Organellar relationships in the Golgi region of the pancreatic beta cell line, HIT-T15, visualized by high resolution electron tomography
Organellar relationships in the Golgi region of the pancreatic beta cell line, HIT-T15, visualized by high resolution electron tomography
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DOI：
10.1073/pnas.051631998
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发表时间：
2001-02-27
影响因子：
11.1
通讯作者：
McIntosh, JR
中科院分区：
文献类型：
--
作者：
Marsh, BJ;Mastronarde, DN;McIntosh, JR
本文在三维空间（3D）中，以6 nm的分辨率，对哺乳动物胰岛素分泌细胞的细胞质密集区中所有可见细胞器的位置关系进行了分析。一个快速冷冻/冷冻取代的HIT-T15细胞，其中包括一个大部分的高尔基带的一部分，在3-D重建的电子断层扫描。重建的体积（3.1 × 3.2 × 1.2 μ m（3））允许细胞器之间的相互作用，微管和细胞器膜之间的网站，以准确地定义在3-D和定量分析的空间密度分析。我们的数据证实，在间期哺乳动物细胞中的高尔基体是一个单一的带状细胞器，由一堆堆扁平的池组成，池中有各种大小的开口[Rambourg，A.，克莱蒙特，Y.，& Hermo，L.（1979）Am. J. Anat 154，455-476]。这些数据还表明，内质网（ER）是一个单一的连续隔室，与线粒体，多个transGolgi池和内-溶酶体系统的隔室形成密切接触。ER通过脑池开口从一侧到另一侧穿过高尔基带。微管与顺式高尔基体、内质网和内溶酶体隔室形成紧密的、非随机的关联。尽管在高尔基体区域中细胞器密集堆积，但计算出约66%的重建体积代表细胞质基质。我们在高尔基体区域的细胞器之间的结构关联的亲密度，量化的空间密度分析，在哺乳动物细胞中的膜贩运和细胞器通信的生化机制。
The positional relationships among all of the visible organelles in a densely packed region of cytoplasm from an insulin secreting, cultured mammalian cell have been analyzed in three dimensions (3-D) at approximate to6 nm resolution. Pa rt of a fast frozen/freeze-substituted HIT-T15 cell that included a large portion of the Golgi ribbon was reconstructed in 3-D by electron tomography. The reconstructed Volume(3.1 x 3.2 x 1.2 mum(3)) allowed sites of interaction between organelles, and between microtubules and organellar membranes, to be accurately defined in 3-D and quantitatively analyzed by spatial density analyses. Our data confirm that the Golgi in an interphase mammalian cell is a single, ribbon-like organelle composed of stacks of flattened cisternae punctuated by openings of Various sizes [Rambourg, A., Clermont, Y., & Hermo, L. (1979) Am. J. Anat 154, 455-476]. The data also show that the endoplasmic reticulum (ER) is a single continuous compartment that forms close contacts with mitochondria, multiple trans Golgi cisternae, and compartments of the endo-lysosomal system. This ER traverses the Golgi ribbon from one side to the other via cisternal openings. Microtubules form close, non-random associations with the cis Golgi, the ER, and endo-lysosomal compartments. Despite the dense packing of organelles in this Golgi region, approximate to 66% of the reconstructed Volume is calculated to represent cytoplasmic matrix. We relate the intimacy of structural associations between organelles in the Golgi region, as quantified by spatial density analyses, to biochemical mechanisms for membrane trafficking and organellar communication in mammalian cells.