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- W3176455114 abstract "Gi/o coupled P2Y and μ-opioid receptors inhibit transmitter release in bovine adrenal chromaffin cells. One mechanism involves inhibition of voltage-gated Ca2+ channels (Ca-channels) by G-protein βγ subunits (Gβγ). Evidence also suggests that Gβγ can modulate a late stage of exocytosis independently from Ca2+ entry. To investigate this possibility, we monitored release of catecholamines from individual chromaffin cells using carbon fiber amperometry. We used Ca2+ ionophores to bypass Ca-channels and directly stimulate secretion. Alternatively, cells were patch-clamped at a holding potential of -80mV using standard whole-cell recording and patch-pipette solution containing ~50 μM free Ca2+ which diffused into the cell and directly triggered secretion. Extracellular application of ATP or DAMGO was used to activate P2Y or μ-opioid receptors and endogenous Gi/o -proteins. This significantly reduced the number of amperometric spikes, indicating that fewer vesicular fusion events were occurring. To confirm this was mediated by Gβγ, we expressed GFP-fused Gβ along with Gγ in chromaffin cells. GFP positive transfected cells were identified visually. Gβγ expressing cells displayed fewer amperometric spikes compared to control cells expressing GFP alone. Furthermore, the mean charge of the amperometric spikes was significantly reduced in Gβ γ-expressing cells. Our data suggest that Gβγ can reduce exocytosis downstream of Ca-channel. Supported by AHA predoctoral fellowship (to EY), NIH NS052446 (to KC), and NIH EY010291 (to HH)" @default.
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- W3176455114 date "2007-01-01" @default.
- W3176455114 modified "2023-09-23" @default.
- W3176455114 title "Gβγ mediated modulation of exocytosis downstream of Ca2+ entry in bovine adrenal chromaffin cells" @default.
- W3176455114 doi "https://doi.org/10.1096/fasebj.21.6.lb14" @default.
- W3176455114 hasPublicationYear "2007" @default.
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