Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering, Department of Biology, School of Life Science, Xiamen University, 361005 Xiamen, Fujian, People's Republic of China.
J Mol Histol. 2010 Oct;41(4-5):295-305. doi: 10.1007/s10735-010-9292-4. Epub 2010 Sep 8.
Heterochronical gene lin-41 plays an important role in regulate the timing of many decelopmental events in Caenorhabditis elegans. Mammalian developmental timing is poorly understood even though many tissues are under temporal control during development. The lin-41 homologues in mouse and chick has been isolated and its expression in developing limb buds and branchial arches has been reported by in situ (Kanamoto et al. in Dev Dyn 235:1142-1149, 2006; Lancman et al. in Dev Dyn 234:948-960, 2005; Schulman et al. in Dev Dyn 234:1046-1054, 2005), but the protein expression pattern in mouse adult tissue and embryo remained to be clarified. To help elucidate the expression of C. elegans lin-41 orthorlogs in mouse adult tissue and developmental embryo, lin-41 cDNA fragment was amplified from the mouse embryonic day 9.5(E9.5) mRNA and expressed in E. coli. The transcripts of mlin-41 and the protein level in mouse adult tissues and embryos from 9.5 to 13 days were detected by RT-PCR and western blot. RT-PCR and western blot showed the expression of mLIN-41 was detected in the mouse adult heart, muscle, and small intestine as well as in the day E9.5 to E12 embryos. Immuno-localization of mLIN-41 in the day E10.5 embryo revealed that mLIN-41 was present in the neuro-epithelium and epithelial tissue covering the first and second branchial arch, somites and mesoderm cells, limb buds as well as the gut epithelium. The expression of mLIN-41 represented the tissue-specific expression pattern. Immuno-precipitation combine with MALDI-TOF mass spectrometry was used to identify the potential proteins interacting with LIN-41. Five potential specific proteins were obtained for future identification in mouse.
异时性基因 lin-41 在调控秀丽隐杆线虫许多发育事件的时间方面发挥着重要作用。尽管许多组织在发育过程中受到时间控制，但哺乳动物的发育时间仍未得到很好的理解。已经分离出小鼠和鸡的 lin-41 同源物，并通过原位杂交报告了其在发育中的肢芽和鳃弓中的表达（Kanamoto 等人，Dev Dyn 235:1142-1149, 2006；Lancman 等人，Dev Dyn 234:948-960, 2005；Schulman 等人，Dev Dyn 234:1046-1054, 2005），但在小鼠成年组织和胚胎中的蛋白表达模式仍有待阐明。为了帮助阐明秀丽隐杆线虫 lin-41 同源物在小鼠成年组织和发育胚胎中的表达，我们从小鼠胚胎第 9.5 天（E9.5）mRNA 中扩增了 lin-41 cDNA 片段，并在大肠杆菌中表达。通过 RT-PCR 和 Western blot 检测了 mlin-41 的转录物和从 9.5 天到 13 天的小鼠成年组织和胚胎中的蛋白水平。RT-PCR 和 Western blot 显示，mLIN-41 在小鼠成年心脏、肌肉和小肠以及 E9.5 至 E12 天的胚胎中均有表达。在 E10.5 天胚胎中的 mLIN-41 免疫定位显示，mLIN-41 存在于神经上皮和覆盖第一和第二鳃弓、体节和中胚层细胞、肢芽以及肠道上皮的上皮组织中。mLIN-41 的表达呈现出组织特异性表达模式。免疫沉淀结合 MALDI-TOF 质谱用于鉴定与 LIN-41 相互作用的潜在蛋白质。获得了五个潜在的特定蛋白质，以便在小鼠中进一步鉴定。
