Developing novel instruments and technologies for spatio-temporal and dynamic meas-urements of the intricate cellular effects involving molecular translocation, signal transduction, and molecular interactions inside living cells is essential for the cell and molecular biology science. For the purpose of monitoring and investigating molecular events in living cells at real-time, the surface plasmon resonance based cytosensor (SBCS) for cell cultur-ing and signal monitoring was established, and on the basis of it, the corresponding technology was also established by monitoring and analyzing SPR re-sponses induced in rat C6 glioma cells by phorbol 12-myristate 13-acetate (PMA). The SPR signals induced by PMA in living C6 cells were significantly different from those groups without cells. These re-sponses were strongly dependent on and saturable to the concentrations of PMA, and could be sup-pressed by the specific and potent PKC inhibitors, which indicated that the measured signal could be the reflection of the redistribution of intracellular components near the cell membrane triggered by the activation of PKC. This research provides a quantita-tive and non-invasive technique to study the spa-tio-temporal characteristics of the cellular effects in living cells at real-time. Furthermore, this technology could also be widely used in the basic research as well as applied realms, such as space effects evaluation, environmental safety assessment, bio-logical weapon detection, cellular and molecular re-search, and drug screening.
WANG Chunyan LI Yinghui XIONG Jianghui,TAN Yingjun YU Jianru
Lack of gravity during spaceflight has profound effects on cardiovascular system, but little is known about how the cardiomyocytes respond to microgravity. In the present study, the effects of spaceflight on the structure and function of cultured cardiomyocytes were reported. The primary cultures of neo- natal rat cardiomyocytes were carried on Shenzhou-6 spacecraft and activated at 4 h in orbit. 8 samples were fixed respectively at 4, 48 and 96 h after launching for immunofluorescence of cytoskeleton, and 2 samples remained unfixed to analyze contractile and secretory functions of the cultures. Ground sam- ples were treated in our laboratory in parallel. After 115 h spaceflight, video recordings displayed that the number of spontaneous beating sites in flown samples decreased significantly, and the cells in the beating aggregate contracted in fast frequency without synchrony. Radioimmunoassay of the medium showed that the atrial natriuretic peptide secreted from flown cells reduced by 59.6%. Confocal images demonstrated the time-dependant disassembly of mirotubules versus unchanged distribution and or- ganization of microfilaments. In conclusion, above results indicate reduced function and disorganized cytoskeleton of cardiomyocytes in spaceflight, which might provide some cellular basis for further investigations to probe into the mechanisms underlying space cardiovascular dysfunction.
YANG Fen LI YingHui DING Bai NIE JieLin WANG HongHui ZHANG XiaoYou WANG ChunYan LING ShuKuan NI ChengZhi DAI ZhongQuan TAN YingJun WAN YuMin
