Label-free whole-cell assays: expanding the scope of GPCR screening

Scott, Clay W.; Peters, Matthew F.

doi:10.1016/j.drudis.2010.06.008

Cited by 146 publications

(101 citation statements)

References 64 publications

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“…TM assay's ability to distinguish the activation of G␣ subfamily (G i/o , G s , and G q ) by patterns of ⌬Z changes (19). To show typical different ⌬Z values caused by each ligand to specific types of GPCRs, HEK293 cells expressing -opioid receptors, coupled to G i/o , and C6 cells endogenously expressing ␤-adrenergic receptors, coupled to G s , were used (14,35).…”

Section: Effects Of Amitriptyline On the Changes Of Impedance (⌬Z) Inmentioning

confidence: 99%

Tricyclic Antidepressant Amitriptyline-induced Glial Cell Line-derived Neurotrophic Factor Production Involves Pertussis Toxin-sensitive Gαi/o Activation in Astroglial Cells

Hisaoka-Nakashima

Miyano

Matsumoto

et al. 2015

Journal of Biological Chemistry

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Background: A significant non-neural, monoamine-independent mechanism underlies the antidepressant effect of amitriptyline. Results: Amitriptyline-evoked GDNF production is mediated by pertussis toxin (PTX)-sensitive G␣ i/o . Conclusion: PTX-sensitive G␣ i/o activation is critical for the cascade that underpins the biological effect of amitriptyline. Significance: Further elaboration of the intracellular mechanism of amitriptyline could lead to greater understanding of depression and novel antidepressant treatments.

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Section: Effects Of Amitriptyline On the Changes Of Impedance (⌬Z) Inmentioning

confidence: 99%

Tricyclic Antidepressant Amitriptyline-induced Glial Cell Line-derived Neurotrophic Factor Production Involves Pertussis Toxin-sensitive Gαi/o Activation in Astroglial Cells

Hisaoka-Nakashima

Miyano

Matsumoto

et al. 2015

Journal of Biological Chemistry

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“…Importantly, compared to the orthogonal cell activation assays mentioned above, PC biosensors are able to simultaneously detect a broad range of cellular events downstream of GPCR signaling that lead to changes in cell morphology or adhesion properties. As a consequence, such technology may display a more holistic cellular response that is usually not detected by other conventional assays (28)(29)(30).…”

Section: Discussionmentioning

confidence: 99%

“…Such events usually occur downstream of several cellular activation pathways, specifically those involving GPCR-derived cell responses. By recording activation signals over time, a kinetic and cell/receptor-specific activation profile can be obtained (28)(29)(30)(31). Our data obtained by this novel method and by orthogonal assays provide new evidence for a significant role for C5a desArg in cell activation and point to a new, easy, rapid, and reliable technology for detecting GPCR-mediated cell activation.…”

Section: Introductionmentioning

confidence: 85%

“…Briefly, optical biosensors located at the bottom of a microtiter plate detect changes in dielectric permittivity of attached biomolecules or cells (by measuring the resonant reflection spectrum after illumination by a source of infrared light). In the case of receptor assays, signal shifts may occur due to changes in cell adhesion, cell morphology, or dynamic mass redistribution upon stimulation of the GPCR; the resulting response profile is dependent on the cell type and signaling pathway, among others (28)(29)(30)(31). Selected wells of a 384-well BIND CA2 biosensor plate (SRU Biosystems), containing a cell attachment matrix consisting of collagen, fibronectin and ovalbumin (28), were hydrated with 50 μl of deionized water for 30 min at room temperature.…”

Section: Cell Activation Assaysmentioning

confidence: 99%

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C5aR-dependent cell activation by physiological concentrations of C5a-desArg: Insights from a novel label-free cellular assay

et al. 2012

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The complement anaphylatoxins C3a, C5a, and C5a desArg play critical roles in the induction of inflammation and the modulation of innate and acquired immune responses after binding to their G protein-coupled receptors, C3aR and C5aR. The role of C5a desArg in inducing cell activation has been often neglected, since the affinity of C5a desArg for C5aR has been reported to be much lower than that of C5a. We have used a novel label-free cellular assay to reassess the potential of C5a desArg to induce activation of transfected and primary immune cells. Our results indicate that physiological levels of C5a desArg induce significant levels of cell activation that are even higher than that achieved by stimulating cells with analogous concentrations of C5a. Such activation was strictly dependent on C5aR, since it was completely abrogated by PMX-53, a C5aR antagonist. Pharmacological inhibition of specific G proteins located downstream of C5aR indicated differential involvement of G α proteins upon C5aR engagement by C5a or C5a desArg . Further, mass spectrometric characterization of plasma-derived C5a and C5a desArg provided important insight into the post-translational modification pattern of these anaphylatoxins, which includes glycosylation at Asn 64 and partial cysteinylation at Cys 27 . While the context-specific physiological contribution of C5a desArg has to be further explored, our data suggest that C5a desArg acts as a key molecule in the triggering of local inflammation as well as the maintenance of blood surveillance and homeostatic status.

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“…[1][2][3][4][5][6][7] The biosensor transforms a cellular response into an integrated and kinetic response, termed DMR. 8 However, current RWG biosensors utilize a static system, wherein a ligand is introduced using a pipetting system and cells are continuously exposed to the ligand throughout assays.…”

mentioning

confidence: 99%

Microfluidic resonant waveguide grating biosensor system for whole cell sensing

Zaytseva

Miller

Goral

et al. 2011

Applied Physics Letters

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We report on a fluidic resonant waveguide grating (RWG) biosensor system that enables medium throughput measurements of cellular responses under microfluidics in a 32-well format. Dynamic mass redistribution assays under microfluidics differentiate the cross-desensitization process between the β2-adrenoceptor agonist epinephrine and the adenylate cyclase activator forskolin mediated signaling. This system opens new possibility to study cellular processes that are otherwise difficult to achieve using conventional RWG configurations.

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Label-free whole-cell assays: expanding the scope of GPCR screening

Cited by 146 publications

References 64 publications

Tricyclic Antidepressant Amitriptyline-induced Glial Cell Line-derived Neurotrophic Factor Production Involves Pertussis Toxin-sensitive Gαi/o Activation in Astroglial Cells

Tricyclic Antidepressant Amitriptyline-induced Glial Cell Line-derived Neurotrophic Factor Production Involves Pertussis Toxin-sensitive Gαi/o Activation in Astroglial Cells

C5aR-dependent cell activation by physiological concentrations of C5a-desArg: Insights from a novel label-free cellular assay

Microfluidic resonant waveguide grating biosensor system for whole cell sensing

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