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难治性癫痫患儿新皮质切片中的局部突触回路和癫痫样活动。

Local synaptic circuits and epileptiform activity in slices of neocortex from children with intractable epilepsy.

作者信息

Tasker J G, Peacock W J, Dudek F E

机构信息

Mental Retardation Research Center, University of California School of Medicine, Los Angeles 90024.

出版信息

J Neurophysiol. 1992 Mar;67(3):496-507. doi: 10.1152/jn.1992.67.3.496.

Abstract
  1. Single and dual intracellular recordings were performed in neocortical slices obtained from tissue samples surgically removed from children (8 mo to 15 yr) for the treatment of intractable epilepsy. Electrical stimulation and glutamate microapplication were used to study local synaptic inputs to pyramidal cells. 2. In recordings with potassium-acetate electrodes, activation of presynaptic neocortical neurons with glutamate microdrops did not elicit a clear increase in postsynaptic potentials (PSPs) but did suppress current-evoked repetitive spike firing in recorded neurons. Bicuculline (10 microM) blocked this effect, suggesting it was caused by the activation of presynaptic gamma-aminobutyric acid (GABA) cells. In recordings with KCl electrodes, glutamate microdrops elicited an increase in the frequency and amplitude of depolarizing PSPs. Bicuculline (5-10 microM) blocked the glutamate-evoked PSPs, suggesting they were reversed GABAA-receptor-mediated inhibitory postsynaptic potentials (IPSPs). In one cell recorded with a KCl electrode (total n = 8), current-evoked spike trains elicited afterdischarges of reversed IPSPs, thus revealing a recurrent inhibitory circuit. Therefore local inhibitory synaptic circuits were robust and could be observed in tissue from patients as young as 11 mo. 3. In addition to short-latency (10-25 ms), monosynaptic excitatory postsynaptic potentials (EPSPs), electrical stimulation at low intensities sometimes elicited delayed EPSPs (20-60 ms). When GABAA-receptor-mediated synaptic inhibition was partially reduced in bicuculline (5-10 microM), electrical stimulation evoked large EPSPs at long and variable latencies (100-300 ms). Glutamate microapplication caused an increase in the frequency and amplitude of EPSPs; preliminary results suggest that glutamate microdrops were less likely to evoke EPSPs in tissue from younger patients (8-12 mo) than in slices from patients greater than 4 yr. Evidence for local excitatory synaptic circuits was thus found when synaptic inhibition was partially reduced. 4. After further reduction of inhibition in bicuculline (5-50 microM), electrical stimulation elicited epileptiform bursts. In pairs of simultaneously recorded neurons, bursts were generated synchronously from long-latency EPSPs (100-300 ms) in slices from patients as young as 8 mo. Reflected EPSPs at very long and variable latencies (500-1,100 ms) and repetitive epileptiform bursts could be evoked synchronously in pairs of cells. Glutamate activation of local presynaptic neurons elicited robust epileptiform events in recorded cells. This was seen in slices from patients as young as 16 mo. 5. These data provide physiological evidence for the presence of local inhibitory and excitatory synaptic circuits in human neocortex at least as early as 11 and 8 mo, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 对从8个月至15岁儿童身上手术切除的用于治疗顽固性癫痫的组织样本所获取的新皮质切片进行单细胞和双细胞细胞内记录。采用电刺激和谷氨酸微量应用来研究对锥体细胞的局部突触输入。2. 在使用醋酸钾电极记录时,用谷氨酸微滴激活突触前新皮质神经元并未引起突触后电位(PSP)明显增加,但确实抑制了记录神经元中电流诱发的重复放电。荷包牡丹碱(10微摩尔)可阻断这种效应,表明其由突触前γ-氨基丁酸(GABA)细胞的激活所致。在使用氯化钾电极记录时,谷氨酸微滴引起去极化PSP的频率和幅度增加。荷包牡丹碱(5 - 10微摩尔)可阻断谷氨酸诱发的PSP,表明它们是反转的GABAA受体介导的抑制性突触后电位(IPSP)。在用氯化钾电极记录的一个细胞中(总共n = 8),电流诱发的动作电位序列引发了反转IPSP的后放电,从而揭示了一个反复抑制性回路。因此,局部抑制性突触回路很强大,在年仅11个月大的患者组织中就能观察到。3. 除了短潜伏期(10 - 25毫秒)的单突触兴奋性突触后电位(EPSP)外,低强度电刺激有时会诱发延迟的EPSP(20 - 60毫秒)。当在荷包牡丹碱(5 - 10微摩尔)中GABAA受体介导的突触抑制被部分减弱时,电刺激会在长且可变的潜伏期(100 - 300毫秒)诱发大的EPSP。谷氨酸微量应用导致EPSP的频率和幅度增加;初步结果表明,谷氨酸微滴在8 - 12个月大的年轻患者组织中比在4岁以上患者的切片中诱发EPSP的可能性更小。因此,当突触抑制被部分减弱时,发现了局部兴奋性突触回路的证据。4. 在荷包牡丹碱(5 - 50微摩尔)中进一步减弱抑制后,电刺激诱发癫痫样爆发。在同时记录的成对神经元中,在年仅8个月大的患者切片中,爆发从长潜伏期EPSP(100 - 300毫秒)同步产生。在成对细胞中可以同步诱发极长且可变潜伏期(500 - 1100毫秒)的反射性EPSP和重复性癫痫样爆发。局部突触前神经元的谷氨酸激活在记录细胞中诱发强烈的癫痫样事件。这在年仅16个月大的患者切片中可见。5. 这些数据分别为至少早在11个月和8个月大时人类新皮质中存在局部抑制性和兴奋性突触回路提供了生理学证据。(摘要截选至400字)

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