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  • 1
    ISSN: 1432-041X
    Keywords: Key words Cell fate ; Skeletogenic potential ; Echinoid ; Induction ; Secondary mesenchyme cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  During the normal development of echinoids, an animal cap consisting of 8 mesomeres in a 16-cell stage embryo differentiates exclusively into ectoderm. Micromeres in an embryo at the same stage differentiate into primary mesenchyme cells (PMC) and coelomic pouch constituents. An animal cap and a quartet of micromeres were isolated from a 16-cell stage embryo and recombined to make a chimeric embryo devoid of presumptive endoderm and secondary mesenchyme cells (SMC). The PMC in the chimeric embryo were completely removed at the mesenchyme blastula stage. The PMC-depleted chimeric embryos formed an archenteron derived from the mesomeres. Some secondary mesenchyme-like cells (induced SMC) were released from the archenteron tip. A considerable fraction of the induced SMC formed the typical mesenchyme pattern after migrating into the vegetal region, synthesized skeletogenic mesenchyme cell-surface protein (msp130) and produced the larval skeleton. These findings indicate that induced SMC derived from the presumptive ectoderm have the same nature as natural SMC in both the timing of their release and their skeletogenic potential expressed in the absence of PMC.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 2 (1982), S. 103-113 
    ISSN: 0886-1544
    Keywords: actin ; cleavage ; fluorescein-labeled phalloidin ; microinjection ; phalloidin ; sand dollar eggs ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Effects of microinjection of phalloidin on fertilization and cleavage of sand dollar (Clypeaster japonicus and Scaphechinus mirabilis) eggs were studied. The drug, previously injected into unfertilized eggs, showed no effect on the elevation of the fertilization membrane upon insemination up to an intracellular concentration of 50 μM. However, the movement of the egg pronucleus to the sperm pronucleus was inhibited and the fusion of pronuclei did not occur. The subsequent development no longer took place. When phalloidin was injected into fertilized eggs, the thickness of the cortical layer increased and the microvilli became conspicuous. Both nuclear division and cleavage were inhibited at the intracellular concentration of more than 20 μM, though the latter seemed to be more sensitive to phalloidin than the former.Fluorescein-labeled phalloidin (FL-phalloidin) was injected into eggs in order to investigate F-actin localization by fluorescence microscopy. In both unfertilized and fertilized eggs, FL-phalloidin was localized in the cortical layer within 1 min after injection. It was also localized in the cortical layer as radially oriented rodlike structures when injected into fertilized eggs before the disappearance of the nuclear membrane. No distinct fluorescence was detected in the mitotic apparatus or in the cleavage furrow. FL-phalloidin redistributed gradually into egg cytoplasm. In unfertilized eggs, fluorescent rods were found especially in the egg pronucleus 30 min after injection.
    Additional Material: 6 Ill.
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  • 3
    ISSN: 0886-1544
    Keywords: mitosis ; mitotic apparatus ; monoclonal antibodies ; sand dollar egg ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The heterogeneity of mitotic microtubules in dividing sea urchin eggs was investigated by indirect immunofluorescence using five anti-α-tubulin (YL1/2, DM1A, E3B8, D2D6, and 6-11B-1) and two anti-β-tubulin (E6B6 and DM1B) antibodies. These antibodies were divided into four classes in regard to the different immunofluorescent staining patterns: class I, which strongly stained both the spindle and aster (YL1/2, DM1A, E3B8 and E6B6); class II, which strongly stained the spindle but weakly stained the aster (D2D6); class III, which stained only the aster (DM1B); and class IV, which did not stain the mitotic apparatus (6-11B-1). These results suggest that tubulin isotypes are distributed differently in the sea urchin mitotic microtubules and that α-tubulin isotype(s) recognized by D2D6 is (are) localized mainly in spindle microtubules, whereas β-tubulin isotype(s) recognized by DM1B is (are) found only in astral microtubules.
    Additional Material: 8 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1981), S. 387-397 
    ISSN: 0886-1544
    Keywords: birefringence ; polarizing microscope ; sea urchin egg ; cortex ; mitosis ; cleavage ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Birefringence (BR) at the cell surface of fertilized eggs of the sand-dollar, Clypeaster japonicus, during mitosis and cleavage was determined with a photoelectric BR detection apparatus [Hiramoto et al, 1981a]. The cortex of about 2 μm thickness is birefringent positive with respect to the normal to the cell surface. The hyaline layer is negatively birefringent. The halo-layer consisting of a row of microvilli surrounding the egg is positively birefringent in normal Ca-free sea water, while it is negatively birefringent in Ca-free sea water with high refractive index. The BR of the cortex gradually increases over the entire surface during mitosis until the onset of cleavage. The BR of the cortex at the polar region reaches a maximum shortly after the onset of cleavage and then decreases, while the BR of the cortex at the equatorial region begins to decrease shortly before the onset of cleavage, reaches a minimum shortly after the cleavage starts, and then increases again as the cleavage furrow advances. The coefficient of birefringence of the cortex is about 2.5 × 10-5 at the maximum. The BR change of the cortex during mitosis and cleavage is interpreted as a passive deformation caused by the constriction of the contractile ring as well as an active structural change of the cortex occurring in the dividing cell.
    Additional Material: 5 Ill.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 29 (1994), S. 241-249 
    ISSN: 0886-1544
    Keywords: immunofluorescence ; microinjection ; mitotic apparatus ; monoclonal antibodies ; sand dollar egg ; tubulin isotypes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effect on fixation on the reactivities of mitotic microtubules with monoclonal anti-tubulin antibodies was investigated by the indirect immunofluorescence procedure. All of the seven antibodies used intensely stained mitotic microtubules in sea urchin eggs lysed and fixed with methanol at -20°C, whereas only two of them stained the stabilized microtubules in the lysed eggs before the fixation. The other five did not stain the mitotic microtubules even after microtubule components other than tubulin were removed by treating the lysed eggs with 0.4 M KCl solution containing taxol. These results exclude the possibility that the fixation affects proteins, which interact with microtubules including microtubule-associated proteins (MAPs) and interfere with the binding of monoclonal antibodies with tubulin, and strongly suggest that the fixation directly affects the three-dimensional conformation of tubulin Furthermore, microinjection of these antibodies indicated the results as follows [combining the results reported previously; Oka et al., 1990: Cell Struct. Funct. 15: 373-378]: The antibodies which stained mitotic microtubules stabilized in the lysed eggs induced disassembly of native mitotic microtubules in the living eggs, but those which did not stain the stabilized microtubules did not disassemble the native microtubules. From these results, it is suggested that the monoclonal antibodies which stain microtubules in the eggs lysed but not fixed are useful for microinjection experiments. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 6 (1986), S. 549-559 
    ISSN: 0886-1544
    Keywords: activation ; microinjection ; polar body ; sea urchin eggs ; starfish oocytes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Redistribution of alpha-actinin during fertilization was investigated by means of the microinjection of fluorescently labeled egg alpha-actinin in the sea urchin, Hemicentrotus pulcherrimus. Upon fertilization, labeled alpha-actinin accumulated locally around the sperm binding site, where the fertilization cone formed soon afterwards. The accumulation propagated all over the cortex within 10 sec after a latent period of 10-20 sec. When an egg in Na-free seawater was injected with both alpha-actinin and calcium buffer (intracellular free Ca2+ concentration = 9 μM), the accumulation of alpha-actinin was similar to that in normal seawater, which suggests that the accumulation did not depend on the increase in intracellular pH but only on the increase in the intracellular free Ca2+ concentration. In immature oocytes the accumulation was detected in the cortical region, including the huge protruding cytoplasm where the sperm entered. When labeled egg alpha-actinin was injected into starfish (Asterias amurensis) oocytes followed by insemination, it accumulated in the cortical layer in a manner similar to the case of sea urchin, except that the accumulation in fertilization cones of maturing oocytcs or reception cones of immature oocytes appeared ringlike and rodlike, respectively. Moreover, just after the arrival of the meiotic apparatus, egg alpha-actinin accumulated in the cortical region, where the formation of the polar body was expected. This suggests that the meiotic apparatus somehow induced the differentiation of the cortex so as to form a polar body. It is concluded that the cortical region where alpha-actinin accumulated coincided with the microfilament-rich region. This suggests that alpha-actinin plays a role in forming the cortical meshwork of actin filaments.
    Additional Material: 8 Ill.
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  • 7
    ISSN: 0886-1544
    Keywords: unequal cell division ; spindle positioning ; spindle anchorage ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We investigated the nature of the asymmetric positioning and attachment of Chaetopterus oocyte meiotic spindles to the animal pole cortex by micromanipulation. The manipulated spindle's behavior was analyzed in clarified oocyte fragments using video-enhanced polarized light microscopy. As the spindle was drawn towards the cell interior with a microneedle, the cell surface dimpled inwards adjacent to the outer spindle pole. As the spindle was pulled further inwards, the dimple suddenly receded indicating a rupture of a mechanical link between the cell cortex and outer spindle pole. The spindle paused briefly when released from the microneedle; then it spontaneously migrated back to the original attachment site and reassociated with the cell cortex. Positive birefringent astral fibers were seen running between the outer spindle pole and the cortex during the migration. The velocity of the spindle during its migration tended to increase as it came closer to the cortex. Velocities as high as 1.25 μm/sec. were measured. If removed too far from the attachment site cortex ( 〉 35 μm), the spindle remained stationary until pushed closer to the original attachment site. Spindles, inverted by micromanipulation, migrated and reattached to the cortical site by their former inner pole; thus either spindle pole can seek out and migrate to the original attachment site. However, spindle poles pushed against other cortical regions did not attach demonstrating that there is only one unique, localized attachment site for spindle attachment.
    Additional Material: 9 Ill.
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  • 8
    Publication Date: 1997-03-27
    Print ISSN: 0949-944X
    Electronic ISSN: 1432-041X
    Topics: Biology
    Published by Springer
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