Geng Xiaofang, Xu Tiantian, Niu Zhipeng, Zhou Xiaochun, Zhao Lijun, Xie Zhaohui, Xue Deming, Zhang Fuchun, Xu Cunshuan
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China; State Key Laboratory Cultivation Base for Cell Differentiation Regulation, Henan Normal University, Xinxiang 453007, China.
State Key Laboratory Cultivation Base for Cell Differentiation Regulation, Henan Normal University, Xinxiang 453007, China; College of Life Science, Henan Normal University, Xinxiang 453007, China.
Differentiation. 2014 Nov-Dec;88(4-5):85-96. doi: 10.1016/j.diff.2014.10.002. Epub 2014 Nov 15.
Following amputation, the newt has the remarkable ability to regenerate its limb, and this process involves dedifferentiation, proliferation and differentiation. To investigate the potential proteome during a dynamic network of Chinese fire-bellied newt limb regeneration (CNLR), two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and mass spectrum (MS) were applied to examine changes in the proteome that occurred at 11 time points after amputation. Meanwhile, several proteins were selected to validate their expression levels by Western blot. The results revealed that 1476 proteins had significantly changed as compared to the control group. Gene Ontology annotation and protein network analysis by Ingenuity Pathway Analysis 9.0 (IPA) software suggested that the differentially expressed proteins were involved in 33 kinds of physiological activities including signal transduction, cell proliferation, cell differentiation, etc. Among these proteins, 407 proteins participated in cell differentiation with 212 proteins in the differentiation of skin cell, myocyte, neurocyte, chondrocyte and osteocyte, and 37 proteins participated in signaling pathways of BCC, CRH, CXCR4, GnRH, GPCR and IL1 which regulated cell differentiation and redifferentiation. On the other hand, the signal transduction activity and cell differentiation activity were analyzed by IPA based on the changes in the expression of these proteins. The results showed that BCC, CRH, CXCR4, GnRH, GPCR and IL1 signaling pathways played an important role in regulating the differentiation of skin cell, myocyte, neurocyte, chondrocyte and osteocyte during CNLR.
截肢后，东方蝾螈具有显著的肢体再生能力，这一过程涉及去分化、增殖和分化。为了研究中国林蛙肢体再生（CNLR）动态网络中的潜在蛋白质组，应用二维荧光差异凝胶电泳（2D-DIGE）和质谱（MS）来检测截肢后11个时间点蛋白质组的变化。同时，选择了几种蛋白质通过蛋白质免疫印迹法验证其表达水平。结果显示，与对照组相比，有1476种蛋白质发生了显著变化。通过基因本体注释和Ingenuity Pathway Analysis 9.0（IPA）软件进行的蛋白质网络分析表明，差异表达的蛋白质参与了33种生理活动，包括信号转导、细胞增殖、细胞分化等。在这些蛋白质中，407种蛋白质参与细胞分化，其中212种蛋白质参与皮肤细胞、肌细胞、神经细胞、软骨细胞和骨细胞的分化，37种蛋白质参与了BCC、CRH、CXCR4、GnRH、GPCR和IL1的信号通路，这些信号通路调节细胞分化和再分化。另一方面，基于这些蛋白质表达的变化，通过IPA分析信号转导活性和细胞分化活性。结果表明，BCC、CRH、CXCR4、GnRH、GPCR和IL1信号通路在CNLR过程中对皮肤细胞、肌细胞、神经细胞、软骨细胞和骨细胞的分化调节中起重要作用。
