Demontis Gian Carlo, Moroni Anna, Gravante Biagio, Altomare Claudia, Longoni Biancamaria, Cervetto Luigi, DiFrancesco Dario
Dipartimento di Psichiatria, Neurobiologia, Farmacologia e Biotecnologie, Università di Pisa, Via Bonanno, 6-56126 Pisa, Italy.
J Physiol. 2002 Jul 1;542(Pt 1):89-97. doi: 10.1113/jphysiol.2002.017640.
Gating of voltage-dependent conductances in retinal photoreceptors is the first step of a process leading to the enhancement of the temporal performance of the visual system. The molecular components underlying voltage-dependent gating in rods are presently poorly defined. In the present work we have investigated the isoform composition and the functional characteristics of hyperpolarisation-activated cyclic nucleotide-gated channels (HCN) in rabbit rods. Using immunocytochemistry we show the expression in the inner segment and cell body of the isoform 1 (HCN1). Electrophysiological investigations show that hyperpolarisation-activated currents (I(h)) can be measured only from the cell regions where HCN1 is expressed. Half-activation voltage (-75.0 +/- 0.3 mV) and kinetics (t(1/2) of 101 +/- 8 ms at -110 mV and 20 degrees C) of the I(h) in rods are similar to those of the macroscopic current carried by homomeric rabbit HCN1 channels expressed in HEK 293 cells. The homomeric nature of HCN1 channels in rods is compatible with the observation that cAMP induces a small shift (2.3 +/- 0.8 mV) in the half-activation voltage of I(h). In addition, the observation that within the physiological range of membrane potentials, cAMP does not significantly affect the gain of the current-to-voltage conversion, may reflect the need to protect the first step in the processing of visual signals from changes in cAMP turnover.
视网膜光感受器中电压依赖性电导的门控是导致视觉系统时间性能增强过程的第一步。目前,视杆细胞中电压依赖性门控的分子成分尚不清楚。在本研究中,我们研究了兔视杆细胞中超极化激活的环核苷酸门控通道(HCN)的亚型组成和功能特性。通过免疫细胞化学,我们显示了亚型1(HCN1)在视杆细胞内段和细胞体中的表达。电生理研究表明,超极化激活电流(I(h))只能从表达HCN1的细胞区域测量到。视杆细胞中I(h)的半激活电压(-75.0±0.3 mV)和动力学(在-110 mV和20℃时t(1/2)为101±8 ms)与在HEK 293细胞中表达的同源兔HCN1通道携带的宏观电流相似。视杆细胞中HCN1通道的同源性质与cAMP诱导I(h)半激活电压小幅度偏移(2.3±0.8 mV)的观察结果一致。此外,在膜电位的生理范围内,cAMP不会显著影响电流-电压转换增益的观察结果,可能反映了保护视觉信号处理第一步免受cAMP周转变化影响的必要性。