, now dened as the neural
border in the young gastrula, is mapped in several vertebrate embryos,
and is marked by early molecular markers, the most specic being the
transcription factor pax3/7. The neural border includes progenitors for
the dorsal neural tube, the neural crest, placodes and ectoderm. The
neural border is rst induced by synergy between various secreted signals. It is then stabilized by interactions between several transcription
factors co-expressed in this region, the neural border speciers. Finally,
in coordination with additional secreted signaling pathways, neural
border speciers promote uncommitted neural crest progenitor proliferation on one hand, and induce the neural crest-specic regulators
on the other hand. These studies have provided a novel framework
that will allow building the detailed neural border gene regulatory network in the near future.
Acknowledgments
The authors are grateful to the members of the Monsoro-Burq
laboratory for their critical comments on the manuscript. Our research
is supported by grants from Ligue Contre le Cancer, Association pour
la Recherche contre le Cancer (SFI20101201882), CNRS, Universit
Paris Sud-11 (Attractivit), and Agence Nationale pour la Recherche to
A. M-B. C.M. is supported by fellowships from DIM Stem-Pole/Rgion
Ile-de-France and Universit Paris-Sud-11.
30
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Developmental Biology 366 (2012) 2233

Contents lists available at SciVerse ScienceDirect

Neural crest induction at the neural plate border in vertebrates

C. Milet, A.H. Monsoro-Burq / Developmental Biology 366 (2012) 2233

Ectoderm lateral and anterior

Neural folds corresponding to

Ectoderm of early gastrula

mNC in rostral sections,

Luo et al., 2003

Shen et al., 1997

Sauka-Spengler et al., 2007;

Thomas et al., 2008

Neural fold at st12.5

Not described in human early

Glavic et al., 2004

Myat et al., 1996

Williams et al., 1995

Sauka-Spengler et al., 2007

pNC in early neurula

Anterior region of the

DlxA, B, and C: Ventro-lateral

Feledy et al., 1999

Pera and Kessel, 1999

Robinson and Mahon, 1994

Sauka-Spengler et al., 2007

Not described in chick early

Lateral edges of the neural

Central part of the epiblast,

Gastrula (E6.5), neuroectoderm and underlying

Niss and Leutz, 1998

Bulfone et al., 1993, Byrd and

Not described in chick early

Ectoderm at the end of

Ectoderm except in prospective

Neural fold at st12.5-13

Not described in human early

Not described in human early

Not described in lamprey early

Not described in human early

Not described in mouse early

Not described in human early

Bellefroid et al., 1998

Sanchez-Guardado et al., 2011

Sauka-Spengler et al., 2007

confined dorsally to the lateral

MsxA: Ectoderm of gastrula

Suzuki et al., 1997

Suzuki et al., 1991

Hill et al., 1989

Sauka -Spengler et al., 2007;

dorsal mesoderm in gastrula

not described in chick early

tip of the neural folds at E8,

Sauka-Spengler et al., 2007

Bellefroid et al., 1998

Not described in human early

Yang et al., 1998

Glavic et al., 2004

Not described in human early

NT emigrating cells in culture,

Davidson and Hill, 1991

Lateral domains of the

Dorsal part of the