Production of germline chimeric chickens following the administration of a busulfan emulsion
Production of germline chimeric chickens following the administration of a busulfan emulsion
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DOI：
10.1002/mrd.20218
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发表时间：
2005-04
影响因子：
2.5
通讯作者：
Yonghong Song;S. D’Costa;S. Pardue;J. Petitte
中科院分区：
文献类型：
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作者：
Yonghong Song;S. D’Costa;S. Pardue;J. Petitte
用Busulfan（1,4 -丁二醇二甲基磺酸盐）消耗内源性生殖细胞，以提高鸡种系嵌合体的产量。利用免疫组织化学方法对27期鸡胚胎的原始宝石细胞（PGCs）进行鉴定，比较了两种分娩配方（丁硫丹悬浮液（BS）和溶解丁硫丹乳剂（SBE））内源性PGC消耗的程度。与对照组相比，两种磺胺处理均导致PGCs显著减少。然而，与BS处理相比，SBE导致了更一致和更广泛的PGCs消耗。SBE处理的胚胎用外源PGCs重新繁殖，导致27期胚胎的PGCs增加了三倍。随后，通过将来自barbarplymouth Rock胚胎的雄性性腺PGCs转移到未处理和SBE处理的White Leghorn胚胎中，产生了种系嵌合体。对推定嵌合体与成年斑茅岩鸡进行子代试验，以评价种系嵌合体的生产效率。与未治疗的受体相比，SBE治疗受体的种系嵌合频率增加了5倍。受体胚胎经SBE处理后，来自种系嵌合体的供体后代数量也增加了8倍。这些结果表明，在未孵化的鸡蛋蛋黄中注入一种丁硫丹乳剂可以改善胚胎中内源性PGCs的消耗，提高种系嵌合体的生产效率。摩尔。天线转换开关。开发。70:438-444,2005。©2005 Wiley‐Liss, Inc。
Busulfan (1,4‐butanediol dimethanesulfonate) was used to deplete endogenous germ cells for the enhanced production of chicken germline chimeras. Utilizing immunohistochemical identification of primordial gem cells (PGCs) in Stage 27 chicken embryos, two delivery formulations were compared relative to the degree of endogenous PGC depletion, a busulfan suspension (BS) and a solublized busulfan emulsion (SBE). Both busulfan treatments resulted in a significant reduction in PGCs when compared to controls. However, the SBE resulted in a more consistent and extensive depletion of PGCs than that observed with the BS treatment. Repopulation of SBE‐treated embryos with exogenous PGCs resulted in a threefold increase of PGCs in Stage 27 embryos. Subsequently, germline chimeras were produced by the transfer of male gonadal PGCs from Barred Plymouth Rock embryos into untreated and SBE‐treated White Leghorn embryos. Progeny testing of the presumptive chimeras with adult Barred Plymouth Rock chickens was performed to evaluate the efficiency of germline chimera production. The frequency of germline chimerism in SBE‐treated recipients increased fivefold when compared to untreated recipients. The number of donor‐derived offspring from the germline chimeras also increased eightfold following SBE‐treatment of the recipient embryos. These results demonstrated that the administration of a busulfan emulsion into the egg yolk of unincubated eggs improved the depletion of endogenous PGCs in the embryo and enhanced the efficiency of germline chimera production. Mol. Reprod. Dev. 70: 438–444, 2005. © 2005 Wiley‐Liss, Inc.