Department of Neuroscience, Physiology and Pharmacology, University College London, London WC1E 6BT, United Kingdom.
J Neurosci. 2010 Aug 18;30(33):11068-72. doi: 10.1523/JNEUROSCI.2597-10.2010.
Expressed metabotropic group 1 glutamate mGluR5 receptors and nucleotide P2Y1 receptors (P2Y1Rs) show promiscuous ion channel coupling in sympathetic neurons: their stimulation inhibits M-type [Kv7, K(M)] potassium currents and N-type (Ca(V)2.2) calcium currents (Kammermeier and Ikeda, 1999; Brown et al., 2000). These effects are mediated by G(q) and G(i/o) G-proteins, respectively. Via their C-terminal tetrapeptide, these receptors also bind to the PDZ domain of the scaffold protein NHERF2, which enhances their coupling to G(q)-mediated Ca(2+) signaling (Fam et al., 2005; Paquet et al., 2006b). We investigated whether NHERF2 could modulate coupling to neuronal ion channels. We find that coexpression of NHERF2 in sympathetic neurons (by intranuclear cDNA injections) does not affect the extent of M-type potassium current inhibition produced by either receptor but strongly reduced Ca(V)2.2 inhibition by both P2Y1R and mGluR5 activation. NHERF2 expression had no significant effect on Ca(V)2.2 inhibition by norepinephrine (via alpha(2)-adrenoceptors, which do not bind NHERF2), nor on Ca(V)2.2 inhibition produced by an expressed P2Y1R lacking the NHERF2-binding DTSL motif. Thus, NHERF2 selectively restricts downstream coupling of mGluR5 and P2Y1Rs in neurons to G(q)-mediated responses such as M-current inhibition. Differential distribution of NHERF2 in neurons may therefore determine coupling of mGluR5 receptors and P2Y1 receptors to calcium channels.
表达代谢型谷氨酸 mGluR5 受体和核苷酸 P2Y1 受体 (P2Y1R) 在交感神经元中表现出混杂的离子通道偶联:它们的刺激抑制 M 型 [Kv7,K(M)] 钾电流和 N 型(Ca(V)2.2)钙电流(Kammermeier 和 Ikeda,1999;Brown 等人,2000)。这些作用分别由 G(q) 和 G(i/o) G 蛋白介导。通过它们的 C 末端四肽,这些受体还与支架蛋白 NHERF2 的 PDZ 结构域结合,从而增强它们与 G(q) 介导的 Ca(2+)信号转导的偶联(Fam 等人,2005;Paquet 等人,2006b)。我们研究了 NHERF2 是否可以调节与神经元离子通道的偶联。我们发现,NHERF2 在交感神经元中的共表达(通过核内 cDNA 注射)不会影响两种受体产生的 M 型钾电流抑制的程度,但强烈降低了 P2Y1R 和 mGluR5 激活引起的 Ca(V)2.2 抑制。NHERF2 表达对去甲肾上腺素(通过不与 NHERF2 结合的 alpha(2)-肾上腺素受体)引起的 Ca(V)2.2 抑制以及表达的缺乏 NHERF2 结合 DTSL 基序的 P2Y1R 引起的 Ca(V)2.2 抑制没有显著影响。因此,NHERF2 选择性地限制了神经元中 mGluR5 和 P2Y1R 的下游偶联到 G(q)介导的反应,例如 M 电流抑制。神经元中 NHERF2 的差异分布可能因此决定了 mGluR5 受体和 P2Y1 受体与钙通道的偶联。