Yelavarthi K K, Schmidt C M, Ehlenfeldt R G, Orr H T, Hunt J S
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City 66160-7400.
J Immunol. 1993 Oct 1;151(7):3638-45.
The human MHC class I gene, HLA-G, is unique among members of the class I gene family in that it is nonpolymorphic, and expression is primarily restricted to extraembryonic tissues. To examine regulatory elements that direct tissue- and cell lineage-specific expression of this gene, transgenic mice expressing HLA-G have been established. In this study, in situ hybridization was used to evaluate the cellular distribution of HLA-G mRNA in transgenic placentas. Extraembryonic tissues were obtained at gestation day 12.5 from embryos that had been microinjected with either 6.0 or 5.7 kb of HLA-G genomic DNA and had been transferred into pseudopregnant HLA-G transgenic mice or Swiss mice. The 6.0 kb transgene contained an additional 250 bp at the extreme 5'-end of the upstream region. Genotype of the recipient had no discernable effect on the cellular distribution of HLA-G mRNA. HLA-G mRNA was present in both trophoblast and mesenchymal cells in transgenic placentas carrying 6.0 kb of genomic HLA-G, a pattern strikingly similar to that of HLA-G message distribution in early gestation human placentas. By contrast, in placentas from embryos carrying 5.7 kb HLA-G DNA, specific mRNA was found primarily in mesenchymal cells at the base of the placenta. Thus, the 6.0 genomic fragment contains elements capable of directing HLA-G expression in placentas, and is particularly influential in the trophoblastic cell lineage.
人类主要组织相容性复合体I类基因HLA - G在I类基因家族成员中独具特色,因为它是非多态性的,且表达主要局限于胚外组织。为了研究指导该基因组织和细胞谱系特异性表达的调控元件,已构建了表达HLA - G的转基因小鼠。在本研究中,采用原位杂交技术评估转基因胎盘组织中HLA - G mRNA的细胞分布情况。在妊娠第12.5天,从分别显微注射了6.0 kb或5.7 kb HLA - G基因组DNA并移植到假孕的HLA - G转基因小鼠或瑞士小鼠体内的胚胎中获取胚外组织。6.0 kb的转基因在上游区域的5'端极端位置额外包含250 bp。受体的基因型对HLA - G mRNA的细胞分布没有明显影响。在携带6.0 kb基因组HLA - G的转基因胎盘中,滋养层细胞和间充质细胞中均存在HLA - G mRNA,这一模式与早期妊娠人类胎盘中HLA - G信息分布模式极为相似。相比之下,在携带5.7 kb HLA - G DNA的胚胎的胎盘中,特异性mRNA主要在胎盘基部的间充质细胞中发现。因此,6.0 kb的基因组片段包含能够指导胎盘组织中HLA - G表达的元件,并且对滋养层细胞谱系具有特别的影响。