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Characterisation of inhibitory and excitatory postsynaptic currents of the rat medial superior olive

机译:大鼠内侧上橄榄抑制和兴奋性突触后电流的表征

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摘要

class="enumerated" style="list-style-type:decimal">The medial superior olive (MSO) is part of the binaural auditory pathway, receiving excitatory projections from both cochlear nuclei and an inhibitory input from the ipsilateral medial nucleus of the trapezoid body (MNTB). We characterised the excitatory and inhibitory synaptic currents of MSO neurones in 3- to 14-day-old rats using whole-cell patch-clamp methods in a brain slice preparation.A dual component EPSC was mediated by AMPA and NMDA receptors. The AMPA receptor-mediated EPSC decayed with a time constant of 1.99 ± 0.16 ms (n = 8).Following blockade of glutamate receptors, a monosynaptic strychnine-sensitive response was evoked on stimulation of the MNTB, indicative of a glycine receptor-mediated IPSC. GABAA receptors contributed to IPSCs in rats under 6 days old (bicuculline blocked 30% of the IPSC). In older rats little or no bicuculline-sensitive component was detectable, except in the presence of flunitrazepam. These glycinergic IPSCs showed a reversal potential that varied with changes in [Cl]i, as predicted by the Nernst equation.The IPSC exhibited two developmentally relevant changes. (i) At around postnatal day 6, the GABAA receptor-mediated component declined, leaving a predominant glycine-mediated IPSC. The isolated glycinergic IPSC decayed with time constants of 7.8 ± 0.3 and 38.3 ± 1.7 ms, with the slower component contributing 7.8 ± 0.6% of the peak amplitude (n = 121, 3–11 days old, −70 mV, 25°C). (ii) After day 11 the IPSC fast decay accelerated to 3.9 ± 0.3 ms (n = 12) and the magnitude of the slow component declined to less than 1%.Spontaneous miniature glycinergic IPSCs (mIPSCs) were variable in amplitude and were of large conductance (1.83 ± 0.19 nS, n = 8). The amplitude was unchanged on lowering [Ca2+]o.The time course of evoked and spontaneous miniature glycinergic IPSCs were compared. The 10-90% rise times were 0.7 and 0.6 ms, respectively. The evoked IPSC decayed with a fast time constant of 7.2 ± 0.7 ms, while the mIPSC decayed with a fast time constant of 5.3 ± 0.4 ms in the same seven cells.The glycinergic IPSC decay was voltage dependent with an e-fold change over 118 mV. The temperature dependence of the IPSC decay indicated a Q10 value of 2.1. Picrotoxin and cyanotriphenylborate had little or no effect on IPSCs from 6- to 14-day-old animals, implying homomeric channels are rare.We conclude that the MSO receives excitatory inputs mediated by AMPA and NMDA receptors and a strong glycinergic IPSC which has a significant contribution from GABAA receptors in neonatal rats. Functionally, the IPSC could increase membrane conductance during the decay of binaural glutamatergic EPSCs, thus refining coincidence detection and interaural timing differences.
机译:class =“ enumerated” style =“ list-style-type:decimal”> <!-list-behavior =枚举前缀-word = mark-type = decimal max-label-size = 0-> 内侧上橄榄(MSO)是双耳听觉通路的一部分,从耳蜗核和梯形体同侧内侧核(MNTB)接受抑制性输入。我们采用全细胞膜片钳方法在脑切片制备中表征了3至14日龄大鼠MSO神经元的兴奋性和抑制性突触电流。 AMPA介导双组分EPSC。和NMDA受体。 AMPA受体介导的EPSC的时间常数为1.99±0.16 ms(n = 8)。 在谷氨酸受体被阻滞后,在刺激MNTB时会引起单突触对士的宁敏感。甘氨酸受体介导的IPSC。 GABAA受体对6天龄以下的大鼠的IPSC有贡献(比库林阻断了30%的IPSC)。在老年大鼠中,除了存在氟尼西epa以外,几乎没有或没有检测到双微碱敏感成分。如能斯特方程所预测,这些甘氨酸能的IPSCs的逆转潜力随[Cl -] i的变化而变化。 IPSC表现出两个与发育相关的变化。 (i)在出生后第6天左右,GABAA受体介导的成分下降,剩下主要的甘氨酸介导的IPSC。分离出的甘氨酸能性IPSC的时间常数为7.8±0.3和38.3±1.7 ms,其中较慢的成分贡献了峰值幅度的7.8±0.6%(n = 121,3–11天,-70 mV,25°C) 。 (ii)第11天后,IPSC的快速衰减加速至3.9±0.3 ms(n = 12),而慢速分量的幅度下降至不足1%。 自发的微型甘氨酸IPSC(mIPSC)为振幅可变,并且具有较大的电导率(1.83±0.19 nS,n = 8)。降低[Ca 2 + ] o时幅度没有变化。 比较了诱发的和自发的微型甘氨酸IPSCs的时间过程。 10-90%的上升时间分别为0.7和0.6 ms。诱发的IPSC在相同的七个细胞中以7.2±0.7 ms的快速时间常数衰减,而mIPSC在相同的七个细胞中以5.3±0.4 ms的快速时间常数衰减。 甘氨酸IPSC的衰减与电压有关。超过118 mV的e倍变化。 IPSC衰减的温度依赖性表明Q10值为2.1。微毒素和氰基三苯硼酸酯对6至14日龄动物的IPSC几乎没有影响,甚至没有影响,这表明同质通道很少。 我们得出的结论是,MSO接受AMPA和NMDA受体介导的兴奋性输入,并且在新生大鼠中,GABAA受体具有重要作用,具有强力的甘氨酸IPSC。在功能上,IPSC可以在双耳谷氨酸能EPSC衰变过程中增加膜电导,从而改善巧合检测和耳间时序差异。

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