Rajasekaran A K, Morimoto T, Hanzel D K, Rodriguez-Boulan E, Kreibich G
Department of Cell Biology and Anatomy, Cornell University Medical College, New York, NY 10021.
J Cell Sci. 1993 Jun;105 ( Pt 2):333-45. doi: 10.1242/jcs.105.2.333.
A striking reorganization of the rough endoplasmic reticulum (RER) from a tubulo-vesicular (TV-RER) to a stacked cisternal (SC-RER) configuration was observed when the secretory activity of AR42J cells, a cell line derived from a rat pancreatic acinar carcinoma, was induced by dexamethasone. Treatment with 10 nM dexamethasone resulted in a 6.6-fold increase in the intracellular and a 4.6-fold increase in the secreted amylase activity, respectively. On the basis of the morphometric analysis of thin-section electron micrographs it has been previously reported that this increase in secretory activity is accompanied by a 2.4-fold or 30-fold increase in the size of the RER. We have developed a new biochemical method to determine the size of the RER by quantifying the membrane-bound ribosomes. Using this procedure we did not detect any change in the size of the RER after induction of an active secretory state in AR42J cells. Electron microscopic observation showed the predominance of SC-RER in dexamethasone-treated cells compared to the abundance of TV-RER in control cells. Laser scanning confocal microscopy showed a patchy distribution of ER staining in dexamethasone-treated cells compared to more basal localization in control cells. On the basis of our observations we conclude that in AR42J cells the increase in secretory activity induced by dexamethasone is accompanied by a reorganization of the RER rather than by an increase in ER surface area, as reported by others. Our results suggest that SC-RER is a biosynthetically more efficient form of the RER, which is found predominantly in actively secreting cells.
当用 dexamethasone 诱导源自大鼠胰腺腺泡癌的 AR42J 细胞系的分泌活性时,观察到糙面内质网(RER)从管状小泡状(TV-RER)显著重排为堆叠的扁平囊状(SC-RER)结构。用 10 nM dexamethasone 处理分别导致细胞内淀粉酶活性增加 6.6 倍和分泌的淀粉酶活性增加 4.6 倍。基于薄切片电子显微镜照片的形态计量分析,先前已报道分泌活性的这种增加伴随着 RER 大小增加 2.4 倍或 30 倍。我们开发了一种新的生化方法,通过定量膜结合核糖体来确定 RER 的大小。使用该方法,我们在 AR42J 细胞诱导活跃分泌状态后未检测到 RER 大小有任何变化。电子显微镜观察显示,与对照细胞中丰富的 TV-RER 相比,dexamethasone 处理的细胞中 SC-RER 占优势。激光扫描共聚焦显微镜显示,与对照细胞中更多的基础定位相比,dexamethasone 处理的细胞中 ER 染色呈斑驳分布。基于我们的观察结果,我们得出结论,在 AR42J 细胞中,dexamethasone 诱导的分泌活性增加伴随着 RER 的重排,而不是如其他人所报道的 ER 表面积增加。我们的结果表明,SC-RER 是 RER 的一种生物合成效率更高的形式,主要存在于活跃分泌的细胞中。