Microgravity induces alterations in the functioning of immune cell; however, the underlying mechanisms have not yet been identified. In this study, hemocytes (blood cells) of the blue mussel Mytilus edulis were investigated under altered gravity conditions. The study was conducted on the ground in preparation for the BIOLAB TripleLux-B experiment, which will be performed on the International Space Station (ISS). On-line kinetic measurements of reactive oxygen species (ROS) production during the oxidative burst and thus cellular activity of isolated hemocytes were performed in a photomultiplier (PMT)-clinostat (simulated microgravity) and in the 1g operation mode of the clinostat in hypergravity on the Short-Arm Human Centrifuge (SAHC) as well as during parabolic flights. In addition to studies with isolated hemocytes, the effect of altered gravity conditions on whole animals was investigated. For this purpose, whole mussels were exposed to hypergravity (1.8 g) on a multi-sample incubator centrifuge (MuSIC) or to simulated microgravity in a submersed clinostat. After exposure for 48 h, hemocytes were taken from the mussels and ROS production was measured under 1 g conditions. The results from the parabolic flights and clinostat studies indicate that mussel hemocytes respond to altered gravity in a fast and reversible manner. Hemocytes (after cryo-conservation) exposed to simulated microgravity (μ g), as well as fresh hemocytes from clinorotated animals, showed a decrease in ROS production. Measurements during a permanent exposure of hemocytes to hypergravity (SAHC) show a decrease in ROS production. Hemocytes of mussels measured after the centrifugation of whole mussels did not show an influence to the ROS response at all. Hypergravity during parabolic flights led to a decrease but also to an increase in ROS production in isolated hemocytes, whereas the centrifugation of whole mussels did not influence the ROS response at all. This study is a good example how ground-based facility experiments can be used to prepare for an upcoming ISS experiment, in this case the TRIPLE LUX B experiment.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Adrian, A., Schoppmann, K., Sromicki, J., Brungs, S., von der Wiesche, M., Hock, B., et al.: The oxidative burst reaction in mammalian cells depends on gravity. J. Cell Commun. Signal 11, 98 (2013)
Allen, R.: Phagocytic leukocyte oxygenation activities and chemiluminescence: A kinetic approach to analysis. Method Enzymol 133, 449–93 (1986)
Anderson, R.S., Mora, M.L., Brubacher, L.: Luminol-dependent Chemi-luminescence in Molluscs. In: Stolen, J.S., Fletcher, T.C., Smith, S.A., Zellikoff, J.T., Kaattari, S.L., Anderson, R.S., Söderhäll, K., W.A. (eds.) Techniques in Fish Immunology- 4 Immunology and Pathology of Aquatic Invertebrates, pp. 101–107. Weeks-Perkins, SOS Publications, Fair Haven (1995)
Anken, R., Baur, U., Hilbig, R.: Clinorotation increases the growth of utricular otoliths of developing cichlid fish. Microgravity Sci. Technol 22, 151–154 (2009)
Arumugam, M., Romestand, B., Torreilles, J., Roch, P.: In vitro production of superoxide and nitric oxide (as nitrite and nitrate) by Mytilus galloprovincialis haemocytes upon incubation with PMA or laminarin or during yeast phagocytosis. Eur. J. Cell. Biol 79, 513–519 (2000)
Babior, B.M.: NADPH oxidase: An update. Blood 93(5), 1464–1476 (1999)
Brungs, S., Kolanus, W., Hemmersbach, R.: Impact of altered gravity on the oxidative burst in macrophages. J. Cell Commun. Signal 13, 9 (2015)
Brungs, S., Hauslage, J., Hilbig, R., Hemmersbach, R., Anken, R.: Effects of simulated weightlessness on fish otolith growth: Clinostat versus rotating-wall vessel. Adv. Space Res 48, 792–798 (2011)
Brungs, S.: The influence of altered gravity on the oxidative burst in macrophages, PhD thesis, University of Bonn (2013)
Crawley, M.J.: Statistics: An Introduction using R. John Wiley & Sons, Ltd, Chichester (2005)
Ernst, W., Weigelt, S., Rosenthal, H., Hansen, P.-D.: Testing Bioconcentration of Organic Chemicals with the Common Mussel (Mytilus edulis). In: Nagel, R., Loskill, R. (eds.) Bioaccumulation in Aquatic Systems, pp. 99–131. VCH Verlagsgesellschaft, Weinheim (1991)
Gagné, F., Blaise, C., Fournier, M., Hansen, P.-D.: Effects of selected pharmaceutical products on phagocytic activity in Elliptio complanata mussels. Comp. Biochem. Physiol. C Toxicol. Pharmacol 143(2), 179–186 (2006)
Hansen, P.-D., Bock, R., Brauer, F.: Investigations of phagocytosis concerning the immunological defence mechanism of Mytilus edulis using a sublethal luminescent bacterial assay (Photobacterium phosphoreum). Comp. Biochem. Physiol 100C(1/2), 129–132 (1991)
Hansen, P.-D.: Phagocytosis in Mytilus edulis, a system for understanding the sublethal effects of anthropogenic pollutants and the use of AOX as an integrating parameter for the study of equilibria between chlorinated organics in Dreissena polymorpha following long term exposures. In: Neumann, Jenner (eds.) Limnologie aktuell, the Zebra Mussel Dreissena polymorpha, vol. 4. Gustav Fischer Verlag, Stuttgart, Jena, New York (1992)
Hansen, P.D., Unruh, E.: TRIPLE LUX-B: phagocytosis in mussel hemocytes. In: Proceedings of the 9th European Symposium on Life Sciences Research in Space /26th Annual International Gravitational Physiology Meeting. Cologne Germany (2005)
Horn, A., Ulrich, O., Huber, K., Hemmersbach, R.: PMT (Photomultiplier) Clinostat. MST 23(1) (2010)
Horn, A.: Vorbereitungen für das Biolab Experiment TRIPLE LUX A Hardwareentwicklung, Kalibrierung und biologische Bodenkontrollen. PhD thesis, Otto-von-Guericke-Universität, Magdeburg, 7–71 (2011)
Huber, K.: Phagozytose und oxidative Burst als Biomarker für Immuntoxizität – Der Einfluss von simulierter Schwerelosigkeit auf die Makrophagenzelllinie NR8383, Dissertation Technische Universität München, 90–103 (2009)
Kaur, I., Simons, E.R., Castro, V.A., Ott, M.C., Pierson, D.L.: Changes in neutrophil functions in astronauts. Brain Behav. Immun. 18(5), 443–450 (2004)
Kaur, I., Simons, E.R., Kapadia, A.S., Ott, C.M., Pierson, D.L.: Effect of spaceflight on ability of monocytes to respond to endotoxins of Gram-negative bacteria. Clin. Vaccine Immunol. 15(10), 1523–1528 (2008)
Manduzio, H., Monsinjon, T., Rocher, B., Leboulenger, F., Galap, C.: Characterization of an inducible isoform of the Cu/Zn superoxide dismutase in the Blue Mussel Mytilus edulis. Aquat. Toxicol 64, 73–83 (2003)
Noel, D., Bachere, E., Mialhe, E.: Phagocytosis associated chemiluminescence of heinocyte In Mytilus edulis (Bivalvia). Dev. Comp. Immunol. 17, 483–493 (1993)
Paulsen, K., Thiel, C., Timm, J., Schmidt, P.M., Huber, K., Tauber, S., Hemmersbach, R., Seibt, D., Kroll, H., Grote, K.-H., Zipp, F., Schneider-Stock, R., Cogoli, A., Hilliger, A., Engelmann, F., Ullrich, O.: Microgravity-induced alterations in signal transduction in cells of the immune system. Acta Astronaut 67(9–10), 1116–1125 (2010)
Pipe, R.K., Coles, J.A., Farley, S.R.: Assays for measuring immune response in the mussel mytilus edulis. In: Stolen, J.S., Fletcher, T.C., Smith, S.A., Zellikoff, J.T., S.L. (eds.) Techniques in Fish Immunology-4, 1995. Immunology and Pathology of Aquatic Invertebrates (1995)
Renwrantz, L., Daniel, I., Hansen, P.-D.: Lectin - binding to hemocytes of Mytilus edulis. Dev. Comp. Immunol 9, 203–210 (1985)
Schuber, M., Seibt, D., Anken, R.: BIOLAB on the international space station (ISS): Facility and experiments. Current Biotechnol. 2, 3 (2013)
Siegmund, E.: Untersuchungen von Hämozyten von Mytilus edulis L, Dissertation, Universität Hamburg (2001)
Van Dyke, K., Patel, S., Vallyathan, V.: Luciginin chemoluminescence assay as an adjunctive tool for assessment of various stages of inflammation: A study of quiescent inflammatory cells. J. Biosci 28(1), 115–119 (2003)
Winston, G.W., Moore, M.N., Kirchin, M.A., Soverchia, C.: Production of reactive oxygen species by Hemocytes from the marine mussel. Mytilus edulis: Lysosomal localization and effect of xenobiotics. Comp Biochem Physiol C 113(/2), 221–229 (1996)
Yaqin, K., Lay, B.W., Riani, E., Masud, Z.A., Hansen, P.-D.: Hot Spot biomonitoring of marine pollution effects using cholinergic and immunity biomarkers of tropical green mussel (Perna viridis) of the Indonesian waters. J. Toxicol. Environ. Health Sci. 3(14), 356–365 (2011). Legends
The authors gratefully thank ESA and NOVE-SPACE for the opportunity to test the hypothesis on board the aircraft Zero-G. We thank especially Dr. Hemmersbach (DLR, Biomedical Research Department) and ESA (GBF Project CORA-GBF-2012-002) for studies in gravitational biology (ground based facilities: clinostats and centrifuges). We gratefully thank the Alfred-Wegener Institute (Wadden Sea Station, List/Sylt) for supporting the hemocyte preparations. The TRIPLELUX-B experiments are financially supported by the DLR/BWi (Project No. 50WB0811). The authors gratefully acknowledge the DLR for funding.
Conflict of interest
The authors declare that they have no conflicts of interest.
About this article
Cite this article
Unruh, E., Brungs, S., Langer, S. et al. Comprehensive Study of the Influence of Altered Gravity on the Oxidative Burst of Mussel (Mytilus edulis) Hemocytes. Microgravity Sci. Technol. 28, 275–285 (2016). https://doi.org/10.1007/s12217-015-9438-9
- Reactive oxygen species (ROS)
- Parabolic flight