Studies of the cleavage in the frog egg

Development ◽  
1969 ◽  
Vol 21 (1) ◽  
pp. 119-129
Author(s):  
T. Kubota
Keyword(s):  
Sea Urchin ◽  
Sea Urchins ◽  
Cleavage Furrow ◽  
Mitotic Apparatus ◽  
Animal Pole ◽  
Sea Urchin Eggs ◽  
Rana Nigromaculata ◽  
Vegetal Pole ◽  
Egg Cortex

In sea-urchin eggs, once karyokinesis reaches metaphase or anaphase, the cleavage furrow can be formed even if the mitotic apparatus is destroyed (Swann & Mitchison, 1953) or removed (Hiramoto, 1956). A similar result was obtained in frog eggs (Kubota, 1966). In amphibian eggs a much longer time is available for performing experiments than in sea urchins as the furrow first appears at the animal pole and slowly travels toward the vegetal pole. Taking advantage of this situation, Waddington (1952) and Dan & Kuno-Kojima (1963) performed various kinds of operations to elucidate the roles of the egg cortex and the inner cytoplasm in furrow formation, and Selman & Waddington (1955) also made cytological observations of the process. In the present paper a shift of the inner cytoplasm relative to the cortex and its influence on the course of the furrow was analysed for eggs of the frog Rana nigromaculata.

On ‘the Stationary Surface Ring’ in Heart-Shaped Cleavage

1958 ◽  
Vol 35 (2) ◽  
pp. 400-406
Author(s):  
KATSUMA DAN
Keyword(s):  
Cell Surface ◽  
Sea Urchin ◽  
Mitotic Spindle ◽  
Cleavage Furrow ◽  
Sand Dollar ◽  
Animal Pole ◽  
Sea Urchin Eggs ◽  
Stationary Surface ◽  
Vegetal Pole

1. The eggs of the sand dollar, Astriclypeus manni, and the medusa, Spirocodon saltatrix, were used for the reason that they cleave in heart shape, the cleavage furrow appearing earlier at the animal than at the vegetal pole. 2. By the superposition of drawings showing contours and astral centres as well as the positions of carbon markers on the cell surface, the presence of a pair of stationary circular zones of the cortex can be demonstrated. These remain absolutely stationary through successive stages of cleavage, as was shown to be true of regularly cleaving sea-urchin eggs. 3. The two planes determined by this pair of stationary surface rings tilt toward each other on the animal pole side in linear proportion to the eccentricity of the mitotic spindle within the cell, and the loci of the astral centres tend to slant toward the animal pole. 4. The above phenomena can be explained by the previously proposed theory for heart-shaped cleavage; i.e. the primary cause of heart-shaped cleavage is the eccentric position of the spindle, which in turn causes the rotation of the asters and the bending of the spindle.

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.

Induction of cleavage in nucleated and enucleated frog eggs by injection of isolated sea-urchin mitotic apparatus

1978 ◽  
Vol 31 (1) ◽  
pp. 117-135
Author(s):  
Y. Masui ◽  
A. Forer ◽  
A.M. Zimmerman
Keyword(s):  
Sea Urchin ◽  
Room Temperature ◽  
Rana Pipiens ◽  
Sea Urchins ◽  
Cooperative Interaction ◽  
Nuclear Materials ◽  
Cytological Examination ◽  
Mitotic Apparatus ◽  
Frog Brain ◽  
Hexylene Glycol

Mitotic apparatus (MA) were isolated in glycerol-dimethylsulphoxide solution (MTME) from zygotes of sea urchins (Stronglyocentrotus purpuratus). Freshly isolated MA were stored in 1/10 strength MTME for varying periods of time and were then injected into unfertilized frog (Rana pipiens) eggs. These injections induced 40–60% of the recipient frog eggs to initiate cleavage, resulting in the formation of blastula cell clusters. The cleavage-inducing activity of MA stored in 1/10 MTME at room temperature decreased with time of storage in 1/10 strength MTME, and disappeared by about 6 h. There was no change in the ultrastructure of MA during storage. MA isolated and stored in MTME at room temperature had a constant level of cleavage-inducing activity during the first 48 h of storage, but this activity slowly declined upon further storage; almost no activity was left after 3 weeks. MA isolated in hexylene glycol (HG) and immediately transferred into MTME were compared with MA isolated in MTME; both MA had the same cleavage-inducing activity on the day of isolation, after which the MA isolated in HG quickly lost activity. On the other hand, MA isolated and stored in HG had little cleavage-inducing activity when tested 3 h following isolation. Cleavage-inducing agent (CIA) isolated from frog brains induced cleavage and blastula formation when injected into nucleated frog eggs, but had no such activity when injected into enucleated frog eggs. MA isolated in MTME induced cleavage and blastula formation in enucleated frog eggs as well as in nucleated frog eggs. Cytological examination revealed that blastula cells which developed from MA-injected enucleated eggs contained Feulgennegative nuclei, whereas cells which developed from CIA-injected nucleated eggs contained Feulgen-positive nuclei. These results suggest that sea-urchin nuclear materials participate in mitosis in frog eggs. Isolated MA which had been stored in MTME for 3 weeks and which exhibited little cleavage-inducing activity were injected together with frog brain CIA into either normal or enucleated eggs; normal recipient eggs cleaved with significantly higher frequencies (70%) than those injected with CIA alone (40%). Furthermore, enucleated eggs injected with CIA alone failed to cleave, while those injected with MA and CIA together cleaved with significant frequencies (overall 29%). This result suggests a cooperative interaction between CIA and the inactivated MA to restore the cleavage-inducing activity of MA.

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 IMMUNOCHEMICAL STUDIES OF 22S PROTEIN FROM ISOLATED MITOTIC APPARATUS

1969 ◽  
Vol 41 (2) ◽  
pp. 577-590 ◽  
Author(s):  
Thomas Bibring ◽  
Jane Baxandall
Keyword(s):  
Sea Urchin ◽  
Morphological Features ◽  
Mitotic Apparatus ◽  
Antibody Preparation ◽  
Sea Urchin Eggs ◽  
Microtubule Protein ◽  
Mild Acid

Evidence is presented that the "22S protein" of mitotic apparatus isolated from sea urchin eggs is not microtubule protein. An antibody preparation active against 22S protein is described, and immunochemical studies of the distribution of 22S protein in various cellular fractions and among morphological features of mitotic apparatus are reported. The protein is ubiquitous in the metaphase egg fractions that were tested but is not found in sperm flagella. It is immunologically distinct from proposed microtubule protein isolated from mitotic apparatus by the method of Sakai, and from proposed microtubule protein obtained after extraction with mild acid. It exists in nontubule material of isolated mitotic apparatus but is not detectable in microtubules.

Non-plasmalemmal localisation of the major ganglioside in sea urchin eggs

Zygote ◽  
1993 ◽  
Vol 1 (3) ◽  
pp. 215-223 ◽  
Author(s):  
Hidehiko Shogomori ◽  
Kazuyoshi Chiba ◽  
Hideo Kubo ◽  
Motonori Hoshi
Keyword(s):  
Endoplasmic Reticulum ◽  
Sea Urchin ◽  
Indirect Immunofluorescence ◽  
Immunofluorescence Microscopy ◽  
Mitotic Apparatus ◽  
Sea Urchin Eggs ◽  
Indirect Immunofluorescence Microscopy ◽  
The One ◽  
Major Ganglioside ◽  
Hemicentrotus Pulcherrimus

SummaryM5 ganglioside (NeuGcα2–6Glcβl-' Cer) is the predominant glycosphingolipid in sea urchin eggs. Distribution of M5 ganglioside was studied in unfertilised and fertilised eggs of the sea urchin Hemicentrotus pulcherrimus by indirect immunofluorescence microscopy. In the cortices of unfertilised eggs, anti-M5 antibody strongly stained the submembranous, polygonal and tubular network of endoplasmic reticulum that was revealed by a membrane-staining dye, DiIC18(3). In addition to the cortical network of endoplasmic reticulum, at least two morphologically distinct vesicles were positive to the antibody. In the cortices isolated from fertilised eggs 30 min after insemination, the antibody stained only a similar network of endoplasmic reticulum, presumably the one reconstructed 5–10 min after fertilisation. During mitosis the endoplasmic reticulum is known to aggregate within the asters of the mitotic apparatus. Indeed, the antibody stained the asters and (more strongly) the vesicular components attaching to the periphery of the mitotic apparatus.

scholarly journals Effects of caffeine and other methylxanthines on the development and metabolism of sea urchin eggs. Involvement of NADP and glutathione.

1976 ◽  
Vol 68 (3) ◽  
pp. 440-450 ◽  
Author(s):  
J Nath ◽  
J I Rebhun
Keyword(s):  
Cell Division ◽  
Sea Urchin ◽  
Reductase Activity ◽  
Inhibitory Effect ◽  
Pentose Phosphate ◽  
Nad Kinase ◽  
Mitotic Apparatus ◽  
Sea Urchin Eggs ◽  
Glutathione Reductase Activity

Methylxanthines (MX) inhibit cell division in sea urchin and clam eggs. This inhibitory effect is not mediated via cAMP. MX also inhibit respiration in marine eggs, at concentrations which inhibit cleavage. Studies showed that no changes occurred in ATP and ADP levels in the presence of inhibitory concentrations of MX, indicating an extra-mitochondrial site of action for the drug. Subsequent studies revealed decreased levels of NADP+ and NADPH, when eggs were incubated with inhibitory concentrations of MX, but no change in levels of NAD+ and NADH. MX did not affect the pentose phosphate shunt pathway and did not have any effect on the enzyme NAD+ -kinase. Further studies showed a marked inhibitory effect on the glutathione reductase activity of MX-treated eggs. Reduced glutathione (GSH) could reverse the cleavage inhibitory effect of MX. Moreover, diamide, a thiol-oxidizing agent specific for GSH in living cells, caused inhibition of cell division in sea urchin eggs. Diamide added to eggs containing mitotic apparatus (MA) could prevent cleavage by causing a dissolution of the formed MA. Both MX and diamide inhibit a Ca2+-activated ATPase in whole eggs. The enzyme can be reactivated by sulfhydryl reducing agents added in the assay mixture. In addition, diamide causes an inhibition of microtubule polymerization, reversible with dithioerythritol. All experimental evidence so far suggests that inhibition of mitosis in sea urchin eggs by MX is mediated by perturbations of the in vivo thiol-disulfide status of target systems, with a primary effect on glutathione levels.

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