Direct Current Electric Stimulation Alters the Frequency and the Distribution of Mitotic Cells in Planarians.
Direct Current Electric Stimulation Alters the Frequency and the Distribution of Mitotic Cells in Planarians.
复制标题
DOI：
10.1089/bioe.2020.0026
复制
发表时间：
2020-09
期刊：
影响因子：
2.3
通讯作者：
D. Davidian;Benjamin Ziman;A. Escobar;Néstor J. Oviedo
中科院分区：
文献类型：
--
作者：
D. Davidian;Benjamin Ziman;A. Escobar;Néstor J. Oviedo
背景：使用直流电刺激（DCS）是治疗疾病和增强身体功能的有效策略。因此，DCS治疗是一种有吸引力的生物医学替代方案，但分子基础仍然是未知的。缺乏从分子到生物体水平剖析DCS影响的实验模型是一个重要的警告。在这里，我们介绍了作为一个听话的有机体在体内研究的DCS的Planarian扁形虫Schmidtea meditheea。我们开发了一种实验方法，便于应用直流电刺激整个涡虫体（pDCS）。材料与方法：通过麻醉、琼脂包埋和通过专用热电冷却装置的低温联合治疗实现了Planarian固定。使用拉制玻璃微电极递送用于pDCS的电流。通过恒压电源供应电势。给予pDCS长达6小时，并通过使用视频记录、免疫组织化学和基因表达分析来测量行为和分子效应。结果如下：行为固定的影响是可逆的，和pDCS导致有丝分裂细胞的重新分布沿着中外侧轴的涡虫体。pDCS效应依赖于电场的极性，这导致与pDCS时间相关的有丝分裂密度降低的增加。有丝分裂细胞的变化与干细胞标志物smedwi-1基因表达的明显重新分布一致。结论：在这项工作中提出的固定技术有利于研究，旨在解剖外源性电刺激在成人体内的影响。用DCS治疗可以施用不同的时间，并且在不同的水平上评估结果，包括动物行为、细胞和转录变化。事实上，用pDCS处理可以改变细胞和转录参数，这取决于电场的极性和暴露的持续时间。
Background: The use of direct current electric stimulation (DCS) is an effective strategy to treat disease and enhance body functionality. Thus, treatment with DCS is an attractive biomedical alternative, but the molecular underpinnings remain mostly unknown. The lack of experimental models to dissect the effects of DCS from molecular to organismal levels is an important caveat. Here, we introduce the planarian flatworm Schmidtea mediterranea as a tractable organism for in vivo studies of DCS. We developed an experimental method that facilitates the application of direct current electrical stimulation to the whole planarian body (pDCS). Materials and Methods: Planarian immobilization was achieved by combining treatment with anesthesia, agar embedding, and low temperature via a dedicated thermoelectric cooling unit. Electric currents for pDCS were delivered using pulled glass microelectrodes. The electric potential was supplied through a constant voltage power supply. pDCS was administered up to six hours, and behavioral and molecular effects were measured by using video recordings, immunohistochemistry, and gene expression analysis. Results: The behavioral immobilization effects are reversible, and pDCS resulted in a redistribution of mitotic cells along the mediolateral axis of the planarian body. The pDCS effects were dependent on the polarity of the electric field, which led to either increase in reductions in mitotic densities associated with the time of pDCS. The changes in mitotic cells were consistent with apparent redistribution in gene expression of the stem cell marker smedwi-1. Conclusion: The immobilization technique presented in this work facilitates studies aimed at dissecting the effects of exogenous electric stimulation in the adult body. Treatment with DCS can be administered for varying times, and the consequences evaluated at different levels, including animal behavior, cellular and transcriptional changes. Indeed, treatment with pDCS can alter cellular and transcriptional parameters depending on the polarity of the electric field and duration of the exposure.