Green fluorescent protein labeling of primordial germ cells using a nontransgenic method and its application for germ cell transplantation in salmonidae

Goro Yoshizaki, Yasuko Tago, Yutaka Takeuchi, Etsuko Sawatari, Terumasa Kobayashi, Toshio Takeuchi

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Transplanting primordial germ cells (PGCs) has a number of potential applications in fish bioengineering. Previously, we established a system to visualize live PGCs in the rainbow trout by introducing the green fluorescent protein (Gfp) gene driven by rainbow trout vasa gene regulatory regions. However, for PGC transplantation to be practically useful in aquaculture, visualization of PGCs using a nontransgenic technique is required. In this study, we demonstrate a method for labeling PGCs from various fish species by introducing chimeric RNAs composed of the Gfp coding region and vasa gene 3′-untranslated regions (UTRs); these sequences play a critical role in stabilizing mRNA in zebrafish PGCs. The GFP chimeric RNAs, including vasa 3′-UTR RNAs from rainbow trout, Nibe croaker, and zebrafish, were microinjected into the cytoplasm of fertilized eggs of several Salmonidae species. All the resulting embryos showed specific labeling in PGCs after the somatogenesis stage, which continued to be visible for at least 50 days. To apply this technique to PGC transplantation, PGCs labeled with chimeric RNA were microinjected into the peritoneal cavity of newly hatched salmonid embryos. The GFP labeling was sufficiently long-lived for the initial stage of donor PGC behavior to be followed in the recipient embryos. Importantly, donor PGCs from brown trout and masu salmon were incorporated into xenogeneic genital ridges in recipient rainbow trout. This nontransgenic method for labeling fish PGCs should be extremely useful for applications of PGC transplantation where the resulting progeny are to be released into the environment, such as PGC cryopreservation for fish stocks and surrogate brood stock technology.

Original languageEnglish
Pages (from-to)88-93
Number of pages6
JournalBiology of Reproduction
Volume73
Issue number1
DOIs
Publication statusPublished - Jul 1 2005

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Salmonidae
Cell Transplantation
Green Fluorescent Proteins
Germ Cells
Oncorhynchus mykiss
Fishes
Embryonic Structures
3' Untranslated Regions
Zebrafish
RNA
Genes
Introduced Species
Bioengineering
Aquaculture
Untranslated RNA
Trout
Zygote
Salmon
Nucleic Acid Regulatory Sequences
Cryopreservation

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Green fluorescent protein labeling of primordial germ cells using a nontransgenic method and its application for germ cell transplantation in salmonidae. / Yoshizaki, Goro; Tago, Yasuko; Takeuchi, Yutaka; Sawatari, Etsuko; Kobayashi, Terumasa; Takeuchi, Toshio.

In: Biology of Reproduction, Vol. 73, No. 1, 01.07.2005, p. 88-93.

Research output: Contribution to journalArticle

Yoshizaki, Goro ; Tago, Yasuko ; Takeuchi, Yutaka ; Sawatari, Etsuko ; Kobayashi, Terumasa ; Takeuchi, Toshio. / Green fluorescent protein labeling of primordial germ cells using a nontransgenic method and its application for germ cell transplantation in salmonidae. In: Biology of Reproduction. 2005 ; Vol. 73, No. 1. pp. 88-93.
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