Microinjection of Exogenous DNA into Eggs of Halocynthia roretzi

Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1029)


Exogenous gene expression assays during development, including reporters under the control of 5′ upstream enhancer regions of genes, constitute a powerful technique for understanding the mechanisms of tissue-specific gene expression regulation and determining the characteristics, behaviors, and functions of cells that express these genes. The simple marine chordate Halocynthia roretzi has been used for these transgenic analyses for a long time and is an excellent model system for such studies, especially in comparative analyses with other ascidians. In this study, I describe simple methods for microinjecting H. roretzi eggs with exogenous DNA, such as a promoter construct consisting of a 5′ upstream region and a reporter gene, which are prerequisites for transgenic analyses. I also describe basic knowledge regarding this ascidian species, providing reasons why it is an ideal subject for developmental biology studies.


Halocynthia roretzi Microinjection Ooplasmic segregation Micromanipulator Fertilized eggs Enhancer regions 



I would like to thank Dr. Takaharu Numakunai for providing the image in Fig. 4.1b, Dr. Hiroki Nishida for providing the data on the developmental timetable in Fig. 4.2, and Ms. Kaori Miyaoku and Mr. Tao Zheng for their help in preparing the images in Fig. 4.4a–f.


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© Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  1. 1.Asamushi Research Center for Marine Biology, Graduate School of Life SciencesTohoku UniversityAsamushiJapan

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