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大鼠视交叉上核急性分离神经元中的钠电流和钙电流。

Sodium and calcium currents in acutely dissociated neurons from rat suprachiasmatic nucleus.

作者信息

Huang R C

机构信息

Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.

出版信息

J Neurophysiol. 1993 Oct;70(4):1692-703. doi: 10.1152/jn.1993.70.4.1692.

Abstract
  1. Neurons were acutely dissociated from the suprachiasmatic nucleus (SCN) of adult rats and studied with whole-cell and perforated-patch recordings at room temperature. 2. Acutely dissociated SCN neurons had spherical cell bodies of 12 microns in average diameter. The recorded cells were randomly selected and had either no process (38%), one (41%), two (19%), or three processes (2%). They had a resting potential of about -60 mV, an input resistance of approximately 5 G omega, and a cell capacitance of approximately 7 pF. 3. The dissociated neurons had variable spontaneous firing rates, typically (76%) < 1 Hz. 4. Under current clamp, continuous current injection elicited repetitive action potentials. 1 microM tetrodotoxin (TTX) reduced the amplitudes of the action potentials as well as the firing rate, whereas 200 microM Cd2+ stopped repetitive firing altogether. Action potentials were completely eliminated with Cd2+ and TTX present. These results suggest that both Na+ and Ca2+ contribute to the action potential in these cells. 5. With 200 microM Cd2+ present to block calcium currents, a train of brief depolarizing pulses could still elicit repetitive sodium action potentials, but these became attenuated at stimulating frequencies as low as 1 Hz. 6. Under voltage clamp, the sodium current was activated at about -40 mV and peaked at about -10 mV. It inactivated with a time constant of approximately 0.5 ms at 0 mV, and in steady state the current was half-inactivated at about -60 mV. Recovery of the current from inactivation showed two very different phases with time constants of approximately 30 and 600 ms at -60 mV. The slow phase was probably responsible for the very low firing rate of the sodium action responsible for the very low firing rate of the sodium action potential. 7. In the absence of external sodium, depolarization-activated calcium action potentials were preferentially blocked by 20 microM Cd2+, whereas a posthyperpolarizing depolarizing (or anode break) was preferentially reduced by 100 microM Ni2+. These differential effects hinted at the presence of both low-threshold and high-threshold calcium currents in these cells. 8. Voltage-clamp experiments confirmed the presence of a low-threshold, transient calcium current that was activated by depolarizations above -70 mV. It inactivated with a time constant of approximately 25 ms between -50 and -30 mV. Steady-state inactivation was half-complete at about -90 mV and complete at about -70 mV.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 从成年大鼠的视交叉上核(SCN)中急性分离出神经元,并在室温下用全细胞和穿孔膜片钳记录技术进行研究。2. 急性分离的SCN神经元具有平均直径为12微米的球形细胞体。记录的细胞是随机选择的,没有突起的占38%,有一个突起的占41%,有两个突起的占19%,有三个突起的占2%。它们的静息电位约为 -60 mV,输入电阻约为5 GΩ,细胞电容约为7 pF。3. 分离的神经元具有可变的自发放电率,通常(76%)<1 Hz。4. 在电流钳制下,持续的电流注入引发重复动作电位。1 μM河豚毒素(TTX)降低了动作电位的幅度以及放电率,而200 μM Cd2+ 则完全停止了重复放电。当同时存在Cd2+ 和TTX时,动作电位被完全消除。这些结果表明,Na+ 和Ca2+ 都参与了这些细胞的动作电位形成。5. 在存在200 μM Cd2+ 以阻断钙电流的情况下,一串短暂的去极化脉冲仍可引发重复的钠动作电位,但这些动作电位在低至1 Hz的刺激频率下就会衰减。6. 在电压钳制下,钠电流在约 -40 mV时被激活,在约 -10 mV时达到峰值。在0 mV时,它以约0.5 ms的时间常数失活,在稳态下,电流在约 -60 mV时半失活。电流从失活状态恢复显示出两个非常不同的阶段,在 -60 mV时时间常数分别约为30和600 ms。慢相可能是钠动作电位放电率极低的原因。7. 在没有外部钠的情况下,去极化激活的钙动作电位优先被20 μM Cd2+ 阻断,而后超极化去极化(或阳极断裂)则优先被100 μM Ni2+ 降低。这些不同的效应暗示这些细胞中存在低阈值和高阈值钙电流。8. 电压钳实验证实存在一种低阈值的瞬时钙电流,它在高于 -70 mV的去极化时被激活。在 -50至 -30 mV之间,它以约25 ms的时间常数失活。稳态失活在约 -90 mV时完成一半,在约 -70 mV时完全完成。(摘要截断于400字)

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