TY - JOUR
T1 - δ-opioid-induced liberation of Gβγ mobilizes Ca2+ stores in NG108- 15 cells
AU - Yoon, Shin Hee
AU - Lo, Tak Man
AU - Loh, Horace H
AU - Thayer, Stanley A
PY - 1999
Y1 - 1999
N2 - Activation of δ-opioid receptors in NG108-15 cells releases Ca2+ from an intracellular store through activation of a pertussis toxin-sensitive G protein. We tested the hypothesis that activation of δ-opioid receptors mobilizes inositol 1,4,5-trisphosphate (IP3)-sensitive Ca2+ stores via liberation of Gβγ. Fura-2-based digital imaging was used to study the mechanism of opioid-induced increases in [Ca2+](i) in NG108-15 cells. Exposure to D-Ala2-D-Leu5 enkephalin (100 nM) for 90 s induced increases in [Ca2+](i) that were blocked by microinjection of the IP3 receptor antagonist heparin (pipette concentration = 100 mg/ml) but not by sham injection. Microinjection of a peptide that binds Gβγ (QEHA, 1 mM) decreased the D-Ala2-D-Leu5 enkephalin-evoked response. Microinjection of an inactive peptide (SKEE, 1 mM) that does not bind to Gβγ failed to inhibit the opioid-induced increase in [Ca2+](i). Microinjection of a peptide (QLKK, 15 mM) that binds to free Gα(q) blocked the increase evoked by 3 nM bradykinin, but microinjection of an inactive peptide (ADRK, 15 mM) did not. Microinjection of QLKK did not significantly affect the opioid- induced increase in [Ca2+](i). Collectively, these data demonstrate that activation of δ-opioid receptors induces the release of Ca2+ from IP(3)- sensitive stores in NG108-15 cells through activation of the βγ subunits of inhibitory G proteins.
AB - Activation of δ-opioid receptors in NG108-15 cells releases Ca2+ from an intracellular store through activation of a pertussis toxin-sensitive G protein. We tested the hypothesis that activation of δ-opioid receptors mobilizes inositol 1,4,5-trisphosphate (IP3)-sensitive Ca2+ stores via liberation of Gβγ. Fura-2-based digital imaging was used to study the mechanism of opioid-induced increases in [Ca2+](i) in NG108-15 cells. Exposure to D-Ala2-D-Leu5 enkephalin (100 nM) for 90 s induced increases in [Ca2+](i) that were blocked by microinjection of the IP3 receptor antagonist heparin (pipette concentration = 100 mg/ml) but not by sham injection. Microinjection of a peptide that binds Gβγ (QEHA, 1 mM) decreased the D-Ala2-D-Leu5 enkephalin-evoked response. Microinjection of an inactive peptide (SKEE, 1 mM) that does not bind to Gβγ failed to inhibit the opioid-induced increase in [Ca2+](i). Microinjection of a peptide (QLKK, 15 mM) that binds to free Gα(q) blocked the increase evoked by 3 nM bradykinin, but microinjection of an inactive peptide (ADRK, 15 mM) did not. Microinjection of QLKK did not significantly affect the opioid- induced increase in [Ca2+](i). Collectively, these data demonstrate that activation of δ-opioid receptors induces the release of Ca2+ from IP(3)- sensitive stores in NG108-15 cells through activation of the βγ subunits of inhibitory G proteins.
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U2 - 10.1124/mol.56.5.902
DO - 10.1124/mol.56.5.902
M3 - Article
C2 - 10531393
AN - SCOPUS:0013657555
SN - 0026-895X
VL - 56
SP - 902
EP - 908
JO - Molecular Pharmacology
JF - Molecular Pharmacology
IS - 5
ER -