Altered expression of spatially regulated embryonic genes in the progeny of separated sea urchin blastomeres

Development ◽  
1989 ◽  
Vol 106 (3) ◽  
pp. 567-579 ◽  
Author(s):  
D.L. Hurley ◽  
L.M. Angerer ◽  
R.C. Angerer
Keyword(s):  
Cell Fate ◽  
Sea Urchin ◽  
Sea Water ◽  
Negative Control ◽  
Normal Embryo ◽  
Cell Stage ◽  
Mrna Accumulation ◽  
Altered Expression ◽  
Intact Embryo ◽  
Dissociated Cells

We have examined the importance of the extracellular environment on the ability of separated cells of sea urchin embryos (Strongylocentrotus purpuratus) to carry out patterns of mRNA accumulation and decay characteristic of intact embryos. Embryos were dissociated into individual blastomeres at 16-cell stage and maintained in calcium-free sea water so that daughter cells continuously separated. Levels of eleven different mRNAs in these cells were compared to those in control embryos when the latter reached mesenchyme blastula stage, by which time cells in major regions of the intact embryo have assumed distinctive patterns of message accumulation. Abrogation of interactions among cells resulted in marked differences in accumulation and/or turnover of the individual mRNAs, which are expressed with diverse temporal and spatial patterns of prevalence in intact embryos. In general, separated cells are competent to execute initial events of mRNA accumulation and decay that occur uniformly in most or all blastomeres of the intact embryo and are likely to be regulated by maternal molecules. The ability of separated cells to accumulate mRNAs that appear slightly later in development depends upon the presumptive tissue in which a given mRNA is found in the normal embryo. Messages that normally accumulate in cells at the vegetal pole also accumulate in dissociated cells either at nearly normal levels or at increased levels. In one such case, that of actin CyIIa, which is normally restricted to mesenchyme cells, in situ hybridization demonstrates that the fraction of dissociated cells expressing this message is 4- to 5-fold higher than in the normal embryo. In contrast, separated cells accumulate significant levels of a message expressed uniformly in the early ectoderm but are unable to execute accumulation and decay of different messages that distinguish oral and aboral ectodermal regions. These data are consistent with the idea that interactions among cells in the intact embryo are important for both positive and negative control of expression of different genes that are early indicators of the specification of cell fate.

scholarly journals SOME STRUCTURAL AND FUNCTIONAL ASPECTS OF THE MITOTIC APPARATUS IN SEA URCHIN EMBRYOS

1962 ◽  
Vol 14 (3) ◽  
pp. 475-487 ◽  
Author(s):  
Patricia Harris
Keyword(s):  
Sea Urchin ◽  
Salt Concentration ◽  
Sea Water ◽  
Divalent Cations ◽  
Cell Stage ◽  
Mitotic Apparatus ◽  
Central Spindle ◽  
Sea Urchin Embryos ◽  
Cilia Formation ◽  
Dividing Cells

The mitotic figures in dividing cells of sea urchin embryos, from first division to the onset of cilia formation, were studied with regard to the filament system and its relation to kinetochores, chromosomes, and poles, as well as to fixation conditions which would best preserve these structures. With regard to fixation, variations in the salt concentration and pH of the fixative indicated that an extraction effect on the chromosomes noted in earlier work was probably due to a combination of neutral pH and salt concentration equivalent to sea water. The presence of the 15 mµ filaments depended on the presence of either of two stabilizing conditions: pH 6.1 or presence of the salts of sea water, presumably the divalent cations of Ca and Mg. Kinetochores and centrioles were unaffected by the fixative variations. The 15 mµ filaments, reported earlier in the central spindle, are also found in great numbers in the asters of early cleavage divisions. However, with successive divisions and reduction in cell size, the aster disappears at about the 32 to 64 cell stage, and the 15 mµ filaments are entirely associated with the central spindle. This disappearance of the aster suggests that it may be, in fact, merely a specialization of large cells for cytokinesis.

scholarly journals A Cytochemical Study of the Sulfhydryl Groups of Sea Urchin Eggs during the First Cleavage

1958 ◽  
Vol 4 (5) ◽  
pp. 615-620 ◽  
Author(s):  
Naoko Kawamura ◽  
Katsuma Dan
Keyword(s):  
Sea Urchin ◽  
Azo Dye ◽  
Sea Water ◽  
Color Intensity ◽  
Cell Stage ◽  
Cytochemical Study ◽  
Sea Urchin Eggs ◽  
Sh Groups ◽  
Precursor Material ◽  
Hemicentrotus Pulcherrimus

In the eggs of four species of echinoderms, Mespilia globulus, Pseudocentrotus depressus, Hemicentrotus pulcherrimus and Clypeaster japonicus, changes in the distribution of protein-bound SH groups from fertilization to the 2 cell stage have been studied cytochemically by use of a mercaptide-forming azo dye. In the eggs of these species, the color intensity in the cytoplasm increased upon fertilization. The astral centers and spindle during mitosis were stained deeply. When the aster formation was suppressed by ether, hyaline spots appeared in the egg cytoplasm instead of well formed astral centers and these spots were stained by the SH-specific dye. Upon recovery of such eggs in pure sea water, and when cleavage ensued, such spots disappeared and two new astral centers were reorganized. The SH-protein occurring in the centrosphere is considered to be the precursor material for the asters and spindle, and this material is apparently derived from the cytoplasm.

Adenosine induces dormancy in starfish blastulae

Development ◽  
1988 ◽  
Vol 103 (2) ◽  
pp. 345-351
Author(s):  
N. Tsuchimori ◽  
S. Miyashiro ◽  
H. Shibai ◽  
S. Ikegami
Keyword(s):  
Epithelial Cells ◽  
Sea Water ◽  
Intracellular Concentration ◽  
Normal Embryo ◽  
Embryonic Cells ◽  
Macromolecular Synthesis ◽  
Cell Stage ◽  
External Application ◽  
Dna And Rna ◽  
Asterina Pectinifera

External application of 50 micrograms ml-1 adenosine inhibits development of the starfish Asterina pectinifera at the 256-cell stage when all the embryonic cells differentiate to epithelial cells. Intracellular concentration of adenosine in the adenosine-treated embryo is 2.7 times higher than those of the normal embryo whereas the contents of ATP, ADP, AMP and adenosine 3′,5′-monophosphate are the same for both embryos. Adenosine causes more than 95% reduction in the rate of protein, DNA and RNA syntheses. By returning the embryo to normal sea water, macromolecular synthesis restarts and the embryo develops to the bipinnaria stage.

Nuclear beta-catenin is required to specify vegetal cell fates in the sea urchin embryo

Development ◽  
1999 ◽  
Vol 126 (2) ◽  
pp. 345-357 ◽  
Author(s):  
C.Y. Logan ◽  
J.R. Miller ◽  
M.J. Ferkowicz ◽  
D.R. McClay
Keyword(s):  
Cell Fate ◽  
Sea Urchin ◽  
Beta Catenin ◽  
Cell Fate Specification ◽  
Cell Fates ◽  
Cell Stage ◽  
Animal Pole ◽  
Fate Specification ◽  
The Relationship ◽  
Vegetal Cell

Beta-catenin is thought to mediate cell fate specification events by localizing to the nucleus where it modulates gene expression. To ask whether beta-catenin is involved in cell fate specification during sea urchin embryogenesis, we analyzed the distribution of nuclear beta-catenin in both normal and experimentally manipulated embryos. In unperturbed embryos, beta-catenin accumulates in nuclei that include the precursors of the endoderm and mesoderm, suggesting that it plays a role in vegetal specification. Using pharmacological, embryological and molecular approaches, we determined the function of beta-catenin in vegetal development by examining the relationship between the pattern of nuclear beta-catenin and the formation of endodermal and mesodermal tissues. Treatment of embryos with LiCl, a known vegetalizing agent, caused both an enhancement in the levels of nuclear beta-catenin and an expansion in the pattern of nuclear beta-catenin that coincided with an increase in endoderm and mesoderm. Conversely, overexpression of a sea urchin cadherin blocked the accumulation of nuclear beta-catenin and consequently inhibited the formation of endodermal and mesodermal tissues including micromere-derived skeletogenic mesenchyme. In addition, nuclear beta-catenin-deficient micromeres failed to induce a secondary axis when transplanted to the animal pole of uninjected host embryos, indicating that nuclear beta-catenin also plays a role in the production of micromere-derived signals. To examine further the relationship between nuclear beta-catenin in vegetal nuclei and micromere signaling, we performed both transplantations and deletions of micromeres at the 16-cell stage and demonstrated that the accumulation of beta-catenin in vegetal nuclei does not require micromere-derived cues. Moreover, we demonstrate that cell autonomous signals appear to regulate the pattern of nuclear beta-catenin since dissociated blastomeres possessed nuclear beta-catenin in approximately the same proportion as that seen in intact embryos. Together, these data show that the accumulation of beta-catenin in nuclei of vegetal cells is regulated cell autonomously and that this localization is required for the establishment of all vegetal cell fates and the production of micromere-derived signals.

Altering cell fates in sea urchin embryos by overexpressing SpOtx, an orthodenticle-related protein

Development ◽  
1996 ◽  
Vol 122 (5) ◽  
pp. 1489-1498 ◽  
Author(s):  
C.A. Mao ◽  
A.H. Wikramanayake ◽  
L. Gan ◽  
C.K. Chuang ◽  
R.G. Summers ◽  
...  
Keyword(s):  
Cell Fate ◽  
Sea Urchin ◽  
Biological Effects ◽  
Gene Activation ◽  
Related Protein ◽  
Cell Fates ◽  
Cell Stage ◽  
Morphological Alterations ◽  
Transactivation Domains ◽  
Sea Urchin Embryo

While many general features of cell fate specification in the sea urchin embryo are understood, specific factors associated with these events remain unidentified. SpOtx, an orthodenticle-related protein, has been implicated as a transcriptional activator of the aboral ectoderm-specific Spec2a gene. Here, we present evidence that SpOtx has the potential to alter cell fates. SpOtx was found in the cytoplasm of early cleavage stage embryos and was translocated into nuclei between the 60- and 120-cell stage, coincident with Spec gene activation. Eggs injected with SpOtx mRNA developed into epithelial balls of aboral ectoderm suggesting that SpOtx redirected nonaboral ectoderm cells to an aboral ectoderm fate. At least three distinct domains on SpOtx, the homeobox and regions in the N-terminal and C-terminal halves of the protein, were required for the morphological alterations. These same N-terminal and C-terminal regions were shown to be transactivation domains in a yeast transactivation assay, indicating that the biological effects of overexpressing SpOtx were due to its action as a transcription factor. Our results suggest that SpOtx is involved in aboral ectoderm differentiation by activating aboral ectoderm-specific genes and that modulating its expression can lead to changes in cell fate.

SpSoxB1, a maternally encoded transcription factor asymmetrically distributed among early sea urchin blastomeres

Development ◽  
1999 ◽  
Vol 126 (23) ◽  
pp. 5473-5483 ◽  
Author(s):  
A.P. Kenny ◽  
D. Kozlowski ◽  
D.W. Oleksyn ◽  
L.M. Angerer ◽  
R.C. Angerer
Keyword(s):  
Transcription Factor ◽  
Sea Urchin ◽  
Protein Distribution ◽  
Blastula Stage ◽  
Cell Stage ◽  
Mrna Accumulation ◽  
Cis Element ◽  
Sea Urchin Embryo ◽  
Asymmetrically Distributed

We have identified a Sox family transcription factor, SpSoxB1, that is asymmetrically distributed among blastomeres of the sea urchin embryo during cleavage, beginning at 4th cleavage. SpSoxB1 interacts with a cis element that is essential for transcription of SpAN, a gene that is activated cell autonomously and expressed asymmetrically along the animal-vegetal axis. In vitro translated SpSoxB1 forms a specific complex with this cis element whose mobility is identical to that formed by a protein in nuclear extracts. An anti-SpSoxB1 rabbit polyclonal antiserum specifically supershifts this DNA-protein complex and recognizes a single protein on immunoblots of nuclear proteins that comigrates with in vitro translated SpSoxB1. Developmental immunoblots of total proteins at selected early developmental stages, as well as EMSA of egg and 16-cell stage proteins, show that SpSoxB1 is present at low levels in unfertilized eggs and progressively accumulates during cleavage. SpSoxB1 maternal transcripts are uniformly distributed in the unfertilized egg and the protein accumulates to similar, high concentrations in all nuclei of 4- and 8-cell embryos. However, at fourth cleavage, the micromeres, which are partitioned by asymmetric division of the vegetal 4 blastomeres, have reduced nuclear levels of the protein, while high levels persist in their sister macromeres and in the mesomeres. During cleavage, the uniform maternal SpSoxB1 transcript distribution is replaced by a zygotic nonvegetal pattern that reinforces the asymmetric SpSoxB1 protein distribution and reflects the corresponding domain of SpAN mRNA accumulation at early blastula stage (approximately 150 cells). The vegetal region lacking nuclear SpSoxB1 gradually expands so that, after blastula stage, only cells in differentiating ectoderm accumulate this protein in their nuclei. The results reported here support a model in which SpSoxB1 is a major regulator of the initial phase of asymmetric transcription of SpAN in the nonvegetal domain by virtue of its distribution at 4th cleavage and is subsequently an important spatial determinant of expression in the early blastula. This factor is the earliest known spatially restricted regulator of transcription along the animal-vegetal axis of the sea urchin embryo.

scholarly journals A BMP pathway regulates cell fate allocation along the sea urchin animal-vegetal embryonic axis

Development ◽  
2000 ◽  
Vol 127 (5) ◽  
pp. 1105-1114 ◽  
Author(s):  
L.M. Angerer ◽  
D.W. Oleksyn ◽  
C.Y. Logan ◽  
D.R. McClay ◽  
L. Dale ◽  
...  
Keyword(s):  
Cell Fate ◽  
Sea Urchin ◽  
Animal Cell ◽  
Bmp Signaling ◽  
Epithelial Tissue ◽  
Cell Fates ◽  
Cell Stage ◽  
Bmp Pathway ◽  
The One ◽  
Dose Dependent

To examine whether a BMP signaling pathway functions in specification of cell fates in sea urchin embryos, we have cloned sea urchin BMP2/4, analyzed its expression in time and space in developing embryos and assayed the developmental consequences of changing its concentration through mRNA injection experiments. These studies show that BMP4 mRNAs accumulate transiently during blastula stages, beginning around the 200-cell stage, 14 hours postfertilization. Soon after the hatching blastula stage, BMP2/4 transcripts can be detected in presumptive ectoderm, where they are enriched on the oral side. Injection of BMP2/4 mRNA at the one-cell stage causes a dose-dependent suppression of commitment of cells to vegetal fates and ectoderm differentiates almost exclusively as a squamous epithelial tissue. In contrast, NOGGIN, an antagonist of BMP2/4, enhances differentiation of endoderm, a vegetal tissue, and promotes differentiation of cells characteristic of the ciliated band, which contains neurogenic ectoderm. These findings support a model in which the balance of BMP2/4 signals produced by animal cell progeny and opposing vegetalizing signals sent during cleavage stages regulate the position of the ectoderm/ endoderm boundary. In addition, BMP2/4 levels influence the decision within ectoderm between epidermal and nonepidermal differentiation.

HpEtsimplicated in primary mesenchyme cell differentiation of the sea urchin (Hemicentrotus pulcherrimus) embryo

Zygote ◽  
1999 ◽  
Vol 8 (S1) ◽  
pp. S33-S34 ◽  
Author(s):  
Daisuke Kurokawa ◽  
Takashi Kitajima ◽  
Keiko Mitsunaga-Nakatsubo ◽  
Shonan Amemiya ◽  
Hiraku Shimada ◽  
...  
Keyword(s):  
Cell Fate ◽  
Sea Urchin ◽  
Sea Urchins ◽  
Marker Genes ◽  
Asymmetric Distribution ◽  
Specific Marker ◽  
Cell Stage ◽  
Mesenchyme Cell ◽  
Maternal Determinants ◽  
And Function

In sea urchin embryogenesis it has been suggested that the initial territories are specified by a combination of the asymmetric distribution of cytoplasmic determinants and cell-cell interactions. At the 60-cell stage blastomeres clonally originated from founder cells divide the embryo into five distinct territories: small micromeres, large micromeres, vegetal plate, oral ectoderm and aboral ectoderm. The territories are identified by the expression of specific marker genes and their cell lineages (Davidson, 1989, 1991). The large micromeres are thought to play a role as an organiser and initiate a cascade of signal transduction toward overlying cells (Davidson, 1989). In this model the large micromeres induce the overlying veg2 tier, specifying the vegetal plate (Ransick & Davidson, 1993, 1995). The veg2 tier then induces the overlying cells, which include gut cells and cells of the prospective ectodermal territories (Wikramanayakeet al., 1995; Wikramanayake & Klein, 1997). Thus, the large micromeres, which are the prospective primary mesenchyme cells (PMCs), play a key role in cell fate specification and axis determination during sea urchin embryogenesis. Previous data suggested that the large micromeres are autonomously specified to become PMCs by maternally inherited determinants (Okazaki, 1975; Kitajima & Okazaki, 1980). An important question in sea urchins embryogenesis is the identity and function of the proposed maternal determinants.

Ca(2+) in specification of vegetal cell fate in early sea urchin embryos

2001 ◽  
Vol 204 (5) ◽  
pp. 823-834
Author(s):  
I. Yazaki
Keyword(s):  
Cell Fate ◽  
Sea Urchin ◽  
Small Cells ◽  
Beta Catenin ◽  
Cell Stage ◽  
Sea Urchin Embryos ◽  
Mesoderm Specification ◽  
New Findings ◽  
Vegetal Pole ◽  
Vegetal Cell

In sea urchin embryos, the first specification of cell fate occurs at the fourth cleavage, when small cells (the micromeres) are formed at the vegetal pole. The fate of other blastomeres is dependent on the receipt of cell signals originating from the micromeres. The micromeres are fated to become skeletogenic cells and show the ability to induce the endoderm (the archenteron) in the neighbouring cells during the 16- to 60-cell stage. Several molecules involved in signaling pathways, i.e. Notch for mesoderm specification, bone morphogenic protein (BMP) for ectoderm specification and beta-catenin for endoderm specification, are spatially and temporally expressed during development. In the micromeres, beta-catenin increases and subsequently localizes to the nuclei under the regulation of TCF, a nuclear binding partner of beta-catenin, until the 60-cell stage. However, the mechanisms activating these signaling substances are still unclear. In this article, I demonstrate some specific properties of the membrane and cytoplasm of micromeres including new findings on intracellular Ca(2+) concentration, and propose a mechanism by which the functional micromeres are autonoumously formed. The possible roles of these in the specification of vegetal cell fate in early development are discussed.

scholarly journals SYNTHESIS AND STORAGE OF MICROTUBULE PROTEINS BY SEA URCHIN EMBRYOS

1971 ◽  
Vol 50 (2) ◽  
pp. 516-528 ◽  
Author(s):  
Rudolf A. Raff ◽  
Gerald Greenhouse ◽  
Kenneth W. Gross ◽  
Paul R. Gross
Keyword(s):  
Amino Acids ◽  
Sea Urchin ◽  
Messenger Rna ◽  
Binding Activity ◽  
Total Soluble Protein ◽  
Cell Stage ◽  
Lytechinus Pictus ◽  
Sea Urchin Embryos ◽  
Tritiated Amino Acids ◽  
Vinblastine Sulfate

Studies employing colchicine binding, precipitation with vinblastine sulfate, and acrylamide gel electrophoresis confirm earlier proposals that Arbacia punctulata and Lytechinus pictus eggs and embryos contain a store of microtubule proteins. Treatment of 150,000 g supernatants from sea urchin homogenates with vinblastine sulfate precipitates about 5% of the total soluble protein, and 75% of the colchicine-binding activity. Electrophoretic examination of the precipitate reveals two very prominent bands. These have migration rates identical to those of the A and B microtubule proteins of cilia. These proteins can be made radioactive at the 16 cell stage and at hatching by pulse labeling with tritiated amino acids. By labeling for 1 hr with leucine-3H in early cleavage, then culturing embryos in the presence of unlabeled leucine, removal of newly synthesized microtubule proteins from the soluble pool can be demonstrated. Incorporation of labeled amino acids into microtubule proteins is not affected by culturing embryos continuously in 20 µg/ml of actinomycin D. Microtubule proteins appear, therefore, to be synthesized on "maternal" messenger RNA. This provides the first protein encoded by stored or "masked" mRNA in sea urchin embryos to be identified.

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