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doi:10.22028/D291-43999
Titel: | Kinetics of the thapsigargin-induced Ca2+ mobilisation: A quantitative analysis in the HEK-293 cell line |
VerfasserIn: | Pick, Tillman Gamayun, Igor Tinschert, René Cavalié, Adolfo |
Sprache: | Englisch |
Titel: | Frontiers in physiology |
Bandnummer: | 14 |
Seiten: | 1-19 |
Verlag/Plattform: | Frontiers |
Erscheinungsjahr: | 2023 |
Freie Schlagwörter: | Ca2+ homeostasis Ca2+ leak store-operated Ca2+ entry Ca2+ imaging thapsigargin |
DDC-Sachgruppe: | 610 Medizin, Gesundheit |
Dokumenttyp: | Journalartikel / Zeitschriftenartikel |
Abstract: | Thapsigargin (TG) inhibits the sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) pump and, when applied acutely, it initiates a Ca2+ mobilisation that begins with the loss of Ca2+ from the endoplasmic reticulum (ER) and culminates with store-operated Ca2+ entry (SOCE) from the extracellular space. Using the popular model cell line HEK-293, we quantified TG-induced changes in cytosolic and ER Ca2+ levels using FURA-2 and the FRET-based ER Ca2+ sensor D1ER, respectively. Our analysis predicts an ER Ca2+ leak of 5-6 µM⋅s-1 for the typical basal ER Ca2+ level of 335-407 µM in HEK-293 cells. The resulting cytosolic Ca2+ transients reached peak amplitudes of 0.6-1.0 µM in the absence of external Ca2+ and were amplified by SOCE that amounted to 28-30 nM⋅s-1 in 1 mM external Ca2+. Additionally, cytosolic Ca2+ transients were shaped by a Ca2+ clearance of 10-13 nM⋅s-1. Using puromycin (PURO), which enhances the ER Ca2+ leak, we show that TG-induced cytosolic Ca2+ transients are directly related to ER Ca2+ levels and to the ER Ca2+ leak. A one-compartment model incorporating ER Ca2+ leak and cytosolic Ca2+ clearance accounted satisfactorily for the basic features of TG-induced Ca2+ transients and underpinned the rule that an increase in amplitude associated with shortening of TG-induced cytosolic Ca2+ transients most likely reflects an increase in ER Ca2+ leak. |
DOI der Erstveröffentlichung: | 10.3389/fphys.2023.1127545 |
URL der Erstveröffentlichung: | https://www.frontiersin.org/journals/physiology/articles/10.3389/fphys.2023.1127545/full |
Link zu diesem Datensatz: | urn:nbn:de:bsz:291--ds-439993 hdl:20.500.11880/39375 http://dx.doi.org/10.22028/D291-43999 |
ISSN: | 1664-042X |
Datum des Eintrags: | 14-Jan-2025 |
Fakultät: | M - Medizinische Fakultät |
Fachrichtung: | M - Experimentelle und Klinische Pharmakologie und Toxikologie |
Professur: | M - Prof. Dr. Adolfo Cavalié |
Sammlung: | SciDok - Der Wissenschaftsserver der Universität des Saarlandes |
Dateien zu diesem Datensatz:
Datei | Beschreibung | Größe | Format | |
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fphys-14-1127545.pdf | 4,12 MB | Adobe PDF | Öffnen/Anzeigen |
Diese Ressource wurde unter folgender Copyright-Bestimmung veröffentlicht: Lizenz von Creative Commons