Modeling of Ca2+ transients initiated by GPCR agonists in mesenchymal stromal cells

Kaimachnikov, Nikolai P.; Kotova, Polina D.; Kochkina, Ekaterina N.; Rogachevskaja, Olga A.; Хохлов, А. А.; Bystrova, M. F.; Kolesnikov, Stanislav S.

doi:10.1016/j.bbadva.2021.100012

Cited by 4 publications

(9 citation statements)

References 60 publications

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“…This "all-or-nothing" fashion was also characteristic of agonistinduced Ca 2+ responses in cells of several other types [29][30][31][32]. Such a step-like dose dependence should be intrinsic to agonist transduction that involves CICR, the mechanism capable of producing a large and global Ca 2+ signal of universal shape, regardless of agonist concentrations [31]. The involvement of IP 3 R-mediated CICR in the generation of ACh responses (Figure 1E) was verified by the observation that 0.5 s and 1.5 s UV pulses elicited ACh response-like Ca 2+ transients that were similar kinetically and by magnitude (Figure 1F,G), although the former should have uncaged a nearly three times smaller amount of IP 3 .…”

Section: Ach Responses Of Wt-hek Cellsmentioning

confidence: 70%

“…At concentrations below the threshold of 150-200 nM, ACh insignificantly affected intracellular Ca 2+ in WT-HEK cells but the agonist elicited Ca 2+ transients of similar magnitudes at a variety of higher doses (Figure 1E). This "all-or-nothing" fashion was also characteristic of agonistinduced Ca 2+ responses in cells of several other types [29][30][31][32]. Such a step-like dose dependence should be intrinsic to agonist transduction that involves CICR, the mechanism capable of producing a large and global Ca 2+ signal of universal shape, regardless of agonist concentrations [31].…”

Section: Ach Responses Of Wt-hek Cellsmentioning

confidence: 78%

“…capable of producing a large and global Ca 2+ signal of universal shape, regardless of agonist concentrations [31]. The involvement of IP3R-mediated CICR in the generation of ACh responses (Figure 1E) was verified by the observation that 0.5 s and 1.5 s UV pulses elicited ACh response-like Ca 2+ transients that were similar kinetically and by magnitude (Figure 1F,G), although the former should have uncaged a nearly three times smaller amount of IP3.…”

Section: Ach Responses Of Wt-hek Cellsmentioning

confidence: 99%

“…Previously, we studied Ca 2+ signaling induced by GPCR agonists, including ATP, UTP, adenosine, ACh, 5-HT, and glutamate, in cells of diverse types [29][30][31][32]. The assayed cells universally responded to Ca 2+ -mobilizing agonists in an "all-or-nothing" manner: They either were irresponsive to a particular agonist at subthreshold concentrations or generated quite similar Ca 2+ signals at different agonist doses above the threshold (see Figure 1 below).…”

Section: Introductionmentioning

confidence: 99%

“…The assayed cells universally responded to Ca 2+ -mobilizing agonists in an "all-or-nothing" manner: They either were irresponsive to a particular agonist at subthreshold concentrations or generated quite similar Ca 2+ signals at different agonist doses above the threshold (see Figure 1 below). The body of evidence suggests that being a trigger-like self-driven process, CICR finalized agonist transduction by forming cellular responses of a virtually universal shape, regardless of agonist doses [31]. Given that assayed cells express multiple IP 3 R isoforms, it remains unclear whether the "all-or-nothing" responsiveness is mediated by a specific IP 3 R subtype or whether each IP 3 R isoform can endow agonist-induced Ca 2+ signaling in cells with this feature.…”

Section: Introductionmentioning

confidence: 99%

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Agonist-Induced Ca2+ Signaling in HEK-293-Derived Cells Expressing a Single IP3 Receptor Isoform

Kochkina,

Kopylova,

Rogachevskaja

et al. 2024

Cells

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In mammals, three genes encode IP3 receptors (IP3Rs), which are involved in agonist-induced Ca2+ signaling in cells of apparently all types. Using the CRISPR/Cas9 approach for disruption of two out of three IP3R genes in HEK-293 cells, we generated three monoclonal cell lines, IP3R1-HEK, IP3R2-HEK, and IP3R3-HEK, with the single functional isoform, IP3R1, IP3R2, and IP3R3, respectively. All engineered cells responded to ACh with Ca2+ transients in an “all-or-nothing” manner, suggesting that each IP3R isotype was capable of mediating CICR. The sensitivity of cells to ACh strongly correlated with the affinity of IP3 binding to an IP3R isoform they expressed. Based on a mathematical model of intracellular Ca2+ signals induced by thapsigargin, a SERCA inhibitor, we developed an approach for estimating relative Ca2+ permeability of Ca2+ store and showed that all three IP3R isoforms contributed to Ca2+ leakage from ER. The relative Ca2+ permeabilities of Ca2+ stores in IP3R1-HEK, IP3R2-HEK, and IP3R3-HEK cells were evaluated as 1:1.75:0.45. Using the genetically encoded sensor R-CEPIA1er for monitoring Ca2+ signals in ER, engineered cells were ranged by resting levels of stored Ca2+ as IP3R3-HEK ≥ IP3R1-HEK > IP3R2-HEK. The developed cell lines could be helpful for further assaying activity, regulation, and pharmacology of individual IP3R isoforms.

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Section: Ach Responses Of Wt-hek Cellsmentioning

confidence: 70%

Section: Ach Responses Of Wt-hek Cellsmentioning

confidence: 78%

Section: Ach Responses Of Wt-hek Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Agonist-Induced Ca2+ Signaling in HEK-293-Derived Cells Expressing a Single IP3 Receptor Isoform

Kochkina,

Kopylova,

Rogachevskaja

et al. 2024

Cells

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Responsiveness of HEK 293 Cells to Norepinephrine Analyzed by the On-line Monitoring of Intracellular Ca2+ and cAMP

Kochkina

Rogachevskaja

Kotova

2021

Biochem. Moscow Suppl. Ser. A

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Two Types of Cellular Serotonin Biosensors

Rogachevskaja,

Cherkashin,

Kopylova

et al. 2023

Biologičeskie membrany

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The method of cellular biosensors, which allows local detection of substances secreted by single cells, is traditionally used for studying the peripheral taste system. This method is based on cells that can detect secreted molecules with surface receptors coupled to the mobilization of intracellular Ca2+. Thus, the neurotransmitter release can be tracked on-line by Ca2+ signals generated by a cell-biosensor. A specific feature of such cellular biosensors is that Са2+ signals induced by GPCR ligands may be generated in the “all-or-nothing” manner due to the involvement of trigger-like mechanism, that is Ca2+-induced Ca2+ release (CICR). Although such a sensor validates the fact of neurotransmitter secretion, it does not allow studying regulatory circuits controlling the quantity of the released substance. This implies that biosensors, which are based on distinct intracellular signaling pathways, such as for cAMP signaling, could be more informative. Here we generated two types of serotonin biosensors. One was based on CHO cells expressing the recombinant 5-HT2C receptor coupled by the phospholipase pathway to Ca2+ mobilization. Another involved HEK-293 cells that expressed both 5-HT2C receptor coupled to adenylyl cyclase and the fluorescent protein Pink Flamindo serving as genetically encoded cAMP sensor. Although both cellular biosensors allow detection of nanomolar serotonin, 5-НТ2С cells generated Ca2+ responses in the “all-or-nothing” manner. In contrast, in 5-HT4 and Pink Flamindo expressing cells, serotonin-triggered cAMP responses gradually rose with agonist concentration in the range of 0.3–30 nM. It thus appears that the 5-НТ4/Pink Flamindo biosensor is suitable for studying regulatory mechanism of serotonin secretion.

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Modeling of Ca2+ transients initiated by GPCR agonists in mesenchymal stromal cells

Cited by 4 publications

References 60 publications

Agonist-Induced Ca2+ Signaling in HEK-293-Derived Cells Expressing a Single IP3 Receptor Isoform

Agonist-Induced Ca2+ Signaling in HEK-293-Derived Cells Expressing a Single IP3 Receptor Isoform

Responsiveness of HEK 293 Cells to Norepinephrine Analyzed by the On-line Monitoring of Intracellular Ca2+ and cAMP

Two Types of Cellular Serotonin Biosensors

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