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


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



electroporation, neural tube, presegmented mesoderm, somites


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).


Buckingham M. Skeletal muscle formation in vertebrates. Curr Opin Genet Dev. 2001;11(4):440-8. Epub 2001/07/13. PubMed PMID: 11448631.

Christ B, Huang, R., and Scaal, M. Formation and differentiation of the avian sclerotome. Anat Embryol (Berl). 2004;208:333-50.

Bryson-Richardson RJ, Currie PD. The genetics of vertebrate myogenesis. Nature reviews Genetics. 2008;9(8):632-46. Epub 2008/07/19. doi: 10.1038/nrg2369. PubMed PMID: 18636072.

Molkentin JD, Olson EN. Defining the regulatory networks for muscle development. Curr Opin Genet Dev. 1996;6(4):445-53. Epub 1996/08/01. PubMed PMID: 8791524.

Andermatt I, Wilson N, Stoeckli ET. In ovo electroporation of miRNA-based-plasmids to investigate gene function in the developing neural tube. Methods in molecular biology (Clifton, NJ). 2014;1101:353-68. Epub 2013/11/16. doi: 10.1007/978-1-62703-721-1_17. PubMed PMID: 24233790.

Hu X, Wang Z, Wu H, Jiang W, Hu R. Ras ssDNA aptamer inhibits vascular smooth muscle cell proliferation and migration through MAPK and PI3K pathways. Int J Mol Med. 2015;35(5):1355-61. Epub 2015/03/18. doi: 10.3892/ijmm.2015.2139. PubMed PMID: 25778421.

Murai H, Tadokoro R, Sakai K, Takahashi Y. In ovo gene manipulation of melanocytes and their adjacent keratinocytes during skin pigmentation of chicken embryos. Dev Growth Differ. 2015;57(3):232-41. Epub 2015/03/06. doi: 10.1111/dgd.12201. PubMed PMID: 25739909.

Lopez-Sanchez C, Franco D, Bonet F, Garcia-Lopez V, Aranega A, Garcia-Martinez V. Negative Fgf8-Bmp2 feed-back is regulated by miR-130 during early cardiac specification. Dev Biol. 2015;406(1):63-73. Epub 2015/07/15. doi: 10.1016/j.ydbio.2015.07.007. PubMed PMID: 26165600.

Rao M, Baraban JH, Rajaii F, Sockanathan S. In vivo comparative study of RNAi methodologies by in ovo electroporation in the chick embryo. Dev Dyn. 2004;231(3):592-600. Epub 2004/09/18. doi: 10.1002/dvdy.20161. PubMed PMID: 15376322.

Norris A, Streit A. Morpholinos: studying gene function in the chick. Methods. 2014;66(3):454-65. Epub 2013/11/05. doi: 10.1016/j.ymeth.2013.10.009. PubMed PMID: 24184187; PubMed Central PMCID: PMCPMC3989378.

Gandhi S, Piacentino ML, Vieceli FM, Bronner ME. Optimization of CRISPR/Cas9 genome editing for loss-of-function in the early chick embryo. Dev Biol. 2017;432(1):86-97. Epub 2017/11/19. doi: 10.1016/j.ydbio.2017.08.036. PubMed PMID: 29150011; PubMed Central PMCID: PMCPMC5728388.

Shirazi Fard S, Blixt M, Hallbook F. Whole Retinal Explants from Chicken Embryos for Electroporation and Chemical Reagent Treatments. J Vis Exp. 2015;(103). Epub 2015/10/21. doi: 10.3791/53202. PubMed PMID: 26485513; PubMed Central PMCID: PMCPMC4692626.

Goljanek-Whysall K, Sweetman D, Abu-Elmagd M, Chapnik E, Dalmay T, Hornstein E, et al. MicroRNA regulation of the paired-box transcription factor Pax3 confers robustness to developmental timing of myogenesis. Proceedings of the National Academy of Sciences of the United States of America. 2011;108(29):11936-41. Epub 2011/07/07. doi: 10.1073/pnas.1105362108. PubMed PMID: 21730146; PubMed Central PMCID: PMCPmc3141954.

Wang H, Bonnet A, Delfini MC, Kawakami K, Takahashi Y, Duprez D. Stable, conditional, and muscle-fiber-specific expression of electroporated transgenes in chick limb muscle cells. Dev Dyn. 2011;240(5):1223-32. Epub 2011/04/22. doi: 10.1002/dvdy.22498. PubMed PMID: 21509896.

Croteau LP, Kania A. Optimisation of in ovo electroporation of the chick neural tube. Journal of neuroscience methods. 2011;201(2):381-4. Epub 2011/08/30. doi: 10.1016/j.jneumeth.2011.08.012. PubMed PMID: 21871488.

GEISHA. Whole mount In situ hybridization protocol for mRNA detection. 2011, p 1-6

Hamburger V, Hamilton, HL. A series of normal stages in the development of the chick embryo. Journal of Morphology. 1951;88(1):49-92.

Abu-Elmagd M, Ishii, Y., Cheung, M., Rex,M. Le Roudec,D. Scotting,P.J. cSox3 expression and neurogenesis in the epibranchial placodes. Developmental Biology. 2001;237(2):258–69.

Abu-Elmagd M, Robson, L., Sweetman, D., Hadley, J., Francis-West, P., Münsterberg, A. Wnt/Lef1 signaling acts via Pitx2 to regulate somite myogenesis. Developmental Biology. 2010;337:211–9.

Tripathi V, Ishii, Y., Abu-Elmagd, M., Scotting, P. The surface ectoderm of the chick embryo exhibits dynamic variation in its response to neurogenic signals. Int J Dev Biol. 2009;53:1023-33.




How to Cite

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: