The retinoblastoma-related p130 protein is a member of a conserved family, consisting of Rb, p107 and p130, which are believed to play important roles in cell-cycle control and cellular differentiation. We have generated a null mutation in p130 by gene targeting and crossed the null allele into Balb/cJ and C57BL/6J strains of mice. In an enriched Balb/cJ genetic background, p130(−/−) embryos displayed arrested growth and died between embryonic days 11 and 13. Histological analysis revealed varying degrees of disorganization in neural and dermamyotomal structures. Immunohistochemistry with antibody reactive with Islet-1 indicated markedly reduced numbers of neurons in the spinal cord and dorsal root ganglia. Immunohistochemistry with antibody reactive with desmin indicated a similar reduction in the number of differentiated myocytes in the myotome. The myocardium of mutant embryos was abnormally thin and resembled an earlier staged two-chambered heart consisting of the bulbus cordis and the ventricular chamber. TUNEL analysis indicated the presence of extensive apoptosis in various tissues including the neural tube, the brain, the dermomyotome, but not the heart. Immunohistochemistry with antibody reactive with PCNA revealed increased cellular proliferation in the neural tube and the brain, and decreased proliferation in the heart. The placentas of p130(−/−) embryos did not display elevated apoptosis and were indistinguishable from wild type suggesting that the phenotype was not due to placental failure. Following a single cross with the C57BL/6 mice, p130(−/−) animals were derived that were viable and fertile. These results indicate that p130 in a Balb/cJ genetic background plays an essential role that is required for normal development. Moreover, our experiments establish that second-site modifier genes exist that have an epistatic relationship with p130.

REFERENCES

Bonyadi
M.
,
Rusholme
S. A.
,
Cousins
F. M.
,
Su
H. C.
,
Biron
C. A.
,
Farrall
M.
,
Akhurst
R. J.
(
1997
)
Mapping of a major genetic modifier of embryonic lethality in TGF beta 1 knockout mice.
Nat. Genet
15
,
207
11
Brehm
A.
,
Miska
E. A.
,
McCance
D. J.
,
Reid
J. L.
,
Bannister
A. J.
,
Kouzarides
T.
(
1998
)
Retinoblastoma protein recruits histone deacetylase to repress transcription.
Nature
391
,
597
601
Chen
P. L.
,
Riley
D. J.
,
Chen-Kiang
S.
,
Lee
W. H.
(
1996
)
Retinoblastoma protein directly interacts with and activates the transcription factor NF-IL6.
Proc. Natl. Acad. Sci. USA
93
,
465
9
Clarke
A. R.
,
Maandag
E. R.
,
van Roon
M.
,
van der Lugt
N. M.
,
van der Valk
M.
,
Hooper
M. L.
,
Berns
A.
,
te Riele
H.
(
1992
)
Requirement for a functional Rb-1 gene in murine development.
Nature
359
,
328
30
Cobrinik
D.
,
Lee
M. H.
,
Hannon
G.
,
Mulligan
G.
,
Bronson
R. T.
,
Dyson
N.
,
Harlow
E.
,
Beach
D.
,
Weinberg
R. A.
,
Jacks
T.
(
1996
)
Shared role of the pRB-related p130 and p107 proteins in limb development.
Genes Dev
10
,
1633
44
Corbeil
H. B.
,
Whyte
P.
,
Branton
P. E.
(
1995
)
Characterization of transcription factor E2F complexes during muscle and neuronal differentiation.
Oncogene
11
,
909
20
Ericson
J.
,
Thor
S.
,
Edlund
T.
,
Jessell
T. M.
,
Yamada
T.
(
1992
)
Early stages of motor neuron differentiation revealed by expression of homeobox gene Islet-1.
Science
256
,
1555
60
Flink
I. L.
,
Oana
S.
,
Maitra
N.
,
Bahl
J. J.
,
Morkin
E.
(
1998
)
Changes in E2F complexes containing retinoblastoma protein family members and increased cyclin-dependent kinase inhibitor activities during terminal differentiation of cardiomyocytes.
J. Mol. Cell Cardiol
30
,
563
78
Gavrieli
Y.
,
Sherman
Y.
,
Ben-Sasson
S. A.
(
1992
)
Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation.
J. Cell Biol
119
,
493
501
George
E. L.
,
Georges-Labouesse
E. N.
,
Patel-King
R. S.
,
Rayburn
H.
,
Hynes
R. O.
(
1993
)
Defects in mesoderm, neural tube and vascular development in mouse embryos lacking fibronectin.
Development
119
,
1079
91
Herrera
R. E.
,
Sah
V. P.
,
Williams
B. O.
,
Makela
T. P.
,
Weinberg
R. A.
,
Jacks
T.
(
1996
)
Altered cell cycle kinetics, gene expression, and G1 restriction point regulation in Rb-deficient fibroblasts.
Mol. Cell Biol
16
,
2402
7
Hiebert
S. W.
,
Packham
G.
,
Strom
D. K.
,
Haffner
R.
,
Oren
M.
,
Zambetti
G.
,
Cleveland
J. L.
(
1995
)
E2F-1:DP-1 induces p53 and overrides survival factors to trigger apoptosis.
Mol. Cell Biol
15
,
6864
6874
Hirsinger
E.
,
Duprez
D.
,
Jouve
C.
,
Malapert
P.
,
Cooke
J.
,
Pourquie
O.
(
1997
)
Noggin acts downstream of Wnt and Sonic Hedgehog to antagonize avian somite patterning.
Development
124
,
4605
4614
Howes
K. A.
,
Ransom
N.
,
Papermaster
D. S.
,
Lasudry
J. G. H.
,
Albert
D. M.
,
Windle
J. J.
(
1994
)
Apoptosis or retinoblastoma: alternative fates of photoreceptors expressing the HPV-16 E7 gene in the presence or absence of p53.
Genes Dev
8
,
1300
1310
Hurford
R. K.
Jr.
,
Cobrinik
D.
,
Lee
M. H.
,
Dyson
N.
(
1997
)
pRB and p107/p130 are required for the regulated expression of different sets of E2F responsive genes.
Genes Dev
11
,
1447
63
Jacks
T.
,
Fazeli
A.
,
Schmitt
E. M.
,
Bronson
R. T.
,
Goodell
M. A.
,
Weinberg
R. A.
(
1992
)
Effects of an Rb mutation in the mouse.
Nature
359
,
295
300
Kablar
B.
,
Krastel
K.
,
Ying
C.
,
Asakura
A.
,
Tapscott
S. J.
,
Rudnicki
M. A.
(
1997
)
MyoD and Myf-5 differentially regulate the development of limb versus trunk skeletal muscle.
Development
124
,
4729
38
LeCouter
J. E.
,
Whyte
P. F.
,
Rudnicki
M. A.
(
1996
)
Cloning andexpression of the Rb-related mouse p130 mRNA.
Oncogene
12
,
1433
40
Lee
E. Y.
,
Chang
C. Y.
,
Hu
N.
,
Wang
Y. C.
,
Lai
C. C.
,
Herrup
K.
,
Lee
W. H.
,
Bradley
A.
(
1992
)
Mice deficient for Rb are nonviable and show defects in neurogenesis and haematopoiesis.
Nature
359
,
288
94
Lee
E. Y.
,
Hu
N.
,
Yuan
S. S.
,
Cox
L. A.
,
Bradley
A.
,
Lee
W. H.
,
Herrup
K.
(
1994
)
Dual roles of the retinoblastoma protein in cell cycle regulation and neuron differentiation.
Genes Dev
8
,
2008
21
Lee
M. H.
,
Williams
B. O.
,
Mulligan
G.
,
Mukai
S.
,
Bronson
R. T.
,
Dyson
N.
,
Harlow
E.
,
Jacks
T.
(
1996
)
Targeted disruption of p107: functional overlap between p107 and Rb.
Genes Dev
10
,
1621
32
Liu
J. P.
,
Baker
J.
,
Perkins
A. S.
,
Robertson
E. J.
,
Efstratiadis
A.
(
1993
)
Mice carrying null mutations of the genes encoding insulin-like growth factor I (Igf-1) and type 1 IGF receptor (Igf1r).
Cell
75
,
59
72
Luo
R. X.
,
Postigo
A. A.
,
Dean
D. C.
(
1998
)
Rb interacts with histone deacetylase to repress transcription.
Cell
92
,
463
73
Maandag
E. C.
,
van der Valk
M.
,
Vlaar
M.
,
Feltkamp
C.
,
O'Brien
J.
,
van Roon
M.
,
van der Lugt
N.
,
Berns
A.
,
te Riele
H.
(
1994
)
Developmental rescue of an embryonic-lethal mutation in the retinoblastoma gene in chimeric mice.
EMBO J
13
,
4260
8
Macleod
K. F.
,
Hu
Y.
,
Jacks
T.
(
1996
)
Loss of Rb activates both p53-dependent and independent cell death pathways in the developing mouse nervous system.
EMBO J
15
,
6178
88
Magnaghi-Jaulin
L.
,
Groisman
R.
,
Naguibneva
I.
,
Robin
P.
,
Lorain
S.
,
Le Villain
J. P.
,
Troalen
F.
,
Trouche
D.
,
Harel-Bellan
A.
(
1998
)
Retinoblastoma protein represses transcription by recruiting a histone deacetylase.
Nature
391
,
601
5
Marcelle
C.
,
Stark
M. R.
,
Bronner-Fraser
M.
(
1997
)
Coordinate actions of BMPs, Wnts, Shh and noggin mediate patterning of the dorsal somite.
Development
124
,
3955
3963
McBurney
M. W.
,
Sutherland
L. C.
,
Adra
C. N.
,
Leclair
B.
,
Rudnicki
M. A.
,
Jardine
K.
(
1991
)
The mouse Pgk-1 gene promoter contains an upstream activator sequence.
Nucleic Acids Res
19
,
5755
61
Megeney
L. A.
,
Kablar
B.
,
Garrett
K.
,
Anderson
J. E.
,
Rudnicki
M. A.
(
1996
)
MyoD is required for myogenic stem cell function in adult skeletal muscle.
Genes Dev
10
,
1173
83
Morgenbesser
S. D.
,
Williams
B. O.
,
Jacks
T.
,
DePinho
R. A.
(
1994
)
p53-dependent apoptosis produced by Rb-deficiency in the developing mouse lens.
Nature
371
,
72
4
Muller
R.
(
1995
)
Transcriptional regulation during the mammalian cell cycle.
Trends Genet
11
,
173
8
Munsterberg
A. E.
,
Kitajewski
J.
,
Bumcrot
D. A.
,
McMahon
A. P.
,
Lassar
A. B.
(
1995
)
Combinatorial signaling by Sonic hedgehog and Wnt family members induces myogenic bHLH gene expression in the somite.
Genes Dev
9
,
2911
22
Nevins
J. R.
,
Leone
G.
,
DeGregori
J.
,
Jakoi
L.
(
1997
)
Role of the Rb/E2F pathway in cell growth control.
J. Cell Physiol
173
,
233
6
Pan
H.
,
Griep
A. E.
(
1994
)
Altered cell cycle regulation in the lens of HPV-16 E6 or E7 transgenic mice: implications for tumor suppressor gene function in development.
Genes Dev
8
,
1285
1299
Phillips
A. C.
,
Bates
S.
,
Ryan
K. M.
,
Helin
K.
,
Vousden
K. H.
(
1997
)
Induction of DNA systhesis and apoptosis are separable functions of E2F-1.
Genes Dev
11
,
1853
1863
Proetzel
G.
,
Pawlowski
S. A.
,
Wiles
M. V.
,
Yin
M.
,
Boivin
G. P.
,
Howles
P. N.
,
Ding
J.
,
Ferguson
M. W.
,
Doetschman
T.
(
1995
)
Transforming growth factor-beta 3 is required for secondary palate fusion.
Nat. Genet
11
,
409
14
Puri
P. L.
,
Balsano
C.
,
Burgio
V. L.
,
Chirillo
P.
,
Natoli
G.
,
Ricci
L.
,
Mattei
E.
,
Graessmann
A.
,
Levrero
M.
(
1997
)
MyoD prevents cyclinA/cdk2 containing E2F complexes formation in terminally differentiated myocytes.
Oncogene
14
,
1171
84
Qin
X. Q.
,
Livingston
D. M.
,
Kaelin
W. G.
Jr.
,
Adams
P. D.
(
1994
)
Deregulated transcription factor E2F-1 expression leads to S-phase entry and p53-mediated apoptosis.
Proc. Natl. Acad. Sci. USA
91
,
10918
10922
Raschella
G.
,
Tanno
B.
,
Bonetto
F.
,
Amendola
R.
,
Battista
T.
,
De Luca
A.
,
Giordano
A.
,
Paggi
M. G.
(
1997
)
Retinoblastoma-related protein pRb2/p130 and its binding to the B-myb promoter increase during human neuroblastoma differentiation.
J. Cell Biochem
67
,
297
303
Reshef
R.
,
Maroto
M.
,
Lassar
A. B.
(
1998
)
Regulation of dorsalsomitic cell fates: BMPs and Noggin control the timing and pattern of myogenic factor expression.
Genes Dev
12
,
290
303
Rohrer
D. K.
,
Desai
K. H.
,
Jasper
J. R.
,
Stevens
M. E.
,
Regula
D. P.
Jr.
,
Barsh
G. S.
,
Bernstein
D.
,
Kobilka
B. K.
(
1996
)
Targeted disruption of the mouse beta1-adrenergic receptor gene: developmental and cardiovascular effects.
Proc. Natl Acad. Sci. USA
93
,
7375
80
Rozmahel
R.
,
Wilschanski
M.
,
Matin
A.
,
Plyte
S.
,
Oliver
M.
,
Auerbach
W.
,
Moore
A.
,
Forstner
J.
,
Durie
P.
,
Nadeau
J.
, et al. 
(
1996
)
Modulation of disease severity in cystic fibrosis transmembrane conductance regulator deficient mice by a secondary genetic factor.
Nat. Genet
12
,
280
7
Rudnicki
M. A.
,
Braun
T.
,
Hinuma
S.
,
Jaenisch
R.
(
1992
)
Inactivation of MyoD in mice leads to up-regulation of the myogenic HLH gene Myf-5 and results in apparently normal muscle development.
Cell
71
,
383
90
Shan
B.
,
Lee
W. H.
(
1994
)
Deregulated expression of E2F-1 induces S-phase entry and leads to apoptosis.
Mol. Cell Biol
14
,
8166
73
Shin
E. K.
,
Shin
A.
,
Paulding
C.
,
Schaffhausen
B.
,
Yee
A. S.
(
1995
)
Multiple change in E2F function and regulation occur upon muscle differentiation.
Mol. Cell Biol
15
,
2252
62
Sibilia
M.
,
Wagner
E. F.
(
1995
)
Strain-dependent epithelial defects in mice lacking the EGF receptor.
Science
269
,
234
8
Tanabe
Y.
,
Roelink
H.
,
Jessell
T. M.
(
1995
)
Induction of motor neurons by Sonic hedgehog is independent of floor plate differentiation.
Current Biol
5
,
651
8
Threadgill
D. W.
,
Dlugosz
A. A.
,
Hansen
L. A.
,
Tennenbaum
T.
,
Lichti
U.
,
Yee
D.
,
LaMantia
C.
,
Mourton
T.
,
Herrup
K.
,
Harris
R. C.
, et al. 
(
1995
)
Targeted disruption of mouse EGF receptor: effect of genetic background on mutant phenotype.
Science
269
,
230
4
Tsuchida
T.
,
Ensini
M.
,
Morton
S. B.
,
Baldassare
M.
,
Edlund
T.
,
Jessell
T. M.
,
Pfaff
S. L.
(
1994
)
Topographic organization of embryonic motor neurons defined by expression of LIM homeobox genes.
Cell
79
,
957
970
Vairo
G.
,
Livingston
D. M.
,
Ginsberg
D.
(
1995
)
Functional interaction between E2F-4 and p130: evidence for distinct mechanisms underlying growth suppression by different retinoblastoma protein family members.
Genes Dev
9
,
869
81
Wang
J.
,
Guo
K.
,
Wills
K. N.
,
Walsh
K.
(
1997
)
Rb functions to inhibit apoptosis during myocyte differentiation.
Cancer Res
57
,
351
4
Weinberg
R. A.
(
1995
)
The retinoblastoma protein and cell cycle control.
Cell
81
,
323
30
Whyte
P.
(
1995
)
The retinoblastoma protein and its relatives.
Semin. Cancer Biol
6
,
83
90
Wiggan
O.
,
Taniguchi-Sidle
A.
,
Hamel
P. A.
(
1997
)
Interaction of the pRB-family proteins with factors containing paired-like homeodomains.
Oncogene
16
,
227
236
Williams
B. O.
,
Schmitt
E. M.
,
Remington
L.
,
Bronson
R. T.
,
Albert
D. M.
,
Weinberg
R. A.
,
Jacks
T.
(
1994
)
Extensive contribution of Rb-deficient cells to adult chimeric mice with limited histopathological consequences.
EMBO J
13
,
4251
9
Yamada
T.
,
Pfaff
S. L.
,
Edlund
T.
,
Jessell
T. M.
(
1993
)
Control of cell pattern in the neural tube: motor neuron induction by diffusible factors from notochord and floor plate.
Cell
73
,
673
86
Yamada
T.
,
Placzek
M.
,
Tanaka
H.
,
Dodd
J.
,
Jessell
T. M.
(
1991
)
Control of cell pattern in the developing nervous system: polarizing activity of the floor plate and notochord.
Cell
64
,
635
47
Zacksenhaus
E.
,
Jiang
Z.
,
Chung
D.
,
Marth
J. D.
,
Phillips
R. A.
,
Gallie
B. L.
(
1996
)
pRb controls proliferation, differentiation, and death of skeletal muscle cells and other lineages during embryogenesis.
Genes Dev
10
,
3051
3064
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