Global and temporal regulation of gene expression in Xenopus kidney cells in response to presumed microgravity generated by 3D clinostats.
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.
Kitamoto
Junko
J
Department of Life Sciences (Biology), Graduate School of Arts and Sciences, University of Tokyo.
Fukui
Akimasa
A
Asashima
Makoto
M
D000199:Actins / Q000502:physiology; D000818:Animals; D015820:Cadherins / Q000502:physiology; D002460:Cell Line; D002468:Cell Physiological Phenomena; D005786:Gene Expression Regulation / Q000502:physiology*; D018471:Hypergravity; D007668:Kidney / Q000166:cytology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D012399:Rotation; D013997:Time Factors; D018474:Weightlessness Simulation; D014982:Xenopus laevis
D000199:Actins; D015820:Cadherins; D012333:RNA, Messenger