Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-22T20:07:49.607Z Has data issue: false hasContentIssue false
  • English
  • Français

F-actin is required for spindle anchoring and rotation in Xenopus oocytes: a re-examination of the effects of cytochalasin B on oocyte maturation

Published online by Cambridge University Press:  26 September 2008

David Lynn Gard ,
Byeong-Jik Cha  and
Amy Diane Roeder
David Lynn Gard*
Affiliation:
Department of Biology, University of Utah, Salt Lake City, Utah, USA.
Byeong-Jik Cha
Affiliation:
Department of Biology, University of Utah, Salt Lake City, Utah, USA.
Amy Diane Roeder
Affiliation:
Department of Biology, University of Utah, Salt Lake City, Utah, USA.
*
D.L. Gard, Department of Biology, University of Utah, Salt Lake City, UT 84112, USA. Telephone: 801-581-7365. Fax: 801-581-4668.
Get access Rights & Permissions [Opens in a new window]

Summary

We used confocal immunofluorescence microscopy to examine spindle migration, morphology and orientation during the maturation of Xenopus oocytes, in the presence or absence of cytochalasin B (CB), an inhibitor of actin assembly. Treatment with CB during maturation (10–50 μg/ml beginning 0–3h prior to addition of progesterone) disrupted the normal organisation of the novel MTOC and transient microtubule array (MTOC-TMA complex) that serves as the immediate precursor of the first meiotic spindle, suggesting that F-actin plays an important role in the assembly or maintenance of this complex. However, CB treatment did not block translocation of the MTOC-TMA complex to the oocyte cortex, suggesting that MTOC-TMA translocation is not dependent on an actin-based mechanism. Bipolar spindles were observed in CB-treated oocytes fixed during both M1 and M2. However, rotation of the M1 and M2 spindles into an orientation orthogonal to the oocyte surface was inhibited by CB. Rhodamine-phalloidin revealed a concentration of F-actin at the site of M1 spindle attachment, further suggesting that cortical actin is required for anchoring and rotation of the meiotic spindles. Finally, the incidence of M1 monasters was significantly increased in CB-treated oocytes, suggesting that interactions between the nascent M1 spindle and cortex are dependent on F-actin.

Keywords

Confocal microscopyCytoskeletonF-actinMeiosis
Type
Article
Information
Zygote , Volume 3 , Issue 1 , February 1995 , pp. 17 - 26
Copyright
Copyright © Cambridge University Press 1995

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Bajer, A. (1972). Spindle dynamics and chromosome movements. Int. Rev. Cytol. Suppl. 3, 1–271.Google Scholar
Beetschen, J.-C. & Gautier, J. (1989). In Developmental Biology of the Axolotl, ed Armstrong, J.B. & Malacinski, G.M., pp. 25–35. New York: Oxford University Press.Google Scholar
Blose, S.H., Meltzer, D.I. & Feramisco, J.R. (1984). 10-nm filaments are induced to collapse in living cells microinjected with monoclonal and polyclonal antibodies against tubulin. J. Cell Biol. 98, 847–58.CrossRefGoogle ScholarPubMed
Brachet, J., Hanocq, F. & VanGansen, P. (1970). A cytochemical and ultrastructurall analysis of in vitro maturation in amphibian oocytes. Dev. Biol. 21, 157–95.CrossRefGoogle ScholarPubMed
Chruch, K., Niklas, R.B. & Lin, H.-P.P (1986). Micromanipulated bivalents can trigger mini-spindle formation in Drosophila melanogaster spermatocyte cytoplasm. J. Cell Biol. 103, 2765–73.Google Scholar
Clark, T.G. & Merriam, R.W. (1978). Actin in Xenopus oocytes. I. Polymerization and gelation in vitro. J. Cell. Biol. 77, 427–38.CrossRefGoogle Scholar
Dumont, J.N. (1972). Oogenesis in Xenopus laevis (Daudin)I. stages of oocyte development in laboratory maintained animals. J. Morphol. 136, 153–80.CrossRefGoogle ScholarPubMed
Fernandez, J., Olea, N., Tellez, V. & Matte, C. (1991). Structure and development of the egg of the glossiphonid leech Theromyzon rude: reorganization of the fertilized egg during completion of the first meiotic division. Dev. Biol. 137, 142–54.CrossRefGoogle Scholar
Franke, W.W., Rathke, P.C., Seib, E., Trendeleburg, M.F., Osborn, M. & Weber, K. (1976). Distribution and mode of arragement of microfilamentous structures and actin in the cortex of the amphibian oocyte. Cytobiologie. 14, 111–30.Google Scholar
Gard, D.L. (1991). Organization, nucleation, and acetylation of microtubules in Xenopus laevis oocytes: a study by confocal immunofluorescence microscopy. Dev. Biol. 143, 346–62.CrossRefGoogle ScholarPubMed
Gard, David L. (1992 a). Microtubule organization during maturation of Xenopus oocytes: assembly and rotation of the meiotic spindles. Dev. Biol. 151, 516–30.CrossRefGoogle ScholarPubMed
Gard, David L. (1992 b). Microtubule organization and spindle assembly during meiotic maturation of Xenopus oocytes. J. Cell. Biol. 115, 46a. (abstract).Google Scholar
Gard, David L. (1993). Ectopic spindle assembly during maturation of Xenopus oocytes: evidence for functional polarization of the oocyte cortex. Dev. Biol. 159, 298–310.CrossRefGoogle ScholarPubMed
Gard, David L. (1995). Axis formation during amphibian oogenesis: re-evaluating the role of the cytoskeleton. Curr. Top. Dev. Biol. 30, 213–50.Google Scholar
Huchon, D., Crozet, N., Cantenot, N. & Ozon, R. (1981). Germinal vesicle breakdown in the Xenopus laevis oocyte: description of a transient microtubular structure. Reprod. Nutr. Dev. 21, 135–48.CrossRefGoogle ScholarPubMed
Huchon, D., Jessus, C., Thibier, C. & Ozon, R. (1988). Pressence of microtubules in isolated cortices of prophase I and metaphase II oocytes in Xenopus laevis. Cell Tissue Res. 154, 415–20.Google Scholar
Hyman, A.A. (1989). Centrosome movement in the early divisions of Caenorhabditis elegans: a cortical site determining centrosome position. J. Cell Biol. 109, 1185–93.CrossRefGoogle ScholarPubMed
Hyman, A.A. & White, J.G. (1987). Determination of cell division axes in the early embryogenesis of Caenorhabditis elegans. J. Cell Biol. 105, 2123–35.CrossRefGoogle ScholarPubMed
Jessus, C., Huchon, D. & Ozon, R. (1986). Distribution of microtubules during the breakdown of the nuclear evelope of the Xenopus oocyte: an immunocytochemical study. Biol. Cell. 56, 113–20.CrossRefGoogle Scholar
Jessus, C., Thibier, C. & Ozon, R. (1987). Levels of microtubules during meiotic maturation of the Xenopus oocyte. J. Cell Sci. 87, 705–12.CrossRefGoogle ScholarPubMed
Klymkowski, M.W., Maynell, L.A. & Polson, A. (1987). Polar asymmetry in the organization of the cortical cytokeratin system of Xenopus laevis oocytes and embryos. Development 100, 543–57.CrossRefGoogle Scholar
Klymkowsky, M.W., Maynell, L.A. & Nislow, C. (1991). Cytokeratin phosphorylation, cytokeratin filament severing, and the solubilization of the maternal mRNA Vg1. J. Cell Biol. 114, 787–97.CrossRefGoogle ScholarPubMed
Kropf, D.L., Maddock, A. & Gard, D.L. (1990). Microtubule distribution and function in early pelvetia development. J. Cell Sci. 97, 545–52.CrossRefGoogle Scholar
Lutz, D.A., Hamaguchi, Y. & Inoue, S. (1988). Micromanipulation studies of the asymmetric positioning of the maturation spindle in Chaetopterus sp. oocytes. I. Anchorage of the spindle to the cortex and migration of a displaced spindle. Cell Motil. Cytoskel. 11, 83–96.CrossRefGoogle Scholar
Merriam, R.W. & Clark, T.G. (1978). Actin in Xenopus oocytes. II. Intracellurlar distribution and polymerizability. J. Cell Biol. 77, 439–47.CrossRefGoogle ScholarPubMed
Palecek, J., Habrova, V., Nedvidek, J. & Romanovsky, A. (1985). Dynamics of tubulin structures in Xenopus laevis oogenesis. J. Embryol. Exp. Morphol. 87, 75–86.Google ScholarPubMed
Roeder, A.D. & Gard, D.L. (1994). Confocal microscopy of F-actin distribution in Xenopus oocytes. Zygote 2, 111–24.CrossRefGoogle ScholarPubMed
Ryabova, L.V., Betina, M.I. & Vassetzky, S.G. (1986). Influence of cytochalasin B on oocyte maturation in Xenopus laevis. Cell Diff. 19, 89–96.CrossRefGoogle ScholarPubMed
Steffen, W., Fuge, H., Dietz, R., Bastmeyer, M. & Muller, G. (1986). Aster-free spindle poles in insect spermatocytes: evidence for chromosome-induced spindle formation. J. Cell Biol. 102, 1679–87.CrossRefGoogle ScholarPubMed
Tang, P., Sharpe, C.R., Mohun, T.J. & Wylie, C.C. (1988). Vimentin expression in oocytes, eggs, and early embryos of Xenopus laevis. Development 103, 279–87.CrossRefGoogle ScholarPubMed
Torpey, N.P., Heasman, J. & Wylie, C.C. (1992). Distinct distribution of vimentin and cytokeratin in Xenopus oocytes and early embryos. J. Cell Sci. 101, 151–60.CrossRefGoogle ScholarPubMed
Wylie, C.C., Brown, D., Godsave, S.F., Quarmby, J. & Heasman, J. (1985). The cytoskeleton of Xenopus oocytes and its role in development. J. Embryol. Exp. Morphol. 89, (Suppl.), 1–15.Google ScholarPubMed