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大鼠外侧膝状核急性分离神经元中两种钙电流的鉴定。

Identification of two calcium currents in acutely dissociated neurons from the rat lateral geniculate nucleus.

作者信息

Hernández-Cruz A, Pape H C

机构信息

Department of Neurobiology and Behavior, State University of New York, Stony Brook 11794.

出版信息

J Neurophysiol. 1989 Jun;61(6):1270-83. doi: 10.1152/jn.1989.61.6.1270.

Abstract
  1. Intracellular recording in the in vitro slice preparation and whole-cell, patch-clamp recording of acutely dissociated neurons from the rat lateral geniculate nucleus (LGN) were combined to study the Ca currents underlying their electrical responses. In slices from young animals (postnatal days 13-16), we found that dorsal LGN neurons have responses similar to those of adult preparations, including the presence of a low-threshold Ca spike (LTS). After enzymatic isolation of LGN neurons from the same animals, the firing properties appeared well preserved, as indicated by whole-cell, current-clamp recordings from dissociated multipolar cells (presumably geniculocortical relay neurons). 2. Two types of Ca currents were identified in voltage-clamped, isolated LGN neurons on the basis of their voltage dependency, pharmacology, and selectivity properties. These two currents resemble the low-voltage-activated (LVA) and high-voltage-activated (HVA) Ca channels found in rat sensory neurons (9). 3. The LVA current component required negative potentials (less than -80 mV) to deinactivate completely, started to activate around -60 mV and reached a plateau level around -25 mV. It peaked within 30-6 ms and decayed with a single time constant of approximately 24 ms at -20 mV. Its inactivation curve ranged from -100 to -40 mV, with a half-inactivation near -60 mV. The HVA current component could be isolated by holding the membrane potential positive to -60 mV, activated at potentials positive to -30 mV and peaked around +5 mV. The time-to-peak ranged from 30 to 6 ms in the voltage range from -30 to +35 mV and decayed very slowly with sustained depolarizing pulses (time constant ranged between 1,600 and 40 ms over the same voltage range). 4. The inactivation of LVA Ca current during depolarizing voltage steps was consistent with a voltage-dependent process. The recovery from inactivation after short (100 ms), inactivating prepulses displayed two exponential phases. The slower phase was predominant under conditions that induce large current flow through the membrane, suggesting a Ca-mediated mechanism. 5. The LVA current was preferentially blocked by 50 microM Ni2+, leaving the HVA currents almost unaltered. Fifty micromolars Cd2+, in contrast, seemed more effective in blocking the HVA component of the Ca current.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 将体外脑片制备中的细胞内记录与大鼠外侧膝状体核(LGN)急性解离神经元的全细胞膜片钳记录相结合,以研究其电反应背后的钙电流。在幼龄动物(出生后第13 - 16天)的脑片中,我们发现背侧LGN神经元的反应与成年动物制备的相似,包括存在低阈值钙峰(LTS)。在用酶从相同动物分离LGN神经元后,放电特性似乎保存良好,从解离的多极细胞(可能是膝状体 - 皮质中继神经元)进行的全细胞电流钳记录表明了这一点。2. 根据电压依赖性、药理学和选择性特性,在电压钳制的分离LGN神经元中鉴定出两种类型的钙电流。这两种电流类似于在大鼠感觉神经元中发现的低电压激活(LVA)和高电压激活(HVA)钙通道(9)。3. LVA电流成分需要负电位(小于 - 80 mV)才能完全去失活,在约 - 60 mV开始激活,并在约 - 25 mV达到平台水平。它在30 - 6 ms内达到峰值,并在 - 20 mV时以约24 ms的单一时间常数衰减。其失活曲线范围为 - 100至 - 40 mV,半失活在 - 60 mV附近。通过将膜电位保持在正于 - 60 mV,可以分离出HVA电流成分,它在正于 - 30 mV的电位时激活,并在约 + 5 mV达到峰值。在 - 30至 + 35 mV的电压范围内,峰值时间为30至6 ms,并且在持续去极化脉冲下衰减非常缓慢(在相同电压范围内时间常数在1600至40 ms之间)。4. 在去极化电压阶跃期间LVA钙电流的失活与电压依赖性过程一致。在短(100 ms)失活预脉冲后的失活恢复显示出两个指数阶段。在诱导大电流通过膜的条件下,较慢的阶段占主导,表明是一种钙介导的机制。5. LVA电流优先被50 microM Ni2 +阻断,而HVA电流几乎不受影响。相比之下,50 microM Cd2 +似乎更有效地阻断钙电流的HVA成分。(摘要截断于400字)

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