Int. J. Dev. Biol. 59: 487 - 496 (2015)

https://doi.org/10.1387/ijdb.150363cx

Vol 59, Issue 10-11-12

iTRAQ-based proteomic analysis of adaptive response in the regenerating limb of the Cynops orientalis newt

Developmental Expression Pattern | Published: 15 January 2016

Xiao-Fang Geng^1,2,3, Jian-Lin Guo^2,3,4, Xia-Yan Zang^2,3,4, Jing-Yan Sun^2,3,4, Peng-Fei Li^2,3,4, Fu-Chun Zhang¹* and Cun-Shuan Xu^2,3,4*

¹Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi, ² State Key Laboratory Cultivation Base for Cell Differentiation Regulation, Henan Normal University, Xinxiang, ³Henan Engineering Laboratory for Bioengineering and Drug Development, Henan Normal University, Xinxiang and ⁴College of Life Science, Henan Normal University, Xinxiang, China

Abstract

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, Ca²⁺ 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.

Keywords

Cynops orientalis, limb regeneration, proteomic, stress response, cell death