Targeted Gq-GPCR activation drives ER-dependent calcium oscillations in chondrocytes.

Autor: McDonough RC; Department of Biomedical Engineering, University of Delaware, United States. Electronic address: ryancmcd@udel.edu., Gilbert RM; Department of Biomedical Engineering, University of Delaware, United States. Electronic address: rgilbert@udel.edu., Gleghorn JP; Department of Biomedical Engineering, University of Delaware, United States. Electronic address: gleghorn@udel.edu., Price C; Department of Biomedical Engineering, University of Delaware, United States. Electronic address: cprice@udel.edu.
Jazyk: angličtina
Zdroj: Cell calcium [Cell Calcium] 2021 Mar; Vol. 94, pp. 102363. Date of Electronic Publication: 2021 Jan 30.
DOI: 10.1016/j.ceca.2021.102363
Abstrakt: The temporal dynamics of calcium signaling are critical regulators of chondrocyte homeostasis and chondrogenesis. Calcium oscillations regulate differentiation and anabolic processes in chondrocytes and their precursors. Attempts to control chondrocyte calcium signaling have been achieved through mechanical perturbations and synthetic ion channel modulators. However, such stimuli can lack both local and global specificity and precision when evoking calcium signals. Synthetic signaling platforms can more precisely and selectively activate calcium signaling, enabling improved dissection of the roles of intracellular calcium ([Ca 2+ ] i ) in chondrocyte behavior. One such platform is hM3Dq, a chemogenetic DREADD (Designer Receptors Exclusively Activated by Designer Drugs) that activates calcium signaling via the Gα q -PLCβ-IP 3 -ER pathway upon administration of clozapine N-oxide (CNO). We previously described the first-use of hM3Dq to precisely mediate targeted, synthetic calcium signals in chondrocyte-like ATDC5 cells. Here, we generated stably expressing hM3Dq-ATDC5 cells to investigate the dynamics of Gα q -GPCR calcium signaling in depth. CNO drove robust calcium responses in a temperature- and concentration-dependent (1 pM-100 μM) manner and elicited elevated levels of oscillatory calcium signaling above 10 nM. hM3Dq-mediated calcium oscillations in ATDC5 cells were reliant on ER calcium stores for both initiation and sustenance, and the downregulation and recovery dynamics of hM3Dq after CNO stimulation align with traditionally reported GPCR recycling kinetics. This study successfully generated a stable hM3Dq cell line to precisely drive Gα q -GPCR-mediated and ER-dependent oscillatory calcium signaling in ATDC5 cells and established a novel tool to elucidate the role that GPCR-mediated calcium signaling plays in chondrocyte biology, cartilage pathology, and cartilage tissue engineering.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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