Retrograde Analysis of Calcium Signaling by CaMPARI2 Shows Cytosolic Calcium in Chondrocytes Is Unaffected by Parabolic Flights
Andreas Hammer,
Geraldine Cerretti,
Dario A. Ricciardi,
David Schiffmann,
Simon Maranda,
Raphael Kummer,
Christoph Zumbühl,
Karin F. Rattenbacher-Kiser,
Silvan von Arx,
Sebastian Ammann,
Frederic Strobl,
Rayene Berkane,
Alexandra Stolz,
Ernst H. K. Stelzer,
Marcel Egli,
Enrico Schleiff,
Simon L. Wuest,
Maik Böhmer
Affiliations
Andreas Hammer
Institute for Molecular Biosciences, Johann Wolfgang Goethe Universität, 60438 Frankfurt am Main, Germany
Geraldine Cerretti
Space Biology Group, Institute of Medical Engineering, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6052 Hergiswil, Switzerland
Dario A. Ricciardi
Institute for Molecular Biosciences, Johann Wolfgang Goethe Universität, 60438 Frankfurt am Main, Germany
David Schiffmann
Institute of Mechanical Engineering and Energy Technology, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6048 Horw, Switzerland
Simon Maranda
Institute of Mechanical Engineering and Energy Technology, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6048 Horw, Switzerland
Raphael Kummer
Institute of Electrical Engineering, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6048 Horw, Switzerland
Christoph Zumbühl
Institute of Electrical Engineering, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6048 Horw, Switzerland
Karin F. Rattenbacher-Kiser
Space Biology Group, Institute of Medical Engineering, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6052 Hergiswil, Switzerland
Silvan von Arx
Institute of Mechanical Engineering and Energy Technology, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6048 Horw, Switzerland
Sebastian Ammann
Institute of Mechanical Engineering and Energy Technology, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6048 Horw, Switzerland
Frederic Strobl
Buchmann Institute for Molecular Life Sciences (BMLS), Johann Wolfgang Goethe Universität, 60438 Frankfurt am Main, Germany
Rayene Berkane
Buchmann Institute for Molecular Life Sciences (BMLS), Johann Wolfgang Goethe Universität, 60438 Frankfurt am Main, Germany
Alexandra Stolz
Buchmann Institute for Molecular Life Sciences (BMLS), Johann Wolfgang Goethe Universität, 60438 Frankfurt am Main, Germany
Ernst H. K. Stelzer
Buchmann Institute for Molecular Life Sciences (BMLS), Johann Wolfgang Goethe Universität, 60438 Frankfurt am Main, Germany
Marcel Egli
Space Biology Group, Institute of Medical Engineering, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6052 Hergiswil, Switzerland
Enrico Schleiff
Institute for Molecular Biosciences, Johann Wolfgang Goethe Universität, 60438 Frankfurt am Main, Germany
Simon L. Wuest
Space Biology Group, Institute of Medical Engineering, School of Engineering and Architecture, Lucerne University of Applied Sciences and Arts, 6052 Hergiswil, Switzerland
Maik Böhmer
Institute for Molecular Biosciences, Johann Wolfgang Goethe Universität, 60438 Frankfurt am Main, Germany
Calcium (Ca2+) elevation is an essential secondary messenger in many cellular processes, including disease progression and adaptation to external stimuli, e.g., gravitational load. Therefore, mapping and quantifying Ca2+ signaling with a high spatiotemporal resolution is a key challenge. However, particularly on microgravity platforms, experiment time is limited, allowing only a small number of replicates. Furthermore, experiment hardware is exposed to changes in gravity levels, causing experimental artifacts unless appropriately controlled. We introduce a new experimental setup based on the fluorescent Ca2+ reporter CaMPARI2, onboard LED arrays, and subsequent microscopic analysis on the ground. This setup allows for higher throughput and accuracy due to its retrograde nature. The excellent performance of CaMPARI2 was demonstrated with human chondrocytes during the 75th ESA parabolic flight campaign. CaMPARI2 revealed a strong Ca2+ response triggered by histamine but was not affected by the alternating gravitational load of a parabolic flight.
