Gamper Nikita, Reznikov Vitaliy, Yamada Yoichi, Yang Jian, Shapiro Mark S
Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229, USA.
J Neurosci. 2004 Dec 1;24(48):10980-92. doi: 10.1523/JNEUROSCI.3869-04.2004.
Modulation of voltage-gated Ca2+ channels via G-protein-coupled receptors is a prime mechanism regulating neurotransmitter release and synaptic plasticity. Despite extensive studies, the molecular mechanism underlying Gq/11-mediated modulation remains unclear. We found cloned and native N-type Ca2+ channels to be regulated by phosphatidylinositol [correction] 4,5-bisphosphate (PIP2). In inside-out oocyte patches, PIP2 greatly attenuated or reversed the observed rundown of expressed channels. In sympathetic neurons, muscarinic M1 ACh receptor suppression of the Ca2+ current (ICa) was temporally correlated with PIP2 hydrolysis, blunted by PIP2 in whole-cell pipettes, attenuated by expression of PIP2-sequestering proteins, and became irreversible when PIP2 synthesis was blocked. We also probed mechanisms of receptor specificity. Although bradykinin also induced PIP2 hydrolysis, it did not inhibit ICa. However, bradykinin receptors became nearly as effective as M1 receptors when PIP2 synthesis, IP3 receptors, or the activity of neuronal Ca2+ sensor-1 were blocked, suggesting that bradykinin receptor-induced intracellular Ca2+ increases stimulate PIP2 synthesis, compensating for PIP2 hydrolysis. We suggest that differential use of PIP2 signals underlies specificity of Gq/11-coupled receptor actions on the channels
通过G蛋白偶联受体对电压门控Ca2+通道进行调节是调控神经递质释放和突触可塑性的主要机制。尽管已有广泛研究,但Gq/11介导的调节作用背后的分子机制仍不清楚。我们发现克隆的和天然的N型Ca2+通道受磷脂酰肌醇4,5-二磷酸(PIP2)调控。在膜内面向外的卵母细胞片膜中,PIP2极大地减弱或逆转了所观察到的表达通道的电流衰减。在交感神经元中,毒蕈碱型M1 ACh受体对Ca2+电流(ICa)的抑制与PIP2水解在时间上相关,全细胞膜片钳吸管中的PIP2可使其减弱,PIP2螯合蛋白的表达可使其衰减,当PIP2合成被阻断时则变得不可逆。我们还探究了受体特异性的机制。尽管缓激肽也诱导PIP2水解,但它并不抑制ICa。然而,当PIP2合成、IP3受体或神经元Ca2+传感器-1的活性被阻断时,缓激肽受体的作用几乎与M1受体一样有效,这表明缓激肽受体诱导的细胞内Ca2+增加刺激了PIP2合成,补偿了PIP2水解。我们认为,PIP2信号的差异利用是Gq/11偶联受体对通道作用特异性的基础。