Electroablation: A method for neurectomy and localized tissue injury

José Moya-Díaz, Oscar A. Peña, Mario Sánchez, Daniela A. Ureta, Nicole G. Reynaert, Consuelo Anguita-Salinas, Gonzalo Marín, Miguel L. Allende*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

12 Citations (Scopus)

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.

Original languageEnglish
Article number7
JournalBMC Developmental Biology
Volume14
Issue number1
DOIs
Publication statusPublished - 16 Feb 2014

Bibliographical note

Funding Information:
We thank Joao Botelho for helpful discussions, Catalina Lafourcade and Víctor Guzmán for expert fish care and Florencio Espinoza for technical help. Zebrafish strains were kindly provided by Alex Nechiporuk, David Raible, Christine Dambly-Chaudière, Darren Gilmour, Phillip Crosier, Stephen Renshaw, Graham Lieschke, Brant Weinstein and Vladimir Korzh. This work was supported by grants to MA from FONDAP (15090007), FONDECYT (1110275) and ICGEB (CRP/CHI11-01) and to GM from FONDECYT (1110281).

Keywords

  • Axotomy
  • Inflammation
  • Neurectomy
  • Regeneration
  • Tissue ablation
  • Zebrafish

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