Isope Philippe, Murphy Timothy H
Kinsmen Laboratory and Brain Research Centre, University of British Columbia, Vancouver, BC, Canada V6T 1Z3.
J Physiol. 2005 Jan 1;562(Pt 1):257-69. doi: 10.1113/jphysiol.2004.074211. Epub 2004 Oct 28.
The functional role of low voltage activated (LVA) calcium channels in the cerebellar Purkinje cell dendritic tree is not completely understood. Since the localization of these channels will influence their possible roles in dendritic integration and induction of plasticity, we set out to characterize the LVA calcium current in Purkinje cell dendrites in acute cerebellar slices of young rats. Using a combination of electrophysiological recordings and two-photon laser scanning microscopy, we show that LVA calcium current recorded at the soma can be correlated with voltage-dependent calcium transients in Purkinje cell dendritic spines. Blocking sodium and potassium conductances allowed us to isolate and characterize a fast inactivating inward current activated positive to -55 mV. Activation and steady-state inactivation kinetics, voltage-dependent deactivation kinetics, and pharmacological experiments (using omega-agatoxin-IVA, mibefradil and nickel) show that this current is carried by T-type calcium channels. Furthermore, the LVA calcium transient observed in the dendritic spines of the Purkinje cell is well correlated with the current recorded at the soma, suggesting that T-type calcium channels are the main component of the LVA calcium input in spines. The fast rising phase of the calcium transient in spines and the absence of delay between the onset in the spine and the parent dendrite show that T-type calcium channels are present both in spines and dendrites of the Purkinje cell.
低电压激活(LVA)钙通道在小脑浦肯野细胞树突中的功能作用尚未完全明确。由于这些通道的定位会影响它们在树突整合和可塑性诱导中的潜在作用,我们着手在幼鼠急性小脑切片中对浦肯野细胞树突中的LVA钙电流进行表征。通过结合电生理记录和双光子激光扫描显微镜,我们发现,在胞体记录到的LVA钙电流与浦肯野细胞树突棘中电压依赖性钙瞬变相关。阻断钠电导和钾电导使我们能够分离并表征一种在-55 mV时被激活的快速失活内向电流。激活和稳态失活动力学、电压依赖性失活动力学以及药理学实验(使用ω-芋螺毒素-IVA、米贝拉地尔和镍)表明,该电流由T型钙通道携带。此外,在浦肯野细胞树突棘中观察到的LVA钙瞬变与在胞体记录到的电流高度相关,这表明T型钙通道是树突棘中LVA钙输入的主要成分。树突棘中钙瞬变的快速上升阶段以及树突棘与母树突起始之间无延迟表明,T型钙通道同时存在于浦肯野细胞的树突棘和树突中。