Squecco Roberta, Bencini Chiara, Piperio Claudia, Francini Fabio
Department of Physiological Sciences, University of Florence, Viale G.B. Morgagni 63, I-50134 Florence, Italy.
J Physiol. 2004 Feb 15;555(Pt 1):137-52. doi: 10.1113/jphysiol.2003.051730. Epub 2003 Dec 5.
The dihydropyridine receptors (DHPRs)/L-type Ca2+ channels of skeletal muscle are coupled with ryanodine receptors/Ca2+ release channels (RyRs/CRCs) located in the sarcoplasmic reticulum (SR). The DHPR is the voltage sensor for excitation-contraction (EC) coupling and the charge movement component q gamma has been implicated as the signal linking DHPR voltage sensing to Ca2+ release from the coupled RyR. Recently, a new charge component, qh, has been described and related to L-type Ca2+ channel gating. Evidence has also been provided that the coupled RyR/CRC can modulate DHPR functions via a retrograde signal. Our aim was to investigate whether the newly described qh is also involved in the reciprocal interaction or cross-talk between DHPR/L-type Ca2+ channel and RyR/CRC. To this end we interfered with DHPR/L-type Ca2+ channel function using nifedipine and 1-alkanols (heptanol and octanol), and with RyR/CRC function using ryanodine and ruthenium red (RR). Intramembrane charge movement (ICM) and L-type Ca2+ current (ICa) were measured in single cut fibres of the frog using the double-Vaseline-gap technique. Our records showed that nifedipine reduced the amount of q gamma and qh moved by approximately 90% and approximately 55%, respectively, whereas 1-alkanols completely abolished them. Ryanodine and RR shifted the transition voltages of q gamma and qh and of the maximal conductance of ICa by approximately 4-9 mV towards positive potentials. All these interventions spared q beta. These results support the hypothesis that only q gamma; and qh arise from the movement of charged particles within the DHPR/L-type Ca2+ channel and that these charge components together with ICa are affected by a retrograde signal from RyR/CRC.
骨骼肌中的二氢吡啶受体(DHPRs)/L型Ca2+通道与位于肌浆网(SR)中的兰尼碱受体/Ca2+释放通道(RyRs/CRCs)相偶联。DHPR是兴奋-收缩(EC)偶联的电压传感器,电荷移动成分qγ被认为是将DHPR电压传感与偶联的RyR释放Ca2+相联系的信号。最近,一种新的电荷成分qh被描述并与L型Ca2+通道门控相关。也有证据表明偶联的RyR/CRC可通过逆行信号调节DHPR功能。我们的目的是研究新描述的qh是否也参与DHPR/L型Ca2+通道与RyR/CRC之间的相互作用或串扰。为此,我们使用硝苯地平和1-链烷醇(庚醇和辛醇)干扰DHPR/L型Ca2+通道功能,使用兰尼碱和钌红(RR)干扰RyR/CRC功能。使用双凡士林间隙技术在青蛙的单根切断纤维中测量膜内电荷移动(ICM)和L型Ca2+电流(ICa)。我们的记录显示,硝苯地平使qγ和qh移动的量分别减少了约90%和约55%,而1-链烷醇则完全消除了它们。兰尼碱和RR使qγ和qh以及ICa的最大电导的转变电压向正电位方向移动了约4-9 mV。所有这些干预均未影响qβ。这些结果支持以下假设,即只有qγ和qh源自DHPR/L型Ca2+通道内带电粒子的移动,并且这些电荷成分与ICa一起受到来自RyR/CRC的逆行信号的影响。