Identification and characterization of primordial germ cells in a vocal learning Neoaves species, the zebra finch
Abstract
The zebra finch has been used as a valuable vocal learning animal model for human spoken language. It is representative of vocal learning songbirds specifically, which comprise half of all bird species, and of Neoaves broadly, which comprise 95% of all bird species. Although transgenesis in the zebra finch has been accomplished, it is with a very low efficiency of germ-line transmission and far from the efficiency with a more genetically tractable but vocal nonlearning species, the chicken (a Galloanseriformes). To improve germ-line transmission in the zebra finch, we identified and characterized its primordial germ cells (PGCs) and compared them with chicken. We found striking differences between the 2 species, including that zebra finch PGCs were more numerous, more widely distributed in early embryos before colonization into the gonads, had slower timing of colonization, and had a different developmental gene-expression program. We improved conditions for isolating and culturing zebra finch PGCs in vitro and were able to transfect them with gene-expression vectors and incorporate them into the gonads of host embryos. Our findings demonstrate important differences in the PGCs of the zebra finch and advance the first stage of creating PGC-mediated germ-line transgenics of a vocal learning species.
Additional Information
© 2019 FASEB. Received for publication March 25, 2019; Accepted for publication September 10, 2019; Published Online: 11 Oct 2019. The authors thank the members of the J.Y.H. and E.D.J. laboratories for suggestions and inspirational discussions. The authors also thank Carlos Lois (California Institute of Technology) for useful discussions on the experiments and feedback on the manuscript. This work was supported by the Creative Research Initiatives Program (Center for Avian Germ Cell Modulation and Cloning) from a National Research Foundation of Korea (NRF) grant, funded by the Korean government [Ministry of Science and Information and Communications Technology (MSIP) 2015R1A3A2033826 to J.Y.H.], and the National Science Foundation (NSF) Enabling Discovery through Genomic Tools (EDGE; Award 1645199) and Howard Hughes Medical Insitute (HHMI) funds to E.D.J., Carlos Lois, C.V.M., and T.A.F.V. The authors declare no conflicts of interest. Author Contributions: E. D. Jarvis and J. Y. Han participated in study design and coordination; K. M. Jung participated in the design of the study, carried out the experiments, and wrote the first draft of the manuscript; Y. M. Kim participated in the design of the study and was involved in data interpretation and writing final versions of the manuscript; A. L. Keyte, O. Fedrigo, and B. Haase were involved in data analysis and writing final versions of te manuscript; M. T. Biegler, D. Rengaraj, H. J. Lee, C. V. Mello, and T. A. F. Velho were involved in data interpretation and writing final versions of the manuscript; and all authors have read and approved the final manuscript.Attached Files
Supplemental Material - fj.201900760rr.sf1.docx
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Supplemental Material - fj.201900760rr.st1.docx
Supplemental Material - fj.201900760rr.st2.docx
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Additional details
- Eprint ID
- 99389
- DOI
- 10.1096/fj.201900760rr
- Resolver ID
- CaltechAUTHORS:20191022-110323726
- 2015R1A3A2033826
- National Research Foundation of Korea
- Ministry of Science and Information and Communications Technology (Korea)
- IOS-1645199
- NSF
- Howard Hughes Medical Institute (HHMI)
- Created
-
2019-10-22Created from EPrint's datestamp field
- Updated
-
2021-11-16Created from EPrint's last_modified field