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硬骨鱼Mauthner细胞甘氨酸能突触处的单一电导变化:电压钳制与药理学分析

Unitary conductance changes at teleost Mauthner cell glycinergic synapses: a voltage-clamp and pharmacologic analysis.

作者信息

Faber D S, Korn H

机构信息

Department of Physiology, State University of New York, Buffalo 14214.

出版信息

J Neurophysiol. 1988 Dec;60(6):1982-99. doi: 10.1152/jn.1988.60.6.1982.

Abstract
  1. The magnitude and kinetics of inhibitory postsynaptic currents (IPSCs) evoked in the goldfish Mauthner (M-) cell by intracellular stimulation of identified presynaptic interneurons (unitary responses) and by activation of the recurrent collateral network were determined with single-and double electrode voltage-clamp techniques. 2. The peak magnitude of the inhibitory conductance changes were 5610 +/- 4800 nS (mean +/- SD; n = 13) for the collateral response, and 144 +/- 44 nS (n = 7) for the unitary IPSCs. These synaptic conductances, which are due to the opening of Cl- channels, were independent of the degree of Cl- -loading of the M-cell. 3. The peak amplitude of the collateral inhibitory postsynaptic potential (IPSP) was a constant fraction (0.52 +/- 0.06) of the driving force, which was determined from current-voltage plots for both types of IPSCs and ranged from 10 to 37 mV. These findings confirm indirect measurements from previous current-clamp studies and validate the normalization procedure used to previously calculate synaptic conductances from IPSP amplitudes, a method that therefore may be applicable to other central neurons. 4. At the resting membrane potential, the rise time of the unitary IPSCs was 0.34 +/- 0.07 ms (n = 18), whereas their decay was exponential, with a time constant of 5.7 +/- 1.1 ms (n = 16). 5. Iontophoretic and intramuscular applications of the glycine antagonist strychnine reduced or blocked M-cell inhibitory responses, without altering the excitability of the presynaptic neurons, or the driving force. 6. Amplitude fluctuations of unitary IPSPs recorded during partial blockade by strychnine were analyzed according to a binomial model of quantal transmitter release. In one experimental series, comparison of the binomial parameters before and after applying the antagonist indicated that only quantal size, q, was reduced, whereas n, the number of available release units, and p, the probability of release, were unaffected by strychnine. In a second series, the individual presynaptic cells were injected with horseradish peroxidase (HRP), and it was found that the correlation between n and the number of stained presynaptic boutons and, therefore, of active zones, was maintained in the presence of the drug. No evidence was found for silent synapses in these conditions. 7. The quantal conductance, gq, was estimated from the binomially derived quantal size, in millivolts, and the voltage-clamp measurements of the IPSP driving force and M-cell input conductance. gq averaged 21.5 nS in control conditions and 12.3 nS in the presence of strychnine.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 采用单电极和双电极电压钳技术,测定了金鱼Mauthner(M-)细胞中,经细胞内刺激特定突触前中间神经元(单位反应)以及激活回返侧支网络所诱发的抑制性突触后电流(IPSCs)的大小和动力学。2. 侧支反应的抑制性电导变化峰值大小为5610±4800 nS(平均值±标准差;n = 13),单位IPSCs的为144±44 nS(n = 7)。这些由于Cl-通道开放产生的突触电导,与M-细胞的Cl-负载程度无关。3. 侧支抑制性突触后电位(IPSP)的峰值幅度是驱动力的一个恒定分数(0.52±0.06),该驱动力由两种IPSCs的电流-电压图确定,范围为10至37 mV。这些发现证实了先前电流钳研究的间接测量结果,并验证了先前用于从IPSP幅度计算突触电导的归一化程序,因此该方法可能适用于其他中枢神经元。4. 在静息膜电位时,单位IPSCs的上升时间为0.34±0.07 ms(n = 18),而其衰减呈指数形式,时间常数为5.7±1.1 ms(n = 16)。5. 甘氨酸拮抗剂士的宁的离子电泳和肌肉内应用减少或阻断了M-细胞的抑制反应,而不改变突触前神经元的兴奋性或驱动力。6. 根据量子递质释放的二项式模型,分析了士的宁部分阻断期间记录的单位IPSPs的幅度波动。在一个实验系列中,比较应用拮抗剂前后的二项式参数表明,只有量子大小q减小,而可用释放单位数量n和释放概率p不受士的宁影响。在第二个系列中,向单个突触前细胞注射辣根过氧化物酶(HRP),发现在药物存在的情况下,n与染色突触前终扣数量以及因此与活性区数量之间的相关性得以维持。在这些条件下未发现沉默突触的证据。7. 根据以毫伏为单位的二项式推导量子大小以及IPSP驱动力和M-细胞输入电导的电压钳测量值,估计量子电导gq。在对照条件下,gq平均为21.5 nS,在士的宁存在时为12.3 nS。(摘要截断于400字)

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