scholarly journals An Intronic cis-Regulatory Element Is Crucial for the Alpha Tubulin Pl-Tuba1a Gene Activation in the Ciliary Band and Animal Pole Neurogenic Domains during Sea Urchin Development

PLoS ONE ◽  
2017 ◽  
Vol 12 (1) ◽  
pp. e0170969 ◽  
Author(s):  
Salvatore Costa ◽  
Aldo Nicosia ◽  
Angela Cuttitta ◽  
Fabrizio Gianguzza ◽  
Maria Antonietta Ragusa
Keyword(s):  
Sea Urchin ◽  
Regulatory Element ◽  
Gene Activation ◽  
Ciliary Band ◽  
Animal Pole ◽  
Alpha Tubulin

GSK3beta/shaggy mediates patterning along the animal-vegetal axis of the sea urchin embryo

Development ◽  
1998 ◽  
Vol 125 (13) ◽  
pp. 2489-2498 ◽  
Author(s):  
F. Emily-Fenouil ◽  
C. Ghiglione ◽  
G. Lhomond ◽  
T. Lepage ◽  
C. Gache
Keyword(s):  
Sea Urchin ◽  
Wnt Pathway ◽  
Dominant Negative ◽  
Early Gene ◽  
Expression Domain ◽  
Animal Pole ◽  
Axial Patterning ◽  
Sea Urchin Embryo ◽  
Vegetal Pole ◽  
Dominant Negative Activity

In the sea urchin embryo, the animal-vegetal axis is defined before fertilization and different embryonic territories are established along this axis by mechanisms which are largely unknown. Significantly, the boundaries of these territories can be shifted by treatment with various reagents including zinc and lithium. We have isolated and characterized a sea urchin homolog of GSK3beta/shaggy, a lithium-sensitive kinase which is a component of the Wnt pathway and known to be involved in axial patterning in other embryos including Xenopus. The effects of overexpressing the normal and mutant forms of GSK3beta derived either from sea urchin or Xenopus were analyzed by observation of the morphology of 48 hour embryos (pluteus stage) and by monitoring spatial expression of the hatching enzyme (HE) gene, a very early gene whose expression is restricted to an animal domain with a sharp border roughly coinciding with the future ectoderm / endoderm boundary. Inactive forms of GSK3beta predicted to have a dominant-negative activity, vegetalized the embryo and decreased the size of the HE expression domain, apparently by shifting the boundary towards the animal pole. These effects are similar to, but even stronger than, those of lithium. Conversely, overexpression of wild-type GSK3beta animalized the embryo and caused the HE domain to enlarge towards the vegetal pole. Unlike zinc treatment, GSK3beta overexpression thus appeared to provoke a true animalization, through extension of the presumptive ectoderm territory. These results indicate that in sea urchin embryos the level of GSKbeta activity controls the position of the boundary between the presumptive ectoderm and endoderm territories and thus, the relative extent of these tissue layers in late embryos. GSK3beta and probably other downstream components of the Wnt pathway thus mediate patterning both along the primary AV axis of the sea urchin embryo and along the dorsal-ventral axis in Xenopus, suggesting a conserved basis for axial patterning between invertebrate and vertebrate in deuterostomes.

Centrosomal components immunologically related to tektins from ciliary and flagellar microtubules

1994 ◽  
Vol 107 (8) ◽  
pp. 2095-2105 ◽  
Author(s):  
W. Steffen ◽  
E.A. Fajer ◽  
R.W. Linck
Keyword(s):  
Cell Cycle ◽  
Sea Urchin ◽  
Cho Cells ◽  
Mammalian Cells ◽  
Polyclonal Antibodies ◽  
Polyacrylamide Gel Electrophoresis ◽  
Immunoblot Analysis ◽  
Surf Clam ◽  
Alpha Tubulin ◽  
Sea Urchin Sperm

Centrosomes are critical for the nucleation and organization of the microtubule cytoskeleton during both interphase and cell division. Using antibodies raised against sea urchin sperm flagellar microtubule proteins, we characterize here the presence and behavior of certain components associated with centrosomes of the surf clam Spisula solidissima and cultured mammalian cells. A Sarkosyl detergent-resistant fraction of axonemal microtubules was isolated from sea urchin sperm flagella and used to produce monoclonal antibodies, 16 of which were specific- or cross-specific for the major polypeptides associated with this microtubule fraction: tektins A, B and C, acetylated alpha-tubulin, and 77 and 83 kDa polypeptides. By 2-D isoelectric focussing/SDS polyacrylamide gel electrophoresis the tektins separate into several polypeptide spots. Identical spots were recognized by monoclonal and polyclonal antibodies against a given tektin, indicating that the different polypeptide spots are isoforms or modified versions of the same protein. Four independently derived monoclonal anti-tektins were found to stain centrosomes of S. solidissima oocytes and CHO and HeLa cells, by immunofluorescence microscopy. In particular, the centrosome staining of one monoclonal antibody specific for tektin B (tekB3) was cell-cycle-dependent for CHO cells, i.e. staining was observed only from early prometaphase until late anaphase. By immuno-electron microscopy tekB3 specifically labeled material surrounding the centrosome, whereas a polyclonal anti-tektin B recognized centrioles as well as the centrosomal material throughout the cell cycle. Finally, by immunoblot analysis tekB3 stained polypeptides of 48–50 kDa in isolated spindles and centrosomes from CHO cells.

Specification of endoderm and mesoderm in the sea urchin

Zygote ◽  
1999 ◽  
Vol 8 (S1) ◽  
pp. S41-S41 ◽  
Author(s):  
David R. McClay
Keyword(s):  
Sea Urchin ◽  
Wnt Pathway ◽  
Special Significance ◽  
Nuclear Entry ◽  
Cell Stage ◽  
Animal Pole ◽  
Secondary Axis ◽  
Sea Urchin Embryo ◽  
Vegetal Pole

It has long been recognized that micromeres have special significance in early specification events in the sea urchin embryo. Micromeres have the ability to induce a secondary axis if transferred to the animal pole at the 16-cell stage of sea urchin embryos (Hörstadius, 1939). Without micromeres an isolated animal hemisphere develops into an ectodermal ball called a dauer blastula. Addition of micromeres to an animal half rescues a normal pluteus larva, including endoderm (Hörstadius, 1939). Despite these well-known experiments, however, neither the molecular basis of that induction nor the endogenous inductive role of micromeres in development was known. In recent experiments we learned that if one eliminates micromeres from the vegetal pole at the 16-cell stage the resulting embryo makes no secondary mesenchyme. Earlier it had been found that β-catenin is crucial for specification events that lead to mesoderm and endoderm (Wikra-manayake et al., 1998; Emily-Fenouil et al., 1998; Logan et al., 1999). We noticed that at the 16-cell stage β-catenin enters the nuclei of micromeres, then enters the nuclei of macromeres at the 32-cell stage (Logan et al., 1999). Since nuclear entry of β-catenin is known to be important for its signalling function in the Wnt pathway, we asked whether β-catenin functions in the micromere induction pathway.

scholarly journals Stress response gene activation protects sea urchin embryos exposed to X-rays

2011 ◽  
Vol 16 (6) ◽  
pp. 681-687 ◽  
Author(s):  
Rosa Bonaventura ◽  
Francesca Zito ◽  
Caterina Costa ◽  
Salvatore Giarrusso ◽  
Filippo Celi ◽  
...  
Keyword(s):  
Stress Response ◽  
Sea Urchin ◽  
Gene Activation ◽  
X Rays ◽  
Response Gene ◽  
Sea Urchin Embryos ◽  
Stress Response Gene

scholarly journals Transcriptional Activation of the Decidual/Trophoblast Prolactin-Related Protein Gene1

Endocrinology ◽  
1999 ◽  
Vol 140 (9) ◽  
pp. 4032-4039 ◽  
Author(s):  
Kyle E. Orwig ◽  
Michael J. Soares
Keyword(s):  
Transcriptional Activation ◽  
Promoter Activity ◽  
Mutational Analysis ◽  
Regulatory Element ◽  
Gene Activation ◽  
Cell Formation ◽  
Regulatory Elements ◽  
Related Protein ◽  
Trophoblast Cells ◽  
Decidual Cells

Abstract The decidual/trophoblast PRL-related protein (d/tPRP) is dually expressed by decidual and trophoblast cells during pregnancy. We have characterized the proximal d/tPRP promoter responsible for directing d/tPRP expression in decidual and trophoblast cells. We have demonstrated that the proximal 93 bp of d/tPRP 5′-flanking DNA are sufficient to direct luciferase gene expression in primary decidual and Rcho-1 trophoblast cells, but not in fibroblast, undifferentiated uterine stromal cells or trophoblast cells of a labyrinthine lineage. The 93-bp d/tPRP promoter was also sufficient to direct differentiation-dependent expression in trophoblast giant cells. Mutational analysis demonstrated the differential importance of activating protein-1 and Ets regulatory elements (located within the proximal 93 bp of d/tPRP 5′-flanking DNA) for activation of the d/tPRP promoter in decidual vs. trophoblast cells. Disruption of the activating protein-1 regulatory element inhibited d/tPRP promoter activity by more than 95% in decidual cells, and approximately 80% trophoblast cells. Disruption of the Ets regulatory element reduced d/tPRP promoter activity by approximately 50% in decidual cells, while inactivating the d/tPRP promoter in trophoblast cells. Protein interactions with the trophoblast Ets regulatory element were shown to be cell type specific and to change during trophoblast giant cell formation. In conclusion, a 93-bp region of the d/tPRP promoter is shown to contain regulatory elements sufficient for gene activation in decidual and trophoblast cells.

scholarly journals Identification of a Ty1 regulatory sequence responsive to STE7 and STE12.

1988 ◽  
Vol 8 (6) ◽  
pp. 2545-2554 ◽  
Author(s):  
M Company ◽  
C Adler ◽  
B Errede
Keyword(s):  
Gene Expression ◽  
Complex Formation ◽  
Reporter Gene ◽  
Regulatory Element ◽  
Gene Activation ◽  
Cell Types ◽  
Reporter Gene Expression ◽  
Regulatory Sequence ◽  
Gene Products ◽  
Cell Extracts

Ty1 activation of gene expression observed in haploid cell types of Saccharomyces cerevisiae requires the STE7 and STE12 gene products. An activator sequence within Ty1 that is responsive to these two regulators has been defined. Complex formation between a factor in whole-cell extracts and the DNA regulatory element showed the same dependence on the STE7 and STE12 gene products as did reporter gene expression. Base pair substitutions within the binding site abolished the ability to form the factor-DNA complex and to activate gene expression. The correlation between complex formation and reporter gene expression indicates that factor binding to the cis-acting element is essential for gene activation. Because the predicted protein for the STE7 gene product is homologous to protein kinases, we suggest that protein phosphorylation may directly or indirectly regulate formation of this DNA-protein complex.

scholarly journals The sea urchin animal pole domain is a Six3-dependent neurogenic patterning center

Development ◽  
2009 ◽  
Vol 136 (7) ◽  
pp. 1179-1189 ◽  
Author(s):  
Z. Wei ◽  
J. Yaguchi ◽  
S. Yaguchi ◽  
R. C. Angerer ◽  
L. M. Angerer
Keyword(s):  
Sea Urchin ◽  
Animal Pole

scholarly journals Functional gap junctions are not required for muscle gene activation by induction in Xenopus embryos.

1987 ◽  
Vol 104 (3) ◽  
pp. 557-564 ◽  
Author(s):  
A Warner ◽  
J B Gurdon
Keyword(s):  
Gap Junctions ◽  
Gap Junction ◽  
Close Contact ◽  
Gene Activation ◽  
Gene Probe ◽  
Animal Cells ◽  
Animal Pole ◽  
Junction Protein ◽  
Pole Cells ◽  
Muscle Gene

Muscle gene expression is known to be induced in animal pole cells of a Xenopus blastula after 2-3 h of close contact with vegetal pole cells. We tested whether this induction requires functional gap junctions between vegetal and animal portions of an animal-vegetal conjugate. Muscle gene transcription was assayed with a muscle-specific actin gene probe and the presence or absence of communication through gap junctions was determined electrophysiologically. Antibodies to gap junction protein were shown to block gap junction communication for the whole of the induction time, but did not prevent successful induction of muscle gene activation. The outcome was the same whether communication between inducing vegetal cells and responding animal cells was blocked by introducing antibodies into vegetal cells alone or into animal cells alone. We conclude that gap junctions are not required for this example of embryonic induction.

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