Gingras Marie-Claude, Kharitidi Dmitri, Chénard Valérie, Uetani Noriko, Bouchard Maxime, Tremblay Michel L, Pause Arnim
Goodman Cancer Centre and Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Int J Dev Biol. 2009;53(7):1069-74. doi: 10.1387/ijdb.082820mg.
The putative tyrosine phosphatase HD-PTP, encoded by the protein-tyrosine-phosphatase-n23 (Ptpn23) gene, has been described as a tumor suppressor candidate gene. However, its physiological roles and detailed expression profiles are poorly defined. To investigate HD-PTP functions, we generated a mouse model in which the Ptpn23 locus was disrupted by an in-frame insertion of a beta-galactosidase-neomycin-phosphotransferase II (beta-geo) cassette. This insertion leads to the expression of a catalytically inactive truncated protein preserving only the uncharacterized N-terminal BRO1-like domain in fusion with beta-geo under the control of the endogenous promoter. Here we report that homozygous gene deletion is lethal around embryonic day 9.5, suggesting that Ptpn23 is an essential requirement for early stages of embryonic development. Taking advantage of the beta-galactosidase insertion into the Ptpn23 locus, we define the precise Ptpn23 expression pattern by performing X-gal staining at different stages of mouse development. Our results show that Ptpn23 is expressed early during mouse development and that its expression is maintained in adult tissues, markedly in the epithelial cells of many organs.
由蛋白酪氨酸磷酸酶n23(Ptpn23)基因编码的假定酪氨酸磷酸酶HD-PTP,已被描述为一种肿瘤抑制候选基因。然而,其生理作用和详细的表达谱尚不清楚。为了研究HD-PTP的功能,我们构建了一个小鼠模型,其中Ptpn23基因座被β-半乳糖苷酶-新霉素-磷酸转移酶II(β-geo)盒的读框内插入所破坏。这种插入导致表达一种催化失活的截短蛋白,该蛋白仅保留未表征的N端BRO1样结构域,并与β-geo在内源启动子的控制下融合。我们在此报告,纯合基因缺失在胚胎第9.5天左右是致死的,这表明Ptpn23是胚胎发育早期阶段的必要条件。利用插入到Ptpn23基因座中的β-半乳糖苷酶,我们通过在小鼠发育的不同阶段进行X-gal染色来确定精确的Ptpn23表达模式。我们的结果表明,Ptpn23在小鼠发育早期表达,并且其表达在成年组织中维持,在许多器官的上皮细胞中明显表达。