Shin Masashi, Hirokawa Kae, Fujiwara Kunio
Department of Applied Life Science, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan.
Histochem Cell Biol. 2006 Apr;125(4):369-75. doi: 10.1007/s00418-005-0079-8. Epub 2005 Oct 8.
Polyamines (PAs) are ubiquitous polycationic metabolites in the eukaryotic and prokaryotic cells and are believed to be intimately involved in the regulation of DNA, RNA, and protein biosynthesis. However, the subcellular localization of PAs has not yet been fully elucidated in a variety of cell types. In the present study, a pre-embedding indirect immunoperoxidase approach was used to define the fine structural localization of PAs in the gastrointestinal tract of rat, which was fixed with glutaraldehyde and the monoclonal antibody ASPM-29 specific for spermine (Spm) and spermidine (Spd). Examination by a transmission electron microscopy showed that the peroxidase end products were commonly and predominantly localized in the free and attached ribosomes of the rough endoplasmic reticulum (rER) in the active protein- or peptide-secreting cells, and in rapidly proliferating cells including the gastric chief cells, mucous neck cells, and intestinal crypt cells. The nuclei, mitochondria, and secretory vesicles were devoid of PAs. Of note is the new finding that PAs are also located even on the small number of ribosomes in the cytoplasm of the parietal cells and of the villus-tip cells, because these were the cell types that were found to be almost PA-negative at the light microscopic level. These results seem to be completely consistent with those recently obtained for rat neurons. Thus, the present study generalized the subcellular localization of PAs on the ribosomes, and demonstrated that PAs are one of the components of biologically active ribosomes, possibly in any type of cell, that are closely involved in the translation processes of protein biosynthesis.
多胺(PAs)是真核细胞和原核细胞中普遍存在的多阳离子代谢产物,被认为与DNA、RNA和蛋白质生物合成的调节密切相关。然而,在多种细胞类型中,PAs的亚细胞定位尚未完全阐明。在本研究中,采用包埋前间接免疫过氧化物酶法来确定PAs在大鼠胃肠道中的精细结构定位,大鼠胃肠道用戊二醛固定,并使用对精胺(Spm)和亚精胺(Spd)具有特异性的单克隆抗体ASPM - 29。透射电子显微镜检查显示,过氧化物酶终产物通常主要定位于活跃的蛋白质或肽分泌细胞中粗面内质网(rER)的游离核糖体和附着核糖体上,以及包括胃主细胞、黏液颈细胞和肠隐窝细胞在内的快速增殖细胞中。细胞核、线粒体和分泌小泡中不含PAs。值得注意的是,新发现PAs甚至也位于壁细胞和绒毛顶端细胞细胞质中的少数核糖体上,因为在光学显微镜水平上这些细胞类型几乎是PAs阴性的。这些结果似乎与最近在大鼠神经元中获得的结果完全一致。因此,本研究概括了PAs在核糖体上的亚细胞定位,并证明PAs是生物活性核糖体的组成成分之一,可能存在于任何类型的细胞中,且与蛋白质生物合成的翻译过程密切相关。