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Open Access Highly Accessed Methodology article

Electroablation: a method for neurectomy and localized tissue injury

José Moya-Díaz1, Oscar A Peña1, Mario Sánchez1, Daniela A Ureta1, Nicole G Reynaert1, Consuelo Anguita-Salinas12, Gonzalo Marín34 and Miguel L Allende1*

Author Affiliations

1 FONDAP Center for Genome Regulation, Facultad de Ciencias, Universidad de Chile, Casilla 653, Santiago, Chile

2 Departamento de Ciencias Biológicas, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Santiago, Chile

3 Laboratorio de Neurobiología y Biología del Conocer, Facultad de Ciencias, Universidad de Chile, Santiago, Chile

4 Facultad de Medicina, Universidad Finis Terrae, Santiago, Chile

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BMC Developmental Biology 2014, 14:7  doi:10.1186/1471-213X-14-7

Published: 16 February 2014

Abstract

Background

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.

Results

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.

Conclusions

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.

Keywords:
Axotomy; Neurectomy; Tissue ablation; Regeneration; Inflammation; Zebrafish