Huebner E, Gutzeit H
Department of Zoology, University of Manitoba, Manitoba, Canada.
Tissue Cell. 1986;18(5):753-64. doi: 10.1016/0040-8166(86)90075-3.
Fluorescent staining of Rhodnius prolixus ovarioles with rhodaminylphalloidin revealed an elaborate interconnecting meshwork of F-actin encasing the microtubule rich core. The thick actin struts have branches extending partially into the nurse cell lobes extending from the syncytical core. Basally the network ends at the nurse cell-pre-follicular interface and does not extend into the trophic cords. Light microscopy, TEM and SEM show the dense fibrous struts have numerous branch points creating a porous cylindrical collar at the core periphery. The filaments are organized in a dense felt-like anisotropic meshwork rather in parallel arrays. This morphology coupled with the stability of the entire structure in extraction procedures suggests the actin filaments are crosslinked, presumably by associated proteins. Detergent extraction experiments indicate isolation of the mesh is possible. Comparison of Nomarski DIC images with polarizing microscopy images of the same preparations show the F-actin struts are not birefringent while the microtubules of the core are highly birefringent and resistant to nocodazole. Indirect immunocytochemical staining of sections for actin confirmed the F-actin distribution visualized with phalloidin and contrasted markedly with staining pattern for tubulin. The discovery of this prominent actin meshwork must now be incorporated into models attempting to explain the mechanisms underlying polarized nurse cell-oocyte transport.
用罗丹明鬼笔环肽对红带锥蝽卵巢小管进行荧光染色,结果显示有一个精细的相互连接的F - 肌动蛋白网络,包裹着富含微管的核心。粗壮的肌动蛋白支柱有分支,部分延伸到从合胞体核心延伸出的滋养细胞叶中。在基部,该网络在滋养细胞 - 卵泡前界面处终止,不延伸到营养索中。光学显微镜、透射电子显微镜和扫描电子显微镜显示,致密的纤维支柱有许多分支点,在核心周边形成一个多孔的圆柱形环。这些细丝以密集的毡状各向异性网络形式组织,而非平行排列。这种形态以及整个结构在提取过程中的稳定性表明,肌动蛋白细丝可能通过相关蛋白质交联在一起。去污剂提取实验表明可以分离出该网络。将诺马斯基微分干涉对比(Nomarski DIC)图像与相同制剂的偏光显微镜图像进行比较,结果显示F - 肌动蛋白支柱不具有双折射性,而核心的微管具有高度双折射性且对诺考达唑有抗性。对肌动蛋白切片进行间接免疫细胞化学染色,证实了用鬼笔环肽观察到的F - 肌动蛋白分布,且与微管蛋白的染色模式形成明显对比。现在必须将这一显著的肌动蛋白网络的发现纳入试图解释极化滋养细胞 - 卵母细胞运输潜在机制的模型中。