The transcription factor HNF3beta is required in visceral endoderm for normal primitive streak morphogenesis

Development ◽  
1998 ◽  
Vol 125 (16) ◽  
pp. 3015-3025 ◽  
Author(s):  
D. Dufort ◽  
L. Schwartz ◽  
K. Harpal ◽  
J. Rossant
Keyword(s):  
Transcription Factor ◽  
Primitive Streak ◽  
Chimeric Mouse ◽  
Visceral Endoderm ◽  
Wild Type ◽  
Targeted Mutation ◽  
Notochord Cell ◽  
Cell Aggregations ◽  
Left Right Asymmetry ◽  
Embryonic Ectoderm

During early embryogenesis, the transcription factor HNF3beta is expressed in visceral and definitive endoderm, node, notochord and floorplate. A targeted mutation in the HNF3β gene results in the lack of a definitive node and notochord. Furthermore, lack of HNF3beta results in failure of proper primitive streak elongation. To address whether HNF3beta is required in visceral endoderm, we have used tetraploid embryo-ES cell aggregations to generate chimeric mouse embryos with wild-type visceral endoderm and homozygous mutant HNF3beta embryonic ectoderm or vice versa. Replacing the visceral endoderm of mutant HNF3beta embryos rescued proper primitive streak elongation and, conversely, mutant visceral endoderm imposed a severe embryonic-extraembryonic constriction on wild-type embryonic ectoderm. Restoration of normal streak morphogenesis was not sufficient to allow formation of the node and notochord in HNF3beta mutant embryos. Thus, our results demonstrate that HNF3beta has two separate roles in primitive streak formation. One is to act within the visceral endoderm to promote proper streak morphogenesis. The second is autonomous to the node and its precursors and involves specification of node and notochord cell fates. HNF3beta mutant embryos rescued for the embryonic-extraembryonic constriction developed further than mutant embryos, allowing examination of later roles for HNF3beta. We show that such mutant embryos lack foregut and midgut endoderm. In addition, left-right asymmetry is affected in the mutant embryos.

Lim1 is required in both primitive streak-derived tissues and visceral endoderm for head formation in the mouse

Development ◽  
1999 ◽  
Vol 126 (22) ◽  
pp. 4925-4932 ◽  
Author(s):  
W. Shawlot ◽  
M. Wakamiya ◽  
K.M. Kwan ◽  
A. Kania ◽  
T.M. Jessell ◽  
...  
Keyword(s):  
Marker Gene ◽  
Embryonic Stem ◽  
Homeobox Gene ◽  
Primitive Streak ◽  
Mouse Embryos ◽  
Chimeric Mouse ◽  
Visceral Endoderm ◽  
Wild Type ◽  
Targeted Mutation ◽  
Head Formation

Lim1 is a homeobox gene expressed in the extraembryonic anterior visceral endoderm and in primitive streak-derived tissues of early mouse embryos. Mice homozygous for a targeted mutation of Lim1 lack head structures anterior to rhombomere 3 in the hindbrain. To determine in which tissues Lim1 is required for head formation and its mode of action, we have generated chimeric mouse embryos and performed tissue layer recombination explant assays. In chimeric embryos in which the visceral endoderm was composed of predominantly wild-type cells, we found that Lim1(−)(/)(−) cells were able to contribute to the anterior mesendoderm of embryonic day 7.5 chimeric embryos but that embryonic day 9.5 chimeric embryos displayed a range of head defects. In addition, early somite stage chimeras generated by injecting Lim1(−)(/)(−) embryonic stem cells into wild-type tetraploid blastocysts lacked forebrain and midbrain neural tissue. Furthermore, in explant recombination assays, anterior mesendoderm from Lim1(−)(/)(−) embryos was unable to maintain the expression of the anterior neural marker gene Otx2 in wild-type ectoderm. In complementary experiments, embryonic day 9.5 chimeric embryos in which the visceral endoderm was composed of predominantly Lim1(−)(/)(−) cells and the embryo proper of largely wild-type cells, also phenocopied the Lim1(−)(/)(−) headless phenotype. These results indicate that Lim1 is required in both primitive streak-derived tissues and visceral endoderm for head formation and that its inactivation in these tissues produces cell non-autonomous defects. We discuss a double assurance model in which Lim1 regulates sequential signaling events required for head formation in the mouse.

Insufficient VEGFA activity in yolk sac endoderm compromises haematopoietic and endothelial differentiation

Development ◽  
2002 ◽  
Vol 129 (8) ◽  
pp. 1881-1892 ◽  
Author(s):  
Annette Damert ◽  
Lucile Miquerol ◽  
Marina Gertsenstein ◽  
Werner Risau ◽  
Andras Nagy
Keyword(s):  
Yolk Sac ◽  
Endothelial Differentiation ◽  
Vascular Endothelial ◽  
Visceral Endoderm ◽  
Wild Type ◽  
Hypomorphic Allele ◽  
Targeted Mutation ◽  
Blood Formation ◽  
Opposite Situation ◽  
Endothelial Growth Factor

Vascular endothelial growth factor A (VEGFA) plays a pivotal role in the first steps of endothelial and haematopoietic development in the yolk sac, as well as in the establishment of the cardiovascular system of the embryo. At the onset of gastrulation, VEGFA is primarily expressed in the yolk sac visceral endoderm and in the yolk sac mesothelium. We report the generation and analysis of a Vegf hypomorphic allele, Vegflo. Animals heterozygous for the targeted mutation are viable. Homozygous embryos, however, die at 9.0 dpc because of severe abnormalities in the yolk sac vasculature and deficiencies in the development of the dorsal aortae. We find that providing ‘Vegf wild-type’ visceral endoderm to the hypomorphic embryos restores normal blood and endothelial differentiation in the yolk sac, but does not rescue the phenotype in the embryo proper. In the opposite situation, however, when Vegf hypomorphic visceral endoderm is provided to a wild-type embryo, the ‘Vegf wild-type’ yolk sac mesoderm is not sufficient to support proper vessel formation and haematopoietic differentiation in this extra-embryonic membrane. These findings demonstrate that VEGFA expression in the visceral endoderm is absolutely required for the normal expansion and organisation of both the endothelial and haematopoietic lineages in the early sites of vessel and blood formation. However, normal VEGFA expression in the yolk sac mesoderm alone is not sufficient for supporting the proper development of the early vascular and haematopoietic system.

Identification and characterization of a novel, evolutionarily conserved gene disrupted by the murine H beta 58 embryonic lethal transgene insertion

Development ◽  
1992 ◽  
Vol 115 (1) ◽  
pp. 277-288 ◽  
Author(s):  
J.J. Lee ◽  
G. Radice ◽  
C.P. Perkins ◽  
F. Costantini
Keyword(s):  
Transcription Unit ◽  
Cdna Clones ◽  
Visceral Endoderm ◽  
Wild Type ◽  
Transgenic Mouse Line ◽  
Mouse Tissues ◽  
Post Implantation ◽  
Embryonic Ectoderm ◽  
Transgene Insertion

The H beta 58 transgenic mouse line carries a recessive insertional mutation that results in developmental abnormalities beginning at day 7.5 p.c. and embryonic arrest at about day 9.5. In this paper, we describe the characterization of a novel gene encoded at the H beta 58 locus, whose disruption appears to be responsible for the mutant phenotype. The wild-type H beta 58 gene encodes a single 2.7 kb mRNA during embryonic and fetal development, and in many adult somatic tissues. In the mutant locus, this transcription unit is split by the transgene insertion, and one of its coding exons is deleted. Consistent with the physical disruption of the gene, the level of the H beta 58 mRNA in heterozygous mutant mouse tissues was half the normal level, indicating that the mutant allele fails to encode a stable mRNA. In situ hybridization studies revealed that expression of the wild-type H beta 58 gene begins in the oocyte, and continues throughout pre- and post-implantation embryogenesis, despite the fact that homozygous mutant embryos develop successfully through the egg cylinder stage (day 6.5 p.c.). In the early post-implantation embryo, expression of the normal H beta 58 gene is relatively low in the embryonic ectoderm, the tissue displaying the earliest phenotypic effects of the mutation, and highest in the visceral endoderm. We therefore propose that the effects of the mutation on the embryonic ectoderm may be exerted indirectly, via the visceral endoderm. Sequence analysis of H beta 58 cDNA clones revealed no homology between the 38 × 10(3) M(r) H beta 58 protein and other known proteins. However, the H beta 58 gene displayed extremely strong conservation between mammals and birds (greater than 96% amino acid identity), although it appeared less conserved in amphibians and invertebrates.

Ontogeny of hyaluronan secretion during early mouse development

Development ◽  
1993 ◽  
Vol 117 (2) ◽  
pp. 483-492 ◽  
Author(s):  
J.J. Brown ◽  
V.E. Papaioannou
Keyword(s):  
Primitive Streak ◽  
Synthetic Activity ◽  
Visceral Endoderm ◽  
Mouse Development ◽  
Cell Lineages ◽  
Early Mouse ◽  
Serum Substitute ◽  
Embryonic Ectoderm

The ontogeny of hyaluronan (HA) secretion during early mouse embryogenesis has been investigated using a biotin-labelled HA-binding complex from cartilage proteoglycan. HA is first secreted by visceral endoderm cells of the early egg cylinder on day 5.5 post coitum (p.c.), predominantly into the expanding yolk cavity. On day 6.5 p.c., HA is present in both the yolk and proamniotic cavities, but pericellular staining is restricted to the visceral endoderm and a population of embryonic ectoderm cells at the antimesometrial end of the proamniotic cavity. By the primitive streak stage, HA is secreted into the ectoplacental, exocoelomic, amniotic and yolk cavities, whilst the only cells exhibiting pericellular staining are those of the embryonic and extraembryonic mesoderm, including the allantois. Comparisons of HA-staining patterns of cultured whole blastocysts, microdissected trophectoderm fragments and immunosurgically isolated inner cell masses, revealed no trophoblast-associated HA secretion during outgrowth in vitro but significant synthetic activity by the endodermal derivatives of differentiating inner cell masses. To identify the cell lineages responsible for secretion of HA into the embryonic cavities and to investigate the origin of the HA observed around migrating mesoderm cells, day 7.5 p.c. primitive streak stage conceptuses were dissected into their various embryonic and extraembryonic cell lineages. HA secretion was observed after short-term suspension culture of mesoderm, embryonic ectoderm and embryonic endoderm, but was undetectable in fragments of ectoplacental cone, parietal yolk sac (primary giant trophoblast and parietal endoderm), extraembryonic ectoderm or extraembryonic endoderm. The level of synthesis by the HA-positive tissues was markedly enhanced by culture in medium containing serum, compared with that obtained following culture in medium supplemented with a defined serum substitute containing insulin, transferrin, selenous acid and linoleic acid. This suggests that additional growth factors, present in serum but absent from the serum substitute, are required for optimal HA synthesis by the HA-secreting tissues in vitro, and probably also in vivo. The implications of these events for implantation and the development of peri- and early post-implantation mouse embryos are discussed, and a new role for HA in the initial formation and expansion of the embryonic cavities is proposed.

Targeted mutagenesis of the transcription factor GATA-4 gene in mouse embryonic stem cells disrupts visceral endoderm differentiation in vitro

Development ◽  
1995 ◽  
Vol 121 (11) ◽  
pp. 3877-3888 ◽  
Author(s):  
C. Soudais ◽  
M. Bielinska ◽  
M. Heikinheimo ◽  
C.A. MacArthur ◽  
N. Narita ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Es Cells ◽  
Embryonic Stem ◽  
Yolk Sac ◽  
Embryoid Bodies ◽  
Targeted Mutagenesis ◽  
Clonal Variation ◽  
Chimeric Mouse ◽  
Visceral Endoderm

Transcription factor GATA-4 belongs to a family of zinc finger proteins involved in lineage determination. GATA-4 is first expressed in yolk sac endoderm of the developing mouse and later in cardiac tissue, gut epithelium and gonads. To delineate the role of this transcription factor in differentiation and early development, we studied embryoid bodies derived from mouse embryonic stem (ES) cells in which both copies of the Gata-4 gene were disrupted. Light and electron microscopy demonstrated that embryoid bodies formed from wild-type and heterozygous deficient ES cells were covered with a layer of visceral yolk sac endoderm, whereas no yolk sac endoderm was evident on the surface of the homozygous deficient embryoid bodies. Independently selected homozygous deficient cell lines displayed this distinctive phenotype, suggesting that it was not an artifact of clonal variation. Biochemical markers of visceral endoderm formation, such as alpha-feto-protein, hepatocyte nuclear factor-4 and binding sites for Dolichos biflorus agglutinin, were absent from the homozygous deficient embryoid bodies. Examination of other differentiation markers in the mutant embryoid bodies, studies of ES cell-derived teratocarcinomas and chimeric mouse analysis demonstrated that GATA-4-deficient ES cells have the capacity to differentiate along other lineages. We conclude that, under in vitro conditions, disruption of the Gata-4 gene results in a specific block in visceral endoderm formation. These homozygous deficient cells should yield insights into the regulation of yolk sac endoderm development and the factors expressed by visceral endoderm that influence differentiation of adjoining ectoderm/mesoderm.

Winged helix transcription factor Foxb1 is essential for access of mammillothalamic axons to the thalamus

Development ◽  
2000 ◽  
Vol 127 (5) ◽  
pp. 1029-1038 ◽  
Author(s):  
G. Alvarez-Bolado ◽  
X. Zhou ◽  
A.K. Voss ◽  
T. Thomas ◽  
P. Gruss
Keyword(s):  
Transcription Factor ◽  
Transcriptional Activity ◽  
Wild Type ◽  
Winged Helix ◽  
Targeted Mutation ◽  
Ventricular Cells ◽  
Winged Helix Transcription Factor ◽  
Mutant Cells ◽  
Correct Expression

Our aim was to study the mechanisms of brain histogenesis. As a model, we have used the role of winged helix transcription factor gene Foxb1 in the emergence of a very specific morphological trait of the diencephalon, the mammillary axonal complex. Foxb1 is expressed in a large hypothalamic neuronal group (the mammillary body), which gives origin to a major axonal bundle with branches to thalamus, tectum and tegmentum. We have generated mice carrying a targeted mutation of Foxb1 plus the tau-lacZ reporter. In these mutants, a subpopulation of dorsal thalamic ventricular cells “thalamic palisade” show abnormal persistence of Foxb1 transcriptional activity; the thalamic branch of the mammillary axonal complex is not able to grow past these cells and enter the thalamus. The other two branches of the mammillary axonal complex (to tectum and tegmentum) are unaffected by the mutation. Most of the neurons that originate the mammillothalamic axons suffer apoptosis after navigational failure. Analysis of chimeric brains with wild-type and Foxb1 mutant cells suggests that correct expression of Foxb1 in the thalamic palisade is sufficient to rescue the normal phenotype. Our results indicate that Foxb1 is essential for diencephalic histogenesis and that it exerts its effects by controlling access to the target by one particular axonal branch.

scholarly journals The Transcription Factor RFX3 Directs Nodal Cilium Development and Left-Right Asymmetry Specification

2004 ◽  
Vol 24 (10) ◽  
pp. 4417-4427 ◽  
Author(s):  
E. Bonnafe ◽  
M. Touka ◽  
A. AitLounis ◽  
D. Baas ◽  
E. Barras ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Immune System ◽  
Intraflagellar Transport ◽  
Body Axis ◽  
High Rate ◽  
Ciliated Cells ◽  
Wild Type ◽  
Left Right Asymmetry ◽  
First Time ◽  
Deficient Mice

ABSTRACT There are five members of the RFX family of transcription factors in mammals. While RFX5 plays a well-defined role in the immune system, the functions of RFX1 to RFX4 remain largely unknown. We have generated mice with a deletion of the Rfx3 gene. RFX3-deficient mice exhibit frequent left-right (LR) asymmetry defects leading to a high rate of embryonic lethality and situs inversus in surviving adults. In vertebrates, specification of the LR body axis is controlled by monocilia in the embryonic node, and defects in nodal cilia consequently result in abnormal LR patterning. Consistent with this, Rfx3 is expressed in ciliated cells of the node and RFX3-deficient mice exhibit a pronounced defect in nodal cilia. In contrast to the case for wild-type embryos, for which we document for the first time a twofold increase in the length of nodal cilia during development, the cilia are present but remain markedly stunted in mutant embryos. Finally, we show that RFX3 regulates the expression of D2lic, the mouse orthologue of a Caenorhabditis elegans gene that is implicated in intraflagellar transport, a process required for the assembly and maintenance of cilia. In conclusion, RFX3 is essential for the differentiation of nodal monocilia and hence for LR body axis determination.

scholarly journals Requirement for β-Catenin in Anterior-Posterior Axis Formation in Mice

2000 ◽  
Vol 148 (3) ◽  
pp. 567-578 ◽  
Author(s):  
Joerg Huelsken ◽  
Regina Vogel ◽  
Volker Brinkmann ◽  
Bettina Erdmann ◽  
Carmen Birchmeier ◽  
...  
Keyword(s):  
Wnt Pathway ◽  
Wnt Signaling Pathway ◽  
Primitive Streak ◽  
Mouse Embryos ◽  
Axis Formation ◽  
Deficient Mouse ◽  
Visceral Endoderm ◽  
Embryonic Ectoderm ◽  
Anterior Posterior ◽  
Anterior Posterior Axis

The anterior-posterior axis of the mouse embryo is defined before formation of the primitive streak, and axis specification and subsequent anterior development involves signaling from both embryonic ectoderm and visceral endoderm. Τhe Wnt signaling pathway is essential for various developmental processes, but a role in anterior-posterior axis formation in the mouse has not been previously established. β-Catenin is a central player in the Wnt pathway and in cadherin-mediated cell adhesion. We generated β-catenin–deficient mouse embryos and observed a defect in anterior-posterior axis formation at embryonic day 5.5, as visualized by the absence of Hex and Hesx1 and the mislocation of cerberus-like and Lim1 expression. Subsequently, no mesoderm and head structures are generated. Intercellular adhesion is maintained since plakoglobin substitutes for β-catenin. Our data demonstrate that β-catenin function is essential in anterior-posterior axis formation in the mouse, and experiments with chimeric embryos show that this function is required in the embryonic ectoderm.

Chemotaxis and Transcription Factor Activation of Wild-Type and CCR6-Transfected RLE-6TN

Pneumologie ◽  
2012 ◽  
Vol 66 (11) ◽  
Author(s):  
K Hoehne ◽  
H Eibel ◽  
M Grimm ◽  
M Idzko ◽  
J Müller-Quernheim ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Wild Type ◽  
Transcription Factor Activation

scholarly journals Transcriptional Activation in Yeast Cells Lacking Transcription Factor IIA

Genetics ◽  
1999 ◽  
Vol 153 (4) ◽  
pp. 1573-1581 ◽  
Author(s):  
Susanna Chou ◽  
Sukalyan Chatterjee ◽  
Mark Lee ◽  
Kevin Struhl
Keyword(s):  
Transcription Factor ◽  
Transcriptional Activation ◽  
Large Subunit ◽  
Yeast Cells ◽  
Specific Cell ◽  
Wild Type ◽  
Activation Domains ◽  
Cycle Arrest ◽  
General Transcription Factor

Abstract The general transcription factor IIA (TFIIA) forms a complex with TFIID at the TATA promoter element, and it inhibits the function of several negative regulators of the TATA-binding protein (TBP) subunit of TFIID. Biochemical experiments suggest that TFIIA is important in the response to transcriptional activators because activation domains can interact with TFIIA, increase recruitment of TFIID and TFIIA to the promoter, and promote isomerization of the TFIID-TFIIA-TATA complex. Here, we describe a double-shut-off approach to deplete yeast cells of Toa1, the large subunit of TFIIA, to <1% of the wild-type level. Interestingly, such TFIIA-depleted cells are essentially unaffected for activation by heat shock factor, Ace1, and Gal4-VP16. However, depletion of TFIIA causes a general two- to threefold decrease of transcription from most yeast promoters and a specific cell-cycle arrest at the G2-M boundary. These results indicate that transcriptional activation in vivo can occur in the absence of TFIIA.

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