Ontogeny of TCR V beta 1 expression revealed novel invariant alternative transcripts

Abstract

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.