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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.
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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

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