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原代培养中新生大鼠海马锥体神经元的电生理特性

Electrophysiological properties of identified postnatal rat hippocampal pyramidal neurons in primary culture.

作者信息

Yang J, Thio L L, Clifford D B, Zorumski C F

机构信息

Department of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110.

出版信息

Brain Res Dev Brain Res. 1993 Jan 15;71(1):19-26. doi: 10.1016/0165-3806(93)90100-o.

Abstract

Electrophysiological experiments were performed on primary cell cultures of retrogradely labelled postnatal rat hippocampal neurons. Rhodamine microspheres injected into the dorsal fornix clearly labelled the pyramidal cell layer of the hippocampus while excluding the other hippocampal layers and the dentate gyrus. In dissociated cell culture, the labelled cells were easily identified by fluorescence microscopy. Anti-neuron-specific enolase and anti-glial fibrillary acidic protein antibody staining confirmed that the labelled cells were neurons. The input resistance decreased from 2 G omega to 450 M omega, the input capacitance increased from 25 pF to 75 pF, the percentage of cells showing repetitive action potentials increased from 6% to 30%, and both peak GABA and glutamate responses increased over 100% during the 0 to 10 days time period investigated. This increase in chemosensitivity can be accounted for by an increase in cell size without an increase in the specific amino acid gated channel density. The subset of hippocampal neurons identified by the retrograde tracer technique are similar to non-labelled neurons with respect to the electrophysiological and pharmacological variables investigated. Nevertheless, it is likely that identified neurons may possess unique properties not evident in this study and refinement of the dissociated cell culture system using identified neuronal subpopulations may facilitate investigations looking at neuronal interactions such as synapse formation.

摘要

对逆行标记的新生大鼠海马神经元的原代细胞培养物进行了电生理实验。将罗丹明微球注入背侧穹窿,清晰地标记了海马的锥体细胞层,同时排除了其他海马层和齿状回。在解离细胞培养中,通过荧光显微镜很容易识别出标记的细胞。抗神经元特异性烯醇化酶和抗胶质纤维酸性蛋白抗体染色证实标记的细胞是神经元。在0至10天的研究时间段内,输入电阻从2 GΩ降至450 MΩ,输入电容从25 pF增加到75 pF,显示重复动作电位的细胞百分比从6%增加到30%,GABA和谷氨酸的峰值反应均增加超过100%。化学敏感性的这种增加可以通过细胞大小的增加来解释,而特定氨基酸门控通道密度没有增加。就所研究的电生理和药理学变量而言,通过逆行示踪技术鉴定的海马神经元亚群与未标记的神经元相似。然而,已鉴定的神经元可能具有本研究中未明显体现的独特特性,使用已鉴定的神经元亚群改进解离细胞培养系统可能有助于研究神经元相互作用,如突触形成。

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