Electroablation: a method for neurectomy and localized tissue injury.
Electroablation: a method for neurectomy and localized tissue injury.
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DOI：
10.1186/1471-213x-14-7
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发表时间：
2014-02-16
影响因子：
--
通讯作者：
Allende ML
中科院分区：
文献类型：
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作者：
Moya-Díaz J;Peña OA;Sánchez M;Ureta DA;Reynaert NG;Anguita-Salinas C;Marín G;Allende ML
组织损伤已被用于研究不同的生物过程，如再生和炎症。除了基于物理或外科的组织损伤方法外，目前用于局部组织损伤的方法包括激光和双光子损伤，这两种方法具有很高的准确性，但昂贵且难以应用。相比之下，电损伤是一种简单且廉价的技术，它允许在各种情况下对细胞或组织进行可重复性和局部性的损伤。我们描述了一种新的技术，它结合了斑马鱼在活体细胞可视化方面的优势和电损伤方法的优势，以一种简单而通用的方案，允许研究再生和炎症。电脉冲的来源是一个微电极，它可以精确地放置在表达荧光蛋白的特定细胞附近。我们通过破坏不同的细胞类型和结构，展示了这项技术在斑马鱼幼体中的应用。神经切除术可以在周围神经或脊髓中进行，从而可以研究神经纤维的退化和再生。我们还将这种方法应用于单侧线机械感觉神经肥大的消融，显示了这种方法作为研究器官再生的工具的有效性。此外，我们还表明，电损伤可以诱导免疫细胞向受损组织募集，从而可以在体内研究局限性和局部性损伤中炎症过程中的白细胞动力学。最后，我们证明了将电消融作为成年鱼组织损伤和炎症诱导的一种方法是可能的。使用精细微电极的电损伤可以用于神经元的轴突切断，作为一种通用的组织消融工具和诱导强大的炎症反应的方法。我们证明了它在幼体和成年斑马鱼研究中的有效性，但我们预计这项技术也可以很容易地应用于其他生物。我们把这种以电为基础的组织消融方法称为电消融。
Tissue injury has been employed to study diverse biological processes such as regeneration and inflammation. In addition to physical or surgical based methods for tissue injury, current protocols for localized tissue damage include laser and two-photon wounding, which allow a high degree of accuracy, but are expensive and difficult to apply. In contrast, electrical injury is a simple and inexpensive technique, which allows reproducible and localized cell or tissue damage in a variety of contexts. We describe a novel technique that combines the advantages of zebrafish for in vivo visualization of cells with those of electrical injury methods in a simple and versatile protocol which allows the study of regeneration and inflammation. The source of the electrical pulse is a microelectrode that can be placed with precision adjacent to specific cells expressing fluorescent proteins. We demonstrate the use of this technique in zebrafish larvae by damaging different cell types and structures. Neurectomy can be carried out in peripheral nerves or in the spinal cord allowing the study of degeneration and regeneration of nerve fibers. We also apply this method for the ablation of single lateral line mechanosensory neuromasts, showing the utility of this approach as a tool for the study of organ regeneration. In addition, we show that electrical injury induces immune cell recruitment to damaged tissues, allowing in vivo studies of leukocyte dynamics during inflammation within a confined and localized injury. Finally, we show that it is possible to apply electroablation as a method of tissue injury and inflammation induction in adult fish. Electrical injury using a fine microelectrode can be used for axotomy of neurons, as a general tissue ablation tool and as a method to induce a powerful inflammatory response. We demonstrate its utility to studies in both larvae and in adult zebrafish but we expect that this technique can be readily applied to other organisms as well. We have called this method of electrical based tissue ablation, electroablation.