Abstract
In many animal species the meiosis I spindle in oocytes is anastral and lacks centrosomes. Previous studies of Drosophila oocytes failed to detect the native form of the germline-specific γ-tubulin (γTub37C) in meiosis I spindles, and genetic studies have yielded conflicting data regarding the role of γTub37C in the formation of bipolar spindles at meiosis I. Our examination of living and fixed oocytes carrying either a null allele or strong missense mutation in the γtub37C gene demonstrates a role for γTub37C in the positioning of the oocyte nucleus during late prophase, as well as in the formation and maintenance of bipolar spindles in Drosophila oocytes. Prometaphase I spindles in γtub37C mutant oocytes showed wide, non-tapered spindle poles and disrupted positioning. Additionally, chromosomes failed to align properly on the spindle and showed morphological defects. The kinetochores failed to properly co-orient and often lacked proper attachments to the microtubule bundles, suggesting that γTub37C is required to stabilize kinetochore microtubule attachments in anastral spindles. Although spindle bipolarity was sometimes achieved by metaphase I in both γtub37C mutants, the resulting chromosome masses displayed highly disrupted chromosome alignment. Therefore, our data conclusively demonstrate a role for γTub37C in both the formation of the anastral meiosis I spindle and in the proper attachment of kinetochore microtubules. Finally, multispectral imaging demonstrates the presences of native γTub37C along the length of wild-type meiosis I spindles.
Highlights
In mitosis and male meiosis in animals, the establishment of spindle bipolarity is mediated by centrosomes that act as microtubule organizing centers (MTOCs)
Schuh and Ellenberg [2] have presented strong evidence that the spindle in mouse oocytes is formed by the action of a large number of c-tubulincontaining MTOCs that are self-organized from a cytoplasmic microtubule network
In Drosophila oocytes it was not clear whether c-tubulin played a role in bipolar spindle assembly or if it was even present on the meiotic spindle
Summary
In mitosis and male meiosis in animals, the establishment of spindle bipolarity is mediated by centrosomes that act as microtubule organizing centers (MTOCs). Schuh and Ellenberg [2] have presented strong evidence that the spindle in mouse oocytes is formed by the action of a large number of c-tubulincontaining MTOCs that are self-organized from a cytoplasmic microtubule network. These authors propose that the progressive clustering of MTOCs, along with the action of a kinesin-5 motor protein, facilitates the formation of a bipolar spindle. This mechanism of acentrosomal spindle assembly is fully consistent with mammalian studies of c-tubulin during meiosis I that show localization of c-tubulin throughout the meiosis I spindle [3] and with work by Burbank et al [4] demonstrating the existence of the minus ends of the microtubules throughout the meiosis I spindle
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