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脑源性神经营养因子增加抑制性突触,这在从大鼠视觉皮层培养的单个神经元中得到揭示。

Brain-derived neurotrophic factor increases inhibitory synapses, revealed in solitary neurons cultured from rat visual cortex.

作者信息

Palizvan M R, Sohya K, Kohara K, Maruyama A, Yasuda H, Kimura F, Tsumoto T

机构信息

Division of Neurophysiology, Osaka University Graduate School of Medicine (D-14), Suita, Japan.

出版信息

Neuroscience. 2004;126(4):955-66. doi: 10.1016/j.neuroscience.2004.03.053.

Abstract

To elucidate chronic actions of brain-derived neurotrophic factor (BDNF) on GABAergic synapses, we examined effects of a long-term application of BDNF for 10-15 days on autapses (synapses) of solitary GABAergic neurons cultured from rat visual cortex. Solitary neuron preparations were used to exclude a possible contamination of BDNF actions on excitatory neurons in dissociated neuron culture or slice preparations. Neurons were confirmed to be GABAergic pharmacologically with bicuculline, a selective antagonist for GABAA receptors and immunocytochemically with antibody against glutamic acid decarboxylase 65, a GABA synthesizing enzyme. To evaluate GABAergic synaptic function, evoked and/or miniature inhibitory postsynaptic currents (IPSCs) were recorded in the whole-cell voltage-clamp mode. The treatment with BDNF at a concentration of 100 ng/ml enhanced the amplitude of evoked IPSCs and the frequency of miniature IPSCs. In contrast, BDNF did not have a detectable effect on the amplitude of miniature IPSCs and the paired pulse ratio of IPSCs evoked by two, successive activations. To evaluate morphological changes, neurons were immunocytochemically stained with antibodies against microtubule-associated protein 2, to visualize somatodendritic region and synapsin I, to visualize presynaptic sites. The quantitative analysis indicated that BDNF increased the area of soma, the numbers of primary dendrites and dendritic branching points, the total length of dendrites and the number of synaptic sites. Such an action of BDNF was seen in both subgroups of GABAergic neurons, parvalbumin-positive and -negative neurons. To visualize functionally active presynaptic sites, neurons were stained with a styryl dye, FM1-43. BDNF increased the number of stained sites that was correlated with the frequency of miniature IPSCs. These results suggest that the chronic treatment with BDNF promotes dendritic and synaptic development of GABAergic neurons in visual cortex.

摘要

为了阐明脑源性神经营养因子(BDNF)对γ-氨基丁酸能(GABAergic)突触的慢性作用,我们研究了长期应用BDNF 10 - 15天对从大鼠视觉皮层培养的单个GABAergic神经元的自突触(突触)的影响。使用单个神经元制剂以排除BDNF对解离神经元培养物或脑片制剂中兴奋性神经元作用的可能污染。通过使用荷包牡丹碱(一种GABAA受体的选择性拮抗剂)进行药理学确认神经元为GABAergic,并使用针对谷氨酸脱羧酶65(一种GABA合成酶)的抗体进行免疫细胞化学确认。为了评估GABAergic突触功能,在全细胞电压钳模式下记录诱发的和/或微小抑制性突触后电流(IPSCs)。用浓度为100 ng/ml的BDNF处理可增强诱发IPSCs的幅度和微小IPSCs的频率。相比之下,BDNF对微小IPSCs的幅度和两次连续激活诱发的IPSCs的配对脉冲比率没有可检测到的影响。为了评估形态学变化,用针对微管相关蛋白2的抗体对神经元进行免疫细胞化学染色,以可视化胞体树突区域,并用突触素I进行染色,以可视化突触前位点。定量分析表明,BDNF增加了胞体面积、初级树突数量和树突分支点数量、树突总长度以及突触位点数量。BDNF的这种作用在GABAergic神经元的两个亚组中均可见,即小白蛋白阳性和阴性神经元。为了可视化功能活跃的突触前位点,用苯乙烯基染料FM1 - 43对神经元进行染色。BDNF增加了染色位点的数量,这与微小IPSCs的频率相关。这些结果表明,BDNF的慢性处理促进了视觉皮层中GABAergic神经元的树突和突触发育。

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