Geng Xiao-Fang, Guo Jian-Lin, Zang Xia-Yan, Sun Jing-Yan, Li Peng-Fei, Zhang Fu-Chun, Xu Cun-Shuan
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi, China.
Int J Dev Biol. 2015;59(10-12):487-96. doi: 10.1387/ijdb.150363cx.
The newt has the powerful capacity to regenerate lost limbs following amputation, and represents an excellent model organism to study regenerative processes. However, the molecular basis of the adaptive response in the regenerating limb of the Chinese fire-bellied newt Cynops orientalis immediately after amputation remains unclear. To better understand the adaptive response immediately after limb amputation at the protein level, we used isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC-MS/MS methods to analyze changes in the proteome of the regenerating newt limb that occurred 2 h and 8 h after amputation. We identified 152 proteins with more than 1.5-fold change in expression compared to control. GO annotation analysis classified these proteins into several categories such as signaling, Ca(2+) binding and translocation, transcription and translation, immune response, cell death, cytoskeleton, metabolism, etc. Further ingenuity pathway analysis (IPA) showed that several signaling pathways were significantly changed at 2 h and 8 h after amputation, including EIF2 signaling, acute phase response signaling, tight junction signaling and calcium signaling, suggesting these pathways may be closely related to the adaptive response immediately after limb amputation. This work provides novel insights into understanding the molecular processes related to newt limb regeneration immediately after amputation, and a basis for further study of regenerative medicine.
东方蝾螈在截肢后具有强大的再生缺失肢体的能力，是研究再生过程的优秀模式生物。然而，中国林蛙东方蝾螈截肢后再生肢体中适应性反应的分子基础仍不清楚。为了在蛋白质水平上更好地理解肢体截肢后立即出现的适应性反应，我们使用了相对和绝对定量的等压标签（iTRAQ）结合液相色谱-串联质谱（LC-MS/MS）方法，来分析截肢后2小时和8小时再生蝾螈肢体蛋白质组的变化。我们鉴定出152种与对照相比表达变化超过1.5倍的蛋白质。基因本体（GO）注释分析将这些蛋白质分为几个类别，如信号传导、Ca(2+)结合与转运、转录与翻译、免疫反应、细胞死亡、细胞骨架、代谢等。进一步的 Ingenuity 通路分析（IPA）表明，截肢后2小时和8小时有几个信号通路发生了显著变化，包括真核起始因子2（EIF2）信号传导、急性期反应信号传导、紧密连接信号传导和钙信号传导，这表明这些通路可能与肢体截肢后立即出现的适应性反应密切相关。这项工作为理解蝾螈肢体截肢后立即发生的与再生相关的分子过程提供了新的见解，并为再生医学的进一步研究奠定了基础。
