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1.
We describe an embryonic lethal mutation in Xenopus laevis that provokes regression of cleavage furrow formation. The mutant females (designated as af) were obtained by the back-cross of a female with one of her sons. All the fertilized eggs laid by the mutant females, regardless of the wild-type male used in the mating, failed to cleave although each furrow ran at a proper position superficially. Light and electron microscopic observations of the embryos revealed that the cleavage furrows stayed on the surface and cytoplasmic divisions did not take place at all, while nuclear divisions did. Two-dimensional gel-electrophoretic comparisons of af and wild-type embryos demonstrated that two proteins, having estimated molecular masses of about 38 kDa (pI 6.6) and 78 kDa (pI 7.6), were missing in af embryos. Microinjection of clear cytoplasm from a wild-type egg into fertilized af eggs provoked partial surface contraction and cleavage furrow formation in recipient af eggs. The results showed that the af females carry a lethal maternal-effect mutation which causes cleavage furrow regression by being deficient in a few proteins, and that cytoplasm of wild-type eggs can partially rescue the cleavage furrow formation of af eggs by furnishing the corrective material, presumably a product of the normal allele of af. 相似文献
2.
Role of nuclear material in the early cell cycle of Xenopus embryos 总被引:14,自引:0,他引:14
Activated Xenopus eggs show periodic surface contraction waves and oscillations in endogenous protein phosphorylation, MPF, and kinase activities timed with the cleavage cycle of control fertilized eggs. In this paper, we show that in activated eggs lacking the material that originates from the oocyte nucleus, MPF and kinase oscillations occur in the absence of surface contraction waves. Two mitotic phosphoproteins (M116 and M46), previously described by 32P labeling in nucleated eggs, are no longer detected in the enucleated eggs. We conclude that a cytoplasmic temporal control of MPF and kinase activities is likely to be the essential cell cycle oscillator. The oocyte nuclear components normally stored in the cytoplasm of the embryos are not involved in the clock although they appear to be required for the generation of surface contraction waves. 相似文献
3.
Stiffness of the cell was surface was determined in fertilized sea urchin and starfish eggs by measuring the mechanical resistivity of the cell surface against negative pressure applied to a restricted part with a micropipette in contact with the cell surface at its tip (elastimetry). In both sea urchin and starfish eggs, the stiffness of the cell surface changed almost in parallel between the presumptive furrow and polar surfaces before the onset of the first cleavage, and the stiffness of the furrow surface became larger than that of the polar surface when cleavage started, although temporal changes in the stiffness were different between sea urchin and starfish eggs. The stiffness of the cell surface changed almost in parallel between the surfaces at the equator and at the animal pole in starfish eggs before the onset of polar body formation. The stiffness of the cell surface around the forming polar body increased during the formation of the polar body and remained at a high level after the polar body formation. It seems that the stiffness difference responsible for the formation of the contractile ring develops simultaneously with rather than prior to the formation of the cleavage furrow. 相似文献
4.
Transit into interphase of the first mitotic cell cycle in amphibian eggs is a process referred to as activation and is accompanied by an increase in intracellular free calcium [( Ca2+]i), which may be transduced into cytoplasmic events characteristic of interphase by protein kinase C (PKC). To investigate the respective roles of [Ca2+]i and PKC in Xenopus laevis egg activation, the calcium signal was blocked by microinjection of the calcium chelator BAPTA, or the activity of PKC was blocked by PKC inhibitors sphingosine or H7. Eggs were then challenged for activation by treatment with either calcium ionophore A23187 or the PKC activator PMA. BAPTA prevented cortical contraction, cortical granule exocytosis, and cleavage furrow formation in eggs challenged with A23187 but not with PMA. In contrast, sphingosine and H7 inhibited cortical granule exocytosis, cortical contraction, and cleavage furrow formation in eggs challenged with either A23187 or PMA. Measurement of egg [Ca2+]i with calcium-sensitive electrodes demonstrated that PMA treatment does not increase egg [Ca2+]i in BAPTA-injected eggs. Further, PMA does not increase [Ca2+]i in eggs that have not been injected with BAPTA. These results show that PKC acts downstream of the [Ca2+]i increase to induce cytoplasmic events of the first Xenopus mitotic cell cycle. 相似文献
5.
Myosin and actin are necessary for polar lobe formation and resorption in Ilyanassa obsoleta embryos
During the first mitotic divisions many spiralian embryos form a cytoplasmic protrusion at the vegetal pole called the polar
lobe. In the gastropod Ilyanassa obsoleta the polar lobe is constricted by a contractile ring composed of filamentous actin, myosin, and associated proteins, similar
to the contractile ring of the cleavage furrow. To resolve the role of myosin and actin in polar lobe formation and resorption,
we have applied 2,3-butanedione monoxime and Latrunculin B at different stages of the first cleavage to inhibit myosin and
F-actin, respectively. Our results show that myosin is important for both cytokinesis and polar lobe formation. Additionally,
we have found that the resorption of the polar lobe is a two-step process: the first step is passive, driven by the tension
of the actin-cortex and the second step is active, in which the ATP-hydrolysis of myosin/actin interaction supplies the force
to complete the resorption of the polar lobe. We have summarized our results in a scheme of the first cleavage of Ilyanassa obsoleta.
Received: 6 November 1997 / Accepted:15 March 1998 相似文献
6.
Microtuble organization in Xenopus eggs during the first cleavage and its role in cytokinesis 总被引:2,自引:0,他引:2
It has been suggested that the organization of microtubules during mitosis plays an important role in cytokinesis in animal cells. We studied the organization of microtubules during the first cleavage and its role in cytokinesis of Xenopus eggs. First, we examined the immunofluorescent localization of microtubules in Xenopus eggs at various stages during the first cleavage. The astral microtubules that extend from each of the two centrosomes towards the division plane meet and connect with each other at the division plane as cytokinesis proceeds. The microtubular connection thus advances from the animal pole to the vegetal pole, and its leading edge is located approximately beneath the leading edge of the cleavage furrow. Furthermore, an experiment using nocodazole suggests that microtubules have an essential role in advancement of the cleavage furrow, but neither in contraction nor maintenance of the already formed contractile ring which underlies the cleavage furrow membrane. These results suggest that the astral microtubules play an important role in controlling the formation of the contractile ring in Xenopus eggs. 相似文献
7.
How actin filaments (F-actin) and myosin II (myosin) assemble to form the contractile ring was investigated with fission yeast and Xenopus egg. In fission yeast cells, an aster-like structure composed of F-actin cables is formed at the medial cortex of the cell during prophase to metaphase, and a single F-actin cable(s) extends from this structure, which seems to be a structural basis of the contractile ring. In early mitosis, myosin localizes as dots in the medial cortex independently of F-actin. Then they fuse with each other and are packed into a thin contractile ring. At the growing ends of the cleavage furrow of Xenopus eggs, F-actin at first assembles to form patches. Next they fuse with each other to form short F-actin bundles. The short bundles then form long bundles. Myosin seems to be transported by the cortical movement to the growing end and assembles there as spots earlier than F-actin. Actin polymerization into the patches is likely to occur after accumulation of myosin. The myosin spots and the F-actin patches are simultaneously reorganized to form the contractile ring bundles. The idea that a Ca signal triggers cleavage furrow formation was tested with Xenopus eggs during the first cleavage. We could not detect any Ca signals such as a Ca wave, Ca puffs or even Ca blips at the growing end of the cleavage furrow. Furthermore, cleavages are not affected by Ca-chelators injected into the eggs at concentrations sufficient to suppress the Ca waves. Thus we conclude that formation of the contractile ring is not induced by a Ca signal at the growing end of the cleavage furrow. 相似文献
8.
Takashi Shimizu 《Developmental biology》1981,85(1):77-88
Changes in the cortical organization at the animal pole are examined by scanning and transmission electron microscopy in the Tubifex egg undergoing second polar body formation. At very early anaphase of the second meiosis, the egg surface overlying the meiotic apparatus is undulated, but its neighboring surface appears to be smooth. Although a microfilamentous cortical layer is found in the smooth area, the cortical layer of the undulating area is thin and devoid of filamentous structures except for its central part where some filaments are observed. This local differentiation takes place normally in colchicine-treated eggs where the meiotic apparatus is destroyed. Along with the progression of the anaphase movement, the egg surface of the undulating area is, first, uplifted into a cone-shaped cytoplasmic bulge (presumptive polar body); then the height and surface area of the bulge gradually increase. The distal surface of the growing bulge appears to be undulated whereas the sides of the bulge are relatively smooth. Transmission electron microscopy reveals that a thick microfilamentous cortical layer is always localized at the proximal region of this bulge; other regions of the bulge are characterized by a thin cortical layer which is devoid of filamentous structure except for the apical portion of the bulge. Microfilaments at the base of the bulge are perpendicular or oblique to the egg surface. The cortical layer of the egg which is continuous to that of the proximal region of the bulge comprises microfilaments running parallel to the surface. The attainment of the bulge to its full size is followed by the development of the cleavage furrow along its base. The cleavage furrow appears to bisect the spindle midway between its poles. In cytochalasin B-treated eggs, where some cortical microfilaments are detected at the animal pole, a cytoplasmic bulge lower in height and wider in the diameter of its base than the normal one forms at the animal pole; however, it is subsequently resorbed into the egg. The formation of a cleavage furrow is not observed in these eggs. The mechanism of the polar body formation is discussed in the light of the present observations. 相似文献
9.
Fertilized uncleaved eggs of Xenopus laevis were divided into nucleate and non-nucleate egg fragments. Both fragments, together with the whole egg of the same batch, were observed by time-lapse cinematography.
Two kinds of cyclic surface changes, (1) rounding-up and relaxing movements and (2) surface contraction waves, accompanying each cleavage in the whole eggs and the nucleate fragments, were also observed even in the non-nucleate fragments although they do not cleave.
Cleavage intervals of the whole egg and the nucleate fragment were nearly equal, but the rounding-up intervals of the non-nucleate fragment were slightly but definitely longer than the cleavage intervals of the nucleate fragment and the whole egg. 相似文献
Two kinds of cyclic surface changes, (1) rounding-up and relaxing movements and (2) surface contraction waves, accompanying each cleavage in the whole eggs and the nucleate fragments, were also observed even in the non-nucleate fragments although they do not cleave.
Cleavage intervals of the whole egg and the nucleate fragment were nearly equal, but the rounding-up intervals of the non-nucleate fragment were slightly but definitely longer than the cleavage intervals of the nucleate fragment and the whole egg. 相似文献
10.
Video-enhanced light microscopy of the apical and subapical regions of growing hyphae of several fungal species revealed the existence of momentary synchronized motions of subcellular organelles. First discovered in a temperature-sensitive morphological mutant (ramosa-1) of Aspergillus niger, these seemingly spontaneous cytoplasmic contractions were also detected in wild-type hyphae of A. niger, Neurospora crassa, and Trichoderma atroviride. Cytoplasmic contractions in all fungi lasted about 1 s. Although the cytoplasm recovered its motility and appearance, the contraction usually led to drastic changes in Spitzenkörper (apical body) behavior and hyphal morphology, often both. Within 10 s after the contraction, the Spitzenkörper commonly became dislodged from its polar position; sometimes it disassembled into phase-dark and phase-light components; more commonly, it disappeared completely. Whether partial or complete, the dislocation of the Spitzenkörper was always accompanied by a sharp reduction or cessation of growth, and was usually followed by marked morphological changes that included bulbous hyphal tips, bulges in the hyphal profile, and formation of subapical and apical branches. The cytoplasmic contractions are vivid evidence that the most conspicuous cell organelles (membrane-bound) in living hyphae are interconnected via a contractile cytoskeletal network. 相似文献
11.
Barbara Conta Boyer John M. Arnold Michael Landolfa 《Invertebrate reproduction & development.》2013,57(2):157-170
Summary Changes in egg shape and surface morphology during maturation may be related to the localization of cytoplasmic determinants in the embryos of organisms with spiral cleavage. The eggs of the polyclad turbellarian Hoploplana inquilino undergo pronounced shape changes during the meiotic divisions which have been examined with the scanning electron microscope. Unfertilized eggs have a smooth surface that becomes covered with microvilli and microblebs within 10 min of fertilization. First polar body extrusion is accompanied by the asymmetric appearance of large blebs (Blebbing Cycle I) primarily in the animal hemisphere with one quadrant characteristically smoother than the others and bearing fewer blebs. Blebbing Cycle II, which is less pronounced than the first but is still characterized by a relatively un blebbed quadrant of the zygote, coincides with second polar body formation. These asymmetric shape changes in the animal hemisphere during meiosis may possibly correlate with a primitive form of morphogenetic segregation and beginning quadrant specialization in polyclads, the most primitive spiralians with mosaic development. 相似文献
12.
Surface alterations of fertilized surf clam (Spisula solidissima) eggs induced by concanavalin A 总被引:1,自引:0,他引:1
F J Longo 《Developmental biology》1979,68(2):422-439
Fertilized Spisula eggs, incubated in ConA, were examined at periodic intervals to determine the effects of lectin binding on events of fertilization and cleavage. ConA was localized to specific regions of the vitelline layer and plasma membrane by reacting lectin-treated eggs with horseradish peroxidase and diaminobenzidine. In contrast to eggs, little reaction product was associated with the plasma membrane of spermatozoa. Sperm that fused with ConA-treated eggs failed to move into the cortex of the ovum and were observed as bulbous appendages at the surface of the zygote. Reorganization of sperm nuclei was inhibited, and male pronuclei failed to develop. ConA also inhibited polar body formation and cleavage. The maternally derived chromatin underwent meiosis, and the chromosomes normally taken into the first and second polar bodies were retained within the zygote. All of the maternally derived chromatin was organized within four or more female pronuclei which subsequently entered mitosis. The effects of ConA binding on events at the surface of fertilized Spisula eggs were abrogated by α-methyl-d-mannoside; succinyl-ConA only partially inhibited fertilization-related processes. The effects of ConA are discussed in terms of possible cross-linking of surface components of fertilized Spisula eggs which may inhibit deformation of the zygote cortex. 相似文献
13.
Activators of protein kinase C trigger cortical granule exocytosis, cortical contraction, and cleavage furrow formation in Xenopus laevis oocytes and eggs 总被引:4,自引:1,他引:3
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Prophase I oocytes, free of follicle cells, and metaphase II eggs of the amphibian Xenopus laevis were subjected to transient treatments with the protein kinase C activators, phorbol 12-myristate 13-acetate (PMA), phorbol 12,13-didecanoate, and 1-olyeoyl-2-acetyl-sn-glycerol. In both oocytes and eggs, these treatments triggered early events of amphibian development: cortical granule exocytosis, cortical contraction, and cleavage furrow formation. Surprisingly, activation of oocytes occurred in the absence of meiotic resumption, resulting in cells with an oocytelike nucleus and interior cytoplasm, but with a zygotelike cortex. PMA-induced activation of oocytes and eggs did not require external calcium, a prerequisite for normal activation of eggs. PMA-induced activation of eggs was inhibited by retinoic acid, a known inhibitor of protein kinase C. In addition, pretreatment of eggs with retinoic acid prevented activation by mechanical stimulation and inhibited activation by calcium ionophore A23187. The results suggest that protein kinase C activation is an integral component of the Xenopus fertilization pathway. 相似文献
14.
In the paedogenetic Dipteran insect Heteropeza pygmaea it is possible by physical or chemical means to obtain oocyte-nurse chamber complexes lacking the follicular epithelium. Such oocytes nevertheless complete oogenesis and begin embryonic development. Development of these “naked” eggs has been compared to normal egg development by cinematographic analysis. Eggs which are formed without follicular epithelium are completely spherical in shape and the increase in size which normally occurs during cleavage is much less extensive. Naked eggs show shape changes during the first part of cleavage, in which bulgy cytoplasmic protrusions are formed and disappear continuously. Protrusions which are present during the mitotic divisions are partly cleaved. Cleavage folds occur much earlier in naked eggs than in normal eggs. On the other hand, the duration of the mitotic cycles during nuclear multiplication of normal and naked eggs is similar. Development of naked eggs usually continues for some time after blastoderm formation before degeneration sets in. The events taking place prior to embryonic death are difficult to relate to normal gastrulation events. However, in some cases the morphogenetic movements of naked embryos resemble germ band formation of normal embryos. 相似文献
15.
Localized calcium signals along the cleavage furrow of the Xenopus egg are not involved in cytokinesis
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It has been proposed that a localized calcium (Ca) signal at the growing end of the cleavage furrow triggers cleavage furrow formation in large eggs. We have examined the possible role of a Ca signal in cleavage furrow formation in the Xenopus laevis egg during the first cleavage. We were able to detect two kinds of Ca waves along the cleavage furrow. However, the Ca waves appeared after cleavage furrow formation in late stages of the first cleavage. In addition, cleavage was not affected by injection of dibromoBAPTA or EGTA into the eggs at a concentration sufficient to suppress the Ca waves. Furthermore, even smaller classes of Ca release such as Ca puffs and Ca blips do not occur at the growing end of the cleavage furrow. These observations demonstrate that localized Ca signals in the cleavage furrow are not involved in cytokinesis. The two Ca waves have unique characteristics. The first wave propagates only in the region of newly inserted membrane along the cleavage furrow. On the other hand, the second wave propagates along the border of new and old membranes, suggesting that this wave might be involved in adhesion between two blastomeres. 相似文献
16.
Study of equatorial surface activity occurring immediately before furrowing in Arbacia lixula (=pustulosa) eggs was undertaken to learn more about the establishment of the cleavage mechanism. Behavior of echinochrome granules in the egg surface, observed and recorded with a Nikon AFM camera, was used as the indicator of surface events. An hour after fertilization A. lixula eggs were slightly flattened and periodically photographed until the furrow appeared. By measuring regional changes in the concentration of echinochrome granules, we found that a band of equatorial surface approximately 22 μm wide, which comprises about 32% of the uncleaved egg surface, shrinks about 34% and forms a densely pigmented band averaging 15 μm wide. This contraction in the equatorial zone is accompanied by expansion or stretching in the subequatorial surfaces. The possible relation between these events and formation of the microfilamentous contractile ring is discussed. 相似文献
17.
Pérez-Mongiovi D Malmanche N Bousbaa H Sunkel C 《Development (Cambridge, England)》2005,132(20):4509-4520
The spindle checkpoint is a surveillance mechanism that regulates the metaphase-anaphase transition during somatic cell division through inhibition of the APC/C ensuring proper chromosome segregation. We show that the conserved spindle checkpoint protein BubR1 is required during early embryonic development. BubR1 is maternally provided and localises to kinetochores from prophase to metaphase during syncytial divisions similarly to somatic cells. To determine BubR1 function during embryogenesis, we generated a new hypomorphic semi-viable female sterile allele. Mutant females lay eggs containing undetectable levels of BubR1 show early developmental arrest, abnormal syncytial nuclear divisions, defects in chromosome congression, premature sister chromatids separation, irregular chromosome distribution and asynchronous divisions. Nuclei in BubR1 mutant embryos do not arrest in response to spindle damage suggesting that BubR1 performs a checkpoint function during syncytial divisions. Furthermore, we find that in wild-type embryos BubR1 localises to the kinetochores of condensed polar body chromosomes. This localisation is functional because in mutant embryos, polar body chromatin undergoes cycles of condensation-decondensation with additional rounds of DNA replication. Our results suggest that BubR1 is required for normal synchrony and progression of syncytial nuclei through mitosis and to maintain the mitotic arrest of the polar body chromosomes after completion of meiosis. 相似文献
18.
Cyclic assembly-disassembly of cortical microtubules during maturation and early development of starfish oocytes 总被引:7,自引:0,他引:7
An extensive array of cortical microtubules in oocytes of the starfish Pisaster ochraceus undergoes multiple cycles of disappearance and reappearance during maturation and early development. These events were studied in isolated fragments of the oocyte cortex stained with antitubulin antibodies for indirect immunofluorescence. The meshwork of long microtubules is present in the cortex (a) of immature oocytes, i.e., before treatment with the maturation-inducing hormone 1-methyladenine, (b) for 10-20 min after treatment with 1-methyladenine, (c) after formation of the second polar body (in reduced numbers in unfertilized oocytes), and (d) in the intermitotic period between first and second cleavage divisions. The array of cortical microtubules is absent in oocytes (a) undergoing germinal vesicle breakdown, (b) during the two meiotic divisions (polar body divisions), and (c) during mitosis of the first and, perhaps, subsequent cleavage divisions. The cycle of assembly-disassembly of cortical microtubules is synchronized to the cycle of nuclear envelope breakdown and reformation and to the mitotic cycle; specifically, cortical microtubules are present when a nucleus is intact (germinal vesicle, female pronucleus, zygote nucleus, blastomere nucleus) and are absent whenever a meiotic or mitotic spindle is present. These findings are discussed in terms of microtubule organizing centers in eggs, possible triggers for microtubule assembly and disassembly, the eccentric location of the germinal vesicle, and the regulation of oocyte maturation and cell division. 相似文献
19.
Retarded nuclear migration in Drosophila embryos with aberrant F-actin reorganization caused by maternal mutations and by cytochalasin treatment 总被引:7,自引:0,他引:7
Three X-linked mutations of Drosophila melanogaster, gs(1)N26, gs(1)N441 and paralog, had a common maternal-effect phenotype. Mutant embryos show reduced egg contraction that normally occurs at an early cleavage stage in wild-type embryos. In addition, the mutants exhibited retarded nuclear migration while synchronous nuclear divisions were unaffected. The retarded migration causes nuclei to remain in the anterior part of the embryo retaining their spherical distribution even in a late cleavage stage. This consequently results in an extreme delay in nuclear arrival in the posterior periplasm. A mutant phenocopy was induced in wild-type embryos that were treated with cytochalasin B or D at a very early cleavage stage. Remarkable differences were noticed in the organization of cortical F-actin between the mutants and the wild type throughout the cleavage stage: obvious F-actin aggregates were dispersed in the cortex of mutant embryos, in contrast to the wild type where the cortical F-actin layer was smooth and underlying F-actin aggregates were smaller than those in the mutants; the transition of the distribution pattern of F-actin in the yolk mass, from the centralized to the fragmented type, occurred later in the mutants than in wild type. The results suggest that these mutations affect the mechanism underlying establishment and transition of F-actin organization required for normal egg contraction and nuclear migration in the cleavage embryos. 相似文献
20.
Takashi Iwamatsu Munehiro Kikuyama Yukio Hiramoto 《Development, growth & differentiation》1992,34(6):709-717
To investigate whether or not causal relationship exists between the increase in intracellular Ca2+ and other cortical reactions at fertilization in the medaka, Oryzias latipes , intracellular Ca2+ was determined from luminescence of aequorin previously microinjected into cortical cytoplasm in acetone-treated eggs, when they were inseminated or activated by microinjection of Ca2+ . Neither an increase in cytoplasmic calcium nor exocytosis of cortical alveoli occurred in eggs treated with acetone, though other events of fertilization i.e. completion of meiosis, fusion of pronuclei, and accumulation of cortical cytoplasm with intact cortical alveoli in the animal pole region were observed in normal time sequence in these eggs. When denuded eggs were treated with acetone, contraction of the egg and slow resumption of meiosis (extrusion of polar body) were observed without insemination. When denuded eggs were inseminated immediately after acetone-treatment, the number of spermatozoa that penetrated into the egg was greater in the animal hemisphere than in the vegetal hemisphere. These results may indicate that acetone inactivates the egg plasma membrane or its adjacent cortical cytoplasm so that it cannot participate in a propagative increase in intracellular Ca2+ and exocytosis, while it also induces cytoplasmic activation leading to egg contraction, resumption of meiosis and formation of pronuclei. The present results suggest that sperm penetration, resumption of meiosis and ooplasmic segregation are regulated separately from the release of intracellular Ca2+ and exocytosis. 相似文献