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The structure and function of Golgi apparatus as revealed by rapid freezing techique.

快速冷冻技术揭示高尔基体的结构和功能。

基本信息

批准号：
60480103
负责人：
YAMADA Eichi
金额：
$ 2.56万
依托单位：
Fukuoka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1987
项目状态：
已结题

项目摘要

Observations by mamns of rapid freezing technique: The Golgi apparatus appeared as a continuous and complicated network of stack composed of several cisterns in parallel. Number of cisterns in one stack was usually 4-7 in the exocrinosytes of pancreas, and in some preparations, the electron poacity of cisternal lumen increased gradually from cis to trans side and finally became similar to those of secretory granules. However, the so-called rigid lamella showed no increased luminal density although it located in trans side. In other preparation, the cis most cistern showed characteristic morphology. It was a flat sac of constant thickness about 30nm and sometimes arranged in a small circle. The Golgi vesicles were classified into 2 kind due to their size, namely [60nm and [40nm in diameter. The former group was found in an area between rER and cis most cistern, where an homogeneous matrix of fine reticular mashwork ebbedded. The latter was usually noticed from lateral to trans side of the stack. The peripheral margin of the Golgi stack was surrounded by a row of such vesicles, which was most obvious when viewed en face.Morphological changes by agents affecting intracellular processings of secretory proteins: Primary cultured rat hepatocytes were iincubated for 0.5 and 2hrs with either 10^6M Monensin(M), 10mM Methylamine(A), 100<micrn>M Chloroquine(c), 20mM Trisaminomethane(T), or 10 ^6<micrn>g/ml Brefeldin A (B) respectively, and compared their morphological changes with the control. In each case, rpominant changes were niticed only in the Golgi apparatus. With M, A, C, and T, the Golgi apparatus enlarged and their cisterns became dilated increasingly from cis to trans side, while, with B, most of Golgi elements disappeared leaving some vesiclar and tubular structures. The findings were consistant with the biochemical data, and suggest that the structural compomemts of Golgi apparatus maintain balance by membrane recycling mechanism.

快速冷冻技术观察：高尔基体呈连续的、复杂的网状结构，由多个平行排列的池组成。胰腺外分泌体中池腔的电子密度通常为4-7个，有些标本池腔的电子密度由顺向反逐渐增大，最后与分泌颗粒的电子密度相似。然而，所谓的刚性板层，虽然位于反侧，但没有显示管腔密度增加。在其他制备中，顺式池显示出特征性的形态。它是一个厚度约30 nm的扁平囊，有时排列成一个小圆圈。高尔基体囊泡根据大小分为两类，直径分别为[60 nm]和[40 nm]。前一组位于rER和顺式大池之间，在那里有一个均匀的细网状混合物基质。后者通常是注意到从横向到横向侧的堆栈。影响细胞内分泌蛋白加工的物质的形态学变化：原代培养的大鼠肝细胞分别与106 μ g/ml莫能菌素（M）、10 mM甲胺（A）、100 μ g/ml<micrn>氯喹（c）、20 mM三氨基甲烷（T）和106 μ <micrn>g/ml布雷菲德菌素A（B）孵育0.5和2小时，并与对照组比较。在每一种情况下，只在高尔基体中发现了显著的变化。M、A、C和T的高尔基体增大，池由顺向反逐渐扩大，而B的大部分高尔基体消失，只留下一些泡状和管状结构。这一结果与生物化学数据相一致，提示高尔基体结构成分通过膜循环机制维持平衡。