Three-dimensional structural characterization of centrosomes from early Drosophila embryos
Three-dimensional structural characterization of centrosomes from early Drosophila embryos
复制标题
早期果蝇胚胎中心体的三维结构特征
作者：
M. Moritz;M. Braunfeld;Jennifer C. Fung;J. Sedat;Bruce Alberts;D. Agard
对中心体的近中心粒物质（PCM）内微管（MT）成核位点的机制和结构的理解一直是难以捉摸的。这部分是由于难以获得大量的中心体进行分析，以及与传统的EM技术获得可解释的结构数据的问题。我们描述了一个协议，分离出大量的功能中心体从早期果蝇胚胎。使用自动电子断层扫描，我们已经开始了三维结构的表征，这些完整的中心体与再生MT。中心体重建到约6-8 nm分辨率显示在MT的负端没有大的结构，这表明如果MT成核材料物理接触MT，它必须紧密符合负端的形状。虽然许多MT起源于中心粒附近，但在整个PCM中发现了MT负端，甚至接近其外边界。MTs纵横交错的PCM，这表明成核网站在许多不同的方向取向。没有MT的中心体的重建表明，有一个重组的PCM MT再生后，此外，环状结构，具有类似的直径作为MT是明显的中心体没有MT的PCM，可能是MT成核网站。
An understanding of the mechanism and structure of microtubule (MT)- nucleating sites within the pericentriolar material (PCM) of the centrosome has been elusive. This is partly due to the difficulty in obtaining large quantities of centrosomes for analysis, as well as to the problem of attaining interpretable structural data with conventional EM techniques. We describe a protocol for isolating a large quantity of functional centrosomes from early Drosophila embryos. Using automated electron tomography, we have begun a three-dimensional structural characterization of these intact centrosomes with and without regrown MTs. Reconstructions of the centrosomes to approximately 6-8 nm resolution revealed no large structures at the minus ends of MTs, suggesting that if MT-nucleating material physically contacts the MTs, it must conform closely to the shape of the minus end. While many MTs originate near the centrioles, MT minus ends were found throughout the PCM, and even close to its outer boundary. The MTs criss-crossed the PCM, suggesting that nucleating sites are oriented in many different directions. Reconstructions of centrosomes without MTs suggest that there is a reorganization of the PCM upon MT regrowth; moreover, ring-like structures that have a similar diameter as MTs are apparent in the PCM of centrosomes without MTs, and may be MT- nucleating sites.
影响因子：
2.2
作者：
KOSTER, AJ;CHEN, H;AGARD, DA
通讯作者：
AGARD, DA
影响因子：
10.9
作者：
Smith，Adam L.;Wang，Phillip
通讯作者：
Wang，Phillip
活果蝇胚胎中微管和肌动蛋白丝的行为。
DOI：
10.1242/dev.103.4.675
发表时间：
1988
期刊：
Development (Cambridge, England)
影响因子：
--
作者：
Kellogg,DR;Mitchison,TJ;Alberts,BM
通讯作者：
Alberts,BM