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Retrograde Analysis of Calcium Signaling by CaMPARI2 Shows Cytosolic Calcium in Chondrocytes Is Unaffected by Parabolic Flights

Name: Retrograde Analysis of Calcium Signaling by CaMPARI2 Shows Cytosolic Calcium in Chondrocytes Is Unaffected by Parabolic Flights
Keywords: CaMPARI, articular chondrocytes, cytosolic free calcium, QH301-705.5, parabolic flight, CaMPARI; articular chondrocytes; cytosolic free calcium; gravity; parabolic flight; high throughput screening, Biology (General), Article, gravity, high throughput screening

descriptionPublicationkeyboard_double_arrow_right Article 08 Jan 2022 Switzerland Zenodo

Authors: Hammer, Andreas; Cerretti, Geraldine; Ricciardi, Dario A.; Schiffmann, David; Maranda, Simon; Kummer, Raphael; Zumbühl, Christoph; +11 Authors

doi: 10.5281/zenodo.5902965 , 10.5281/zenodo.5902964

pmc: PMC8773224

pmid: 35052817

Retrograde Analysis of Calcium Signaling by CaMPARI2 Shows Cytosolic Calcium in Chondrocytes Is Unaffected by Parabolic Flights

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- Related research
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Abstract

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.

+ ID der Publikation: hslu_88043 + Art des Beitrages: Wissenschaftliche Medien + Jahrgang: 2022 + Sprache: Englisch + Letzte Aktualisierung: 2022-01-25 16:01:32

Country

Switzerland

Related Organizations

Zentral und Hochschulbibliothek Luzern
Switzerland

Keywords

CaMPARI, articular chondrocytes, cytosolic free calcium, QH301-705.5, parabolic flight, CaMPARI; articular chondrocytes; cytosolic free calcium; gravity; parabolic flight; high throughput screening, Biology (General), Article, gravity, high throughput screening

77 references, page 1 of 8

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3. Kekenes-Huskey, P.M.; Metzger, V.T.; Grant, B.J.; Andrew McCammon, J. Calcium Binding and Allosteric Signaling Mechanisms for the Sarcoplasmic Reticulum Ca2+ ATPase. Protein Sci. 2012, 21, 1429-1443. [CrossRef] [PubMed] [OpenAIRE]

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6. Lee, E.H. Ca2+ Channels and Skeletal Muscle Diseases. Prog. Biophys. Mol. Biol. 2010, 103, 35-43. [CrossRef]

7. Bruton, J.; Cheng, A.J.; Westerblad, H. Measuring Ca2+ in Living Cells. In Calcium Signaling. Advances in Experimental Medicine and Biology; Islam, M., Ed.; Springer: Cham, Switzerland, 2020; Volume 1131, pp. 7-26. [CrossRef]

8. Grynkiewicz, G.; Poenie, M.; Tsien, R.Y. A New Generation of Ca2+ Indicators with Greatly Improved Fluorescence Properties. J. Biol. Chem. 1985, 260, 3440-3450. [CrossRef] [OpenAIRE]

9. Zhao, Y.; Araki, S.; Wu, J.; Teramoto, T.; Chang, Y.-F.; Nakano, M.; Abdelfattah, A.S.; Fujiwara, M.; Ishihara, T.; Nagai, T.; et al. An Expanded Palette of Genetically Encoded Ca2+ Indicators. Science 2011, 333, 1888-1891. [CrossRef] [PubMed]

10. Chen, T.-W.; Wardill, T.J.; Sun, Y.; Pulver, S.R.; Renninger, S.L.; Baohan, A.; Schreiter, E.R.; Kerr, R.A.; Orger, M.B.; Jayaraman, V.; et al. Ultrasensitive Fluorescent Proteins for Imaging Neuronal Activity. Nature 2013, 499, 295-300. [CrossRef] [OpenAIRE]

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	citations This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).	1
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Retrograde Analysis of Calcium Signaling by CaMPARI2 Shows Cytosolic Calcium in Chondrocytes Is Unaffected by Parabolic Flights

Retrograde Analysis of Calcium Signaling by CaMPARI2 Shows Cytosolic Calcium in Chondrocytes Is Unaffected by Parabolic Flights

77 references, page 1 of 8

3 Research products, page 1 of 1

Activity labeling in vivo using CaMPARI2 reveals electrophysiological differences between neurons with high and low firing rate set points

Activity labeling in vivo using CaMPARI2 reveals intrinsic and synaptic differences between neurons with high and low firing rate set points.

Improved methods for marking active neuron populations.