XMAP230 is required for the assembly and organization of acetylated microtubules and spindles in Xenopus oocytes and eggs.
We used affinity-purified polyclonal antibodies to characterize the distribution and function of XMAP230, a heat-stable microtubule-associated protein isolated from Xenopus eggs, during oogenesis. Immunoblots revealed that XMAP230 was present throughout oogenesis and early development, but was most abundant in late stage oocytes, eggs, and early embryos. Immunofluorescence microscopy revealed that XMAP230 was associated with microtubules in oogonia, post-mitotic stage 0 oocytes, early stage I oocytes, and during stage IV-VI of oogenesis. However, staining of microtubules by anti-XMAP230 was not detectable during late stage I through stage III. In stage VI oocytes, anti-XMAP230 stained a large subset of microtubules that were also stained with monoclonal antibodies specific for acetylated (&agr ;)-tubulin. During oocyte maturation, XMAP230 was associated with the transient microtubule array that serves as the precursor of the first meiotic spindle, as well as both first and second meiotic spindles. The extensive array of cytoplasmic microtubules present throughout maturation was not detectably stained by anti-XMAP230. Microinjection of anti-XMAP230 locally disrupted the organization and acetylation of microtubules in stage VI oocytes, and reduced the re-acetylation of microtubules during recovery from cold-induced microtubule disassembly. Subsequent maturation of oocytes injected with anti-XMAP230 resulted in defects in the assembly of the transient microtubules array and first meiotic spindle. These observations suggest that XMAP230 is required for the stabilization and organization of cytoplasmic and spindle microtubules in Xenopus oocytes and eggs.
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B J
BJ
Department of Biology, University of Utah, Salt Lake City, UT 84112, USA.
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D000107:Acetylation; D000818:Animals; D000906:Antibodies / Q000008:administration & dosage; D005260:Female; D066298:In Vitro Techniques; D008540:Meiosis; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D009866:Oogenesis; D010063:Ovum / Q000378:metabolism*; D014981:Xenopus
D000906:Antibodies; D008869:Microtubule-Associated Proteins