ORGANIZATION OF WOLBACHIA-PIPIENTIS IN THE DROSOPHILA FERTILIZED EGG AND EMBRYO REVEALED BY AN ANTI-WOLBACHIA MONOCLONAL-ANTIBODY
ORGANIZATION OF WOLBACHIA-PIPIENTIS IN THE DROSOPHILA FERTILIZED EGG AND EMBRYO REVEALED BY AN ANTI-WOLBACHIA MONOCLONAL-ANTIBODY
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DOI：
10.1016/0925-4773(95)00372-x
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发表时间：
1995-06-01
影响因子：
2.6
通讯作者：
KARR, TL
中科院分区：
文献类型：
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作者：
KOSE, H;KARR, TL
果蝇中的细胞质不相容性 (CI) 与沃尔巴克氏体的存在有关，沃尔巴克氏体是一种在许多昆虫物种中发现的细胞内微生物。为了研究沃尔巴克氏体在卵和胚胎中的细胞内定位，我们从果蝇胚胎中纯化了细菌，随后生成了针对沃尔巴克氏体的特异性单克隆抗体（Mab Wol-1）。使用 Wol-1 的间接免疫荧光染色表明，在有丝分裂过程中，沃尔巴克氏体位于纺锤体极酸性中心体附近。使用抗微管蛋白和抗沃尔巴克氏体抗体的双标记免疫荧光实验表明，沃尔巴克氏体在整个细胞周期中与中心体微管共定位。七个观察结果暗示了细菌与中心体组织的微管之间的直接相互作用：（1）在整个有丝分裂周期中，沃尔巴克氏体的位置和运动精确地模仿中心体的行为，并且显然与中心体组织的微管相关； (2) 沃尔巴克氏体在有丝分裂过程中均匀地分离到每个纺锤体极； (3)沃尔巴克氏体在有丝分裂过程中不与纺锤体微管结合； (4) 位于卵皮层的沃尔巴克氏体在胚盘形成过程中定位于由微管组织的细胞质区域； (5)缺乏中心体但含有相关微管的极体核不含沃尔巴克氏体； (6) 在分化和中心体丢失后，沃尔巴克氏体不再与卵黄核结合； (7) 在极细胞形成过程中，当极细胞形成时，沃尔巴克氏体与细胞核顶端侧的中心体共定位。定量数据表明，尽管在同一时期发生了快速的核增殖和随后的细胞增殖，但在前胚层时期没有发生沃尔巴克氏体生长。这表明沃尔巴克氏体受到宿主严格的生长调节，表明宿主因素在调节卵中沃尔巴克氏体的生长中发挥作用。对在该卵中观察到的宿主和共生体之间广泛的整体相互作用进行进一步的细胞和分子研究，应该为宿主/共生体的进化和细胞质不相容性的细胞生物学提供重要的新见解。
Cytoplasmic incompatibility (CI) in Drosophila is related to the presence of Wolbachia, an intracelluar microorganism found in many species of insects. In order to study the intracellular localization of Wolbachia in eggs and embryos, we have purified the bacteria from fly embryos and subsequently generated a monoclonal antibody (Mab Wol-1) specific for Wolbachia. Indirect immunofluorescence staining using Wol-1 reveals that during mitosis, Wolbachia are localized near spindle poles acid centrosomes. Double label immunofluorescence experiments using anti-tubulin and anti-Wolbachia antibodies show that Wolbachia co-localize with centrosomal microtubules throughout the cell cycle. Direct interactions between the bacteria and centrosome-organized microtubules are implied from seven observations: (1) throughout the mitotic cycle, the position and movement of Wolbachia precisely mimic the behavior of the centrosome and apparently associates with centrosome-organized microtubules; (2) Wolbachia segregate equally to each spindle pole during mitosis; (3) Wolbachia do not associate with spindle microtubules during mitosis; (4) Wolbachia located in the egg cortex localize to the domains of cytoplasm organized by microtubules during blastoderm formation; (5) polar body nuclei that lack centrosomes but contain associated microtubules do not contain Wolbachia; (6) Wolbachia no longer associate with yolk nuclei, following differentiation and loss of centrosomes; (7) during pole cell formation, Wolbachia co-localize with the centrosome on the apical side of the nucleus as pole cells form. Quantitative data indicates that no Wolbachia growth occurs during the preblastoderm period even though rapid nuclear, and subsequent cellular, proliferation takes place during this same period. This indicates that Wolbachia are under strict growth regulation by the host suggesting that host factors play a role in regulating growth of Wolbachia in the egg. Further cellular and molecular studies of the extensive, global interactions between host and symbiont observed in this egg should provide important new insights into the evolution of host/symbiosis and the cell biology of cytoplasmic incompatibility.