Use of chick neural tube for optimizing the PSM and epithelial somites electroporation parameters: A detailed protocol

Authors

  • Muhammad Abu-Elmagd Center of Excellence in Genomic Medicine Research, King Abdulaziz University

DOI:

https://doi.org/10.14440/jbm.2018.253

Keywords:

electroporation, neural tube, presegmented mesoderm, somites

Abstract

Somite myogenesis is one of the crucial early embryonic events that lead to the formation of muscular tissue. A complex of dynamic gene regulatory networks masters this event. To understand and analyze these networks, there remains a genuine need for the use of a reproducible and highly efficient gene transfer technique. In vivo electroporation has proven to be amongst the best approaches in achieving a high level of gene transfer. However, unoptimized electroporation conditions can directly cause varying degrees of cellular damage which may induce abnormal embryonic development as well as changes in the endogenous gene expression. Presegmented mesoderm and epithelial somites are not easy to electroporate. Chick neural tube has served in many functional studies as an ideal experimental model organ which is both robust and easily manipulated. In the current detailed protocol, the neural tube was used as a tool to optimize the electroporation conditions which were subsequently applied in the electroporation of the presegmented mesoderm and epithelial somites. The protocol highlights important notes and hints that enable reproducible results and could be applied in the in vivo electroporation of other chick embryo tissues.

Author Biography

Muhammad Abu-Elmagd, Center of Excellence in Genomic Medicine Research, King Abdulaziz University

I am an Associate Professor at the Centre of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, Saudi Arabia. I am also the Head of the Functional and Developmental Genomics Research Unit at CEGMR. My research focuses mainly on studying gene expression and regulation during early embryonic development aiming at understanding the molecular mechanisms controlling the early events of development. In addition, I analyse the regulatory network of genes that have a dual role in embryonic development and human genetic diseases especially cancer. In addition to my research, I have carried out a number of editorial roles among which I am an editor to the npj Genomic Medicine, a journal I have established in partnership with Nature Publishing Group (NPG).

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Published

2018-06-07

How to Cite

1.
Abu-Elmagd M. Use of chick neural tube for optimizing the PSM and epithelial somites electroporation parameters: A detailed protocol. J Biol Methods [Internet]. 2018Jun.7 [cited 2022Aug.11];5(2):e93. Available from: https://jbmethods.org/jbm/article/view/253

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Section

Protocols