Effect of soft X-ray irradiation on the migratory ability of primordial germ cells in chickens.
1. The present study was conducted to elucidate the effect of soft X-ray irradiation on the migratory ability of primordial germ cells (PGCs) to the germinal ridges of chicken embryos. 2. PGCs (Barred Plymouth Rock, BPR) were isolated from embryonic blood and irradiated with soft X-rays for 1-10 min. Then, the PGCs were transfected in vitro with GFP gene by lipofection. The manipulated PGCs were transferred to recipient embryos (White Leghorn, WL) and migration to the germinal ridges was analysed by examining GFP gene expression in the gonads of recipient embryos under UV light at x40 magnifications. The expression of GFP gene was detected in all the gonads of recipient embryos examined up to 10.5 d of culture. 3. Migration of PGCs irradiated with soft X-rays to the germinal ridges was also confirmed by detecting a single nucleotide polymorphism in the D-loop region of the mitochondrial DNA of BPR and WL chickens. Freshly collected PGCs (BPR) were transferred to the bloodstream of recipient embryos (WL). The fate of the transferred donor PGCs was traced by detecting the single nucleotide polymorphism in the D-loop region of the mitochondrial DNA in BPR and WL used in this study. Transferred donor PGC-derived cells were detected in all the gonads of 17-d cultured embryos by PCR. 4. The results suggest that PGCs irradiated with soft X-rays still retain the ability to migrate to the germinal ridges of recipient embryos.
Naito
M
M
Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Tsukuba, Japan. mnaito@affrc.go.jp
Harumi
T
T
Minematsu
T
T
Tajima
A
A
Kuwana
T
T
D000818:Animals; D002465:Cell Movement / Q000528:radiation effects*; D002642:Chick Embryo / Q000166:cytology* / Q000502:physiology / Q000528:radiation effects; D004272:DNA, Mitochondrial / Q000032:analysis; D005260:Female; D005854:Germ Cells / Q000502:physiology / Q000528:radiation effects*; D006066:Gonads / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000032:analysis; D008297:Male; D011110:Polymorphism, Genetic; D014162:Transfection
D004272:DNA, Mitochondrial; D049452:Green Fluorescent Proteins