Kitamoto Junko, Fukui Akimasa, Asashima Makoto
Department of Life Sciences (Biology), Graduate School of Arts and Sciences, University of Tokyo.
Biol Sci Space. 2004 Nov;18(3):152-3.
We documented changes in morphology and gene expression of the renal epithelial cell line A6 derived from Xenopus leavis adult kidney induced by long-term culturing with three dimensional clinostats. An oligo microarray analysis on A6 cells showed that mRNA levels of 52 out of 8091 genes were significantly altered in response to clinorotation. Upregulation or downregulation of gene expression became evident on day 8 and day 10 while there was no significant change on day 5. However, on day 15, expression of 18 out of 52 genes resumed to the levels similar to its original levels while remaining 33 genes maintained altered levels of expression. Quantitative analyses of gene expression by real-time PCR confirmed that changes in mRNA levels of selected genes were found only under clinorotation but not under hypergravity (7 G) and ground control (1 G). Morphological changes including loss of dome-like structures, disassembly of E-cadherin adherence junctions and disassembly of cortical actin were also observed over 10 days of culturing with clinorotation. The results revealed genes which expression was altered specifically in A6 cells cultured under clinorotation.
我们记录了非洲爪蟾成年肾脏来源的肾上皮细胞系A6在三维回转器上长期培养所诱导的形态学和基因表达变化。对A6细胞进行的寡核苷酸微阵列分析表明,在8091个基因中,有52个基因的mRNA水平在回转培养后发生了显著变化。基因表达的上调或下调在第8天和第10天变得明显,而在第5天没有显著变化。然而,在第15天,52个基因中的18个基因的表达恢复到了与其原始水平相似的水平,而其余33个基因则维持了改变后的表达水平。通过实时PCR对基因表达进行定量分析证实,所选基因的mRNA水平变化仅在回转培养下出现,而在超重力(7G)和地面对照(1G)下未出现。在用回转器培养10天的过程中,还观察到了形态学变化,包括穹顶状结构的丧失、E-钙黏蛋白黏附连接的解体以及皮层肌动蛋白的解体。结果揭示了在回转培养条件下A6细胞中表达发生特异性改变的基因。