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"XMAP230 Is Required for the Organization of Cortical Microtubules and Patterning of the Dorsoventral Axis in FertilizedXenopusEggs"

David Gard

Summary

The dorsoventral axis ofXenopusembryos is specified by a rotation of the egg cortex relative to the underlying yolky cytoplasm. This cortical rotation, which occurs during the first cell cycle after fertilization, is dependent upon an array of parallel microtubules in the subcortical cytoplasm. We have used confocal immunofluorescent microscopy and microinjection of affinity-purified anti-XMAP230 antibody to address the role of XMAP230, one of three highmolecular-weight microtubule-associated proteins (MAPs) inXenopuseggs, in the assembly and organization of the cortical microtubule array and specification of the dorsoventral axis. Confocal immunofluorescence microscopy revealed that XMAP230 was associated with cortical microtubules shortly after their appearance in the subcortical cytoplasm. XMAP230 staining became more prominent as microtubules were aligned and bundled during the cortical rotation. Loss of XMAP230 appeared to precede disassembly of cortical microtubules at the end of the first cell cycle. Deeper within the cytoplasm, XMAP230 was associated with microtubules early in the assembly of the sperm aster. However, later in the first cell cycle, XMAP230 was associated with microtubules (MTs) of the first mitotic spindle, spindle asters, and the cortical MTs, but not with microtubule remnants of the sperm aster. Microinjection of anti-XMAP230 antibody locally disrupted the assembly and organization of microtubules in the cortex of activated or fertilized eggs and resulted in defects in the dorsoventral patterning of embryos. These results indicate that the assembly and/or organization of cortical microtubules in fertilizedXenopuseggs and subsequent specification of the dorsoventral axis are dependent upon XMAP230.