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软X射线照射对鸡原始生殖细胞迁移能力的影响。

Effect of soft X-ray irradiation on the migratory ability of primordial germ cells in chickens.

作者信息

Naito M, Harumi T, Minematsu T, Tajima A, Kuwana T

机构信息

Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Tsukuba, Japan.

出版信息

Br Poult Sci. 2007 Apr;48(2):121-6. doi: 10.1080/00071660701261294.

Abstract
  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.
摘要
  1. 本研究旨在阐明软X射线照射对鸡胚原始生殖细胞(PGCs)向生殖嵴迁移能力的影响。2. 从胚胎血液中分离出PGCs(横斑普利茅斯岩鸡,BPR),并用软X射线照射1 - 10分钟。然后,通过脂质体转染法将绿色荧光蛋白(GFP)基因体外转染到PGCs中。将处理后的PGCs转移到受体胚胎(白来航鸡,WL)中,并通过在40倍放大倍数的紫外光下检测受体胚胎性腺中GFP基因的表达来分析其向生殖嵴的迁移情况。在培养至10.5天的所有检测受体胚胎的性腺中均检测到了GFP基因的表达。3. 通过检测BPR和WL鸡线粒体DNA的D环区域中的单核苷酸多态性,也证实了经软X射线照射的PGCs向生殖嵴的迁移。将新鲜收集的PGCs(BPR)转移到受体胚胎(WL)的血液中。通过检测本研究中使用的BPR和WL线粒体DNA的D环区域中的单核苷酸多态性来追踪转移的供体PGCs的命运。通过聚合酶链反应(PCR)在17天培养胚胎的所有性腺中检测到了转移的供体PGC来源的细胞。4. 结果表明,经软X射线照射的PGCs仍保留向受体胚胎生殖嵴迁移的能力。

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