Bacon L D, Zajchowski L, Clark M E, Etches R J
USDA, Agricultural Research Service, Avian Disease and Oncology Laboratory, East Lansing, Michigan 48823, USA.
Poult Sci. 2002 Oct;81(10):1427-38. doi: 10.1093/ps/81.10.1427.
Chimeric chickens were evaluated as an intermediate for development of transgenic chickens. The transfer of Barred Plymouth Rock (BR) blastodermal cells into White Leghorn (WL) embryos results in BR-->WL chimeras, and some breeder males generate over 30% germline transmission of the BR genotype to offspring based on a feather-color trait. The objectives of the current study were to 1) identify the MHC (B haplotypes) in resident BR and WL lines, 2) establish that B antigens could be detected and quantified in red blood cells (RBC) of chimeras, 3) establish if there is a correlation in chimeras between percentage of RBC with donor B antigens and percentage germline transmission, and 4) evaluate if the MHC genotype influences chimera development. The RBC agglutination data indicated three B haplotypes were present in each line. The B2-like, and B19-like genes were unique to the WL line, and B13-like and B-15-like genes were unique to the BR line, whereas a B21-like gene was present in both lines. In adult BR-->WL chimeras, as well as 10- to 14 d-old WL-->WL chimeras, donor-type B antigens were detectable and quantifiable on RBC using flow cytometry. In BR-->WL chimeras, the percentage germline transmission was significantly correlated with the percentage of RBC with donor B antigen, as well as percentage of black feathers in the plumage. In a retrospective study using previously developed BR-->WL chimeras, the level of chimerism and germline transmission was higher in B*21/*21 type recipients, but this was not statistically significant in two prospective studies. It was concluded that MHC antigens on RBC can be used for identifying, quantifying, and selecting chicken chimeras developed by the transfer of blastodermal cells.
嵌合鸡被评估为转基因鸡发育的中间体。将芦花洛克(BR)胚盘细胞转移到白来航(WL)胚胎中会产生BR→WL嵌合体,一些种用雄性基于羽毛颜色性状可将BR基因型的超过30%种系传递给后代。本研究的目的是:1)鉴定BR和WL品系中的主要组织相容性复合体(B单倍型);2)确定嵌合体红细胞(RBC)中是否能检测和定量B抗原;3)确定嵌合体中带有供体B抗原的RBC百分比与种系传递百分比之间是否存在相关性;4)评估MHC基因型是否影响嵌合体发育。RBC凝集数据表明每个品系中存在三种B单倍型。B2样基因和B19样基因是WL品系特有的,B13样基因和B-15样基因是BR品系特有的,而B21样基因在两个品系中都存在。在成年BR→WL嵌合体以及10至14日龄的WL→WL嵌合体中,使用流式细胞术可在RBC上检测和定量供体型B抗原。在BR→WL嵌合体中,种系传递百分比与带有供体B抗原的RBC百分比以及羽毛中黑色羽毛的百分比显著相关。在一项使用先前开发的BR→WL嵌合体的回顾性研究中,B*21/*21型受体的嵌合度和种系传递水平较高,但在两项前瞻性研究中这一结果无统计学意义。得出的结论是,RBC上的MHC抗原可用于鉴定、定量和选择通过胚盘细胞转移产生的鸡嵌合体。