Kotsias Basilio A, Salim Mohammad, Peracchia Lillian L, Peracchia Camillo
Department of Pharmacology and Physiology, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, NY 14642-8711, USA.
J Membr Biol. 2006;214(1):1-8. doi: 10.1007/s00232-006-0064-8. Epub 2007 Jun 2.
The cystic fibrosis transmembrane regulator (CFTR) is a Cl(-) channel known to influence other channels, including connexin (Cx) channels. To study the functional interaction between CFTR and gap junction channels, we coexpressed in Xenopus oocytes CFTR and either Cx45, Cx40, Cx32 or Cx50 and monitored junctional conductance (G (j)) and its sensitivity to transjunctional voltage (V (j)) by the dual voltage-clamp method. Application of forskolin induced a Cl(-) current; increased G (j) approximately 750%, 560%, 64% and 8% in Cx45, Cx40, Cx32 and Cx50, respectively; and decreased sensitivity to V (j ) gating, monitored by a change in the ratio between G (j) steady state and G (j) peak (G (j)SS/G (j)PK) at the pulse. In oocyte pairs expressing just Cx45 in one oocyte (#1) and both Cx45 and CFTR in the other (#2), with negative pulses applied to oocyte #1 forskolin application still increased G (j) and decreased the sensitivity to V (j) gating, indicating that CFTR activation is effective even when it affects only one of the two hemichannels and that the G (j) and V (j) changes are not artifacts of decreased membrane resistance in the pulsed oocyte. COOH-terminus truncation reduced the forskolin effect on Cx40 (Cx40TR) but not on Cx32 (Cx32TR) channels. The data suggest a cross-talk between CFTR and a variety of gap junction channels. Cytoskeletal scaffolding proteins and/or other intermediate cytoplasmic proteins are likely to play a role in CFTR-Cx interaction.
囊性纤维化跨膜调节因子(CFTR)是一种氯离子通道,已知其可影响包括连接蛋白(Cx)通道在内的其他通道。为了研究CFTR与缝隙连接通道之间的功能相互作用,我们在非洲爪蟾卵母细胞中共表达CFTR和Cx45、Cx40、Cx32或Cx50,并用双电压钳法监测连接电导(G(j))及其对跨连接电压(V(j))的敏感性。应用福斯可林可诱导氯离子电流;分别使Cx45、Cx40、Cx32和Cx50中的G(j)增加约750%、560%、64%和8%;并降低对V(j)门控的敏感性,通过脉冲时G(j)稳态与G(j)峰值之比(G(j)SS/G(j)PK)的变化来监测。在一个卵母细胞(#1)中仅表达Cx45而另一个卵母细胞(#2)中同时表达Cx45和CFTR的卵母细胞对中,对卵母细胞#1施加负脉冲时,应用福斯可林仍可增加G(j)并降低对V(j)门控的敏感性,这表明即使CFTR仅影响两个半通道中的一个,其激活也是有效的,并且G(j)和V(j)的变化不是脉冲卵母细胞膜电阻降低的假象。COOH末端截短降低了福斯可林对Cx40(Cx40TR)通道的作用,但对Cx32(Cx32TR)通道没有影响。数据表明CFTR与多种缝隙连接通道之间存在相互作用。细胞骨架支架蛋白和/或其他中间细胞质蛋白可能在CFTR-Cx相互作用中发挥作用。