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大鼠视皮层中投射至上丘的神经元的非钙依赖性去极化激活钾电流

Calcium-independent depolarization-activated potassium currents in superior colliculus-projecting rat visual cortical neurons.

作者信息

Albert J L, Nerbonne J M

机构信息

Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.

出版信息

J Neurophysiol. 1995 Jun;73(6):2163-78. doi: 10.1152/jn.1995.73.6.2163.

Abstract
  1. K+ conductances were characterized in isolated, identified superior colliculus-projecting (SCP) rat visual cortical neurons. SCP neurons were identified in vitro under epifluorescence illumination after in vivo retrograde labeling with rhodamine-labeled microspheres or "beads." For experiments, SCP neurons were isolated from the primary visual cortex of postnatal day 7 to 16 (P7-P16) Long Evans rat pups after bead injections into the ipsilateral superior colliculus at p5. 2. Recording conditions were optimized to allow the characterization of Ca2+ -independent K+ conductances. SCP cells that were largely devoid of processes were selected for recording, and experiments were completed 2-30 h after cell isolation. Ca2+ -independent, depolarization-activated K+ currents were routinely recorded during 200-ms voltage steps to potentials positive to -50 mV from a holding potential of -70 mV. 3. Peak outward current densities and the relative amplitudes of the peak and plateau outward currents evoked during 200-ms voltage steps varied among SCP cells. Although cells were isolated from animals at different ages (P7-P16) and maintained for varying times in vitro (2-30 h), no correlations were found between the variations in peak current densities or peak to plateau current ratios and the age of the animal from which the cell was isolated or the length of time the cell was maintained in vitro before recording. 4. Pharmacological experiments revealed the coexpression of three K+ current components in SCP cells that could be separated on the basis of differing sensitivities to the K+ channel blockers, 4-aminopyridine (4-AP) and tetraethylammonium (TEA). Varying the concentration of 4-AP, for example, facilitated the separation of two rapidly activating K+ currents similar to A (IA) and D(ID) type currents in other cells. ID in SCP neurons is blocked by micromolar concentrations of 4-AP, whereas micromolar concentrations of 4-AP are required to effect complete block of IA in these cells. The current component remaining in the presence of high concentrations (5-10 mM) of 4-AP is slowly activating outward K+ current, similar to delayed rectifier (IK) currents in other cells. IK in SCP neurons is blocked by micromolar concentrations of TEA. 5. Activation of IA, ID, and IK in SCP neurons is voltage dependent, although the three current components display distinct time- and voltage-dependent properties. For example, although both IA and ID begin to activate at approximately -50 mV, IA activates two to three times faster than ID. In addition, the threshold for activation of IK (-30 mV) is approximately 20 mV depolarized from that of IA (or ID), and the voltage dependence of IK activation is steeper than that of IA and ID.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 在分离出的、已鉴定的投射至上丘的大鼠视觉皮层神经元(SCP)中对钾离子电导进行了特性描述。在用罗丹明标记的微球或“珠子”进行体内逆行标记后,在落射荧光照明下于体外鉴定出SCP神经元。对于实验,在出生后第5天向同侧上丘注射珠子后,从出生后第7至16天(P7 - P16)的Long Evans大鼠幼崽的初级视觉皮层中分离出SCP神经元。2. 优化记录条件以对与钙离子无关的钾离子电导进行特性描述。选择基本没有突起的SCP细胞进行记录,实验在细胞分离后2 - 30小时内完成。从 - 70 mV的钳制电位开始,在200毫秒的电压阶跃至 - 50 mV以上的电位期间,常规记录与钙离子无关的、去极化激活的钾离子电流。3. 在200毫秒电压阶跃期间诱发的峰值外向电流密度以及峰值和平台期外向电流的相对幅度在SCP细胞之间有所不同。尽管细胞是从不同年龄(P7 - P16)的动物中分离出来的,并且在体外维持了不同的时间(2 - 30小时),但在峰值电流密度或峰值与平台期电流比值的变化与分离细胞的动物年龄或记录前细胞在体外维持的时间长度之间未发现相关性。4. 药理学实验揭示了SCP细胞中三种钾离子电流成分的共表达,这三种成分可根据对钾离子通道阻滞剂(即4 - 氨基吡啶(4 - AP)和四乙铵(TEA))的不同敏感性进行区分。例如,改变4 - AP的浓度有助于分离出两种快速激活的钾离子电流,类似于其他细胞中的A(IA)型和D(ID)型电流。SCP神经元中的ID可被微摩尔浓度的4 - AP阻断,而在这些细胞中需要微摩尔浓度的4 - AP才能完全阻断IA。在高浓度(5 - 10 mM)的4 - AP存在下剩余的电流成分是缓慢激活的外向钾离子电流,类似于其他细胞中的延迟整流器(IK)电流。SCP神经元中的IK可被微摩尔浓度的TEA阻断。5. SCP神经元中IA、ID和IK的激活是电压依赖性的,尽管这三种电流成分表现出不同的时间和电压依赖性特性。例如,尽管IA和ID都在大约 - 50 mV开始激活,但IA的激活速度比ID快两到三倍。此外,IK激活的阈值( - 30 mV)比IA(或ID)的阈值大约去极化20 mV,并且IK激活的电压依赖性比IA和ID更陡峭。(摘要截断于400字)

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