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通过早期胚盘细胞移植从棕色和白色来航鸡获得的体细胞和生殖系嵌合体鸡。

Somatic and germline chicken chimeras obtained from brown and white Leghorns by transfer of early blastodermal cells.

作者信息

Thoraval P, Lasserre F, Coudert F, Dambrine G

机构信息

Laboratoire de Virologie et d'Oncologie Aviaire, Institut National de la Recherche Agronomique, Nouzilly, France.

出版信息

Poult Sci. 1994 Dec;73(12):1897-905. doi: 10.3382/ps.0731897.

Abstract

Stage X blastodermal cells were isolated from freshly laid unincubated Brown Leghorn chicken eggs. Five hundred cells from Stage X Brown Leghorn embryos were injected into the subgerminal cavity of White Leghorn unincubated embryos exposed to 550 rad of gamma irradiation from a cesium-137 source. Of 712 White Leghorn embryos that were irradiated and injected with Brown Leghorn blastodermal cells, 52 (7.3%) survived to hatching. Somatic chimerism was examined in the melanocyte population and erythroid lineage. The presence of brown feathers indicating donor cell contribution to melanocyte pigmentation was observed in 23 (44%) out of the 52 hatched chicks. Analysis of blood DNA was performed using a probe that revealed an endogenous retroviral gag fragment specific for the donor genome. Three out of these 23 chimeric chickens exhibited the gag-specific fragment. To test germline chimerism, chickens that reached sexual maturity were mated with Brown Leghorns. Three somatically chimeric hens produced Brown Leghorn progeny at a rate of 30.7, 9.2, and 2.9% respectively, thus proving donor cell contribution to the germline differentiation. Chimeric chickens obtained after injection of nonirradiated embryos exhibited a lower extent of chimerism at the feather level and did not show any chimerism in the erythroid lineage and the germline, thus demonstrating the value of the use of compromised recipient embryos to produce chimeras in chickens. Nevertheless, the extent of somatic chimerism could not be used to predict the germline chimerism.

摘要

从刚产下的未孵化的棕色来航鸡鸡蛋中分离出X期囊胚细胞。将来自X期棕色来航鸡胚胎的500个细胞注射到经铯-137源550拉德γ射线照射的未孵化的白色来航鸡胚胎的胚下腔中。在712个接受照射并注射了棕色来航鸡囊胚细胞的白色来航鸡胚胎中,有52个(7.3%)存活至孵化。在黑素细胞群体和红细胞系中检测了体细胞嵌合现象。在52只孵化出的小鸡中,有23只(44%)观察到有棕色羽毛,这表明供体细胞对黑素细胞色素沉着有贡献。使用揭示供体基因组特有的内源性逆转录病毒gag片段的探针进行血液DNA分析。这23只嵌合鸡中有3只表现出gag特异性片段。为了测试种系嵌合现象,将达到性成熟的鸡与棕色来航鸡交配。3只体细胞嵌合母鸡分别以30.7%、9.2%和2.9%的比例产生了棕色来航鸡后代,从而证明了供体细胞对种系分化有贡献。注射未照射胚胎后获得的嵌合鸡在羽毛水平上的嵌合程度较低,在红细胞系和种系中未表现出任何嵌合现象,从而证明了使用受损受体胚胎在鸡中产生嵌合体的价值。然而,体细胞嵌合的程度不能用于预测种系嵌合。

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