PUBLICATION
Visualization of primordial germ cells in the fertilized pelagic eggs of the barfin flounder Verasper moseri
- Authors
- Goto, R., Saito, T., Kawakami, Y., Kitauchi, T., Takagi, M., Todo, T., Arai, K., Yamaha, E.
- ID
- ZDB-PUB-160213-18
- Date
- 2015
- Source
- The International journal of developmental biology 59: 465-470 (Journal)
- Registered Authors
- Keywords
- teleost, PGC, flounder, pelagic egg, microinjection
- MeSH Terms
-
- Molecular Sequence Data
- Amino Acid Sequence
- In Situ Hybridization
- Zebrafish/growth & development
- Zebrafish/metabolism
- Animals
- 3' Untranslated Regions/genetics
- Germ Cells/cytology*
- Germ Cells/metabolism
- Embryonic Development/physiology*
- Sequence Homology, Amino Acid
- Flounder/embryology*
- Embryo, Nonmammalian/cytology*
- Embryo, Nonmammalian/metabolism
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- Zygote/cytology*
- Zygote/metabolism
- PubMed
- 26864487 Full text @ Int. J. Dev. Biol.
Citation
Goto, R., Saito, T., Kawakami, Y., Kitauchi, T., Takagi, M., Todo, T., Arai, K., Yamaha, E. (2015) Visualization of primordial germ cells in the fertilized pelagic eggs of the barfin flounder Verasper moseri. The International journal of developmental biology. 59:465-470.
Abstract
Primordial germ cells (PGCs) appear during early embryogenesis and differentiate into gametes through oogenesis or spermatogenesis. Teleost PGCs can be visualized by injecting RNA transcribed from the fusion product of a fluorescent protein gene attached to the 3' untranslated region (3'UTR) of zebrafish nanos3 (zf-nos3). Although this method has been widely applied to teleost PGCs, the visualization of PGCs in pelagic species that have eggs with a hard chorion is more problematic due to the technical difficulty of microinjection into their eggs. In this study, we developed a reliable method for microinjection of fertilized eggs in a pelagic species, the barfin flounder. Using a microneedle with a constriction "brake", we were able to introduce gfp-nos3 3'UTR mRNA into embryos and to determine the origin and migration route of PGCs. We also isolated the barfin flounder nos3 (bf-nos3) gene to compare its 3'UTR sequence with that of zebrafish. The 3'UTR of the bf-nos3 sequence was longer than that of zf-nos3. However, PGCs were also visualized after injection of gfp-bf-nos3 3'UTR mRNA both in zebrafish and barfin flounder. These results suggest that the function of nos3 is conserved between these species regardless of the sequence differences. The method developed here for labeling PGCs with gfp-nos3 mRNA will provide a means to study PGC development in the embryos of a wide range of marine fish species.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping