Schüle Birgitt, Li Hong Hua, Fisch-Kohl Claudia, Purmann Carolin, Francke Uta
Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
Am J Hum Genet. 2007 Sep;81(3):492-506. doi: 10.1086/520063. Epub 2007 Aug 2.
Mutations in MECP2 and Mecp2 (encoding methyl-CpG binding protein 2 [MeCP2]) cause distinct neurological phenotypes in humans and mice, respectively, but the molecular pathology is unclear. Recent literature claimed that the developmental homeobox gene DLX5 is imprinted and that its imprinting status is modulated by MeCP2, leading to biallelic expression in Rett syndrome and twofold overexpression of Dlx5 and Dlx6 in Mecp2-null mice. The conclusion that DLX5 is a direct target of MeCP2 has implications for research on the molecular bases of Rett syndrome, autism, and genomic imprinting. Attempting to replicate the reported data, we evaluated allele-specific expression of DLX5 and DLX6 in mouse x human somatic cell hybrids, lymphoblastoid cell lines, and frontal cortex from controls and individuals with MECP2 mutations. We identified novel single-nucleotide polymorphisms in DLX5 and DLX6, enabling the first imprinting studies of DLX6. We found that DLX5 and DLX6 are biallelically expressed in somatic cell hybrids and in human cell lines and brain, with no differences between affected and control samples. We also determined expression levels of Dlx5 and Dlx6 in forebrain from seven male Mecp2-mutant mice and eight wild-type littermates by real-time quantitative reverse-transcriptase polymerase chain reaction assays. Expression of Dlx5 and Dlx6, as well as of the imprinted gene Peg3, in mouse forebrain was highly variable, with no consistent differences between Mecp2-null mutants and controls. We conclude that DLX5 and DLX6 are not imprinted in humans and are not likely to be direct targets of MeCP2 modulation. In contrast, the imprinting status of PEG3 and PEG10 is maintained in MeCP2-deficient tissues. Our results confirm that MeCP2 plays no role in the maintenance of genomic imprinting and add PEG3 and PEG10 to the list of studied imprinted genes.
MECP2和Mecp2(编码甲基化CpG结合蛋白2 [MeCP2])中的突变分别在人类和小鼠中导致不同的神经表型,但分子病理学尚不清楚。最近的文献称,发育同源框基因DLX5是印记基因,其印记状态受MeCP2调节,导致雷特综合征中双等位基因表达,以及Mecp2基因敲除小鼠中Dlx5和Dlx6的表达量翻倍。DLX5是MeCP2的直接靶点这一结论对雷特综合征、自闭症和基因组印记的分子基础研究具有重要意义。为了重复已报道的数据,我们评估了DLX5和DLX6在小鼠×人类体细胞杂种、淋巴母细胞系以及来自对照和患有MECP2突变个体的额叶皮质中的等位基因特异性表达。我们在DLX5和DLX6中鉴定出了新的单核苷酸多态性,从而能够首次对DLX6进行印记研究。我们发现,DLX5和DLX6在体细胞杂种、人类细胞系和大脑中均为双等位基因表达,患病样本与对照样本之间没有差异。我们还通过实时定量逆转录聚合酶链反应分析,测定了7只雄性Mecp2突变小鼠和8只野生型同窝小鼠前脑中Dlx5和Dlx6的表达水平。小鼠前脑中Dlx5和Dlx6以及印记基因Peg3的表达高度可变,Mecp2基因敲除突变体与对照之间没有一致的差异。我们得出结论,DLX5和DLX6在人类中不是印记基因,不太可能是MeCP2调节的直接靶点。相比之下,PEG3和PEG10的印记状态在MeCP2缺陷组织中得以维持。我们的结果证实,MeCP2在基因组印记的维持中不起作用,并将PEG3和PEG10添加到已研究的印记基因列表中。