Hermo L, Green H, Clermont Y
Department of Anatomy, McGill University, Montreal, Canada.
Anat Rec. 1991 Feb;229(2):159-76. doi: 10.1002/ar.1092290203.
The initial segment of the epididymis of rats, fixed with glutaraldehyde, was postfixed with reduced osmium, a technique that clearly delineates the membranes of cisternae of the endoplasmic reticulum (ER) and the various elements of the Golgi apparatus, or with tannic acid to enhance the coats of vesicles and ribosomes on ER cisternae. The material was also treated to demonstrate various phosphatase activities (NADPase, TPPase, CMPase, G-6-Pase) or impregnated with osmium tetroxide. In osmium-impregnated material, the Golgi apparatus of the epithelial principal cells of the initial segment appeared in the light microscope as a branching, anastomosing ribbon forming a large network in the supranuclear region. In the electron microscope, ER were of two types: the heavily granulated, flattened, rough ER seen in the infranuclear and juxtanuclear regions and the distended, tubular, sparsely granulated ER, showing only few ribosomes, seen interlaced with the Golgi ribbon in the supranuclear region and at the apical pole of the cell. Of particular interest in this cell was the fact that the sparsely granulated ER approximated the Golgi stack on both its cis- and trans-faces. On the cis-face of the Golgi stack, the sparsely granulated ER cisternae showed the usual finger- or bud-like protrusions directed toward the cis element of the Golgi stack and around which numerous small 80 nm vesicles or membranous tubules were clustered. The Golgi stack consisted of the following elements in a cis-trans axis: the cis osmiophilic element, a first saccule slightly dilated, saccules two to four (S2-S4), which were NADPase-positive, and saccules five to seven and the eight Golgi element, which were TPPase-positive. On the trans-aspect of the Golgi stacks, several (up to four) CMPase-positive trans-Golgi networks were observed often in close apposition to the sparsely granulated ER cisternae. One of the trans-Golgi networks showed a "peeling-off" configuration, i.e. part of it was closely apposed to the overlying Golgi element of the stack, whereas the remaining part was separated from the stack by a space occupied by a cisterna of sparsely granulated ER. The other trans-Golgi networks were completely separated from the stack and were often seen sandwiched between sparsely granulated ER cisternae. Thus, ER cisternae showed extensive areas of close apposition but no continuity with the trans-Golgi networks. Although the saccules of the Golgi stacks showed NADPase and/or TPPase activity, the trans-Golgi networks displayed CMPase activity, thus facilitating their identification from the closely associated unreactive sparsely granulated ER cisternae.(ABSTRACT TRUNCATED AT 400 WORDS)
用戊二醛固定大鼠附睾起始段后,用还原锇进行后固定,该技术能清晰勾勒出内质网(ER)池膜和高尔基体的各种成分,或用单宁酸处理以增强ER池上囊泡和核糖体的被膜。该材料还经过处理以显示各种磷酸酶活性(NADP酶、TPP酶、CMP酶、G-6-P酶)或用四氧化锇浸渍。在经锇浸渍的材料中,附睾起始段上皮主细胞的高尔基体在光学显微镜下呈分支、吻合的带状,在核上区形成一个大网络。在电子显微镜下,ER有两种类型:在核下和近核区可见的颗粒密集、扁平的粗面ER,以及在核上区和细胞顶端极与高尔基体带交织在一起的扩张的、管状的、颗粒稀疏的ER,仅显示少数核糖体。该细胞特别有趣的是,颗粒稀疏的ER在其顺面和反面都靠近高尔基体堆叠。在高尔基体堆叠的顺面,颗粒稀疏的ER池显示出通常指向高尔基体堆叠顺侧元件的指状或芽状突起,周围聚集着许多80纳米的小囊泡或膜性小管。高尔基体堆叠在顺-反轴上由以下成分组成:顺嗜锇元件、第一个略扩张的囊泡、囊泡二至四(S2-S4),它们是NADP酶阳性,以及囊泡五至七和第八个高尔基体元件,它们是TPP酶阳性。在高尔基体堆叠的反面,经常观察到几个(最多四个)CMP酶阳性的反高尔基体网络,它们常与颗粒稀疏的ER池紧密相邻。其中一个反高尔基体网络呈“剥离”构型,即其一部分与堆叠的上覆高尔基体元件紧密相邻,而其余部分则通过颗粒稀疏的ER池占据的空间与堆叠分离。其他反高尔基体网络与堆叠完全分离,常夹在颗粒稀疏的ER池之间。因此,ER池显示出广泛的紧密相邻区域,但与反高尔基体网络没有连续性。尽管高尔基体堆叠的囊泡显示出NADP酶和/或TPP酶活性,但反高尔基体网络显示出CMP酶活性,从而便于将它们与紧密相关的无反应性颗粒稀疏的ER池区分开来。(摘要截于400字)