Dunon D, Schwager J, Dangy J P, Imhof B A
Laboratory of Experimental Biology, URA-CNRS 1135, Pierre and Marie Curie University, Paris, France.
J Immunol. 1995 Feb 1;154(3):1256-64.
Analysis of TCR-beta gene recombination and expression was performed by quantitative PCR amplification technique throughout chicken embryogenesis and development. Our data demonstrated that TCR V beta 1 promoters were turned on by day 10 of embryogenesis, 2 days before detection of TCR-beta gene recombination. The V to D recombination step was first detected by day 11 of embryogenesis whereas DJ and V(D)J rearranged genes were detected 1 day later, on day 12 of embryogenesis. Thus, transcription of unrearranged TCR-beta genes in chickens precedes the expression of V(D)J recombinase activity as in mammals. In contrast, although TCR-beta rearrangement starts with the D to J recombination step in mammals, it can start either by the VD or the DJ step in chickens. Furthermore, reverse transcriptase-PCR amplification of TCR-beta transcripts revealed the presence of two kinds of alternative transcripts. These novel alternatively spliced products appeared in thymocytes from embryonic thymus during colonization periods and were absent in transformed T cell lines. Splicing sites are located in the middle of V beta 1 segments and lead to delta V beta 1-C beta and delta V beta 1-D beta-J beta-C beta transcripts. delta V beta 1-C beta transcripts might lead to synthesis of invariant truncated TCR beta-chains containing the aminoterminal portion of the V beta 1 region followed by the C beta region. Because this type of splicing can be generated by using all known V beta 1 members, these invariant forms could play a role in thymocyte development.
通过定量PCR扩增技术,在整个鸡胚胎发生和发育过程中对TCR-β基因重组和表达进行了分析。我们的数据表明,在胚胎发生的第10天,即检测到TCR-β基因重组的2天前,TCR Vβ1启动子被激活。在胚胎发生的第11天首次检测到V到D的重组步骤,而DJ和V(D)J重排基因在1天后,即胚胎发生的第12天被检测到。因此,鸡中未重排的TCR-β基因的转录先于V(D)J重组酶活性的表达,这与哺乳动物的情况相同。相比之下,虽然TCR-β重排在哺乳动物中从D到J的重组步骤开始,但在鸡中它可以从VD或DJ步骤开始。此外,TCR-β转录本的逆转录PCR扩增揭示了两种替代转录本的存在。这些新的可变剪接产物出现在胚胎胸腺定植期的胸腺细胞中,而在转化的T细胞系中不存在。剪接位点位于Vβ1区段的中间,导致δVβ1-Cβ和δVβ1-Dβ-Jβ-Cβ转录本。δVβ1-Cβ转录本可能导致合成包含Vβ1区域氨基末端部分和Cβ区域的不变截短TCRβ链。因为这种类型的剪接可以通过使用所有已知的Vβ1成员产生,这些不变形式可能在胸腺细胞发育中起作用。