| |  Online-Ressource |
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| Verfasst von: | Molitor, Elena von [VerfasserIn]  |
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| | Nürnberg, Elina [VerfasserIn] |
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| | Ertongur-Fauth, Torsten [VerfasserIn] |
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| | Scholz, Paul [VerfasserIn] |
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| | Riedel, Katja [VerfasserIn] |
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| | Hafner, Mathias [VerfasserIn] |
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| | Rudolf, Rüdiger [VerfasserIn] |
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| | Cesetti, Tiziana [VerfasserIn] |
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| Titel: | Analysis of calcium signaling in live human tongue cell 3D-cultures upon tastant perfusion |
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| Verf.angabe: | Elena von Molitor, Elina Nürnberg, Torsten Ertongur-Fauth, Paul Scholz, Katja Riedel, Mathias Hafner, Rüdiger Rudolf, Tiziana Cesetti |
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| E-Jahr: | 2020 |
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| Jahr: | 23 January 2020 |
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| Umfang: | 13 S. |
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| Fussnoten: | Gesehen am 16.06.2020 |
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| Titel Quelle: | Enthalten in: Cell calcium |
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| Ort Quelle: | Edinburgh [u.a.] : Churchill Livingstone, 1980 |
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| Jahr Quelle: | 2020 |
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| Band/Heft Quelle: | 87(2020) Artikel-Nummer 102164, 13 Seiten |
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| ISSN Quelle: | 1532-1991 |
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| Abstract: | Bridging the gap between two-dimensional cell cultures and complex in vivo tissues, three-dimensional cell culture models are of increasing interest in the fields of cell biology and pharmacology. However, present challenges hamper live cell imaging of three-dimensional cell cultures. These include (i) the stabilization of these structures under perfusion conditions, (ii) the recording of many z-planes at high spatio-temporal resolution, (iii) and the data analysis that ranges in complexity from whole specimens to single cells. Here, we addressed these issues for the time-lapse analysis of Ca2+ signaling in spheroids composed of human tongue-derived HTC-8 cells upon perfusion of gustatory substances. Live cell imaging setups for confocal and light sheet microscopy were developed that allow simple and robust spheroid stabilization and high-resolution microscopy with perfusion. Visualization of spheroids made of HTC-8 cells expressing the G-GECO fluorescent Ca2+ sensor revealed Ca2+ transients that showed similar kinetics but different amplitudes upon perfusion of bitter compounds Salicine and Saccharin. Dose-dependent responses to Saccharin required extracellular Ca2+. From the border towards the center of spheroids, compound-induced Ca2+ signals were progressively delayed and decreased in amplitude. Stimulation with ATP led to strong Ca2+ transients that were faster than those evoked by the bitter compounds and blockade of purinergic receptors with Suramin abutted the response to Saccharin, suggesting that ATP mediates a positive autocrine and paracrine feedback. Imaging of ATP-induced Ca2+ transients with light sheet microscopy allowed acquisition over a z-depth of 100 μm without losing spatial and temporal resolution. In summary, the presented approaches permit the study of fast cellular signaling in three-dimensional cultures upon compound perfusion. |
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| DOI: | doi:10.1016/j.ceca.2020.102164 |
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| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: https://dx.doi.org/10.1016/j.ceca.2020.102164 |
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| | Volltext: http://www.sciencedirect.com/science/article/pii/S0143416020300063 |
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| | DOI: https://doi.org/10.1016/j.ceca.2020.102164 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | Human tongue cells |
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| | Light sheet fluorescence microscopy |
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| | Live calcium imaging |
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| | Perfusion |
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| | Saccharin |
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| | Spheroids |
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| K10plus-PPN: | 170067868X |
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| Verknüpfungen: | → Zeitschrift |
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Analysis of calcium signaling in live human tongue cell 3D-cultures upon tastant perfusion / Molitor, Elena von [VerfasserIn]; 23 January 2020 (Online-Ressource)
