Decrease in PIP(2) channel interactions is the final common mechanism involved in PKC- and arachidonic acid-mediated inhibitions of GABA(B)-activated K+ current.

We showed in our previous study that in hippocampal CA1 neurons the stimulation of muscarinic receptors inhibited the GIRK current (I(GIRK)) via a PLC/PKC pathway, whereas group I metabotropic glutamate receptors (mGluR) inhibited I(GIRK) via a PLA(2)/arachidonic acid pathway. In this study, we present evidence that receptor-mediated signalling pathways activated by the two G(q)-coupled receptors (G(q)PCRs) converge on the inhibition of GIRK channel-PIP(2) interaction. I(GIRK) was activated in acutely isolated hippocampal CA1 neurons by repetitive application of baclofen, a GABA(B) receptor agonist, with a 2-3 min interval. When both CCh and DHPG were pretreated before the second I(GIRK) activation, the magnitude of the second I(GIRK) was 52.2 +/- 2.5% of the first I(GIRK), which was not significantly different from the magnitude of inhibition by CCh or DHPG alone. This result shows that the effects of muscarinic receptor and group I mGluR stimulation on I(GIRK) are not additive but occlusive, suggesting that each pathway may converge to a common mechanism that finally regulates I(GIRK). To test the involvement of PIP(2) in this mechanism, the effect of CCh and DHPG on I(GIRK) was tested in cells loaded with exogenous PIP(2). The inhibition of I(GIRK) by CCh or DHPG was almost completely abolished in PIP(2)-loaded cells. We confirmed that the inhibition of I(GIRK) by direct application of phorbol ester or arachidonic acid was also completely reversed in PIP(2)-loaded cells. These results indicate that the decrease in PIP(2)-channel interactions is the final common mechanism responsible for G(q)PCR-induced inhibitions of I(GIRK) mediated by PKC and arachidonic acid.