Vollmer G, Layer P G
Max-Planck-Institut für Entwicklungsbiologie, Tübingen, Federal Republic of Germany.
Cell Tissue Res. 1987 Dec;250(3):481-7. doi: 10.1007/BF00218938.
Dissociated single cells from chicken retina or tectum kept in rotation-mediated cell culture aggregate, proliferate and establish a certain degree of histotypical cell-to-cell relationships ("sorting out"), but these systems never form highly laminated aggregates ("nonstratified" R- and T-aggregates). In contrast, a mixture of retinal plus pigment epithelial cells forms highly "stratified" aggregates ("RPE-aggregates", see Vollmer et al. 1984). The present comparative study of "stratified" and "nonstratified" aggregates enables us to investigate the process of cell proliferation uncoupled from that of tissue stratification. Here we try to relate these two basic neurogenetic processes with patterns of expression of cholinesterases (AChE, BChE) during formation of both types of aggregates. During early aggregate formation, in both "stratified" and "nonstratified" aggregates an increased butyrylcholinesterase activity is observed close to mitotically active cells. Quantitatively both phenomena show their maxima after 2-3 days in culture. In contrast, AChE-expression in all systems increases with incubation time. In nonproliferative areas, in the center of RPE-aggregates, the formation of plexiform layers is characterized initially by weak BChE- and then strong AChE-activity. These areas correspond with the inner (IPL) and outer (OPL) plexiform layers of the retina in vivo. Although by sucrose gradient centrifugation we find that the 6S- and the fiber-associated 11S-molecules of AChE are present in all types of aggregates, during the culture period the ratio of 11S/6S-forms increases only in RPE-aggregates, which again indicates the advanced degree of differentiation within these aggregates.(ABSTRACT TRUNCATED AT 250 WORDS)
从鸡视网膜或视顶盖解离得到的单细胞,在旋转介导的细胞培养中会聚集、增殖,并建立一定程度的组织典型细胞间关系(“分选”),但这些系统从未形成高度分层的聚集体(“非分层”的R聚集体和T聚集体)。相比之下,视网膜细胞与色素上皮细胞的混合物会形成高度“分层”的聚集体(“RPE聚集体”,见Vollmer等人,1984年)。目前对“分层”和“非分层”聚集体的比较研究,使我们能够研究细胞增殖过程与组织分层过程的解偶联。在此,我们试图将这两个基本的神经发生过程与两种聚集体形成过程中胆碱酯酶(乙酰胆碱酯酶、丁酰胆碱酯酶)的表达模式联系起来。在聚集体形成早期,在“分层”和“非分层”聚集体中,靠近有丝分裂活跃细胞处均观察到丁酰胆碱酯酶活性增加。从数量上看,这两种现象在培养2-3天后达到最大值。相比之下,所有系统中乙酰胆碱酯酶的表达均随孵育时间增加。在非增殖区域,即RPE聚集体的中心,神经纤维层的形成最初以弱丁酰胆碱酯酶活性为特征,随后以强乙酰胆碱酯酶活性为特征。这些区域与体内视网膜的内(内网层)外(外网层)神经纤维层相对应。尽管通过蔗糖梯度离心我们发现,乙酰胆碱酯酶的6S分子和与纤维相关的11S分子存在于所有类型的聚集体中,但在培养期间,11S/6S形式的比例仅在RPE聚集体中增加,这再次表明这些聚集体内的分化程度更高。(摘要截选至250词)