Analysis of G-protein-activated inward rectifying K+ (GIRK) channel currents upon GABA(B) receptor activation in rat supraoptic neurons

作者：Harayama Nobuya; Kayano Tomohiko; Moriya Taiki; Kitamura Naoki; Shibuya Izumi^*; Tanaka Yamamoto Keiko; Uezono Yasuhito; Ueta Yoichi; Sata Takeyoshi

来源：Brain Research, 2014, 1591: 1-13.

DOI：10.1016/j.brainres.2014.10.022

• Supraoptic nucleus
• GIRK channels
• GABA(B) receptors
• Gi/o
• Patch clamp
• RT-PCR

While magnocellular neurons in the supraoptic nucleus (SON) possess rich Gi/o-mediated mechanisms, molecular and cellular properties of G-protein-activated inwardly rectifying K+ (GIRK) channels have been controversial. Here, properties of GIRK channels are examined by RT-PCR and whole-cell patch-clamp techniques in rat SON neurons. Patch clamp experiments showed that the selective GABA(B) agonist, baclofen, enhanced currents in a high K+ condition. The baclofen-enhanced currents exhibited evident inward rectification and were blocked by the selective GABA(B) antagonist, CGP55845A, the IRK channel blocker, Ba2+, and the selective GIRK channel blocker, tertiapin, indicating that baclofen activates GIRK channels via GABA(B) receptors. The GIRK currents were abolished by N-ethylmaleimide pretreatment, and prolonged by GTP gamma S inclusion in the patch pipette, suggesting that Gi/o proteins are involved. RT-PCR analysis revealed mRNAs for all four GIRK 1-4 channels and for both GABA(B)R1 and GABA(B)R2 receptors in rat SON. However, the concentration-dependency of the baclofen-induced activation of GIRK currents had an EC50 of 110 mu M, which is about 100 times higher than that of baclofen-induced inhibition of voltage-dependent Ca2+ channels. Moreover, baclofen caused no significant changes in the membrane potential and the firing rate. These results suggest that although GIRK channels can be activated by GABA(B) receptors via the Gi/o pathway, this occurs at high agonist concentrations, and thus may not be a physiological mechanism regulating the function of SON neurons. This property that the membrane potential receives little influence from GIRK currents seems to be uncommon for CNS neurons possessing rich Gi/o-coupled receptors, and could be a special feature of rat SON neurons.

• 出版日期2014-12-3