Reboreda Antonio, Sánchez Estela, Romero Marcos, Lamas J Antonio
Section of Physiology, Department of Functional Biology, Faculty of Sciences, University of Vigo, Lagoas-Marcosende, 36200 Vigo, Spain.
J Physiol. 2003 Aug 15;551(Pt 1):191-205. doi: 10.1113/jphysiol.2003.039917. Epub 2003 Jul 4.
The basis of rhythmic activity observed at the dorsal column nuclei (DCN) is still open to debate. This study has investigated the electrophysiological properties of isolated DCN neurones deprived of any synaptic influence, using the perforated-patch technique. About half of the DCN neurones (64/130) were spontaneously active. More than half of the spontaneous neurones (36/64) showed a low threshold membrane oscillation (LTO) with a mean frequency of 11.4 Hz (range: 4.3-22.1 Hz, n = 20; I = 0). Cells showing LTOs also invariably showed a rhythmic 1.2 Hz clustering activity (groups of 2-5 action potentials separated by silent LTO periods). Also, more than one-third of the silent neurones presented clustering activity, always accompanied by LTOs, when slightly depolarised. The frequency of LTOs was voltage dependent and could be abolished by TTX (0.5 microM) and riluzole (30 microM), suggesting the participation of a sodium current. LTOs were also abolished by TEA (15 mM), which transformed clustering into tonic activity. In voltage clamp, most DCN neurones (85%) showed a TTX-/riluzole-sensitive persistent sodium current (INa,p), which activated at about -60 mV and had a half-maximum activation at -49.8 mV. An M-like, non-inactivating outward current was present in 95% of DCN neurones, and TEA (15 mM) inhibited this current by 73.7 %. The non-inactivating outward current was also inhibited by barium (1 mM) and linopirdine (10 microM), which suggests its M-like nature; both drugs failed to block the LTOs, but induced a reduction in their frequency by 56 and 20%, respectively. These results demonstrate for the first time that DCN neurones have a complex and intrinsically driven clustering discharge pattern, accompanied by subthreshold membrane oscillations. Subthreshold oscillations rely on the interplay of a persistent sodium current and a non-inactivating TEA-sensitive outward current.
在背柱核(DCN)观察到的节律性活动的基础仍存在争议。本研究使用穿孔膜片钳技术,研究了去除任何突触影响的离体DCN神经元的电生理特性。大约一半的DCN神经元(64/130)自发活动。超过一半的自发神经元(36/64)表现出低阈值膜振荡(LTO),平均频率为11.4 Hz(范围:4.3 - 22.1 Hz,n = 20;I = 0)。表现出LTO的细胞也总是表现出节律性的1.2 Hz簇状活动(由沉默的LTO期分隔的2 - 5个动作电位组)。此外,超过三分之一的沉默神经元在轻微去极化时表现出簇状活动,且总是伴有LTO。LTO的频率依赖于电压,可被TTX(0.5 microM)和利鲁唑(30 microM)消除,提示有钠电流参与。LTO也可被TEA(15 mM)消除,后者将簇状活动转变为强直活动。在电压钳实验中,大多数DCN神经元(85%)表现出对TTX/利鲁唑敏感的持续性钠电流(INa,p),该电流在约 - 60 mV时激活,在 - 49.8 mV时达到最大激活的一半。95%的DCN神经元存在一种M样、非失活外向电流,TEA(15 mM)可抑制该电流73.7%。非失活外向电流也可被钡(1 mM)和利诺吡啶(10 microM)抑制,提示其M样性质;两种药物均未能阻断LTO,但分别使LTO频率降低56%和20%。这些结果首次证明DCN神经元具有复杂的、内在驱动的簇状放电模式,并伴有阈下膜振荡。阈下振荡依赖于持续性钠电流和非失活的TEA敏感外向电流的相互作用。