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Rapid alterations of cell cycle control proteins in human T lymphocytes in microgravity.

Thiel CS, Paulsen K, Bradacs G, Lust K, Tauber S, Dumrese C, Hilliger A, Schoppmann K, Biskup J, Gölz N, Sang C, Ziegler U, Grote KH, Zipp F, Zhuang F, Engelmann F, Hemmersbach R, Cogoli A, Ullrich O - Cell Commun. Signal (2012)

Bottom Line: In our study we aimed to identify rapidly reacting gravity-responsive mechanisms in mammalian cells in order to understand if and how altered gravity is translated into a cellular response.Additionally, during 2D clinorotation, Tyr-15-phosphorylation occurred later and was shorter than in the 1 g controls.The histone acetyltransferase (HAT) inhibitor curcumin was able to abrogate microgravity-induced p21 mRNA expression, whereas expression was enhanced by a histone deacetylase (HDAC) inhibitor.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Anatomy, Faculty of Medicine, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland. oliver.ullrich@anatom.uzh.ch.

ABSTRACT
In our study we aimed to identify rapidly reacting gravity-responsive mechanisms in mammalian cells in order to understand if and how altered gravity is translated into a cellular response. In a combination of experiments using "functional weightlessness" provided by 2D-clinostats and real microgravity provided by several parabolic flight campaigns and compared to in-flight-1g-controls, we identified rapid gravity-responsive reactions inside the cell cycle regulatory machinery of human T lymphocytes. In response to 2D clinorotation, we detected an enhanced expression of p21 Waf1/Cip1 protein within minutes, less cdc25C protein expression and enhanced Ser147-phosphorylation of cyclinB1 after CD3/CD28 stimulation. Additionally, during 2D clinorotation, Tyr-15-phosphorylation occurred later and was shorter than in the 1 g controls. In CD3/CD28-stimulated primary human T cells, mRNA expression of the cell cycle arrest protein p21 increased 4.1-fold after 20s real microgravity in primary CD4+ T cells and 2.9-fold in Jurkat T cells, compared to 1 g in-flight controls after CD3/CD28 stimulation. The histone acetyltransferase (HAT) inhibitor curcumin was able to abrogate microgravity-induced p21 mRNA expression, whereas expression was enhanced by a histone deacetylase (HDAC) inhibitor. Therefore, we suppose that cell cycle progression in human T lymphocytes requires Earth gravity and that the disturbed expression of cell cycle regulatory proteins could contribute to the breakdown of the human immune system in space.

No MeSH data available.


Related in: MedlinePlus

In-flight-experiment system during parabolic flights on board the Airbus A300 ZERO-G. a: Experiment hardware structure which consists of an incubator rack to store the cell containers temporarily before the experiment at 37°C (1), an experiment rack, in which all active aggregates are accommodated for the execution of the experiment and where the living cells are handled during altered gravity (2) and a cooling rack to store temporarily all cell containers after the injection of the stop/fixation liquid at 4°C until landing (3). b. Structure of the working rack, rear side. In range A (4°C) are three separate hose pumps, which pump one of the two possible stop liquids (e.g. ethanol or 4% formaldehyde) into the cell containers. The pumps are activated and regulated by the control unit inside range C, which also carries all electrical connections and fuse elements. All liquids are sucked under exclusion of air. Inside range B (37°C) are three bags with activator liquids, connected to three separate hose pumps, which pump the activator solution into the cell container upon activation of the individual experiment. c. Structure of the working rack, front side. Range D is the actual working space with three cell containers in parallel, which are connected to the activator and fixation liquid bag. Range D is also waterproof and the door position is controlled by a safety switch (sensor). Activation of the experiment is only possible when the front door is closed. d. Double-wall liquid-proof cell container. A maximum of 60 container can be accommodated during one flight. 1 = plastic container, 2 = air valve, 3 = internal sterile cell culture bag (Nutrimix 0.25l), 4 = connector 1 (activator liquid), 5 = connector 2 (stop/fixation liquid), 6 = connector 3 (filling port for cells, performed preflight), 7 = plastic flange. e. Fluid loop diagram for one cell container. The entire experiment system has three identical fluid loops.
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Figure 4: In-flight-experiment system during parabolic flights on board the Airbus A300 ZERO-G. a: Experiment hardware structure which consists of an incubator rack to store the cell containers temporarily before the experiment at 37°C (1), an experiment rack, in which all active aggregates are accommodated for the execution of the experiment and where the living cells are handled during altered gravity (2) and a cooling rack to store temporarily all cell containers after the injection of the stop/fixation liquid at 4°C until landing (3). b. Structure of the working rack, rear side. In range A (4°C) are three separate hose pumps, which pump one of the two possible stop liquids (e.g. ethanol or 4% formaldehyde) into the cell containers. The pumps are activated and regulated by the control unit inside range C, which also carries all electrical connections and fuse elements. All liquids are sucked under exclusion of air. Inside range B (37°C) are three bags with activator liquids, connected to three separate hose pumps, which pump the activator solution into the cell container upon activation of the individual experiment. c. Structure of the working rack, front side. Range D is the actual working space with three cell containers in parallel, which are connected to the activator and fixation liquid bag. Range D is also waterproof and the door position is controlled by a safety switch (sensor). Activation of the experiment is only possible when the front door is closed. d. Double-wall liquid-proof cell container. A maximum of 60 container can be accommodated during one flight. 1 = plastic container, 2 = air valve, 3 = internal sterile cell culture bag (Nutrimix 0.25l), 4 = connector 1 (activator liquid), 5 = connector 2 (stop/fixation liquid), 6 = connector 3 (filling port for cells, performed preflight), 7 = plastic flange. e. Fluid loop diagram for one cell container. The entire experiment system has three identical fluid loops.

Mentions: The experiment hardware structure is demonstrated in Figure 4 (please refer to the Figure 4 legend and to the additional file 1). It consists of an incubator rack to temporarily store the cell containers before the experiment at 37°C, an experiment rack in which all active aggregates are accommodated for the execution of the experiment and where the living cells are handled during altered gravity and a cooling rack to store temporarily all cell containers after the injection of the stop/fixation liquid at 4°C until landing.


Rapid alterations of cell cycle control proteins in human T lymphocytes in microgravity.

Thiel CS, Paulsen K, Bradacs G, Lust K, Tauber S, Dumrese C, Hilliger A, Schoppmann K, Biskup J, Gölz N, Sang C, Ziegler U, Grote KH, Zipp F, Zhuang F, Engelmann F, Hemmersbach R, Cogoli A, Ullrich O - Cell Commun. Signal (2012)

In-flight-experiment system during parabolic flights on board the Airbus A300 ZERO-G. a: Experiment hardware structure which consists of an incubator rack to store the cell containers temporarily before the experiment at 37°C (1), an experiment rack, in which all active aggregates are accommodated for the execution of the experiment and where the living cells are handled during altered gravity (2) and a cooling rack to store temporarily all cell containers after the injection of the stop/fixation liquid at 4°C until landing (3). b. Structure of the working rack, rear side. In range A (4°C) are three separate hose pumps, which pump one of the two possible stop liquids (e.g. ethanol or 4% formaldehyde) into the cell containers. The pumps are activated and regulated by the control unit inside range C, which also carries all electrical connections and fuse elements. All liquids are sucked under exclusion of air. Inside range B (37°C) are three bags with activator liquids, connected to three separate hose pumps, which pump the activator solution into the cell container upon activation of the individual experiment. c. Structure of the working rack, front side. Range D is the actual working space with three cell containers in parallel, which are connected to the activator and fixation liquid bag. Range D is also waterproof and the door position is controlled by a safety switch (sensor). Activation of the experiment is only possible when the front door is closed. d. Double-wall liquid-proof cell container. A maximum of 60 container can be accommodated during one flight. 1 = plastic container, 2 = air valve, 3 = internal sterile cell culture bag (Nutrimix 0.25l), 4 = connector 1 (activator liquid), 5 = connector 2 (stop/fixation liquid), 6 = connector 3 (filling port for cells, performed preflight), 7 = plastic flange. e. Fluid loop diagram for one cell container. The entire experiment system has three identical fluid loops.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3275513&req=5

Figure 4: In-flight-experiment system during parabolic flights on board the Airbus A300 ZERO-G. a: Experiment hardware structure which consists of an incubator rack to store the cell containers temporarily before the experiment at 37°C (1), an experiment rack, in which all active aggregates are accommodated for the execution of the experiment and where the living cells are handled during altered gravity (2) and a cooling rack to store temporarily all cell containers after the injection of the stop/fixation liquid at 4°C until landing (3). b. Structure of the working rack, rear side. In range A (4°C) are three separate hose pumps, which pump one of the two possible stop liquids (e.g. ethanol or 4% formaldehyde) into the cell containers. The pumps are activated and regulated by the control unit inside range C, which also carries all electrical connections and fuse elements. All liquids are sucked under exclusion of air. Inside range B (37°C) are three bags with activator liquids, connected to three separate hose pumps, which pump the activator solution into the cell container upon activation of the individual experiment. c. Structure of the working rack, front side. Range D is the actual working space with three cell containers in parallel, which are connected to the activator and fixation liquid bag. Range D is also waterproof and the door position is controlled by a safety switch (sensor). Activation of the experiment is only possible when the front door is closed. d. Double-wall liquid-proof cell container. A maximum of 60 container can be accommodated during one flight. 1 = plastic container, 2 = air valve, 3 = internal sterile cell culture bag (Nutrimix 0.25l), 4 = connector 1 (activator liquid), 5 = connector 2 (stop/fixation liquid), 6 = connector 3 (filling port for cells, performed preflight), 7 = plastic flange. e. Fluid loop diagram for one cell container. The entire experiment system has three identical fluid loops.
Mentions: The experiment hardware structure is demonstrated in Figure 4 (please refer to the Figure 4 legend and to the additional file 1). It consists of an incubator rack to temporarily store the cell containers before the experiment at 37°C, an experiment rack in which all active aggregates are accommodated for the execution of the experiment and where the living cells are handled during altered gravity and a cooling rack to store temporarily all cell containers after the injection of the stop/fixation liquid at 4°C until landing.

Bottom Line: In our study we aimed to identify rapidly reacting gravity-responsive mechanisms in mammalian cells in order to understand if and how altered gravity is translated into a cellular response.Additionally, during 2D clinorotation, Tyr-15-phosphorylation occurred later and was shorter than in the 1 g controls.The histone acetyltransferase (HAT) inhibitor curcumin was able to abrogate microgravity-induced p21 mRNA expression, whereas expression was enhanced by a histone deacetylase (HDAC) inhibitor.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Anatomy, Faculty of Medicine, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland. oliver.ullrich@anatom.uzh.ch.

ABSTRACT
In our study we aimed to identify rapidly reacting gravity-responsive mechanisms in mammalian cells in order to understand if and how altered gravity is translated into a cellular response. In a combination of experiments using "functional weightlessness" provided by 2D-clinostats and real microgravity provided by several parabolic flight campaigns and compared to in-flight-1g-controls, we identified rapid gravity-responsive reactions inside the cell cycle regulatory machinery of human T lymphocytes. In response to 2D clinorotation, we detected an enhanced expression of p21 Waf1/Cip1 protein within minutes, less cdc25C protein expression and enhanced Ser147-phosphorylation of cyclinB1 after CD3/CD28 stimulation. Additionally, during 2D clinorotation, Tyr-15-phosphorylation occurred later and was shorter than in the 1 g controls. In CD3/CD28-stimulated primary human T cells, mRNA expression of the cell cycle arrest protein p21 increased 4.1-fold after 20s real microgravity in primary CD4+ T cells and 2.9-fold in Jurkat T cells, compared to 1 g in-flight controls after CD3/CD28 stimulation. The histone acetyltransferase (HAT) inhibitor curcumin was able to abrogate microgravity-induced p21 mRNA expression, whereas expression was enhanced by a histone deacetylase (HDAC) inhibitor. Therefore, we suppose that cell cycle progression in human T lymphocytes requires Earth gravity and that the disturbed expression of cell cycle regulatory proteins could contribute to the breakdown of the human immune system in space.

No MeSH data available.


Related in: MedlinePlus