Recent gene targeting studies have revealed unexpected roles for endothelins in the development of neural crest-derived tissues. Endothelin converting enzyme-1 (ECE-1) catalyzes the proteolytic activation of big endothelin-1 to endothelin-1(ET-1) in vitro. However, the importance of ECE-1 cleavage in the multiple endothelin pathways in vivo is unknown. Here we generated a targeted null mutation in the mouse ECE-1 gene. ECE-1−/− term embryos exhibited craniofacial and cardiac abnormalities virtually identical to the defects seen in ET-1 and endothelin A receptor (ETA)-deficient embryos. Epidermal melanocytes as well as enteric neurons of the distal gut were also absent in ECE-1−/− embryos, reproducing the developmental phenotype seen in ET-3−/− and endothelin B receptor (ETB)−/− mice. Surprisingly, large amounts of mature ET-1 peptide are found in ECE-1−/− embryos, indicating that non-ECE-1 protease(s) can activate ET-1 at certain sites. However, these enzymes cannot produce sufficient mature endothelin at the locations crucial for normal embryonic development. These findings reveal that ECE-1 is a bona fide activating protease for both big ET-1 and big ET-3 in vivo, and that the cell-cell communication pathways represented by the ET-1/ECE-1/ETA axis and the ET-3/ECE-1/ETB axis are each involved in the development of distinct subsets of neural crest cell lineages. Mutations in ECE-1 may cause developmental defects in humans, such as Hirschsprung disease, velocardiofacial syndrome and related neurocristopathies.

REFERENCES

Arai
H.
,
Hori
S.
,
Aramori
I.
,
Ohkubo
H.
,
Nakanishi
S.
(
1990
)
Cloning and expression of a cDNA encoding an endothelin receptor.
Nature
348
,
730
732
Baynash
A. G.
,
Hosoda
K.
,
Giaid
A.
,
Richardson
J. A.
,
Emoto
N.
,
Hammer
R. E.
,
Yanagisawa
M.
(
1994
)
Interaction of endothelin-3 with endothelin-B receptor is essential for development of epidermal melanocytes and enteric neurons.
Cell
79
,
1277
1285
Benjamin
I. J.
,
Shelton
J.
,
Garry
D. J.
,
Richardson
J. A.
(
1997
)
Temporospatial expression of the small HSP/B-crystallin in cardiac and skeletal muscle during mouse development.
Dev. Dyn
208
,
75
84
Brannan
C. I.
,
Perkins
A. S.
,
Vogal
K. S.
,
Ratner
N.
,
Nordlund
M. L.
,
Reid
S. W.
,
Buchberg
A. M.
,
Jenkins
N. A.
,
Parada
L. F.
,
Copeland
N. G.
(
1994
)
Targeted disruption of the neurofibromatosis type-1 gene leads to developmental abnormalities in heart and various neural crest-derived tissues.
Gene Dev
8
,
1019
1029
Choi
D.-S.
,
Ward
S. J.
,
Messaddeq
N.
,
Launay
J.-M.
,
Maroteaux
L.
(
1997
)
5-HT2B receptor-mediated serotonin morphogenetic functions in mouse cranial neural crest and myocardiac cells.
Development
124
,
1745
1755
Conway
S. J.
,
Henderson
D. J.
,
Copp
A. J.
(
1997
)
Pax3 is required for cardiac neural crest migration in the mouse: evidence from the sploth (SP2h) mutant.
Development
124
,
505
514
Clouthier
D. E.
,
Hosoda
K.
,
Richardson
J. A.
,
Williams
S. C.
,
Yanagisawa
H.
,
Kuwaki
T.
,
Kumada
M.
,
Hammer
R. E.
,
Yanagisawa
M.
(
1998
)
Cranial and cardiac neural crest defects in endothelin-A receptor-deficient mice.
Development
125
,
813
824
Donovan
M. J.
,
Hahn
R.
,
Tessarollo
L.
,
Hempstead
B. L.
(
1996
)
Identification of an essential nonneuronal function of neurotrophin 3 in mammalian cardiac development.
Nature Genet
14
,
210
213
Durbec
P. L.
,
Larsson-Blumberg
L. B.
,
Schuchardt
A.
,
Constantini
F.
,
Pachnis
V.
(
1996
)
Common origin and developmental dependence on c-ret of subsets of enteric and sympathetic neuroblasts.
Development
122
,
349
358
Edery
P.
,
Attie
T.
,
Amiel
J.
,
Pelet
A.
,
Eng
C.
,
Hofstra
R. M. W.
,
Martelli
H.
,
Bidaud
C.
,
Munnich
A.
,
Lyonnet
S.
(
1996
)
Mutation of the endothelin-3 gene in the Waardenburg-Hirschsprung disease (Shah-Waardenburg syndrome).
Nature Genet
12
,
442
444
Emoto
N.
,
Yanagisawa
M.
(
1995
)
Endothelin-converting enzyme-2 is a membrane-bound, phosphoramidon-sensitive metalloprotease with acidic pH optimum.
J. Biol. Chem
270
,
15262
15268
Epstein
M. L.
,
Mikawa
T.
,
Brown
A. M. C.
,
McFarlin
D. R.
(
1994
)
Mapping the origin of the avian enteric nervous system with a retroviral marker.
Dev. Dyn
201
,
236
244
Erickson
C. A.
(
1993
)
From the crest to the periphery: control of pigment cell migration and lineage segregation.
Pigment Cell Res
6
,
336
347
Franz
T.
(
1993
)
The Splotch (Sp1H) and Splotch-delayed (Spd) alleles: differential phenotypic effects on neural crest and limb musculature.
Anat. Embryol
187
,
371
377
Gassmann
M.
,
Casagranda
F.
,
Orioli
D.
,
Simon
H.
,
Lai
C.
,
Klein
R.
,
Lemke
G.
(
1995
)
Aberrant neural and cardiac development in mice lacking the ErbB4 neuregulin receptor.
Nature
378
,
390
394
Hellmich
H. L.
,
Kos
L.
,
Cho
E. S.
,
Mahon
K. A.
,
Zimmer
A.
(
1996
)
Embryonic expression of glial cell-line derived neurotrophic factor (GDNF) suggests multiple developmental roles in neural differentiation and epithelial-mesenchymal interactions.
Mech. Dev
54
,
95
105
Hofstra
R. M. W.
,
Osinga
J.
,
Tan-Sindhunata
G.
,
Wu
Y.
,
Kamsteeg
E.-J.
,
Stulp
R. P.
,
Ravenswaaij-Arts
C. V.
,
Majoor-Krakauer
D.
,
Angrist
M.
,
Chakravarti
A.
,
Meijers
C.
,
Buys
C. H. C. M.
(
1996
)
A homozygous mutation in the endothelin-3 gene associated with a combined Waardenburg type 2 and Hirschsprung phenotype (Shah-Waardenburg syndrome).
Nature Genet
12
,
445
447
Hosoda
K.
,
Hammer
R. E.
,
Richardson
J. A.
,
Baynash
A. G.
,
Cheung
J. C.
,
Giaid
A.
,
Yanagisawa
M.
(
1994
)
Targeted and natural (piebald-lethal) mutations of endothelin-B receptor gene produce megacolon associated with spotted coat color in mice.
Cell
79
,
1267
1276
Inoue
A.
,
Yanagisawa
M.
,
Kimura
S.
,
Kasuya
Y.
,
Miyauchi
T.
,
Goto
K.
,
Masaki
T.
(
1989
)
The human endothelin family: Three structurally and pharmacologically distinct isopeptides predicted by three separate genes.
Proc. Natl. Acad. Sci. USA
86
,
2863
2867
Ishibashi
S.
,
Brown
M. S.
,
Goldstein
J. L.
,
Gerard
R. D.
,
Hammer
R. E.
,
Herz
J.
(
1993
)
Hypercholesterolemia in low density lipoprotein receptor knockout mice and its reversal by adenovirus-mediated gene delivery.
J. Clin. Invest
92
,
883
893
Kapur
R. P.
,
Hoyle
G. W.
,
Mercer
E. H.
,
Brinster
R. L.
,
Palmiter
R. D.
(
1991
)
Some neuronal cell populations express human dopamine-hydroxylase- lacZ transgenes transiently during embryonic development.
Neuron
7
,
717
727
Kirby
M. L.
,
Gale
T. F.
,
Stewart
D. E.
(
1983
)
Neural crest cells contribute to normal aorticopulmonary septation.
Science
220
,
1059
1061
Kirby
M. L.
,
Waldo
K. L.
(
1995
)
Neural crest and cardiovascular patterning.
Circ. Res
77
,
211
215
Kochhar
D. M.
(
1973
)
Limb development in mouse embryos. I. Analysis of teratogenic effects of retinoic acid.
Teratology
7
,
289
298
Köntges
G.
,
Lumsden
A.
(
1996
)
Rhombencephalic neural crest segmentation is preserved throughout craniofacial ontogeny.
Development
122
,
3229
3242
Kuratani
S. C.
,
Kirby
M. L.
(
1991
)
Initial migration and distribution of the cardiac neural crest in the avian embryo: an introduction to the concept of the circumpharyngeal crest.
Am. J. Anat
191
,
215
227
Kurihara
Y.
,
Kurihara
H.
,
Oda
H.
,
Maemura
K.
,
Nagai
R.
,
Ishikawa
T.
,
Yazaki
Y.
(
1995
)
Aortic arch malformations and ventricular septal defect in mice deficient in endothelin-1.
J. Clin. Invest
96
,
293
300
Kurihara
Y.
,
Kurihara
H.
,
Suzuki
H.
,
Kodama
T.
,
Maemura
K.
,
Nagai
R.
,
Oda
H.
,
Kuwaki
T.
,
Cao
W.-H.
,
Kamada
N.
,
Jishage
K.
,
Ouchi
Y.
,
Azuma
S.
,
Toyoda
Y.
,
Ishikawa
T.
,
Kumada
M.
,
Yazaki
Y.
(
1994
)
Elevated blood pressure and craniofacial abnormalities in mice deficient in endothelin-1.
Nature
368
,
703
710
Le Douarin
N. M.
,
Dupin
E.
,
Ziller
C.
(
1994
)
Genetic and epigenetic control in neural crest development.
Curr. Opin. Genet. Dev
4
,
685
695
Le Douarin
N. M.
,
Ziller
C.
,
Couly
G. F.
(
1993
)
Patterning of neural crest derivatives in the avian embryo: In vivo and in vitro studies.
Dev. Biol
159
,
24
49
Le Lievre
C. S.
,
Le Douarin
N. M.
(
1975
)
Mesenchymal derivatives of the neural crest: analysis of chimaeric quail and chick embryos.
J. Embryol. Exp. Morph
34
,
125
154
Lee
K.-F.
,
Simon
H.
,
Chen
H.
,
Bates
B.
,
Hung
M.-C.
,
Hauser
C.
(
1995
)
Requirement for neuregulin receptor erbB2 in neural and cardiac development.
Nature
378
,
394
398
Loring
J. F.
,
Erickson
C. A.
(
1987
)
Neural crest cell migratory pathways in the trunk of the chick embryo.
Dev. Biol
121
,
220
236
Maemura
K.
,
Kurihara
H.
,
Kurihara
Y.
,
Oda
H.
,
Ishikawa
T.
,
Copeland
N. G.
,
Gilbert
D. J.
,
Jenkins
N. A.
,
Yazaki
Y.
(
1996
)
Sequence analysis, chromosomal location, and developmental expression of the mouse preproendothelin-1 gene.
Genomics
31
,
177
184
Matsumoto
H.
,
Suzuki
N.
,
Onda
H.
,
Fujino
M.
(
1989
)
Abundance of endothelin-3 in rat intestine, pituitary glands and brain.
Biochem. Biophys. Res. Commun
164
,
74
80
Mendelsohn
C.
,
Lohnes
D.
,
Decimo
D.
,
Lufkin
T.
,
LeMeur
M.
,
Chambon
P.
,
Mark
M.
(
1994
)
Function of the retinoic acid receptors (RARs) during development(II). Multiple abnormalities at various stages of organogenesis in RAR double mutants.
Development
120
,
2749
2771
Meyer
D.
,
Birchmeier
C.
(
1995
)
Mutiple essential functions of neuregulin in development.
Nature
378
,
386
390
Moore
M. W.
,
Klein
R. D.
,
Fariñas
I.
,
Sauer
H.
,
Armanini
M.
,
Phillips
H.
,
Reichardt
L. F.
,
Ryan
A. M.
,
Carver-Moore
K.
,
Rosenthal
A.
(
1996
)
Renal and neuronal abnormalities in mice lacking GDNF.
Nature
382
,
76
79
Nishibatake
M.
,
Kirby
M. L.
,
Van Mierop
L. H. S.
(
1987
)
Pathogenesis of persistent truncus arteriosus and dextroposed aorta in the chick embryo after neural crest ablation.
Circulation
75
,
255
264
Offermanns
S.
,
Mancino
V.
,
Revel
J.-P.
,
Simon
M. I.
(
1997
)
Vascular system defects and impaired cell chemokinesis as a result of G13deficiency.
Science
275
,
533
536
Oh
S. P.
,
Li
E.
(
1997
)
The signaling pathway mediated by the type IIB activin receptor controls axial patterning and lateral asymmetry in the mouse.
Gene Dev
11
,
1812
1826
Pachnis
V.
,
Mankoo
B.
,
Costantini
F.
(
1993
)
Expression of the c-ret proto-oncogene during mouse embryogenesis.
Development
119
,
1005
1017
Pichel
J. G.
,
Shen
L.
,
Sheng
H. Z.
,
Granholm
A.-C.
,
Drago
J.
,
Grinberg
A.
,
Lee
E. J.
,
Huang
S. P.
,
Saarma
M.
,
Hoffer
B. J.
,
Sariola
H.
,
Westphal
H.
(
1996
)
Defects in enteric innervation and kidney development in mice lacking GDNF.
Nature
382
,
73
76
Puffenberger
E. G.
,
Hosoda
K.
,
Washington
S. S.
,
Nakao
K.
,
deWit
D.
,
Yanagisawa
M.
,
Chakravarti
A.
(
1994
)
A missense mutation of the endothelin-B receptor gene in multigenic Hirschsprung disease.
Cell
79
,
1257
1266
Rawlings
N. D.
,
Barrett
A. J.
(
1993
)
Evolutionary families of peptidases.
Biochem. J
290
,
205
218
Rodriguez
H. A.
,
McGavran
M. H.
(
1969
)
A modified dopa reaction for the diagnosis and investigation of pigment cells.
Am. J. Clin. Pathol
52
,
219
227
Sakurai
T.
,
Yanagisawa
M.
,
Takuwa
Y.
,
Miyazaki
H.
,
Kimura
S.
,
Goto
K.
,
Masaski
T.
(
1990
)
Cloning of a cDNA encoding a non-isopeptide-selective subtype of the endothelin receptor.
Nature
348
,
732
735
Sánchez
M. P.
,
Silos-Santiago
I.
,
Frisen
J.
,
He
B.
,
Lira
S. A.
,
Barbacid
M.
(
1996
)
Renal agenesis and the absence of enteric neurons in mice lacking GDNF.
Nature
382
,
70
73
Schuchardt
A.
,
D'Agati
V.
,
Larsson-Blomberg
L.
,
Costantini
F.
,
Pachnis
V.
(
1994
)
Defects in the kidney and enteric nervous system of mice lacking the tyrosine kinase receptor Ret.
Nature
367
,
380
383
Serbedzija
G. N.
,
Bronner-Fraser
M.
,
Fraser
S. E.
(
1992
)
Vital dye analysis of cranial neural crest cell migration in the mouse embryo.
Development
116
,
297
307
Serbedzija
G. N.
,
McMahon
A. P.
(
1997
)
Analysis of neural crest cell migration in Splotch mice using a neural crest-specific lacZ reporter.
Dev. Biol
185
,
139
147
Shah
N. M.
,
Groves
A. K.
,
Anderson
D. J.
(
1996
)
Alternative neural crest cell fates are instructively promoted by TGFsuperfamily members.
Cell
85
,
331
343
Stemple
D. L.
,
Anderson
D. J.
(
1993
)
Lineage diversification of the neural crest: in vitro investigations.
Dev. Biol
159
,
12
23
Suzuki
N.
,
Matsumoto
H.
,
Kitada
C.
,
Kimura
S.
,
Miyauchi
T.
,
Fujino
M.
(
1990
)
A sandwich-type enzyme immunoassay to detect immunoreactive big-endothelin-1 in plasma.
J. Immunol. Meth
127
,
165
170
Trainor
P. A.
,
Tam
P. P. L.
(
1995
)
Cranial paraxial mesoderm and neural crest cells of the mouse embryo: co-distribution in the craniofacial mesenchyme but distinct segregation in branchial arches.
Development
121
,
2569
2582
Van Mierop
L. H. S.
,
Kutsche
L. M.
(
1984
)
Interruption of the aortic arch and coarctation of the aorta: Pathogenetic relations.
Am. J. Cardiol
54
,
829
834
Wilson
D. I.
,
Burn
J.
,
Scambler
P.
,
Goodship
J.
(
1993
)
DiGeorge syndrome: part of CATCH 22.
J. Med. Genet
30
,
852
856
Xu
D.
,
Emoto
N.
,
Giaid
A.
,
Slaughter
C. A.
,
Kaw
S.
,
de Wit
D.
,
Yanagisawa
M.
(
1994
)
ECE-1: A membrane-bound metalloprotease that catalyzes the proteolytic activation of big endothelin-1.
Cell
78
,
473
485
Yanagisawa
M.
(
1994
)
The endothelin system: A new target for therapeutic intervention.
Circulation
89
,
1320
1322
This content is only available via PDF.